Short title: OsGRF7 shapes rice plant architecture · 2020. 6. 24. · in determining the...

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Short title: OsGRF7 shapes rice plant architecture 1

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Rice GROWTH-REGULATING FACTOR7 Modulates Plant Architecture 3

through Regulating GA and IAA Metabolism 4

Yunping Chena, 2

, Zhiwu Dana, 2

, Feng Gaoa, 3

, Pian Chena, Fengfeng Fan

a & Shaoqing 5

Lia, 4, 5

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a State Key Laboratory of Hybrid Rice, Key Laboratory for Research and Utilization 8

of Heterosis in Indica Rice of Ministry of Agriculture, College of Life Sciences, 9

Wuhan University, Wuhan 430072, China. 10

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One-sentence summary 12

A transcriptional regulator maintains compact plant architecture and decreased leaf 13

angle by regulating gibberellin synthesis and auxin signaling pathways. 14

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Footnotes: 16

Author contributions 17

Y.C. and S.L. conceived and designed the research. Y.C. and Z.D. performed most of 18

the experiments and analyzed the data. F.G. performed overexpression and RNAi 19

vector construction, 5ꞌ RACE. P.C. performed the GRF7-GFP transgenic experiment. 20

Y.C., Z.D., and F.F. performed field experiments. Y.C., Z.D., and S.L. wrote the 21

manuscript. 22

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1 This research was supported by the National Key Research and Development 24

Program (2016YFD0100903), the National Transgenic Research and Development 25

Program (2016ZX08001004-001-002) and the National Natural Science Foundation 26

(31870322) of China. 27

2 These authors contributed equally to this work. 28

Plant Physiology Preview. Published on June 24, 2020, as DOI:10.1104/pp.20.00302

Copyright 2020 by the American Society of Plant Biologists

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3 Present address: Oil Crops Research Institute of the Chinese Academy of 29

Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil 30

Crops, Ministry of Agriculture, Wuhan 430072, China. 31

4 Senior author. 32

5 Author for contact: [email protected]. 33

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Copyright (c) 2020 American Society of Plant Biologists. All rights reserved.


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Abstract 59

Plant-specific GROWTH-REGULATING FACTORs (GRFs) participate in central 60

developmental processes, including leaf and root development; inflorescence, flower 61

and seed formation; senescence; and tolerance to stresses. In rice (Oryza sativa), there 62

are 12 GRFs, but the role of the miR396-OsGRF7 regulatory module remains 63

unknown. Here, we report that OsGRF7 shapes plant architecture via the regulation of 64

auxin and gibberellin metabolism in rice. OsGRF7 is mainly expressed in lamina 65

joints, nodes, internodes, axillary buds, and young inflorescences. Overexpression of 66

OsGRF7 causes a semidwarf and compact plant architecture with an increased culm 67

wall thickness and narrowed leaf angles mediated by shortened cell length, altered cell 68

arrangement, and increased parenchymal cell layers in the culm and adaxial side of 69

the lamina joints. Knock-out and knock-down lines of OsGRF7 exhibit contrasting 70

phenotypes with severe degradation of parenchymal cells in the culm and lamina 71

joints at maturity. Further analysis indicated that OsGRF7 binds the ACRGDA motif 72

in the promoters of Cytochrome P450 gene (OsCYP714B1) and AUXIN RESPONSE 73

FACTOR 12 (OsARF12), which are involved in the gibberellin synthesis and auxin 74

signaling pathways, respectively. Correspondingly, OsGRF7 alters the contents of 75

endogenous gibberellins and auxins and sensitivity to exogenous phytohormones. 76

These findings establish OsGRF7 as a crucial component in the 77

OsmiR396-OsGRF-plant hormone regulatory network that controls rice plant 78

architecture. 79

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Keywords: plant architecture, OsGRF7, transcription factor, rice, gibberellin, auxin 81

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Introduction 90

Plant architecture is a major factor that determines grain productivity in cereal crop 91

species (Wang and Li, 2008). Rice (Oryza sativa L.) is a staple food for more than 92

half of the global population, and rice plant architecture has been continually 93

improved by breeders to achieve high productivity. Rice plant architecture is mainly 94

defined by plant height, the spatial pattern of leaves, and tiller and inflorescence 95

branching patterns (Khush, 1995). Plant height and tiller branching determine the 96

biomass and harvest index, while the spatial pattern of leaves, including leaf shape 97

and angle, influences the photosynthesis rate and therefore the accumulation of 98

carbohydrates (Sinclair and Sheehy, 1999; Springer, 2010; Xing and Zhang, 2010). 99

During plant architecture determination, several factors, especially plant hormones, 100

have been reported to affect the development of the leaf angle, plant height, and tiller 101

number, thus modulating rice plant architecture. 102

Phytohormones regulate many physiological processes that largely influence 103

growth, differentiation, and development (Luo et al., 2016). Auxins and cytokinins 104

mainly control the size and number of plant leaves, culms, inflorescences, and grains 105

by regulating cell size and number (Lavy and Estelle, 2016; Schaller et al., 2015). 106

Auxins also negatively control lamina joint inclination via asymmetric adaxial-abaxial 107

cell division (Zhang et al., 2015). Consistent with these findings, the auxin early 108

response gain-of-function rice mutant leaf inclination1, lc1-D, which encodes the IAA 109

amido synthetase OsGH3-1, showed reduced free auxin levels and enlarged leaf 110

angles due to stimulated cell elongation on the adaxial side of the lamina joints (Zhao 111

et al., 2013). Overexpression of AUXIN RESPONSE FACTOR 19 (OsARF19) resulted 112

in an enlarged leaf angle via increased expression level of GH3s and decreased free 113

IAA content (Zhang et al., 2015). Gibberellins control plant height and tillering by 114

regulating cell elongation and cell division (Gao et al., 2016; Kobayashi et al., 1988; 115

Magome et al., 2013). Blocking the synthesis of gibberellin reduces plant height and 116

therefore increases lodging resistance (Chen et al., 2015; Sasaki et al., 2002). Thus, 117

the integration of multiple phytohormone signaling pathways will appropriately 118 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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coordinate plant architecture development. 119

Increasing evidence indicates that miRNAs also participate in hormone synthesis 120

or signal transduction and plant development (Tang and Chu, 2017). miR396 is one of 121

the most conserved miRNA families in monocots and dicots (Liu et al., 2009). Studies 122

have shown that repressing the expression of OsmiR396 in miR396 mimicry 123

(MIM396) transgenic lines resulted in enlarged panicles (Gao et al., 2015). GRFs, the 124

targets of OsmiR396, are conserved, plant-specific transcription factors. In rice, the 125

GRF family comprises 12 members. All GRF genes have the conserved QLQ and 126

WRC domains in their N-terminal regions. The QLQ domain is essential for 127

protein-protein interaction, and the WRC domain contains a functional nuclear 128

localization signal (Choi et al., 2004). 129

GRFs interact with small cofactors called GRF-INTERACTING FACTORs 130

(GIFs) to form a functional complex to regulate plant growth and development (Kim 131

and Kende, 2004). GIFs have been reported to participate in cell proliferation during 132

leaf development, root meristem homeostasis, and grain size determination (Ercoli et 133

al., 2018; Kim and Kende, 2004; Li et al., 2016). In rice, the GIF family is composed 134

of three members: OsGIF1, OsGIF2, and OsGIF3. OsGRF6 interacts with these three 135

OsGIFs to regulate inflorescence architecture (Gao et al., 2015; Liu et al., 2014), 136

OsGRF10 interacts with OsGIF1 and OsGIF2 to regulate floral organogenesis (Liu et 137

al., 2014), and OsGRF4 interacts with OsGIF1 to regulate grain size (Li et al., 2016). 138

These findings highlight the crucial roles of the OsmiR396-OsGRFs-OsGIFs module 139

in determining the complexity of the regulatory network of rice growth and 140

development. 141

In this study, we report that OsGRF7 and OsmiR396e form a molecular node that 142

regulates plant architecture via hormone-related genes, including OsARF12 (Li et al., 143

2020; Wang et al., 2014) and OsCYP714B1 (Magome et al., 2013), which are 144

involved in the auxin signaling pathway and gibberellin synthesis, respectively. In 145

vivo and in vitro assays indicate that these genes are directly regulated by OsGRF7. 146

OsGRF7 also alters the contents of corresponding endogenous phytohormones and 147

sensitivity to exogenous phytohormones. Our study demonstrates that OsGRF7 is a 148 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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critical regulator of the shaping of plant architecture through GA and IAA signaling 149

networks. 150

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Results 152

OsGRF7 modulates plant architecture in rice 153

A previous study showed that OsmiR396 regulates rice inflorescence branching and 154

grain yield by modulating the expression of OsGRF6 (Gao et al., 2015). Here, we 155

observed that MIM396 transgenic lines presented compact plant architecture. The leaf 156

angle of the MIM396 transgenic lines was reduced by 26.9% at the seedling stage 157

compared with that of the wild-type YuetaiB (YB) (Supplemental Fig. S1A, B). A 158

similar phenotype was also observed for the second and third fully expanded leaves 159

from the top of MIM396 plants at the tillering stage (Supplemental Fig. S1C-E). 160

Corresponding to the significant decrease in the abundance of OsmiR396 family 161

members in MIM396-3 transgenic lines (Supplemental Fig. S1F), and the expression 162

levels of GRF family members, especially OsGRF7, were significantly increased 163

(Supplemental Fig. S1G). 164

To elucidate the functions of OsGRF7, we performed a genetic transformation of 165

OsGRF7 and found that seven out of 13 OsGRF7 overexpression (GRF7OE) lines and 166

four out of six OsGRF7 RNAi (GRF7RNAi) lines showed altered plant architecture 167

(Fig. 1A, B). Compared with that of the WT, the plant height of the GRF7OE lines 168

was reduced by approximately 20% (Fig. 1A; Supplemental Table S1), corresponding 169

to the contraction of each internode in the GRF7OE lines (Supplemental Fig. S2A, B). 170

The flag leaf angles of the WT as well as the GRF7OE-1, GRF7OE-2, 171

GRF7RNAi-1, and GRF7RNAi-2 transgenic lines were 5.5, 1.9, 1.3, 9.5, and 6.3, 172

respectively (Fig. 1B; Supplemental Table S1). Moreover, the plant height, effective 173

panicle, leaf angle, leaf length and width, were strongly correlated with the relative 174

expression levels of OsGRF7 in the transgenic plants (Supplemental Fig. S3), 175

meaning that OsGRF7 regulates plant architecture in a dose-dependent manner. 176

We further crossed GRF7RNAi-1 with MIM396-3 and found that the leaf angles 177

were significantly decreased in the GRF7RNAi-1/MIM396-3 hybrid compared with 178 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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those in the GRF7RNAi-1 (Supplemental Fig. S4A, B). The first fully expanded leaf 179

angles from the top of the main stalk decreased from 17.4 in the GRF7RNAi-1 180

transgenic lines to 12.6 in the GRF7RNAi-1/MIM396-3 hybrid, and the third fully 181

expanded leaf angles decreased from 26.5 in the GRF7RNAi-1 transgenic lines to 182

16.7 in the GRF7RNAi-1/MIM396-3 hybrid (Supplemental Fig. S4C, D), suggesting 183

that OsmiR396-OsGRF7 is involved in leaf angle determination. 184

To further validate the function of OsGRF7 in rice plant architecture 185

determination, 74 independent OsGRF7 knock-out (GRF7KO) lines were generated 186

using CRISPR/Cas9 against two target sites within the second exon of OsGRF7 187

(Supplemental Fig. S5A). After sequence identification, we obtained four independent 188

GRF7KO homologous lines in which OsGRF7 was prematurely terminated 189

(Supplemental Fig. S5A and Supplemental Table S2). Consistent with the leaf angles 190

of the GRF7RNAi transgenic lines, the angles of the second leaves of GRF7KO-2, 191

GRF7KO-14, GRF7KO-64, and GRF7KO-66 mutants were 27.6, 26.1, 32.9, and 192

36.1, respectively (Supplemental Fig. S5B-D), which are significantly larger than 193

those of the WT line (14.8). After analyzing the plant height of GRF7KO lines, we 194

found that the plant height of GRF7KO lines was significantly increased when 195

compared with the WT (Supplemental Fig. S5B, E). These results demonstrate that 196

OsGRF7 is a critical node for plant architecture determination in rice. 197

GRFs are conserved and plant-specific transcription factors. Phylogenetic 198

analysis of the GRFs from rice and Arabidopsis showed that OsGRF7 was closely 199

related to the Arabidopsis homologs AtGRF1 and AtGRF2 (Supplemental Fig. S6). 200

After analyzing the mRNA expression of OsGRFs in the OsGRF7 transgenic lines, we 201

found that when OsGRF7 was downregulated, multiple OsGRFs showed increased 202

expression levels, especially OsGRF4, OsGRF6, OsGRF8, and OsGRF9 203

(Supplemental Fig. S7). At the same time, OsGRF4, OsGRF6, OsGRF8, and OsGRF9, 204

shared similar expression patterns with OsGRF7 (Choi et al., 2004), suggesting that 205

these GRFs may compensate for the loss of OsGRF7 in the GRF7RNAi and GRF7KO 206

lines. 207




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OsGRF7 promotes periclinal division in lamina joints and internodes 209

To clarify the cellular mechanism of OsGRF7 in the control of plant architecture, we 210

observed the microstructure of the uppermost internodes and lamina joints. The 211

results showed that GRF7OE-1 plants had more cell layers in the uppermost internode 212

than WT and GRF7RNAi-1 plants (Fig. 1C). Longitudinal sections revealed that the 213

internode parenchymal cells of the GRF7OE-1 plants were not only shorter but also 214

rounder than those of the WT and GRF7RNAi-1 lines (Fig. 1C, D), and the culm wall 215

thickness of the GRF7OE-1 plants was significantly increased compared with that of 216

the WT (Fig. 1E), corresponding to the short, sturdy, and thick culm of the former 217

(Fig. 1A). 218

Leaf angle increases after the lamina joint emerges from the prior leaf sheath. 219

After analyzing the dynamic lamina joint growth from lamina joint initiation to the 220

maturation of the flag leaf, we found that the GRF7OE-1 transgenic lines showed 221

enlarged vascular bundles during the lamina joint development stage, while 222

GRF7RNAi-1 and GRF7KO-14 showed the opposite phenotype (Supplemental Fig. 223

S8). Additional examination of cross-sections and longitudinal sections of the lamina 224

joints revealed that, compared with the WT and GRF7RNAi-1 plants, the GRF7OE-1 225

plants showed enhanced adaxial cell proliferation and repressed cell elongation (Fig. 226

1F, G), suggesting that overexpression of OsGRF7 promoted the cell periclinal 227

division on the adaxial side of the lamina joint, and thus increased cell layers on the 228

adaxial side that caused narrow lamina joint bending (Fig. 1B), and the compact 229

phenotype of GRF7OE-1 plants. 230

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OsGRF7 is mainly repressed by OsmiR396e 232

GRFs are known to be highly regulated by miR396 (Omidbakhshfard et al., 2015). 233

Here, we found that OsmiR396 could directly cleave OsGRF7 mRNA in vivo at the 234

site within the OsmiR396 pairing region (Fig. 2A). Moreover, we performed transient 235

expression assays of OsGRF7 using the OsmiR396-sensitive construct 35S:OsGRF7 236

and the OsmiR396-resistant construct 35S:OsrGRF7 in rice protoplasts under the 237

background of OsmiR396s (Fig. 2A). RT-qPCR analysis showed that the transcript 238 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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level of OsGRF7 significantly decreased when OsmiR396-sensitive OsGRF7 was 239

coexpressed with OsmiR396s, which contrasted with the slight decrease in OsGRF7 240

under the background of OsrGRF7 together with coexpression of OsmiR396 members 241

(Supplemental Fig. S9). These results demonstrate that OsGRF7 is repressed by 242

OsmiR396. 243

In rice, there are nine miR396s that show various expression profiles (Kozomara 244

and Griffiths-Jones, 2014), implying that each of them may have different functions. 245

To further understand the regulatory activity of OsmiR396 members on OsGRF7, we 246

transiently expressed Ubi:miR396s in GRF7-GFP transgenic rice protoplasts. 247

Immunoblotting assays revealed that at the same OsmiR396 transcript level 248

(Supplemental Fig. S10), the OsGRF7 protein content sharply decreased under the 249

OsmiR396e background (Fig. 2B). Correspondingly, OsmiR396e exhibited the 250

opposite expression pattern of OsGRF7 in different tissues (Fig. 2C), implying that 251

OsmiR396e is mainly responsible for the cleavage of OsGRF7. 252

We further examined the spatial-temporal expression pattern of OsGRF7, and the 253

results showed that OsGRF7 was prominently expressed in the axillary buds, lamina 254

joints, nodes, and internodes as well as highly expressed in young inflorescences and 255

florets (Fig. 2D), corresponding to the RT-qPCR results showing that OsGRF7 was 256

expressed at different levels in various tissues (Fig. 2E). The multi-tissue expression 257

pattern of OsGRF7 was consistent with its pleiotropic roles in controlling plant height, 258

leaf size and angle, and tiller number. 259

Furthermore, transient expression of the GRF7-GFP fusion protein in rice 260

protoplasts showed that OsGRF7 was expressed specifically in the nucleus 261

(Supplemental Fig. S11A), which is consistent with the nuclear localization pattern in 262

GRF7-GFP transgenic lines (Supplemental Fig. S11B), implying that OsGRF7 263

functions in the nucleus. This is in accordance with the transcriptional activity of the 264

OsGRF7 C-terminus (177–430 aa) verified in the yeast expression system 265

(Supplemental Fig. S11C), and the interaction of the OsGRF7 N-terminus with GIFs 266

(Supplemental Fig. S11D, E) such as AtGRF1 and AtGRF2 do in Arabidopsis (Kim 267

and Kende, 2004). These results suggest that OsGRF7 and OsGIFs work together to 268 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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modulate gene expression in the nucleus. 269

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OsGRF7 directly modulates the expression of multiple GA/IAA-related genes 271

To further elucidate how OsGRF7 influences plant architecture, we performed 272

ChIP-seq (chromatin immunoprecipitation followed by high-throughput sequencing) 273

against the GFP antibodies using GRF7-GFP transgenic lines. In general, the more 274

active metabolism of young inflorescences than of the leaves or seedlings makes the 275

ChIP assay more practical; thus, young inflorescences of the GRF7-GFP lines were 276

used to identify the potential targets and binding motifs of OsGRF7. By comparing 277

the ChIP-seq data between YP1 (young inflorescences with a length of ~0.5 cm) and 278

YP2 (young inflorescences with a length of ~2 cm) with the input data, we detected 279

1321 and 2290 OsGRF7-bound peaks, respectively (P < 1.0e-5, one-tailed t-test). Of 280

these, approximately 70% and 63% of the peaks lied in the 5ꞌ-upstream regions of 281

genes in YP1 and YP2, respectively (Fig. 3A), implying that OsGRF7 functions 282

mainly in the regulation of gene expression, consistent with its nuclear localization 283

(Supplemental Fig. S11A, B). These peaks were assigned to the closed genes, 284

therefore giving rise to 1096 and 1822 genes (Supplementary Data S1 and 2), 285

respectively, and were found with 563 overlapping genes that accounted for 51.8% in 286

YP1 and 30.9% in YP2 (Fig. 3B). 287

After we performed a Discriminative Regular Expression Motif Elicitation 288

(DREME) (Bailey, 2011) analysis of the filtered ChIP-seq sequences, an 289

OsGRF7-binding motif, ACRGDA (E-value = 2.6e-21), was predicted (Fig. 3C), 290

which was further validated by an electrophoretic mobility shift assay (EMSA) in 291

vitro (Fig. 3D, E). After analyzing these ChIP-seq identified genes with OsGRF7 292

binding sites individually, we identified 31 functionally known plant 293

architecture-related genes (Supplemental Table S3). These genes could be classified 294

into four categories: plant hormone-related genes, transcription factors, enzymes, and 295

others. Fifteen of them were related to plant hormones, especially OsARF12 and 296

OsCYP714B1, which showed direct regulation by OsGRF7, as verified by ChIP-seq 297

(Fig. 3F). ChIP-qPCR confirmed the in vivo association of OsGRF7 with 298 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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ACRGDA-containing promoter fragments from OsARF12 and OsCYP714B1 (Fig. 299

3G). Similarly, the abundances of mRNAs encoding OsARF12 and OsCYP714B1 300

were relatively enhanced in GRF7OE-1 transgenic lines (Fig. 3H, I). OsGRF7 301

activated the transcription of OsARF12 and OsCYP714B1 by their promoters in 302

transactivation assays (Fig. 3J, K). Finally, EMSA analysis demonstrated the binding 303

of OsGRF7 to ACRGDA-containing promoter fragments from OsARF12 and 304

OsCYP714B1 (Fig. 3L, M). These results demonstrate that these genes are the direct 305

targets of OsGRF7. 306

We further searched the promoter regions of the genes detected by ChIP-seq and 307

found that the promoter regions of IAA-related genes (OsARF3, OsARF4, OsARF8, 308

OsARF9, OsPIN1b, OsPIN1d, and OsPIN8) and GA-related genes (OsSLR1 and 309

OsSLRL1) contained several ACRGDA motifs. RT-qPCR comparisons of WT, 310

GRF7OE-1, GRF7RNAi-1, and GRF7KO-14 indicated that OsGRF7 up-regulated the 311

expression of these IAA/GA-related genes (Fig. 4A). Additionally, ChIP-qPCR also 312

confirmed the activation of OsGRF7 on these genes through the ACRGDA binding 313

site in vivo (Fig. 4B-J), as OsGRF7 greatly enhanced the expression of the luciferase 314

(LUC) reporter gene driven by these promoters (Fig. 4K, L). Taken together, these 315

results support the conclusion that OsGRF7 directly regulates auxin and gibberellin 316

signaling pathway-related genes. 317

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The oscyp714b1 and osarf12 mutant lines are partial phenotypic copies of 319

OsGRF7 transgenic lines 320

To further validate the functions of OsGRF7 regulated genes in plant architecture 321

determination, we collected two mutants with mutations in OsGRF7 direct target 322

genes, oscyp714b1 and osarf12. Of these, the oscyp714b1 mutant, which has a 323

T-DNA insertion in the promoter region of OsCYP714B1 (Supplemental Fig. S12A), 324

presented increased expression of OsCYP714B1 (Supplemental Fig. S12B). A 325

previous report indicated that OsCYP714B1 participates in GA synthesis and that high 326

expression of OsCYP714B1 represses the plant height by affecting cell elongation 327

(Magome et al., 2013). Here, we analyzed the plant height of oscyp714b1 and found 328 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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that it was reduced by approximately 17.9% relative to that of the WT (Supplemental 329

Fig. S12C, D), consistent with the reduced plant height of the GRF7OE lines (Fig. 330

1A). 331

For the osarf12 mutant, a T-DNA insertion in its third intron prevented its 332

transcription (Supplemental Fig. S12E, F). It exhibited an enlarged leaf angle, with 333

the second leaf angle from the top of the main stalk increasing by approximately 73% 334

compared with that of the WT (Supplemental Fig. S12G, H), which was partially 335

similar to that which occurred for the GRF7RNAi lines. These investigations showed 336

that the phenotype of each mutant was partially a copy of the plant architecture of the 337

OsGRF7 transgenic lines, demonstrating that these genes function downstream of 338

OsGRF7 to jointly control plant architecture in rice. 339

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OsGRF7 regulates the contents of endogenous GA and IAA 341

We also analyzed the contents of GA and indole-3-acetic acid (IAA) in 15-d-old 342

seedlings of GRF7OE-1, GRF7RNAi-1, and WT plants. The results showed that the 343

GA1 content was increased in GRF7OE-1 (Fig. 5A). Unfortunately, we did not detect 344

any content of GA4 at this stage (15-d-old seedlings), mainly because the GA1 was 345

predominantly abundant at the vegetative stage, while the level of GA4 was extremely 346

high at the reproductive stage (Kobayashi et al., 1988). In rice, both GA1 and GA4 are 347

bioactive forms derived from GA12, and GA4 is more active than GA1. However, their 348

contents usually exhibit an opposite pattern, and thus, an increase in GA1 leads to a 349

relatively low GA4 content; thus, the dynamic balance between GA1 and GA4 can 350

effectively modulate the plant height (Magome et al., 2013). 351

With respect to auxin, we found that the IAA and methyl indole-3-acetate 352

(ME-IAA) contents of GRF7OE-1 significantly increased (Fig. 5B), which is 353

consistent with the erect leaf angle of those plants (Zhang et al., 2015). 354

Correspondingly, immunohistochemical observations revealed that the adaxial side of 355

the GRF7OE-1 transgenic lines had more auxin than did the GRF7RNAi-1 and 356

GRF7KO-14 lines (Fig. 5C). Auxin is critical in regulating the adaxial/abaxial cell 357

growth of leaves, and the increased auxin content on the adaxial side of the lamina 358 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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joints in the GRF7OE-1 transgenic line promoted cell division, which is consistent 359

with the increased cell layers on the adaxial side of the GRF7OE-1 transgenic line 360

(Fig. 1F, G). These data reveal that OsGRF7 directly regulates the synthesis of 361

endogenous GA and IAA in rice. 362

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OsGRF7 regulates the sensitivity to exogenous IAA and GA treatment 364

We further treated OsGRF7 transgenic lines and investigated their responses to IAA 365

and GA. After a 5-d treatment with different concentrations of IAA, we found that the 366

primary root length of OsGRF7 transgenic lines decreased significantly under 367

different IAA concentrations, while the primary root length of the GRF7RNAi-1 and 368

WT seedlings decreased more than that of the GRF7OE-1 transgenic lines under 1 μM 369

IAA, meaning that the GRF7OE lines are less sensitive to auxin than the WT (Fig. 5D, 370

E). 371

GA treatment caused GRF7OE-1 seedlings to grow rapidly to the height of the 372

WT and GRF7RNAi-1 plants with increasing GA content, although the second sheath 373

length of GRF7OE-1 was shorter than that of the WT (Fig. 5D, F), meaning that GA 374

could rescue the semi-dwarf phenotype of the GRF7OE plants. This is in agreement 375

with the concept that high expression of OsCYP714B1 will lead to an increase in GA1 376

but a reduction in the content of highly active GA4 (Magome et al., 2013) because 377

OsCYP714B1 encodes GA13 oxidase, which converts GA12 to GA1, in rice. In 378

summary, these results demonstrate that GRF7OE lines are less sensitive to 379

exogenous phytohormone treatments. 380

381

DISCUSSION 382

GRFs, the targets of OsmiR396, are members of a conserved, plant-specific gene 383

family and are involved in many plant developmental processes, such as flowering, 384

leaf morphogenesis, root development and seed formation (Baucher et al., 2013; Liu 385

et al., 2009). Increasing numbers of studies have documented that GRFs positively 386

regulate leaf size by promoting cell proliferation and expansion in plants species such 387

as Arabidopsis and rice (Omidbakhshfard et al., 2015; Rodriguez et al., 2009). In 388 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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Arabidopsis, GRFs mainly participate in leaf and cotyledon growth, and cell division 389

mediated by AtGRFs is important for polarized cell differentiation along the 390

adaxial-abaxial axis during leaf morphogenesis (Wang et al., 2011). Overexpression 391

of AtGRF1 and AtGRF2 resulted in stalks and leaves that were shorter and larger than 392

those of the WT (Kim et al., 2003). In contrast, triple insertional null mutant of 393

AtGRF1-AtGRF3 had smaller leaves and cotyledons (Kim et al., 2003), and quadruple 394

mutant of AtGRF1-AtGRF4 displayed much smaller leaves than its parental mutants 395

(Kim and Lee, 2006), revealing that AtGRFs function mainly in plant height and leaf 396

development. Recently, OsGRF1, OsGRF3, OsGRF4, OsGRF6, and OsGRF10 have 397

been characterized to be involved in the regulation of plant development, especially in 398

grain and floret development (Che et al., 2015; Gao et al., 2015; Kuijt et al., 2014; 399

Luo et al., 2005; Tang et al., 2018), while their roles in plant architecture 400

determination are still poorly understood. The OsmiR396-OsGRF module balanced 401

growth and rice blast disease resistance, and overexpression of OsGRFs, including 402

OsGRF6, OsGRF7, OsGRF8, and OsGRF9, enhanced blast resistance. However, the 403

developmental roles of OsGRF7 have not been investigated in detail (Chandran et al., 404

2018). 405

Here, we found that OsGRF7, in combination with OsGIFs (Supplemental Fig. 406

S11C-E), modulated a wide range of rice plant morphological characteristics, 407

including plant height, leaf length and width, and leaf angle (Fig. 1; Supplemental 408

Table S1). This is in agreement with the results of the amino acid sequence-based 409

phylogenetic analysis showing that OsGRF7 was the closest homolog of AtGRF1 and 410

AtGRF2 (Supplemental Fig. S6). So far, 12 members of the OsGRF gene family have 411

been identified in rice (Choi et al., 2004). Previously OsGRF1, OsGRF4, OsGRF6, 412

and OsGRF10 were found to regulate plant height and leaf size (Che et al., 2015; Hu 413

et al., 2015; Li et al., 2018; Lu et al., 2020; Tang et al., 2018). We found that 414

overexpression of OsGRF7 resulted in decreased plant height (Supplemental Fig. S3D) 415

and altered leaf shape, suggesting that this gene promotes cell division and repress 416

cell elongation in the culm and lamina joint (Fig. 1C-F). Supporting this, plant height 417

increased in GRF7KO lines, however, GRF7RNAi transgenic lines did not simply 418 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



15

perform opposite to phenotypes of GRF7OE lines. The apparent conflict between 419

GRF7RNAi and GRF7OE lines might be explained by compensatory changes in 420

expression of other GRF genes when OsGRF7 is downregulated, as evidenced by 421

increased transcript levels of OsGRF4, OsGRF6, OsGRF8, and OsGRF9 422

(Supplemental Fig. S7). Alternatively, we cannot rule out the possibility that the 423

GRF7RNAi constructs had off-target effects, and/or that OsGRF7 negatively regulates 424

other GRF genes. 425

Interestingly, unlike AtGRFs in Arabidopsis, OsGRF7 was expressed mainly 426

around the vascular bundle of leaves and culms (Fig. 1 and Supplemental Fig. S8), 427

which promoted cell expansion and cell proliferation of parenchymal cells through 428

periclinal division, especially on the adaxial side of the lamina joints, leading to 429

reduced leaf angle and culm length in GRF7OE-1 transgenic lines, implying that 430

OsGRF7 plays a vital role in plant architecture establishment. These findings reflect 431

the functional diversification of structural orthologous genes in phylogenetically 432

distant species. 433

Plant hormones regulate many physiological processes that mainly influence 434

growth, differentiation, and development (Luo et al., 2016). Recently, miRNAs have 435

been found to modulate not only a wide range of agronomic characteristics but also 436

many phytohormone-related biological processes (Djami-Tchatchou et al., 2017; Tang 437

and Chu, 2017). These miRNAs function through their targets to modulate 438

downstream genes involved in plant hormone perception and signaling, receptor 439

proteins or even enzyme synthesis (Tang and Chu, 2017). Here, we found that 440

OsGRF7 can directly regulate a series of GA/IAA-related genes (Figs. 3 and 4), and 441

can fine tune the hormone balance of GAs and IAAs (Fig. 5A-C). Hence, they 442

coordinately regulate the development of internodes, leaves, and lamina joints, and 443

ultimately shape rice plant architecture (Fig. 6). A reduced auxin level causes an 444

enhanced leaf angle due to the stimulation of cell elongation and repression of the 445

division of parenchymal cells on the adaxial side, suggesting a negative effect of 446

auxin on leaf inclination (Zhao et al., 2013; Zhou et al., 2017). Indeed, a high auxin 447

level is detected at the lamina joint of GRF7OE-1 transgenic lines (Fig. 5B, C), which 448 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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is consistent with the increased expression of OsGRF7 in the lamina joints (Fig. 2D, 449

E). Combined with the cytological observations, our results showed that auxin is 450

synthesized and undergoes polar transport, resulting in a high local auxin 451

concentration on the adaxial side of the lamina joint, stimulating cell division but 452

repressing cell elongation in GRF7OE-1 transgenic lines (Fig. 5C). These findings 453

greatly expand our understanding of the involvement of the miR396-GRF module in 454

hormone balance, signaling, and plant architecture determination. 455

As such, short plant stature has been the main target for the improvement of 456

lodging resistance in rice breeding because it affords a lower risk to lodging (Ookawa 457

et al., 2010). Here, GRF7OE lines exhibited decreased plant height and sturdy stalks 458

resulting from decreased culm length (Supplemental Fig. S2) and increased culm wall 459

thickness (Fig. 1C and E). The size of the vascular bundles was significantly and 460

negatively correlated with the lodging index, and increasing the size of vascular 461

bundles can improve the lodging resistance in rice (Zhang et al., 2016). Cytological 462

observations revealed that the GRF7OE lines had relatively large vascular bundles in 463

the culms and leaves, and the parenchymal cells in the GRF7RNAi and GRF7KO 464

leaves degraded faster than did those in the WT and GRF7OE lines as the leaves 465

matured (Supplemental Fig. S8), meaning that the leaves and culms of the GRF7RNAi 466

and GRF7KO lines senesced earlier, and the parenchymal tissues became thinner as 467

the plant matured. Consistent with this idea, auxin was concentrated mainly at the 468

lamina joint and vascular bundles, and an apparent local auxin gradient from the 469

cortical tissue inward was observed (Fig. 5C), reflecting that auxin modulates the leaf 470

and culm development and senescence. An optimum level of auxin is required for cell 471

division, differentiation and elongation; vascular development; and organ patterning 472

(Woodward and Bartel, 2005). Auxin can also promote the vascular bundle 473

development to regulate leaf angle (Zhang et al., 2015). These cytological 474

characteristics in the GRF7RNAi and GRF7KO lines will cause the leaves to drop and 475

the plants to lodge when encountering catastrophic weather with strong wind and 476

heavy rain. In contrast, the high auxin content in the GRF7OE-1 transgenic lines 477

increased the vascular bundle size both in the lamina joints and in the internodes (Fig. 478 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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1C, F and Supplemental Fig. S8), resulting in erect leaf angles and sturdy stalks, 479

which can be used in modern breeding for enhancing rice lodging resistance. 480

The leaf angle of rice is an important agronomic trait that constitutes the 481

foundation of plant architecture. Increased leaf angles can expose leaf blades to more 482

light but decreased light capture for the canopy, which is adverse for dense planting. 483

In contrast, erect plant architecture is favored by crop breeders because it improves 484

photosynthetic efficiency and increases plant density, thus improving the 485

accumulation of leaf nitrogen for grain filling and increasing grain yield (Wang et al., 486

2018). The selection of novel genes controlling leaf angle and plant height is vital to 487

further improve rice plant architecture. Our characterization of the OsGRF7 gene 488

provides knowledge of the molecular basis of plant architecture control, and 489

utilization of OsGRF7 could be a favorable option for improving plant architecture 490

and increasing lodging resistance. 491

492

Methods and materials 493

Plant materials and growth conditions 494

Rice Yuetai B (YB) (Oryza sativa L. ssp. indica) was used as the transgenic receptor 495

in this study. The osarf12 and oscyp714b1 mutants (under Dongjin, a Japonica 496

accession) were obtained from the Rice Functional Genomics Express Database of 497

Korea (Jeong et al., 2002). MIM396-3 is the same as that in our previous study (Gao 498

et al., 2015). GRF7OE-1, GRF7OE-2, GRF7RNAi-1, and GRF7RNAi-2 transgenic 499

lines are the same as those in our previous study (Chandran et al., 2018). The T4 500

generation of the OsGRF7 overexpression and RNAi transgenic lines were used for 501

the experiments. All the rice plants used in this study were grown in either Wuhan 502

(May to October), or Hainan (December to April), China during 2013-2018. To 503

investigate the agronomic traits, ten plants were planted in each row, and the middle 504

five plants of each row were harvested to measure the agronomic traits. The 505

plant-to-plant spacing was 16.5 cm within each row and the row-to-row spacing was 506

26.7 cm. Field management practices essentially followed normal agricultural 507

practices. 508 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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509

Phenotype measurement 510

The plant height, effective panicle number, flag leaf length and width, and leaf angle 511

were measured at the maturity stage (more than 95% of grains were matured). The 512

plant height was measured from the lowest node to the flag leaf of the main tiller, and 513

the effective panicle number was counted with the panicle bearing more than 10 fully 514

developed seeds. The flag leaf of the main tiller was used to measure the flag leaf 515

length and width, and the leaf sheath was imaged and measured with Image J 516

software. 517

518

Vector construction and plant transformation 519

The 1.3 kb full-length cDNA of OsGRF7 was amplified from the young inflorescence 520

of YB and recombined into the overexpression vector pH7WG2D (Invitrogen) derived 521

by CAMV 35S promoter. To generate GRF7RNAi transgenic plants, 316 bp cDNA 522

(661 bp to 977 bp) was inserted into a pANIC8A (Arabidopsis Biological Resource 523

Center) vector. For the construction of GRF7-GFP, full-length cDNA of OsGRF7 was 524

recombined into the plant expression vector pGWB5 (Invitrogen). A ~2 kb promoter 525

fragment of OsGRF7 was cloned into the pGWB3 vector (Invitrogen) to create the 526

GRF7pro:GUS reporter gene construct. These vectors were introduced into the 527

Agrobacterium tumefaciens strain EHA105 and transformed into YB. Primers used in 528

this study were listed in Supplemental Data S3. 529

530

Generation of the OsGRF7 knock-out mutant using the CRISPR/Cas9 system 531

The CRISPR/Cas9 knockout plasmid for OsGRF7 was constructed as previously 532

reported (Ma et al., 2015). We generated two sgRNA constructs (designed in the 533

second exon, Supplemental Fig. S5A), in which the sgRNA was driven by the rice U6 534

promoter, and the plant-optimized Cas9 was driven by the Ubi promoter. The 535

integrated sgRNA expression cassettes were amplified and cloned into the 536

CRISPR/Cas9 vector pYLCRISPR/Cas9Pubi-H (Ma et al., 2015). The construct was 537

introduced into the wild-type variety YB. Then the 74 independent transgenic lines 538 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



19

were subjected to PCR amplification and sequencing analysis. The T2 generation of 539

GRF7KO lines were used for the experiments. The genotype of GRF7KO lines was 540

listed in Supplemental Table S2. 541

542

Histological analysis 543

Plant tissues were fixed in FAA solution (3.7% (v/v) formaldehyde, 50% (v/v) ethanol, 544

and 5% (v/v) acetic acid) and embedded in paraplast plus (Sigma-Aldrich). 8 μm thick 545

sections were stained with toluidine blue for light microscopic analysis. Five 546

independent sections were microscopically examined and photographed to measure 547

the cell layers, culm thickness, and cell length; ten independent sections were 548

microscopically examined and photographed to measure the area of the large vascular 549

bundles (Leica). 550

551

5ꞌ RACE analysis 552

Total RNA from YB young inflorescence was isolated with TRIzol reagent 553

(Invitrogen), then ligated to RNA adaptors and synthesized into cDNA using 554

Superscript III according to the manufacturer’s instructions (Invitrogen). 5ꞌ RACE 555

was performed with the GeneRace kit (Invitrogen). 556

557

Transient expression, protein extraction, and immunoblotting 558

CDS of OsrGRF7 was synthesized and ligated downstream of the 35S promoter. For 559

OsmiR396/Os(r)GRF7 coexpression experiments, an equal amount of plasmids (3 μg) 560

containing Ubi:miR396s and 35S:(r)GRF7 were mixed before transformed into rice 561

protoplasts. After transformation, protoplasts were placed at 28 °C for 24 h before 562

RNA extraction. The OsmiR396 precursors were ligated downstream of the ubiquitin 563

promoter with restriction site BamHI in the pCAMBIA1301 vector. Protoplasts 564

isolated from etiolated rice seedlings of GRF7-GFP transgenic lines were transfected 565

with 3 μg Ubi:miR396s, or empty vector plasmids by the 40% (w/v) 566

polyethyleneglycol (PEG), respectively. After incubation for 16 h, the OsGRF7 567 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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protein was detected by western blot analysis using an anti-GFP antibody. Total RNA 568

was extracted to detect the transcript levels of OsmiR396s. 569

570

Total RNA isolation and RT-qPCR 571

Total RNA (2 μg) was extracted and synthesized into cDNA using the Moloney 572

murine leukemia virus reverse transcriptase (Invitrogen) according to the user’s 573

manual. RT-qPCR was performed by Light Cycler 480 II (Roche, Germany) with 574

SYBR Green PCR Mixture (Roche, Germany) according to the manufacturer’s 575

instruction. The OsUBI gene was used as the internal control. Three technical 576

replicates were performed to generate the average value and three biological repeats 577

were performed for each analysis. 578

579

Histochemical GUS staining 580

Different tissues were stained with GUS staining buffer (100 mM sodium phosphate, 581

10 mM EDTA, 0.1% (v/v) Triton X-100, and 1 mM X-Gluc 582

(5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid) (Sigma), pH = 7.0) overnight at 583

37 °C. All samples were observed with an SLR camera (Nikon SMZ-645). Two 584

independent lines were analyzed for GUS staining. 585

586

Subcellular localization and bimolecular fluorescence complementation (BiFC) 587

For subcellular localization, the full-length of OsGRF7 cDNA was cloned into 588

pGWB5 (Invitrogen). For BiFC, the full-length of OsGRF7 cDNA was amplified with 589

restriction sites XbalI and ClaI and cloned into the pSPYNE (N-terminal of YFP), and 590

the full-length of OsGRF7, OsGIF1, OsGIF2, and OsGIF3 cDNA were amplified 591

with restriction sites XbalI and ClaI and inserted into the pSPYCE (C-terminal of YFP) 592

(Walter et al., 2004). Rice protoplasts were isolated as described previously (Yoo et al., 593

2007), with some modifications. After overnight incubation in the darkness, the 594

fluorescent signals and bright-field images of the protoplasts were taken by an 595

FV1000 confocal system (Olympus, Tokyo, Japan). Empty vectors of BiFC constructs 596

were used as a negative control. 597 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



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598

Transactivation activation assay 599

To test the transactivation activity, the full-length or the truncated OsGRF7 with 600

restriction sites BamHI and EcoRI were amplified and cloned into pGBKT7 601

(Clontech). The plasmids were transformed into yeast strain AH109, then 602

transformants were plated on either SD/-Trp or SD/-Trp-His-Leu-Ade medium. 603

604

Yeast two-hybrid assay 605

The full-length cDNA of OsGIF1, OsGIF2, OsGIF3, and OsGRF7 with restriction 606

sites EcoRI and BamHI were cloned into the prey vector pGADT7 (Clontech), and the 607

full-length or truncated cDNA of OsGRF7 with restriction sites BamHI and EcoRI 608

was cloned into the pGBKT7 (Clontech). The prey and bait plasmids were 609

co-transformed into AH109 and plated on SD/-Leu-Trp media for 3 days at 30 °C. 610

Interactions between bait and prey were further tested on SD/-Trp-His-Leu-Ade 611

medium. 612

613

ChIP-seq and bioinformatics analysis 614

The transgenic lines GRF7-GFP were used for ChIP assays according to the method 615

(Feng et al., 2012) with some modifications. Briefly, 5 g < 0.5 cm (YP1) or < 2 cm 616

(YP2) inflorescences were harvested and crosslinked with 1% (v/v) formaldehyde 617

under vacuum for 30 min, then ground into powder in liquid nitrogen. The chromatin 618

complexes were isolated, sonicated, and incubated with the anti-GFP antibodies 619

(Abcam, ab290). The precipitated DNA was recovered and dissolved in 1×TE for later 620

in-depth analysis. 621

Illumina sequencing libraries were constructed with the above-prepared DNA 622

samples by ThruPLEX DNA-seq Kit mainly according to the manufacturer’s 623

instructions. Then, the library containing 270 bp to 330 bp fragments were purified 624

and sequenced with Illumina Hiseq3000 system. Clean reads were mapped to the rice 625

genome using Bowtie (Langmead et al., 2009). MACS (Liu, 2014) was used with a 626

p-value cutoff of 10−5

for binding peaking. The reads per million (RPM) around the 627 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



22

peak summit (± 5 kb) was documented with a 100-nt window size to calculate the 628

average enrichment level of the peak. To predict binding motifs, the flanking 629

sequences of all peak summits (± 50 bp) were filtered by RepeatMasker Web Server 630

(http://www.repeatmasker.org) and the masked sequences were subjected to 631

Discriminative Regular Expression Motif Elicitation (DREME) (Bailey, 2011). 632

633

ChIP-qPCR 634

The prepared DNA in ChIP was quantified using quantitative PCR with their primers. 635

PCR reactions were performed in triplicate for each sample, and the values were 636

normalized to the input sample to obtain the enrichment fold. The fold enrichment 637

was calculated against the OsUBI promoter. No addition of antibodies (NoAbs) was 638

served as a negative control. Three biological replicates were performed for each 639

analysis. 640

641

Transient transactivation assay 642

Approximately 2-kb promoter regions from each of OsARF3, OsARF8, OsPIN1b, 643

OsPIN1d, OsPIN8, and OsSLR1 with restriction sites XhoI and BamHI; OsARF9 and 644

OsSLRL1 with restriction sites XhoI and BglII; OsARF4, OsARF12, and 645

OsCYP714B1 with restriction sites HindIII and BamHI were amplified from YB, and 646

then cloned into a pGreenII-0800-Luc (EK-Bioscience) vector containing the renilla 647

luciferase reporter gene driven by the 35S promoter and the firefly luciferase reporter 648

gene, thus generating the vectors containing specific promoters fused to firefly 649

luciferase. Transient transactivation assays were performed using rice protoplasts, and 650

the Dual-Luciferase Reporter Assay System (Promega) was used to detect the 651

luciferase activity, with the renilla luciferase gene as the internal control. Three 652

biological replicates were performed for each analysis. 653

654

Expression and purification of His-GRF7 fusion proteins in Escherichia coli 655

The full-length cDNA fragment was amplified with restriction sites BamHI and 656

EcoRI and fused into expression vector pCOLD. To extract recombinant protein, 657 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



23

Rosetta (DE3) cells carrying pCOLD-GRF7 plasmid were inoculated at 37 °C to an 658

optical density. Then IPTG (isopropyl-ß-D-thiogalactoside) was added to a final 659

concentration of 0.2 mM, and cultures were grown for 24 h at 15 °C. Cultured cells 660

were harvested, lysed, and centrifuged, then the supernatant was purified by 661

His-Binding-resin (GE Healthcare) using AKTA Prime Plus protein purification 662

system (GE Healthcare). 663

664

Electrophoretic mobility shift assay (EMSA) 665

Oligonucleotides were synthesized and labeled by digoxin. Promoter regions were 666

amplified, labeled by digoxin and purified as the probe, respectively. Then, 200 ng 667

DNA probe, 2 μg of protein (His-GRF7) and 2 μl of 10×binding buffer (100 mM Tris, 668

500 mM KCl, and 10 mM dithiothreitol, pH = 7.5), 1 μl of 50% (v/v) glycerol, 1 μl of 669

1% (v/v) Nonidet P-40, 1 μl of 1 M KCl, 1 μl of 0.5 M EDTA, 1 μl of 1 mg ml-1

poly 670

(dI-dC), and double-distilled water were mixed to a final volume of 20 μl (Liu et al., 671

2014), and reacted for 20 min at 25 °C. Further, electrophoresed on 3.5% (w/v) native 672

polyacrylamide gels and then transferred to N+ nylon membranes (Millipore) in TBE 673

buffer (44.5 mM Tris-borate, 1 mM EDTA) at 200 mA at 4 °C for 1 h. 674

Digoxin-labeled DNA was detected by the CDP-star easy to use kit (Roche). 675

676

Measuring endogenous phytohormones 677

Contents of IAA and GA were analyzed using an LC-ESI-MS/MS system. To detect 678

endogenous GA and IAA, 15 seeds of the OsGRF7 transgenic lines were planted in a 679

container filled with soil with 2 × 2 cm spacing in a phytotron. Fresh 15-d-old 680

seedlings were harvested, weighed and then immediately ground into powder in liquid 681

nitrogen. Since tissue close to the soil may be contaminated, approximately 0.5 cm of 682

tissue near soil was removed with scissors. After being extracted with 1 ml of 80% 683

(v/v) methanol at 4 °C for 12 h, the extract was centrifuged at 12,000 g under 4 °C for 684

15 min. The supernatant was collected and evaporated to dryness under nitrogen gas 685

stream, then reconstituted in 100 ml 95% (v/v) acetonitrile. The supernatant was 686

collected for LC-MS analysis after the solution was centrifuged. Each series of 687 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



24

experiments was performed in biological triplicates. 688

689

Immunohistochemical observation (IHC) of IAA 690

To cross-link IAA, excised lamina joints were prefixed for 2 h in 3% (w/v) EDAC 691

(Sigma-Aldrich) at room temperature and then transferred to FAA at 4 °C for 24 h. 8 692

μm thick sections were deparaffinized and hydrated. Antigen retrieval was carried out 693

by rinsing the slides in 0.1 M sodium citrate buffer and microwave-heated at high 694

power for 5 min. After being washed three times with 10 mM PBS buffer for 10 min, 695

the slides were subsequently incubated in 10 mM PBS containing 0.1% (v/v) Tween 696

20, 1.5% (v/v) glycine, and 5% (w/v) BSA for 45 min at 22 °C. Samples were then 697

rinsed in a regular salt rinse solution [RSRS; 10 mM PBS, 0.88% (w/v) NaCl, 0.1% 698

(v/v) Tween 20 and 0.8% (w/v) BSA] and washed briefly with 10 mM PBS 699

containing 0.8% (w/v) BSA (PBS+BSA) solution. After the application of anti-IAA 700

antibodies to each slide, samples were incubated overnight in a humidity chamber at 701

4 °C. After hybridization, samples were subjected to a series of vigorous washes, 702

twice with a high-salt rinse solution [HSRS; 10 mM PBS, 2.9% (w/v) NaCl, 0.1% 703

(v/v) Tween 20 and 0.1% (w/v) BSA] for 15 min, once with RSRS for 15 min, and 704

briefly with PBS+BSA. The Alexa 488-conjugated goat anti-mouse IgG antibodies 705

were then placed on each slide, and these were incubated for 4–6 h in a humidity 706

chamber at room temperature. After washing with RSRS twice for 15 min, samples 707

were mounted with an anti-fade reagent, covered with a coverslip, and observed under 708

a fluorescent microscope (Sakata et al., 2010). 709

710

Plant hormone treatment 711

For hormone treatment, de-hulled seeds were sowed on the 1/2 MS agar medium 712

containing different phytohormones. After a 5-d growth, the length of the root and the 713

second sheath were analyzed using ImageJ software (NIH). Three biological 714

replicates were performed and five plants for each replicate were collected for data 715

analysis. 716




25

Phylogenetic analysis 718

For phylogenetic analysis of OsGRF7 homologs in Oryza sativa and Arabidopsis, a 719

total of 12 rice GRF protein sequence and 9 Arabidopsis GRF protein sequence were 720

obtained in PLAZA 2.5 (Van Bel et al., 2012). The amino acid sequences were aligned 721

using the CLUSTAL X program. The phylogenetic tree was constructed using MEGA 722

X (Kumar et al., 2018) based on the Maximum Likelihood method with 1,000 723

bootstrap replicates. 724

725

Statistical analysis 726

Statistical analyses were performed using IBM SPSS statistics, version 20.0 (IBM, 727

Armonk, NY, USA). The two-tailed t-test was used for comparing the agronomic 728

traits of each transgenic line with the WT. The correlation analysis was performed 729

with GraphPad Prism 8 to generate the r and P values. 730

731

Accession numbers 732

Sequence from this article can be found in the GeneBank / EMBL databases under the 733

following accession numbers: OsGRF7, Os12g0484900; OsGIF1, Os03g0733600; 734

OsGIF2, Os11g0615200; OsGIF3, Os12g0496900; OsCYP714B1, Os07g0681300; 735

OsARF12, Os04g0671900; OsUBI, Os03g0234200. High-throughput sequencing data 736

created in this study have been deposited in the Gene Expression Omnibus database 737

(GSE109802). 738

739

Supplemental Data 740

Supplemental Figure S1. Phenotypic and molecular analysis of miR396 mimicry 741

(MIM396) transgenic lines. 742

Supplemental Figure S2. Comparison of internode length between WT and OsGRF7 743

transgenic lines. 744

Supplemental Figure S3. The relationship between OsGRF7 expression levels and 745

phenotypic traits of the OsGRF7 transgenic lines. 746

Supplemental Figure S4. Genetic analysis between OsmiR396 and OsGRF7. 747 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



26

Supplemental Figure S5. Phenotype and molecular characterization of OsGRF7 748

knock-out lines. 749

Supplemental Figure S6. Phylogenetic analysis of GRFs in Arabidopsis and rice. 750

Supplemental Figure S7. Expression analysis of OsGRF members in the OsGRF7 751

transgenic lines. 752

Supplemental Figure S8. Cytological dynamics of lamina joint from the initiation to 753

maturation. 754

Supplemental Figure S9. Regulation of OsGRF7 by OsmiR396s. 755

Supplemental Figure S10. The transcription levels of the OsmiR396 family in 756

GRF7-GFP transgenic rice protoplasts were analyzed with RT-PCR. 757

Supplemental Figure S11. Analysis of interactions between OsGRF7 and OsGIFs. 758

Supplemental Figure S12. Architecture and molecular identification of oscyp714b1 759

and osarf12 mutants. 760

Supplemental Table S1. Agronomic traits of OsGRF7 transgenic lines 761

Supplemental Table S2. Genotype analysis of GRF7KO transgenic lines 762

Supplemental Table S3. Functional categories of genes associated with OsGRF7 763

binding sites 764

Supplemental Dataset S1. Genes associated with OsGRF7 binding sites identified by 765

ChIP-seq in YP1 (< 0.5 cm). 766

Supplemental Dataset S2. Genes associated with OsGRF7 binding sites identified by 767

ChIP-seq in YP2 (< 2 cm). 768

Supplemental Dataset S3. Primers used in this paper. 769

770

Acknowledgements 771

We thank Dr. Kun Wang and Zhenying Shi for their critical reading and advice during 772

the preparation of this paper. We also thank Dr. Ai Qin for providing assistance in the 773

bioinformatic analysis. 774

775

Figure legends 776 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



27

Figure 1. Phenotypic characterization. A, Gross morphology of WT, OsGRF7 777

overexpression (GRF7OE-1) and OsGRF7 RNAi (GRF7RNAi-1) plants. Scale bars, 778

15 cm. B, Comparison of the flag leaf angles (up-panel) and the top second leaf angles 779

(down-panel) between OsGRF7 transgenic lines. Scale bars, 1 cm. C, Transverse 780

(up-panel) and longitudinal (down-panel) section of internodes in WT, GRF7OE-1, 781

and GRF7RNAi-1 plants. Scale bars, 200 μm. D, Cell length of the internode. E, Culm 782

wall thickness of OsGRF7 transgenic lines. F, Transverse (up-panel) and longitudinal 783

(down-panel) section of the lamina joint of flag leaves. Scale bars, 200 μm. Red boxes 784

areas in up-panel are enlarged. Red arrows mean adaxial side cell layers measured in 785

the panel G. pc, parenchymal cell; vb, vascular bundle; ad, adaxial surface of the 786

lamina joint. G, Adaxial side cell layers of the transverse sections of lamina joints. 787

Panels D, E, and G: Values are means ± SD of five biological replicates. *** P < 788

0.001 (two-tailed Student’s t-test). 789

790

Figure 2. OsGRF7 is mainly repressed by OsmiR396e and expresses in various 791

tissues. A, The OsGRF7 cleavage site sequence complementary to OsmiR396. The 792

position corresponding to the 5′ ends of the cleaved OsGRF7 mRNA determined by 5′ 793

RACE and the frequency of 5′ RACE clones corresponding to the cleavage site is 794

shown by the arrow. 8/12 means eight of twelve clones have OsmiR396 cleavage site. 795

The mutated sites in OsrGRF7 were marked in red. The asterisks indicate 796

mis-matched sites between OsmiR396 and OsGRF7. The gray box indicates the QLQ 797

domain, the pink box indicates the WRC domain, and the purple box indicates the C 798

terminal of OsGRF7, respectively. B, Protein immunoblotting of OsGRF7 799

coexpressed with OsmiR396s in GRF7-GFP transgenic rice protoplasts. Vector and 800

control mean with or without the empty vector, respectively. Coomassie brilliant blue 801

(CBB) staining was used as loading controls. The relative protein amounts were 802

determined by ImageJ (NIH). C, Relative expression levels of OsGRF7 and 803

OsmiR396s were detected by RT-qPCR in internode, root, inflorescence (10 cm), 804

node, flag leaf, SAM (shoot apical meristem), axillary bud, and lamina joint, 805

respectively. D, GRF7pro:GUS expression patterns in transgenic plants. (1) Small 806 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



28

panicle branch. (2-4) Inflorescence at different stages. (5-6) Spikelet. (7) Root tip. (8) 807

Flag leaf. (9) Node. (10) Internode. (11) Axillary bud. (12) Lamina joint of the 30-d 808

seedling. Scale bars, 2 mm. E, Relative expression levels of OsGRF7 in different 809

tissues analyzed with RT-qPCR. OsUBI was used as an internal reference. Values are 810

means ± SD of three biological replicates. 811

812

Figure 3. ChIP analysis of the OsGRF7 target genes. A, Distribution of OsGRF7 813

binding peaks in the rice genome. B, Genes reproducibly associated with OsGRF7 814

binding sites in YP1 (young inflorescences with a length of ~0.5 cm) or YP2 (young 815

inflorescences with a length of ~2 cm). C, Putative OsGRF7-binding motif predicted 816

by DREME. D, Expression and purification of OsGRF7 protein in E. coli. The red 817

arrowhead indicates the purified OsGRF7 protein. E, EMSA validation of interaction 818

between OsGRF7 and motif ACRGDA (ACAGTA). TAAGTA was used as the control. 819

F, OsGRF7 binding peaks in the promoters of OsCYP714B1 and OsARF12. Blue 820

boxes indicate probe locations. Bar = 1 kb. G, ChIP-qPCR validation of OsGRF7 821

binding sites in the promoters of OsCYP714B1 and OsARF12. The fold enrichment 822

was normalized against the promoter of OsUBI. No addition of antibodies (NoAbs) 823

served as a negative control. H and I, Relative expression levels of the OsCYP714B1 824

and OsARF12 in the lamina joint of OsGRF7 transgenic lines were detected with 825

RT-qPCR. OsUBI was used as an internal reference. J and K, OsGRF7 actives 826

OsCYP714B1 and OsARF12 promoter-luciferase fusion constructs in transient 827

transactivation assays. L and M, EMSA validation of bindings between OsGRF7 and 828

promoters of OsCYP714B1 and OsARF12, respectively. The 2-, 10- and 100- fold 829

unmodified probes were used as competitors. The presence (+) or absence (-) of 830

components in the reaction mixture was indicated. Red triangles indicate the site of 831

ACRGDA motifs on promoter region. Panels G-K: Values are means ± SD of three 832

biological replicates. * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed Student’s 833

t-test). 834

835

Figure 4. OsGRF7 regulates expression of multiple IAA/GA-related genes. A, 836 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



29

Relative expression levels of auxin signaling related and GA synthesis related genes 837

in the lamina joint of the OsGRF7 transgenic lines. OsUBI was used as an internal 838

reference. B to J, GRF7-GFP mediated ChIP-qPCR enrichment (relative to the 839

promoter region of OsUBI) of ACRGDA-containing fragments (marked with 840

arrowhead) from promoters of auxin signaling related genes (ARF3, ARF4, ARF8, 841

ARF9, PIN1b, PIN1d, and PIN8) and GA synthesis related genes (SLR1 and SLRL1). 842

The promoter region of OsUBI was used as a control. K and L, OsGRF7 actives ARF3, 843

ARF4, ARF8, ARF9, PIN1b, PIN1d, PIN8, SLR1, and SLRL1 promoter-luciferase 844

fusion constructs in transient transactivation assays. Panels B-J, L: Values are means 845

± SD of three biological replicates. * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed 846

Student’s t-test). 847

848

Figure 5. Endogenous phytohormone contents and exogenous phytohormone 849

sensitivity test of OsGRF7 transgenic lines. A and B, Quantification of GA (A) and 850

IAA (B) derivatives in OsGRF7 transgenic seedlings detected with LC-MS/MS. 851

Values are means ± SD of three biological replicates. * P < 0.05 (two-tailed Student’s 852

t-test). Abbreviations: ME-IAA, methyl indole-3-acetate; ICA, 853

indole-3-carboxaldehyde; IAA, indole-3-acetic acid; NAA, 1-naphthylacetic acid. 854

F.W., fresh weight. nd indicate not detected (below the detection limit). C, 855

Immunohistochemical observation (IHC) of indole-3-acetic acid at the lamina joint of 856

OsGRF7 transgenic lines. Anti-IAA antibody and Alexa 488-conjugated goat 857

anti-rabbit IgG antibody were used. Scale bars, 100 μm. D, Phenotypes of OsGRF7 858

transgenic seedlings treated by different concentrations of phytohormones. The same 859

control has been used for 0 μM IAA and GA. Scale bars, 2 cm. E, Effects of different 860

concentrations of IAA on primary root length in WT, GRF7OE-1, and GRF7RNAi-1. 861

Values are means ± SD (n = 15, three biological replicates and five plants per 862

replicate). F, Effects of different concentrations of GA3 on the second sheath length in 863

WT, GRF7OE-1, and GRF7RNAi-1 seedlings. Values are means ± SD (n = 15, three 864

biological replicates and five plants per replicate). Different letters denote significant 865

differences (P < 0.05) from Duncan’s multiple range tests. 866 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



30

867

Figure 6. A proposed model for the regulation of rice plant architecture by OsGRF7. 868

In GRF7OE plants, OsGRF7 co-activates with OsGIFs to up-regulate the expressions 869

of downstream hormone-related genes through binding to ACRGDA motifs. Then the 870

synthesis of GA4 is inhibited, and the synthesis of IAA is promoted. Finally, the 871

GRF7OE plants display a semi-dwarf and compact plant architecture. Abbreviations: 872

IAA, indole-3-acetic acid; GA, gibberellin. 873

874

References 875

Bailey T L. (2011). DREME: motif discovery in transcription factor ChIP-seq data. Bioinformatics 876

27:1653-1659. 877

Baucher M, Moussawi J, Vandeputte O M, Monteyne D, Mol A, Perez-Morga D, and El Jaziri M. 878

(2013). A role for the miR396/GRF network in specification of organ type during flower 879

development, as supported by ectopic expression of Populus trichocarpa miR396c in transgenic 880

tobacco. Plant Biol. 15:892-898. 881

Chandran V, Wang H, Gao F, Cao X L, Chen Y P, Li G B, Zhu Y, Yang X M, Zhang L L, Zhao Z 882

X, et al. (2018). miR396-OsGRFs module balances growth and rice blast disease-resistance. Front. 883

Plant Sci. 9:1999. 884

Che R, Tong H, Shi B, Liu Y, Fang S, Liu D, Xiao Y, Hu B, Liu L, Wang H, et al. (2015). Control 885

of grain size and rice yield by GL2-mediated brassinosteroid responses. Nat. Plants 2:15195. 886

Chen X, Lu S, Wang Y, Zhang X, Lv B, Luo L, Xi D, Shen J, Ma H, and Ming F. (2015). OsNAC2 887

encoding a NAC transcription factor that affects plant height through mediating the gibberellic acid 888

pathway in rice. Plant J. 82:302-314. 889

Choi D, Kim J H, and Kende H. (2004). Whole genome analysis of the OsGRF gene family encoding 890

plant-specific putative transcription activators in rice (Oryza sativa L.). Plant and Cell Physiology 891

45:897-904. 892

Djami-Tchatchou A T, Sanan-Mishra N, Ntushelo K, and Dubery I A. (2017). Functional roles of 893

microRNAs in agronomically important plants-potential as targets for crop improvement and 894

protection. Front. Plant Sci. 8:378. 895 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



31

Ercoli M F, Ferela A, Debernardi J M, Perrone A P, Rodriguez R E, and Palatnik J F. (2018). GIF 896

transcriptional co-regulators control root meristem homeostasis. Plant Cell 30:347-359. 897

Feng J, Liu T, Qin B, Zhang Y, and Liu X S. (2012). Identifying ChIP-seq enrichment using MACS. 898

Nat. Protoc. 7:1728-1740. 899

Gao F, Wang K, Liu Y, Chen Y, Chen P, Shi Z, Luo J, Jiang D, Fan F, Zhu Y, et al. (2015). 900

Blocking miR396 increases rice yield by shaping inflorescence architecture. Nat. Plants 2:15196. 901

Gao S, Fang J, Xu F, Wang W, and Chu C. (2016). Rice HOX12 regulates panicle exsertion by 902

directly modulating the expression of ELONGATED UPPERMOST INTERNODE1. Plant Cell 903

28:680-695. 904

Hu J, Wang Y, Fang Y, Zeng L, Xu J, Yu H, Shi Z, Pan J, Zhang D, Kang S, et al. (2015). A rare 905

allele of GS2 enhances grain size and grain yield in rice. Mol. Plant 8:1455-1465. 906

Jeong D H, An S, Kang H G, Moon S, Han J J, Park S, Lee H S, An K, and An G. (2002). T-DNA 907

insertional mutagenesis for activation tagging in rice. Plant Physiol. 130:1636-1644. 908

Khush G S. (1995). Breaking the yield frontier of rice. GeoJournal 35:329–332. 909

Kim J H, Choi D, and Kende H. (2003). The AtGRF family of putative transcriptioon factors is 910

involved in leaf and cotyledon growth in Arabidopsis. Plant J. 36:94-104. 911

Kim J H, and Kende H. (2004). A transcriptional coactivator, AtGIF1, is involved in regulating leaf 912

growth and morphology in Arabidopsis. Proc. Natl. Acad. Sci. USA 101:13374-13379. 913

Kim J H, and Lee B H. (2006). GROWTH-REGULATING FACTOR4 of Arabidopsis thaliana is 914

required for development of leaves, cotyledons, and shoot apical meristem. J. Plant Biol. 915

49:463-468. 916

Kobayashi M, Yamaguchi I, Murofushi N, Ota Y, and Takahashi N. (1988). Fluctuation and 917

localization of endogenous gibberellins in rice. Agricultural and Biological Chemistry 918

52:1189-1194. 919

Kozomara A, and Griffiths-Jones S. (2014). miRBase: annotating high confidence microRNAs using 920

deep sequencing data. Nucleic Acids Res. 42:D68-73. 921

Kuijt S J, Greco R, Agalou A, Shao J, t Hoen C C, Overnas E, Osnato M, Curiale S, Meynard D, 922

van Gulik R, et al. (2014). Interaction between the GROWTH-REGULATING FACTOR and 923

KNOTTED1-LIKE HOMEOBOX families of transcription factors. Plant Physiol. 164:1952-1966. 924

Kumar S, Stecher G, Li M, Knyaz C, and Tamura K. (2018). MEGA X: molecular evolutionary 925 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



32

genetics analysis across computing platforms. Mol Biol Evol 35:1547-1549. 926

Langmead B, Trapnell C, Pop M, and Salzberg S L. (2009). Ultrafast and memory-efficient 927

alignment of short DNA sequences to the human genome. Genome Biol. 10:R25. 928

Lavy M, and Estelle M. (2016). Mechanisms of auxin signaling. Development 143:3226-3229. 929

Li S, Gao F, Xie K, Zeng X, Cao Y, Zeng J, He Z, Ren Y, Li W, Deng Q, et al. (2016). The 930

OsmiR396c-OsGRF4-OsGIF1 regulatory module determines grain size and yield in rice. Plant 931

Biotechnol. J. 14:2134-2146. 932

Li S, Tian Y, Wu K, Ye Y, Yu J, Zhang J, Liu Q, Hu M, Li H, Tong Y, et al. (2018). Modulating 933

plant growth-metabolism coordination for sustainable agriculture. Nature 560:595-600. 934

Li Y, Li J, Chen Z, Wei Y, Qi Y, and Wu C. (2020). OsmiR167a-targeted auxin response factors 935

modulate tiller angle via fine-tuning auxin distribution in rice. Plant Biotechnol. J. 936

10.1111/pbi.13360. 937

Liu D, Song Y, Chen Z, and Yu D. (2009). Ectopic expression of miR396 suppresses GRF target gene 938

expression and alters leaf growth in Arabidopsis. Physiol. Plant. 136:223-236. 939

Liu H, Guo S, Xu Y, Li C, Zhang Z, Zhang D, Xu S, Zhang C, and Chong K. (2014). 940

OsmiR396d-regulated OsGRFs function in floral organogenesis in rice through binding to their 941

targets OsJMJ706 and OsCR4. Plant Physiol. 165:160-174. 942

Liu T. (2014). Use Model-Based Analysis of ChIP-seq (MACS) to analyze short reads generated by 943

sequencing protein-DNA interactions in embryonic stem cells. Methods Mol. Biol. 1150:81-95. 944

Lu Y, Meng Y, Zeng J, Luo Y, Feng Z, Bian L, and Gao S. (2020). Coordination between 945

GROWTH-REGULATING FACTOR1 and GRF-INTERACTING FACTOR1 plays a key role in 946

regulating leaf growth in rice. BMC Plant Biol. 20:200. 947

Luo A D, Liu L, Tang Z S, Bai X Q, Cao S Y, and Chu C C. (2005). Down-regulation of OsGRF1 948

gene in rice rhd1 mutant results in reduced heading date. J. Integr. Plant Biol. 47:745-752. 949

Luo X, Zheng J, Huang R, Huang Y, Wang H, Jiang L, and Fang X. (2016). Phytohormones 950

signaling and crosstalk regulating leaf angle in rice. Plant Cell Rep. 35:2423-2433. 951

Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, et al. (2015). A 952

robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot 953

and dicot plants. Mol. Plant 8:1274-1284. 954

Magome H, Nomura T, Hanada A, Takeda-Kamiya N, Ohnishi T, Shinma Y, Katsumata T, 955 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



33

Kawaide H, Kamiya Y, and Yamaguchi S. (2013). CYP714B1 and CYP714B2 encode gibberellin 956

13-oxidases that reduce gibberellin activity in rice. Proc. Natl. Acad. Sci. USA 110:1947-1952. 957

Omidbakhshfard M A, Proost S, Fujikura U, and Mueller-Roeber B. (2015). Growth-Regulating 958

Factors (GRFs): a small transcription factor family with important functions in plant biology. Mol. 959

Plant 8:998-1010. 960

Ookawa T, Hobo T, Yano M, Murata K, Ando T, Miura H, Asano K, Ochiai Y, Ikeda M, Nishitani 961

R, et al. (2010). New approach for rice improvement using a pleiotropic QTL gene for lodging 962

resistance and yield. Nat. Commun. 1:132. 963

Rodriguez R E, Mecchia M A, Debernardi J M, Schommer C, Weigel D, and Palatnik J F. (2009). 964

Control of cell proliferation in Arabidopsis thaliana by microRNA miR396. Development 965

137:103-112. 966

Sakata T, Oshino T, Miura S, Tomabechi M, Tsunaga Y, Higashitani N, Miyazawa Y, Takahashi H, 967

Watanabe M, and Higashitani A. (2010). Auxins reverse plant male sterility caused by high 968

temperatures. Proc. Natl. Acad. Sci. USA 107:8569-8574. 969

Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, 970

Kobayashi M, Khush G S, et al. (2002). Green revolution: a mutant gibberellin-synthesis gene in 971

rice. Nature 416:701-702. 972

Schaller G E, Bishopp A, and Kieber J J. (2015). The yin-yang of hormones: cytokinin and auxin 973

interactions in plant development. Plant Cell 27:44-63. 974

Sinclair T R, and Sheehy J E. (1999). Erect leaves and photosynthesis in rice. Science 975

283:1456-1457. 976

Springer N. (2010). Shaping a better rice plant. Nat. Genet. 42:475-476. 977

Tang J, and Chu C. (2017). MicroRNAs in crop improvement: fine-tuners for complex traits. Nat. 978

Plants 3:17077. 979

Tang Y, Liu H, Guo S, Wang B, Li Z, Chong K, and Xu Y. (2018). OsmiR396d affects gibberellin 980

and brassinosteroid signaling to regulate plant architecture in rice. Plant Physiol. 176:946-959. 981

Van Bel M, Proost S, Wischnitzki E, Movahedi S, Scheerlinck C, Van de Peer Y, and Vandepoele 982

K. (2012). Dissecting plant genomes with the PLAZA comparative genomics platform. Plant 983

Physiol. 158:590-600. 984

Walter M, Chaban C, Schutze K, Batistic O, Weckermann K, Nake C, Blazevic D, Grefen C, 985 https://plantphysiol.orgDownloaded on December 30, 2020. - Published by



34

Schumacher K, Oecking C, et al. (2004). Visualization of protein interactions in living plant cells 986

using bimolecular fluorescence complementation. Plant J. 40:428-438. 987

Wang B, Smith S M, and Li J. (2018). Genetic regulation of shoot architecture. Annu. Rev. Plant Biol. 988

69:437-468. 989

Wang L, Gu X, Xu D, Wang W, Wang H, Zeng M, Chang Z, Huang H, and Cui X. (2011). 990

miR396-targeted AtGRF transcription factors are required for coordination of cell division and 991

differentiation during leaf development in Arabidopsis. J. Exp. Bot. 62:761-773. 992

Wang S, Zhang S, Sun C, Xu Y, Chen Y, Yu C, Qian Q, Jiang D A, and Qi Y. (2014). Auxin 993

response factor (OsARF12), a novel regulator for phosphate homeostasis in rice (Oryza sativa). 994

New Phytol. 201:91-103. 995

Wang Y, and Li J. (2008). Molecular basis of plant architecture. Annu. Rev. Plant Biol. 59:253-279. 996

Woodward A W, and Bartel B. (2005). Auxin: regulation, action, and interaction. Ann. Bot. 997

95:707-735. 998

Xing Y, and Zhang Q. (2010). Genetic and molecular bases of rice yield. Annu. Rev. Plant Biol. 999

61:421-442. 1000

Yoo S D, Cho Y H, and Sheen J. (2007). Arabidopsis mesophyll protoplasts: a versatile cell system 1001

for transient gene expression analysis. Nat. Protoc. 2:1565-1572. 1002

Zhang S, Wang S, Xu Y, Yu C, Shen C, Qian Q, Geisler M, Jiang de A, and Qi Y. (2015). The auxin 1003

response factor, OsARF19, controls rice leaf angles through positively regulating OsGH3-5 and 1004

OsBRI1. Plant Cell Environ. 38:638-654. 1005

Zhang W, Wu L, Wu X, Ding Y, Li G, Li J, Weng F, Liu Z, Tang S, Ding C, et al. (2016). Lodging 1006

resistance of japonica rice (Oryza Sativa L.): morphological and anatomical traits due to 1007

top-dressing nitrogen application rates. Rice 9:31. 1008

Zhao S Q, Xiang J J, and Xue H W. (2013). Studies on the rice LEAF INCLINATION1 (LC1), an 1009

IAA-amido synthetase, reveal the effects of auxin in leaf inclination control. Mol. Plant 6:174-187. 1010

Zhou L J, Xiao L T, and Xue H W. (2017). Dynamic cytology and transcriptional regulation of rice 1011

lamina joint development. Plant Physiol. 174:1728-1746. 1012

1013



Figure 1

Figure 1. Phenotypic characterization. A, Gross morphology of WT, OsGRF7

overexpression (GRF7OE-1) and OsGRF7 RNAi (GRF7RNAi-1) plants. Scale bars, 15

cm. B, Comparison of the flag leaf angles (up-panel) and the top second leaf angles

(down-panel) between OsGRF7 transgenic lines. Scale bars, 1 cm. C, Transverse (up-

panel) and longitudinal (down-panel) section of internodes in WT, GRF7OE-1, and

GRF7RNAi-1 plants. Scale bars, 200 μm. D, Cell length of the internode. E, Culm wall

thickness of OsGRF7 transgenic lines. F, Transverse (up-panel) and longitudinal

(down-panel) section of the lamina joint of flag leaves. Scale bars, 200 μm. Red boxes

areas in up-panel are enlarged. Red arrows mean adaxial side cell layers measured in

the panel G. pc, parenchymal cell; vb, vascular bundle; ad, adaxial surface of the lamina

joint. G, Adaxial side cell layers of the transverse sections of lamina joints. Panels D,

E, and G: Values are means ± SD of five biological replicates. *** P < 0.001 (two-

tailed Student’s t-test).

WT

GRF7OE-1

GRF7RNAi-1

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400

600

800

1000

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Figure 2

Figure 2. OsGRF7 is mainly repressed by OsmiR396e and expresses in various tissues.

A, The OsGRF7 cleavage site sequence complementary to OsmiR396. The position

corresponding to the 5′ ends of the cleaved OsGRF7 mRNA determined by 5′ RACE

and the frequency of 5′ RACE clones corresponding to the cleavage site is shown by

the arrow. 8/12 means eight of twelve clones have OsmiR396 cleavage site. The

mutated sites in OsrGRF7 were marked in red. The asterisks indicate mis-matched sites

between OsmiR396 and OsGRF7. The gray box indicates the QLQ domain, the pink

box indicates the WRC domain, and the purple box indicates the C terminal of OsGRF7,

respectively. B, Protein immunoblotting of OsGRF7 coexpressed with OsmiR396s in

GRF7-GFP transgenic rice protoplasts. Vector and control mean with or without the

empty vector, respectively. Coomassie brilliant blue (CBB) staining was used as

loading controls. The relative protein amounts were determined by ImageJ (NIH). C,

Relative expression levels of OsGRF7 and OsmiR396s were detected by RT-qPCR in

internode, root, inflorescence (10 cm), node, flag leaf, SAM (shoot apical meristem),

axillary bud, and lamina joint, respectively. D, GRF7pro:GUS expression patterns in

transgenic plants. (1) Small panicle branch. (2-4) Inflorescence at different stages. (5-

6) Spikelet. (7) Root tip. (8) Flag leaf. (9) Node. (10) Internode. (11) Axillary bud. (12)

Lamina joint of the 30-d seedling. Scale bars, 2 mm. E, Relative expression levels of

OsGRF7

OsmiR396e

OsmiR396d

OsmiR396i

OsmiR396c

OsmiR396b

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reference. Values are means ± SD of three biological replicates.



Figure 3

Figure 3. ChIP analysis of the OsGRF7 target genes. A, Distribution of OsGRF7

binding peaks in the rice genome. B, Genes reproducibly associated with OsGRF7

binding sites in YP1 (young inflorescences with a length of ~0.5 cm) or YP2 (young

inflorescences with a length of ~2 cm). C, Putative OsGRF7-binding motif predicted

by DREME. D, Expression and purification of OsGRF7 protein in E. coli. The red

arrowhead indicates the purified OsGRF7 protein. E, EMSA validation of interaction

between OsGRF7 and motif ACRGDA (ACAGTA). TAAGTA was used as the control.

533

1259

563

Genes association with OsGRF7binding sites in YP1 (1096)

Genes association with OsGRF7binding sites in YP2 (1822)

70%

19%

9%

63%

21%

14%2% 2%ChIP-YP1 ChIP-YP2

gene intergenic

upstream upstream

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Probe region-544bp -285bp -1003bp -817bp

M

A B C

D E F

H I

L

intergenic



F, OsGRF7 binding peaks in the promoters of OsCYP714B1 and OsARF12. Blue boxes

indicate probe locations. Bar = 1 kb. G, ChIP-qPCR validation of OsGRF7 binding sites

in the promoters of OsCYP714B1 and OsARF12. The fold enrichment was normalized

against the promoter of OsUBI. No addition of antibodies (NoAbs) served as a negative

control. H and I, Relative expression levels of the OsCYP714B1 and OsARF12 detected

with RT-qPCR. OsUBI was used as an internal reference. J and K, OsGRF7 actives

OsCYP714B1 and OsARF12 promoter-luciferase fusion constructs in transient

transactivation assays. L and M, EMSA validation of bindings between OsGRF7 and

promoters of OsCYP714B1 and OsARF12, respectively. The 2-, 10- and 100- fold

unmodified probes were used as competitors. The presence (+) or absence (-) of

components in the reaction mixture was indicated. Red triangles indicate the site of

ACRGDA motifs on promoter region. Panels G-K: Values are means ± SD of three

biological replicates. * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed Student’s t-

test).



Figure 4

Figure 4. OsGRF7 regulates expression of multiple IAA/GA-related genes. A, Relative

expression levels of auxin signaling related and GA synthesis related genes in the

lamina joint of the OsGRF7 transgenic lines. OsUBI was used as an internal reference.

B to J, GRF7-GFP mediated ChIP-qPCR enrichment (relative to the promoter region

of OsUBI) of ACRGDA-containing fragments (marked with arrowhead) from

promoters of auxin signaling related genes (ARF3, ARF4, ARF8, ARF9, PIN1b, PIN1d,

and PIN8) and GA synthesis related genes (SLR1 and SLRL1). The promoter region of

OsUBI was used as a control. K and L, OsGRF7 actives ARF3, ARF4, ARF8, ARF9,

PIN1b, PIN1d, PIN8, SLR1, and SLRL1 promoter-luciferase fusion constructs in

transient transactivation assays. Panels B-J, L: Values are means ± SD of three

biological replicates. * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed Student’s t-

test).

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Figure 5

Figure 5. Endogenous phytohormone contents and exogenous phytohormone

sensitivity test of OsGRF7 transgenic lines. A and B, Quantification of GA (A) and IAA

(B) derivatives in OsGRF7 transgenic seedlings detected with LC-MS/MS. Values are

means ± SD of three biological replicates. * P < 0.05 (two-tailed Student’s t-test).

Abbreviations: ME-IAA, methyl indole-3-acetate; ICA, indole-3-carboxaldehyde; IAA,

indole-3-acetic acid; NAA, 1-naphthylacetic acid. F.W., fresh weight. nd indicate not

detected (below the detection limit). C, Immunohistochemical observation (IHC) of

indole-3-acetic acid at the lamina joint of OsGRF7 transgenic lines. Anti-IAA antibody

and Alexa 488-conjugated goat anti-rabbit IgG antibody were used. Scale bars, 100 μm.

D, Phenotypes of OsGRF7 transgenic seedlings treated by different concentrations of

phytohormones. The same control has been used for 0 μM IAA and GA. Scale bars, 2

cm. E, Effects of different concentrations of IAA on primary root length in WT,

GRF7OE-1, and GRF7RNAi-1. Values are means ± SD (n = 15, three biological

replicates and five plants per replicate). F, Effects of different concentrations of GA3 on

the second sheath length in WT, GRF7OE-1, and GRF7RNAi-1 seedlings. Values are

means ± SD (n = 15, three biological replicates and five plants per replicate). Different

letters denote significant differences (P < 0.05) from Duncan’s multiple range tests.

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Figure 6

Figure 6. A proposed model for the regulation of rice plant architecture by OsGRF7.

In GRF7OE plants, OsGRF7 co-activates with OsGIFs to up-regulate the expressions

of downstream hormone-related genes through binding to ACRGDA motifs. Then the

synthesis of GA4 is inhibited, and the synthesis of IAA is promoted. Finally, the

GRF7OE plants display a semi-dwarf and compact plant architecture. Abbreviations:

IAA, indole-3-acetic acid; GA, gibberellin.

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GRF7



Parsed CitationsBailey T L. (2011). DREME: motif discovery in transcription factor ChIP-seq data. Bioinformatics 27:1653-1659.

Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Baucher M, Moussawi J, Vandeputte O M, Monteyne D, Mol A, Perez-Morga D, and El Jaziri M. (2013). A role for the miR396/GRFnetwork in specification of organ type during flower development, as supported by ectopic expression of Populus trichocarpa miR396cin transgenic tobacco. Plant Biol. 15:892-898.


Chandran V, Wang H, Gao F, Cao X L, Chen Y P, Li G B, Zhu Y, Yang X M, Zhang L L, Zhao Z X, et al. (2018). miR396-OsGRFs modulebalances growth and rice blast disease-resistance. Front. Plant Sci. 9:1999.


Che R, Tong H, Shi B, Liu Y, Fang S, Liu D, Xiao Y, Hu B, Liu L, Wang H, et al. (2015). Control of grain size and rice yield by GL2-mediated brassinosteroid responses. Nat. Plants 2:15195.


Chen X, Lu S, Wang Y, Zhang X, Lv B, Luo L, Xi D, Shen J, Ma H, and Ming F. (2015). OsNAC2 encoding a NAC transcription factor thataffects plant height through mediating the gibberellic acid pathway in rice. Plant J. 82:302-314.


Choi D, Kim J H, and Kende H. (2004). Whole genome analysis of the OsGRF gene family encoding plant-specific putative transcriptionactivators in rice (Oryza sativa L.). Plant and Cell Physiology 45:897-904.


Djami-Tchatchou A T, Sanan-Mishra N, Ntushelo K, and Dubery I A. (2017). Functional roles of microRNAs in agronomically importantplants-potential as targets for crop improvement and protection. Front. Plant Sci. 8:378.


Ercoli M F, Ferela A, Debernardi J M, Perrone A P, Rodriguez R E, and Palatnik J F. (2018). GIF transcriptional co-regulators controlroot meristem homeostasis. Plant Cell 30:347-359.


Feng J, Liu T, Qin B, Zhang Y, and Liu X S. (2012). Identifying ChIP-seq enrichment using MACS. Nat. Protoc. 7:1728-1740.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Gao F, Wang K, Liu Y, Chen Y, Chen P, Shi Z, Luo J, Jiang D, Fan F, Zhu Y, et al. (2015). Blocking miR396 increases rice yield by shapinginflorescence architecture. Nat. Plants 2:15196.


Gao S, Fang J, Xu F, Wang W, and Chu C. (2016). Rice HOX12 regulates panicle exsertion by directly modulating the expression ofELONGATED UPPERMOST INTERNODE1. Plant Cell 28:680-695.


Hu J, Wang Y, Fang Y, Zeng L, Xu J, Yu H, Shi Z, Pan J, Zhang D, Kang S, et al. (2015). A rare allele of GS2 enhances grain size andgrain yield in rice. Mol. Plant 8:1455-1465.


Jeong D H, An S, Kang H G, Moon S, Han J J, Park S, Lee H S, An K, and An G. (2002). T-DNA insertional mutagenesis for activationtagging in rice. Plant Physiol. 130:1636-1644.


Khush G S. (1995). Breaking the yield frontier of rice. GeoJournal 35:329–332.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Kim J H, Choi D, and Kende H. (2003). The AtGRF family of putative transcriptioon factors is involved in leaf and cotyledon growth inArabidopsis. Plant J. 36:94-104.



https://www.ncbi.nlm.nih.gov/pubmed?term=Bailey T L%2E %282011%29%2E DREME%3A motif discovery in transcription factor ChIP%2Dseq data%2E Bioinformatics&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Bailey&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=DREME: motif discovery in transcription factor ChIP-seq data&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=DREME: motif discovery in transcription factor ChIP-seq data&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Bailey&as_ylo=2011&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Baucher M%2C Moussawi J%2C Vandeputte O M%2C Monteyne D%2C Mol A%2C Perez%2DMorga D%2C and El Jaziri M%2E %282013%29%2E A role for the miR396%2FGRF network in specification of organ type during flower development%2C as supported by ectopic expression of Populus trichocarpa miR396c in transgenic tobacco%2E Plant Biol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Baucher&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=A role for the miR396/GRF network in specification of organ type during flower development, as supported by ectopic expression of Populus trichocarpa miR396c in transgenic tobacco&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=A role for the miR396/GRF network in specification of organ type during flower development, as supported by ectopic expression of Populus trichocarpa miR396c in transgenic tobacco&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Baucher&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Chandran V%2C Wang H%2C Gao F%2C Cao X L%2C Chen Y P%2C Li G B%2C Zhu Y%2C Yang X M%2C Zhang L L%2C Zhao Z X%2C et al%2E %282018%29%2E miR396%2DOsGRFs module balances growth and rice blast disease%2Dresistance%2E Front%2E Plant Sci&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Chandran&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=miR396-OsGRFs module balances growth and rice blast disease-resistance&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Che R%2C Tong H%2C Shi B%2C Liu Y%2C Fang S%2C Liu D%2C Xiao Y%2C Hu B%2C Liu L%2C Wang H%2C et al%2E %282015%29%2E Control of grain size and rice yield by GL2%2Dmediated brassinosteroid responses%2E Nat%2E Plants&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Chen X%2C Lu S%2C Wang Y%2C Zhang X%2C Lv B%2C Luo L%2C Xi D%2C Shen J%2C Ma H%2C and Ming F%2E %282015%29%2E OsNAC2 encoding a NAC transcription factor that affects plant height through mediating the gibberellic acid pathway in rice%2E Plant J&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Choi D%2C Kim J H%2C and Kende H%2E %282004%29%2E Whole genome analysis of the OsGRF gene family encoding plant%2Dspecific putative transcription activators in rice %28Oryza sativa L%2E%29%2E Plant and Cell Physiology&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Choi&hl=en&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Djami%2DTchatchou A T%2C Sanan%2DMishra N%2C Ntushelo K%2C and Dubery I A%2E %282017%29%2E Functional roles of microRNAs in agronomically important plants%2Dpotential as targets for crop improvement and protection%2E Front%2E Plant Sci&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Ercoli M F%2C Ferela A%2C Debernardi J M%2C Perrone A P%2C Rodriguez R E%2C and Palatnik J F%2E %282018%29%2E GIF transcriptional co%2Dregulators control root meristem homeostasis%2E Plant Cell&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Feng J%2C Liu T%2C Qin B%2C Zhang Y%2C and Liu X S%2E %282012%29%2E Identifying ChIP%2Dseq enrichment using MACS%2E Nat%2E Protoc&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Gao F%2C Wang K%2C Liu Y%2C Chen Y%2C Chen P%2C Shi Z%2C Luo J%2C Jiang D%2C Fan F%2C Zhu Y%2C et al%2E %282015%29%2E Blocking miR396 increases rice yield by shaping inflorescence architecture%2E Nat%2E Plants&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Hu J%2C Wang Y%2C Fang Y%2C Zeng L%2C Xu J%2C Yu H%2C Shi Z%2C Pan J%2C Zhang D%2C Kang S%2C et al%2E %282015%29%2E A rare allele of GS2 enhances grain size and grain yield in rice%2E Mol%2E Plant&dopt=abstract

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https://www.ncbi.nlm.nih.gov/pubmed?term=Jeong D H%2C An S%2C Kang H G%2C Moon S%2C Han J J%2C Park S%2C Lee H S%2C An K%2C and An G%2E %282002%29%2E T%2DDNA insertional mutagenesis for activation tagging in rice%2E Plant Physiol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Jeong&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=T-DNA insertional mutagenesis for activation tagging in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=T-DNA insertional mutagenesis for activation tagging in rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Jeong&as_ylo=2002&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Khush G S%2E %281995%29%2E Breaking the yield frontier of rice%2E GeoJournal 35%3A329%26%23x&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Khush&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Breaking the yield frontier of rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Breaking the yield frontier of rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Khush&as_ylo=1995&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Kim J H%2C Choi D%2C and Kende H%2E %282003%29%2E The AtGRF family of putative transcriptioon factors is involved in leaf and cotyledon growth in Arabidopsis%2E Plant J&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kim&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=The AtGRF family of putative transcriptioon factors is involved in leaf and cotyledon growth in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=The AtGRF family of putative transcriptioon factors is involved in leaf and cotyledon growth in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kim&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1


Kim J H, and Kende H. (2004). A transcriptional coactivator, AtGIF1, is involved in regulating leaf growth and morphology inArabidopsis. Proc. Natl. Acad. Sci. USA 101:13374-13379.


Kim J H, and Lee B H. (2006). GROWTH-REGULATING FACTOR4 of Arabidopsis thaliana is required for development of leaves,cotyledons, and shoot apical meristem. J. Plant Biol. 49:463-468.


Kobayashi M, Yamaguchi I, Murofushi N, Ota Y, and Takahashi N. (1988). Fluctuation and localization of endogenous gibberellins inrice. Agricultural and Biological Chemistry 52:1189-1194.


Kozomara A, and Griffiths-Jones S. (2014). miRBase: annotating high confidence microRNAs using deep sequencing data. NucleicAcids Res. 42:D68-73.


Kuijt S J, Greco R, Agalou A, Shao J, t Hoen C C, Overnas E, Osnato M, Curiale S, Meynard D, van Gulik R, et al. (2014). Interactionbetween the GROWTH-REGULATING FACTOR and KNOTTED1-LIKE HOMEOBOX families of transcription factors. Plant Physiol.164:1952-1966.


Kumar S, Stecher G, Li M, Knyaz C, and Tamura K. (2018). MEGA X: molecular evolutionary genetics analysis across computingplatforms. Mol Biol Evol 35:1547-1549.


Langmead B, Trapnell C, Pop M, and Salzberg S L. (2009). Ultrafast and memory-efficient alignment of short DNA sequences to thehuman genome. Genome Biol. 10:R25.


Lavy M, and Estelle M. (2016). Mechanisms of auxin signaling. Development 143:3226-3229.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Li S, Gao F, Xie K, Zeng X, Cao Y, Zeng J, He Z, Ren Y, Li W, Deng Q, et al. (2016). The OsmiR396c-OsGRF4-OsGIF1 regulatory moduledetermines grain size and yield in rice. Plant Biotechnol. J. 14:2134-2146.


Li S, Tian Y, Wu K, Ye Y, Yu J, Zhang J, Liu Q, Hu M, Li H, Tong Y, et al. (2018). Modulating plant growth-metabolism coordination forsustainable agriculture. Nature 560:595-600.


Li Y, Li J, Chen Z, Wei Y, Qi Y, and Wu C. (2020). OsmiR167a-targeted auxin response factors modulate tiller angle via fine-tuning auxindistribution in rice. Plant Biotechnol. J. 10.1111/pbi.13360.


Liu D, Song Y, Chen Z, and Yu D. (2009). Ectopic expression of miR396 suppresses GRF target gene expression and alters leaf growthin Arabidopsis. Physiol. Plant. 136:223-236.


Liu H, Guo S, Xu Y, Li C, Zhang Z, Zhang D, Xu S, Zhang C, and Chong K. (2014). OsmiR396d-regulated OsGRFs function in floralorganogenesis in rice through binding to their targets OsJMJ706 and OsCR4. Plant Physiol. 165:160-174.


Liu T. (2014). Use Model-Based Analysis of ChIP-seq (MACS) to analyze short reads generated by sequencing protein-DNA interactionsin embryonic stem cells. Methods Mol. Biol. 1150:81-95.


Lu Y, Meng Y, Zeng J, Luo Y, Feng Z, Bian L, and Gao S. (2020). Coordination between GROWTH-REGULATING FACTOR1 and GRF-INTERACTING FACTOR1 plays a key role in regulating leaf growth in rice. BMC Plant Biol. 20:200.



https://www.ncbi.nlm.nih.gov/pubmed?term=Kim J H%2C and Kende H%2E %282004%29%2E A transcriptional coactivator%2C AtGIF1%2C is involved in regulating leaf growth and morphology in Arabidopsis%2E Proc%2E Natl%2E Acad%2E Sci%2E USA&dopt=abstract


https://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kim&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Kim J H%2C and Lee B H%2E %282006%29%2E GROWTH%2DREGULATING FACTOR4 of Arabidopsis thaliana is required for development of leaves%2C cotyledons%2C and shoot apical meristem%2E J%2E Plant Biol&dopt=abstract


https://scholar.google.com/scholar?as_q=GROWTH-REGULATING FACTOR4 of Arabidopsis thaliana is required for development of leaves, cotyledons, and shoot apical meristem&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=GROWTH-REGULATING FACTOR4 of Arabidopsis thaliana is required for development of leaves, cotyledons, and shoot apical meristem&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kim&as_ylo=2006&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Kobayashi M%2C Yamaguchi I%2C Murofushi N%2C Ota Y%2C and Takahashi N%2E %281988%29%2E Fluctuation and localization of endogenous gibberellins in rice%2E Agricultural and Biological Chemistry&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kobayashi&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Fluctuation and localization of endogenous gibberellins in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Fluctuation and localization of endogenous gibberellins in rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kobayashi&as_ylo=1988&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Kozomara A%2C and Griffiths%2DJones S%2E %282014%29%2E miRBase%3A annotating high confidence microRNAs using deep sequencing data%2E Nucleic Acids Res%2E 42%3AD&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kozomara&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=miRBase: annotating high confidence microRNAs using deep sequencing data&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=miRBase: annotating high confidence microRNAs using deep sequencing data&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kozomara&as_ylo=2014&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Kuijt S J%2C Greco R%2C Agalou A%2C Shao J%2C t Hoen C C%2C Overnas E%2C Osnato M%2C Curiale S%2C Meynard D%2C van Gulik R%2C et al%2E %282014%29%2E Interaction between the GROWTH%2DREGULATING FACTOR and KNOTTED1%2DLIKE HOMEOBOX families of transcription factors%2E Plant Physiol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kuijt&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Interaction between the GROWTH-REGULATING FACTOR and KNOTTED1-LIKE HOMEOBOX families of transcription factors&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Interaction between the GROWTH-REGULATING FACTOR and KNOTTED1-LIKE HOMEOBOX families of transcription factors&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kuijt&as_ylo=2014&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Kumar S%2C Stecher G%2C Li M%2C Knyaz C%2C and Tamura K%2E %282018%29%2E MEGA X%3A molecular evolutionary genetics analysis across computing platforms%2E Mol Biol Evol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kumar&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=MEGA X: molecular evolutionary genetics analysis across computing platforms&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=MEGA X: molecular evolutionary genetics analysis across computing platforms&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kumar&as_ylo=2018&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Langmead B%2C Trapnell C%2C Pop M%2C and Salzberg S L%2E %282009%29%2E Ultrafast and memory%2Defficient alignment of short DNA sequences to the human genome%2E Genome Biol%2E 10%3AR&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Langmead&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Ultrafast and memory-efficient alignment of short DNA sequences to the human genome&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Ultrafast and memory-efficient alignment of short DNA sequences to the human genome&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Langmead&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Lavy M%2C and Estelle M%2E %282016%29%2E Mechanisms of auxin signaling%2E Development&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lavy&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Mechanisms of auxin signaling&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Mechanisms of auxin signaling&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lavy&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Li S%2C Gao F%2C Xie K%2C Zeng X%2C Cao Y%2C Zeng J%2C He Z%2C Ren Y%2C Li W%2C Deng Q%2C et al%2E %282016%29%2E The OsmiR396c%2DOsGRF4%2DOsGIF1 regulatory module determines grain size and yield in rice%2E Plant Biotechnol%2E J&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Li&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=The OsmiR396c-OsGRF4-OsGIF1 regulatory module determines grain size and yield in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=The OsmiR396c-OsGRF4-OsGIF1 regulatory module determines grain size and yield in rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Li&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Li S%2C Tian Y%2C Wu K%2C Ye Y%2C Yu J%2C Zhang J%2C Liu Q%2C Hu M%2C Li H%2C Tong Y%2C et al%2E %282018%29%2E Modulating plant growth%2Dmetabolism coordination for sustainable agriculture%2E Nature&dopt=abstract


https://scholar.google.com/scholar?as_q=Modulating plant growth-metabolism coordination for sustainable agriculture&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Modulating plant growth-metabolism coordination for sustainable agriculture&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Li&as_ylo=2018&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Li Y%2C Li J%2C Chen Z%2C Wei Y%2C Qi Y%2C and Wu C%2E %282020%29%2E OsmiR167a%2Dtargeted auxin response factors modulate tiller angle via fine%2Dtuning auxin distribution in rice%2E Plant Biotechnol%2E J%2E 10%2E1111%2Fpbi&dopt=abstract


https://scholar.google.com/scholar?as_q=OsmiR167a-targeted auxin response factors modulate tiller angle via fine-tuning auxin distribution in rice.&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=OsmiR167a-targeted auxin response factors modulate tiller angle via fine-tuning auxin distribution in rice.&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Li&as_ylo=2020&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Liu D%2C Song Y%2C Chen Z%2C and Yu D%2E %282009%29%2E Ectopic expression of miR396 suppresses GRF target gene expression and alters leaf growth in Arabidopsis%2E Physiol%2E Plant&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Liu&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Ectopic expression of miR396 suppresses GRF target gene expression and alters leaf growth in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Ectopic expression of miR396 suppresses GRF target gene expression and alters leaf growth in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Liu&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Liu H%2C Guo S%2C Xu Y%2C Li C%2C Zhang Z%2C Zhang D%2C Xu S%2C Zhang C%2C and Chong K%2E %282014%29%2E OsmiR396d%2Dregulated OsGRFs function in floral organogenesis in rice through binding to their targets OsJMJ706 and OsCR4%2E Plant Physiol&dopt=abstract


https://scholar.google.com/scholar?as_q=OsmiR396d-regulated OsGRFs function in floral organogenesis in rice through binding to their targets OsJMJ706 and OsCR4&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=OsmiR396d-regulated OsGRFs function in floral organogenesis in rice through binding to their targets OsJMJ706 and OsCR4&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Liu&as_ylo=2014&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Liu T%2E %282014%29%2E Use Model%2DBased Analysis of ChIP%2Dseq %28MACS%29 to analyze short reads generated by sequencing protein%2DDNA interactions in embryonic stem cells%2E Methods Mol%2E Biol&dopt=abstract


https://scholar.google.com/scholar?as_q=Use Model-Based Analysis of ChIP-seq (MACS) to analyze short reads generated by sequencing protein-DNA interactions in embryonic stem cells&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Use Model-Based Analysis of ChIP-seq (MACS) to analyze short reads generated by sequencing protein-DNA interactions in embryonic stem cells&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Liu&as_ylo=2014&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Lu Y%2C Meng Y%2C Zeng J%2C Luo Y%2C Feng Z%2C Bian L%2C and Gao S%2E %282020%29%2E Coordination between GROWTH%2DREGULATING FACTOR1 and GRF%2DINTERACTING FACTOR1 plays a key role in regulating leaf growth in rice%2E BMC Plant Biol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lu&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Coordination between GROWTH-REGULATING FACTOR1 and GRF-INTERACTING FACTOR1 plays a key role in regulating leaf growth in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Coordination between GROWTH-REGULATING FACTOR1 and GRF-INTERACTING FACTOR1 plays a key role in regulating leaf growth in rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lu&as_ylo=2020&as_allsubj=all&hl=en&c2coff=1


Luo A D, Liu L, Tang Z S, Bai X Q, Cao S Y, and Chu C C. (2005). Down-regulation of OsGRF1 gene in rice rhd1 mutant results inreduced heading date. J. Integr. Plant Biol. 47:745-752.


Luo X, Zheng J, Huang R, Huang Y, Wang H, Jiang L, and Fang X. (2016). Phytohormones signaling and crosstalk regulating leaf anglein rice. Plant Cell Rep. 35:2423-2433.


Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, et al. (2015). A robust CRISPR/Cas9 system for convenient,high-efficiency multiplex genome editing in monocot and dicot plants. Mol. Plant 8:1274-1284.


Magome H, Nomura T, Hanada A, Takeda-Kamiya N, Ohnishi T, Shinma Y, Katsumata T, Kawaide H, Kamiya Y, and Yamaguchi S. (2013).CYP714B1 and CYP714B2 encode gibberellin 13-oxidases that reduce gibberellin activity in rice. Proc. Natl. Acad. Sci. USA 110:1947-1952.


Omidbakhshfard M A, Proost S, Fujikura U, and Mueller-Roeber B. (2015). Growth-Regulating Factors (GRFs): a small transcriptionfactor family with important functions in plant biology. Mol. Plant 8:998-1010.


Ookawa T, Hobo T, Yano M, Murata K, Ando T, Miura H, Asano K, Ochiai Y, Ikeda M, Nishitani R, et al. (2010). New approach for riceimprovement using a pleiotropic QTL gene for lodging resistance and yield. Nat. Commun. 1:132.


Rodriguez R E, Mecchia M A, Debernardi J M, Schommer C, Weigel D, and Palatnik J F. (2009). Control of cell proliferation inArabidopsis thaliana by microRNA miR396. Development 137:103-112.


Sakata T, Oshino T, Miura S, Tomabechi M, Tsunaga Y, Higashitani N, Miyazawa Y, Takahashi H, Watanabe M, and Higashitani A. (2010).Auxins reverse plant male sterility caused by high temperatures. Proc. Natl. Acad. Sci. USA 107:8569-8574.


Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush G S, et al. (2002).Green revolution: a mutant gibberellin-synthesis gene in rice. Nature 416:701-702.


Schaller G E, Bishopp A, and Kieber J J. (2015). The yin-yang of hormones: cytokinin and auxin interactions in plant development. PlantCell 27:44-63.


Sinclair T R, and Sheehy J E. (1999). Erect leaves and photosynthesis in rice. Science 283:1456-1457.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Springer N. (2010). Shaping a better rice plant. Nat. Genet. 42:475-476.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Tang J, and Chu C. (2017). MicroRNAs in crop improvement: fine-tuners for complex traits. Nat. Plants 3:17077.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Tang Y, Liu H, Guo S, Wang B, Li Z, Chong K, and Xu Y. (2018). OsmiR396d affects gibberellin and brassinosteroid signaling to regulateplant architecture in rice. Plant Physiol. 176:946-959.


Van Bel M, Proost S, Wischnitzki E, Movahedi S, Scheerlinck C, Van de Peer Y, and Vandepoele K. (2012). Dissecting plant genomeswith the PLAZA comparative genomics platform. Plant Physiol. 158:590-600.


Walter M, Chaban C, Schutze K, Batistic O, Weckermann K, Nake C, Blazevic D, Grefen C, Schumacher K, Oecking C, et al. (2004).


https://www.ncbi.nlm.nih.gov/pubmed?term=Luo A D%2C Liu L%2C Tang Z S%2C Bai X Q%2C Cao S Y%2C and Chu C C%2E %282005%29%2E Down%2Dregulation of OsGRF1 gene in rice rhd1 mutant results in reduced heading date%2E J%2E Integr%2E Plant Biol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Luo&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Down-regulation of OsGRF1 gene in rice rhd1 mutant results in reduced heading date&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Down-regulation of OsGRF1 gene in rice rhd1 mutant results in reduced heading date&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Luo&as_ylo=2005&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Luo X%2C Zheng J%2C Huang R%2C Huang Y%2C Wang H%2C Jiang L%2C and Fang X%2E %282016%29%2E Phytohormones signaling and crosstalk regulating leaf angle in rice%2E Plant Cell Rep&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Luo&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Phytohormones signaling and crosstalk regulating leaf angle in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Ma X%2C Zhang Q%2C Zhu Q%2C Liu W%2C Chen Y%2C Qiu R%2C Wang B%2C Yang Z%2C Li H%2C Lin Y%2C et al%2E %282015%29%2E A robust CRISPR%2FCas9 system for convenient%2C high%2Defficiency multiplex genome editing in monocot and dicot plants%2E Mol%2E Plant&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Ma&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Magome H%2C Nomura T%2C Hanada A%2C Takeda%2DKamiya N%2C Ohnishi T%2C Shinma Y%2C Katsumata T%2C Kawaide H%2C Kamiya Y%2C and Yamaguchi S%2E %282013%29%2E CYP714B1 and CYP714B2 encode gibberellin 13%2Doxidases that reduce gibberellin activity in rice%2E Proc%2E Natl%2E Acad%2E Sci%2E USA&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Magome&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=CYP714B1 and CYP714B2 encode gibberellin 13-oxidases that reduce gibberellin activity in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Omidbakhshfard M A%2C Proost S%2C Fujikura U%2C and Mueller%2DRoeber B%2E %282015%29%2E Growth%2DRegulating Factors %28GRFs%29%3A a small transcription factor family with important functions in plant biology%2E Mol%2E Plant&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Omidbakhshfard&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Growth-Regulating Factors (GRFs): a small transcription factor family with important functions in plant biology&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Ookawa T%2C Hobo T%2C Yano M%2C Murata K%2C Ando T%2C Miura H%2C Asano K%2C Ochiai Y%2C Ikeda M%2C Nishitani R%2C et al%2E %282010%29%2E New approach for rice improvement using a pleiotropic QTL gene for lodging resistance and yield%2E Nat%2E Commun&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Ookawa&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=New approach for rice improvement using a pleiotropic QTL gene for lodging resistance and yield&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=New approach for rice improvement using a pleiotropic QTL gene for lodging resistance and yield&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Ookawa&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Rodriguez R E%2C Mecchia M A%2C Debernardi J M%2C Schommer C%2C Weigel D%2C and Palatnik J F%2E %282009%29%2E Control of cell proliferation in Arabidopsis thaliana by microRNA miR396%2E Development&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Rodriguez&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Control of cell proliferation in Arabidopsis thaliana by microRNA miR396&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Control of cell proliferation in Arabidopsis thaliana by microRNA miR396&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Rodriguez&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Sakata T%2C Oshino T%2C Miura S%2C Tomabechi M%2C Tsunaga Y%2C Higashitani N%2C Miyazawa Y%2C Takahashi H%2C Watanabe M%2C and Higashitani A%2E %282010%29%2E Auxins reverse plant male sterility caused by high temperatures%2E Proc%2E Natl%2E Acad%2E Sci%2E USA&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Sakata&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Auxins reverse plant male sterility caused by high temperatures&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Auxins reverse plant male sterility caused by high temperatures&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Sakata&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Sasaki A%2C Ashikari M%2C Ueguchi%2DTanaka M%2C Itoh H%2C Nishimura A%2C Swapan D%2C Ishiyama K%2C Saito T%2C Kobayashi M%2C Khush G S%2C et al%2E %282002%29%2E Green revolution%3A a mutant gibberellin%2Dsynthesis gene in rice%2E Nature&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Sasaki&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Green revolution: a mutant gibberellin-synthesis gene in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Green revolution: a mutant gibberellin-synthesis gene in rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Sasaki&as_ylo=2002&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Schaller G E%2C Bishopp A%2C and Kieber J J%2E %282015%29%2E The yin%2Dyang of hormones%3A cytokinin and auxin interactions in plant development%2E Plant Cell&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Schaller&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=The yin-yang of hormones: cytokinin and auxin interactions in plant development&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=The yin-yang of hormones: cytokinin and auxin interactions in plant development&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Schaller&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Sinclair T R%2C and Sheehy J E%2E %281999%29%2E Erect leaves and photosynthesis in rice%2E Science&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Sinclair&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Erect leaves and photosynthesis in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Erect leaves and photosynthesis in rice&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Sinclair&as_ylo=1999&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Springer N%2E %282010%29%2E Shaping a better rice plant%2E Nat%2E Genet&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Springer&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Shaping a better rice plant&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Shaping a better rice plant&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Springer&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Tang J%2C and Chu C%2E %282017%29%2E MicroRNAs in crop improvement%3A fine%2Dtuners for complex traits%2E Nat%2E Plants&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Tang&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=MicroRNAs in crop improvement: fine-tuners for complex traits&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=MicroRNAs in crop improvement: fine-tuners for complex traits&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Tang&as_ylo=2017&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Tang Y%2C Liu H%2C Guo S%2C Wang B%2C Li Z%2C Chong K%2C and Xu Y%2E %282018%29%2E OsmiR396d affects gibberellin and brassinosteroid signaling to regulate plant architecture in rice%2E Plant Physiol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Tang&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=OsmiR396d affects gibberellin and brassinosteroid signaling to regulate plant architecture in rice&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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https://www.ncbi.nlm.nih.gov/pubmed?term=Van Bel M%2C Proost S%2C Wischnitzki E%2C Movahedi S%2C Scheerlinck C%2C Van de Peer Y%2C and Vandepoele K%2E %282012%29%2E Dissecting plant genomes with the PLAZA comparative genomics platform%2E Plant Physiol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Van&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Dissecting plant genomes with the PLAZA comparative genomics platform&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Dissecting plant genomes with the PLAZA comparative genomics platform&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Van&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1


Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation. Plant J. 40:428-438.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Wang B, Smith S M, and Li J. (2018). Genetic regulation of shoot architecture. Annu. Rev. Plant Biol. 69:437-468.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Wang L, Gu X, Xu D, Wang W, Wang H, Zeng M, Chang Z, Huang H, and Cui X. (2011). miR396-targeted AtGRF transcription factors arerequired for coordination of cell division and differentiation during leaf development in Arabidopsis. J. Exp. Bot. 62:761-773.


Wang S, Zhang S, Sun C, Xu Y, Chen Y, Yu C, Qian Q, Jiang D A, and Qi Y. (2014). Auxin response factor (OsARF12), a novel regulatorfor phosphate homeostasis in rice (Oryza sativa). New Phytol. 201:91-103.


Wang Y, and Li J. (2008). Molecular basis of plant architecture. Annu. Rev. Plant Biol. 59:253-279.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Woodward A W, and Bartel B. (2005). Auxin: regulation, action, and interaction. Ann. Bot. 95:707-735.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Xing Y, and Zhang Q. (2010). Genetic and molecular bases of rice yield. Annu. Rev. Plant Biol. 61:421-442.Pubmed: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Yoo S D, Cho Y H, and Sheen J. (2007). Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expressionanalysis. Nat. Protoc. 2:1565-1572.


Zhang S, Wang S, Xu Y, Yu C, Shen C, Qian Q, Geisler M, Jiang de A, and Qi Y. (2015). The auxin response factor, OsARF19, controlsrice leaf angles through positively regulating OsGH3-5 and OsBRI1. Plant Cell Environ. 38:638-654.


Zhang W, Wu L, Wu X, Ding Y, Li G, Li J, Weng F, Liu Z, Tang S, Ding C, et al. (2016). Lodging resistance of japonica rice (Oryza SativaL.): morphological and anatomical traits due to top-dressing nitrogen application rates. Rice 9:31.


Zhao S Q, Xiang J J, and Xue H W. (2013). Studies on the rice LEAF INCLINATION1 (LC1), an IAA-amido synthetase, reveal the effects ofauxin in leaf inclination control. Mol. Plant 6:174-187.


Zhou L J, Xiao L T, and Xue H W. (2017). Dynamic cytology and transcriptional regulation of rice lamina joint development. Plant Physiol.174:1728-1746.



https://www.ncbi.nlm.nih.gov/pubmed?term=Walter M%2C Chaban C%2C Schutze K%2C Batistic O%2C Weckermann K%2C Nake C%2C Blazevic D%2C Grefen C%2C Schumacher K%2C Oecking C%2C et al%2E %282004%29%2E Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation%2E Plant J&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Walter&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Walter&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Wang B%2C Smith S M%2C and Li J%2E %282018%29%2E Genetic regulation of shoot architecture%2E Annu%2E Rev%2E Plant Biol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Wang&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Genetic regulation of shoot architecture&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Genetic regulation of shoot architecture&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wang&as_ylo=2018&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Wang L%2C Gu X%2C Xu D%2C Wang W%2C Wang H%2C Zeng M%2C Chang Z%2C Huang H%2C and Cui X%2E %282011%29%2E miR396%2Dtargeted AtGRF transcription factors are required for coordination of cell division and differentiation during leaf development in Arabidopsis%2E J%2E Exp%2E Bot&dopt=abstract


https://scholar.google.com/scholar?as_q=miR396-targeted AtGRF transcription factors are required for coordination of cell division and differentiation during leaf development in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=miR396-targeted AtGRF transcription factors are required for coordination of cell division and differentiation during leaf development in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wang&as_ylo=2011&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Wang S%2C Zhang S%2C Sun C%2C Xu Y%2C Chen Y%2C Yu C%2C Qian Q%2C Jiang D A%2C and Qi Y%2E %282014%29%2E Auxin response factor %28OsARF12%29%2C a novel regulator for phosphate homeostasis in rice %28Oryza sativa%29%2E New Phytol&dopt=abstract


https://scholar.google.com/scholar?as_q=Auxin response factor (OsARF12), a novel regulator for phosphate homeostasis in rice (Oryza sativa)&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Auxin response factor (OsARF12), a novel regulator for phosphate homeostasis in rice (Oryza sativa)&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wang&as_ylo=2014&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Wang Y%2C and Li J%2E %282008%29%2E Molecular basis of plant architecture%2E Annu%2E Rev%2E Plant Biol&dopt=abstract


https://scholar.google.com/scholar?as_q=Molecular basis of plant architecture&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Molecular basis of plant architecture&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Wang&as_ylo=2008&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Woodward A W%2C and Bartel B%2E %282005%29%2E Auxin%3A regulation%2C action%2C and interaction%2E Ann%2E Bot&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Woodward&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Auxin: regulation, action, and interaction&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Auxin: regulation, action, and interaction&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Woodward&as_ylo=2005&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Xing Y%2C and Zhang Q%2E %282010%29%2E Genetic and molecular bases of rice yield%2E Annu%2E Rev%2E Plant Biol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Xing&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Genetic and molecular bases of rice yield&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Genetic and molecular bases of rice yield&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Xing&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Yoo S D%2C Cho Y H%2C and Sheen J%2E %282007%29%2E Arabidopsis mesophyll protoplasts%3A a versatile cell system for transient gene expression analysis%2E Nat%2E Protoc&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Yoo&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Yoo&as_ylo=2007&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Zhang S%2C Wang S%2C Xu Y%2C Yu C%2C Shen C%2C Qian Q%2C Geisler M%2C Jiang de A%2C and Qi Y%2E %282015%29%2E The auxin response factor%2C OsARF19%2C controls rice leaf angles through positively regulating OsGH3%2D5 and OsBRI1%2E Plant Cell Environ&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zhang&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zhang&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Zhang W%2C Wu L%2C Wu X%2C Ding Y%2C Li G%2C Li J%2C Weng F%2C Liu Z%2C Tang S%2C Ding C%2C et al%2E %282016%29%2E Lodging resistance of japonica rice %28Oryza Sativa L%2E%29%3A morphological and anatomical traits due to top%2Ddressing nitrogen application rates%2E Rice&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zhang&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Lodging resistance of japonica rice (Oryza Sativa L&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Lodging resistance of japonica rice (Oryza Sativa L&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zhang&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Zhao S Q%2C Xiang J J%2C and Xue H W%2E %282013%29%2E Studies on the rice LEAF INCLINATION1 %28LC1%29%2C an IAA%2Damido synthetase%2C reveal the effects of auxin in leaf inclination control%2E Mol%2E Plant&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zhao&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Studies on the rice LEAF INCLINATION1 (LC1), an IAA-amido synthetase, reveal the effects of auxin in leaf inclination control&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Studies on the rice LEAF INCLINATION1 (LC1), an IAA-amido synthetase, reveal the effects of auxin in leaf inclination control&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zhao&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

https://www.ncbi.nlm.nih.gov/pubmed?term=Zhou L J%2C Xiao L T%2C and Xue H W%2E %282017%29%2E Dynamic cytology and transcriptional regulation of rice lamina joint development%2E Plant Physiol&dopt=abstract

https://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zhou&hl=en&c2coff=1

https://scholar.google.com/scholar?as_q=Dynamic cytology and transcriptional regulation of rice lamina joint development&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

https://scholar.google.com/scholar?as_q=Dynamic cytology and transcriptional regulation of rice lamina joint development&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zhou&as_ylo=2017&as_allsubj=all&hl=en&c2coff=1


Short title: OsGRF7 shapes rice plant architecture · 2020. 6. 24. · in determining the...

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