The development and ripening of fleshy fruits

PLB/HRT 865Plant Growth and Development

(Fall 2008)

Dual Functionality of fruits

Secondary metabolites can act as a feeding deterrent

Tewksbury and Nabhan, 2001, Nature 42: 403-404

Fitness impacts and mechanics of fungal infection

Tewksbury J. J. et.al. PNAS 2008;105:11808-11811

© 2008 by The National Academy of Sciences of the USA

Effects of capsaicinoids on Fusarium infection

Tewksbury J. J. et.al. PNAS 2008;105:11808-11811

© 2008 by The National Academy of Sciences of the USA

volatile precursor Cerasiforme nl/g FW/ h

Flora Dade nl/ g FW / h

Odor threshold (ppb)

Cis-3-hexenal Fatty acid 16.28 5.25 0.25

Β-Ionone Carotenoid 0.03 0.02 0.007

Hexanal Fatty acid 27.21 17.16 5

Β-Damascenone carotenoid N/D ND 0.002

1-Penten-3-one Fatty acid 0.21 0.03 1

2-Methylbutanal isoleucine 0.75 0.25 1

3-Methylbutanal leucine 0.67 0.18 0.2

trans-2-hexenal Fatty acid 0.7 0.26 17

isobutylthiozole unknown 0.32 0.8 3.5

1-Nitro-2-phenylethanal phenylalanine 0.018 0.013 2

Trans-2-heptenal Fatty acid 0.16 0.13 13

Phenylacetaldehyde phenylalanine 0.06 0.09 4

6-Methyl-5-hepten-2-one carotenoid 0.99 1.84 2000

Cis-3-hexenol Fatty acid 19.83 13.29 70

2-phenylethanol phenylalanine 0.21 0.32 750

3-methylbutanol leucine 3.83 1.23 120

Methyl salicylate phenylalanine 0.08 0.04 40

Goff and Klee, 2006,Science 311: 815 -819

Fleshy fruits are anatomically diverse:

“Berry” – a pulpy fruit from a single pistil with one or more carpels with several to many seeds

“Drupe” – a simple fruit where the mesocarp tissue becomes thick and fleshy and the endocarp stony

“Pepo” – fruits derived from an inferior ovary that develops from multiple carpels bearing many seeds

“Hesperidium” – fruits of several carpels with a leathery rind and inner pulp juice sacs or vesicles

“Pome” – a simple fruit comprised of several carpels, the edible portion of which is made up of accessory tissue

“aggregate fruits” – a number of small ovaries formed in a Single flower and arranged over the surface of a single Receptacle. Individual ovaries are called fruitlets

“multiple fruits” – matured ovaries of several flowersunited into a single mass. Fruit develops from accessory tissue.

Fleshy fruits are anatomically diverse:

Dry fruits are subdivided into dehiscent and indehiscent classes

Dehiscent fruit include the following subclasses, capsule, follicle, silique and legume

Indehiscent fruits can be subdivided as follows: achenes, caryopsis, nut, samara, schizocarp

Achene – thin walled fruit containing one seed in which the seed coat is free and attached to the pericarp at only one point

Fruit (achene)

Flesh (receptacle)

Fruit of the strawberry is botanically a dried fruit for horticultural purposes it is treated as a fleshy fruit because of the edible receptacle tissue

genetic maps ripening mutants

EST collection

BAC libraries

diploid genetics

self-pollinating and easily out-crossed

dramatic ripening phenotype

easily transformed

large germplasmcollection

microarrays

emerging genomesequence

comparativegenomics

http://zamir.sgn.cornell.edu/Qtl/Html/home.htm

Lippman, Z.B., Semel, Y. and Zamir, D. (2007) Current Opinion in Genetics & Development, 17, 545-552.

Copyright ©2004 American Society of Plant Biologists

Tanksley,S. D. Plant Cell 2004;16:S181-S189

Collage depicting wide variation in size and shape of tomato fruit

Srivastava, A. and Handa, A.K. (2005) Hormonal regulation of tomato fruit development: A molecular perspective. Journal of Plant Growth Regulation, 24, 67-82.

Figure 1. Model for GA action through DELLA protein degradation. In the absence of GA, GA action is repressed by DELLA protein. When GA is present, the GID1 receptor binds GA. The GID1–GA complex then interacts with the DELLA and TVHYNP motifs of the DELLA protein, resulting in the recognition of DELLA protein by the SCFGID2/SLY1 complex (consisting of Skp1, Cullin, F-box protein, and Rbx1). After DELLA protein is polyubiquitinated by the SCFGID2/SLY1 complex, DELLA protein is degraded through the 26S proteasome pathway, and as a consequence GA action is activated. This consecutive reaction is predicted to occur in the nucleus. Abbreviation: Ub, ubiquitin.

Hirano, K., Ueguchi-Tanaka, M. and Matsuoka, M. (2008) GID1-mediated gibberellin signaling in plants. Trends in Plant Science, 13, 192-199.

Marti, C., Orzaez, D., Ellul, P., Moreno, V., Carbonell, J. and Granell, A. (2007) Silencing of DELLA induces facultative parthenocarpy in tomato fruits. Plant Journal, 52, 865-876.

ARF activators bind to auxin-response elements in promoters of auxin-response genes. a, When auxin concentrations are low, Aux/IAA repressors associate with the ARF activators (via domains III and IV, see Fig. 2) and repress expression of the genes. b, When auxin concentrations increase, auxin binds to the TIR1 receptor in the SCFTIR1 complex, leading to recruitment of the Aux/IAA repressors to TIR1. Once recruited to the SCFTIR1 complex, the repressors enter a pathway that leads to their destruction and the subsequent activation of the auxin-response genes.

Guilfoyle, T. (2007) Plant biology - Sticking with auxin. Nature, 446, 621-622.

Copyright ©2005 American Society of Plant Biologists

Wang, H., et al. Plant Cell 2005;17:2676-2692

Fruit Set and Parthenocarpy in AS-IAA9 Lines

Copyright ©2007 American Society of Plant Biologists

Costantini, E., et al. Plant Physiol. 2007;143:1689-1694

Grape bunches harvested from Thompson Seedless control (A, left) and genetically modified (GM; A, right), and Silcora control (B, left) and GM (B, right)

High auxin grapes

Fig. 1. (A) Fruit size extremes in the genus Lycopersicon. On the left is a fruit from the wild tomato species L. pimpinellifolium, which like all other wild tomato species, bears very small fruit. On the right is a fruit from L. esculentum cv Giant Red, bred to produce extremely large tomatoes. (B) Phenotypic effect of the fw2.2 transgene in the cultivar Mogeor. Fruit are from R1 progeny of fw107 segregating for the presence (+) or absence ( ) of cos50 containing the small-fruit allele.

Frary, A., Nesbitt, T.C., Grandillo, S., van der Knaap, E., Cong, B., Liu, J.P., Meller, J., Elber, R., Alpert, K.B. and Tanksley, S.D. (2000) fw2.2: A quantitative trait locus key to the evolution of tomato fruit size. Science, 289, 85-88.

Profiles for developing fruit of TA1143 and TA1144 NILs

Cong B. et.al. PNAS 2002;99:13606-13611

Copyright © 2002, The National Academy of Sciences

Copyright ©2005 American Society of Plant Biologists

Cheniclet, C., et al. Plant Physiol. 2005;139:1984-1994

Relationship between fruit size and ploidy

Gonzalez, N., Gevaudant, F., Hernould, M., Chevalier, C. and Mouras, A. (2007) The cell cycle-associated protein kinase WEE1 regulates cell size in relation to endoreduplication in developing tomato fruit. Plant Journal, 51, 642-655.

Copyright ©2006 American Society of Plant Biologists

Fernandez, L., et al. Plant Physiol. 2006;140:537-547

Morphology of the ovary and berry of the Flb- mutant

Cong, B., Barrero, L.S. and Tanksley, S.D. (2008) Regulatory change in YABBY-like transcription factor led to evolution of extreme fruit size during tomato domestication. Nature Genetics, 40, 800-804.

Molecular cloning of the OVATE QTL

Liu J. et.al. PNAS 2002;99:13302-13306

Copyright © 2002, The National Academy of Sciences

Complementation of pear-shaped phenotypes by OVATE

Liu J. et.al. PNAS 2002;99:13302-13306

Copyright © 2002, The National Academy of Sciences

Xiao, H., Jiang, N., Schaffner, E., Stockinger, E.J. and van der Knaap, E. (2008) A retrotransposon-mediated gene duplication underlies morphological variation of tomato fruit. Science, 319, 1527-1530.

biochemically diverse:

varied chemical compositionspigmentscell wall components – texturesvitamin / organic acid compositionsaroma compounds are distinct

Ripening diversity:on the vinefruits that ripen off the vine – persimmon, avocado

TYPICAL CHANGES THAT OCCUR DURING RIPENING

1) Change in color

2) Softening and associated alteration in texture

3) Synthesis of aroma compounds

4) Altered sugar metabolism

5) Increase susceptibility to pathogen infection

Table 1.  Variation between species in the extent of cell wall modifications occurring during ripeningLoss of Gal and Ara from unripe to ripe: high > 70%, moderate 25–70%, low < 25%. Data compiled from Gross and Sams (1984); Huber (1984); Elkashif and Huber (1988); et al. (1989); Ranwala et al. (1992); Wade et al. (1992); Yoshioka et al. (1992); Huber and O’Donoghue (1993); Redgwell et al. (1997a, b); Paull et al. (1999); Harpster et al. (2002a); Brummell et al. (2004a)

Brummell DA (2006) Cell wall disassembly in ripening fruit. Functional Plant Biology 33, 103–119.

Climacteric versus non-climacteric

versus

Cell enlargement

Ethylene production

Non-Climacteric fruitsClimacteric fruits

Respiration

Gas

pro

ducti

on ra

te Grow

th

Maturation

Ripening

Sene

scen

ce

Growth

Cell division

Transgenic approaches and mutant analysis to inhibit ethylene synthesis and action

Copyright restrictions may apply.

Nishiyama, K. et al. J. Exp. Bot. 2007 58:1281-1290; doi:10.1093/jxb/erl283

Flesh firmness of wild-type and ACO antisense Charentais melons during ripening on the vine (left panel) and ACO antisense fruit after 1 d and 4 d of ethylene treatment (50 ppm) beginning at

46 DAP (right panel)

Copyright ©2007 American Society of Plant Biologists

Schaffer, R. J., et al. Plant Physiol. 2007;144:1899-1912

Volatiles produced by AO3 fruit after exposure to ethylene

Time Point:

Stage Name:

DAP:

Expression Ratio:

1 2 3 4 5 6 7 8 109

1cm MG B B+5 B+10 B+15

7 17 27 39 41 47 52 57

17 7

2717

3927

5257

5247

4743

4139

43

Replicas of Pooled Sample: 3

Cy3/Cy5 Dye-Flip Replicates:

333 3 3 3 3 3

2 2 2 2 2 2 2 2 2

17 DAP 27 DAP

Total Cy3/Cy5Hybridizations: 6 6666

B-1 B+1

42

4241

4241

6 6 6 6

Figure 1: Direct sequential design for expression profiling in ten different stages of tomato fruit development. For each of the nine sequential time-point comparisons, Cy™-labeled cDNAs were hybridized to six independent microarrays using a triple replica dye-flip design (see Methods). Equivalent age fruit tissue was collected from a normal line (cv. Ailsa craig) and a line nearly isogenic and homozygous for the NR mutation. Abbreviations: Cy3™, cyanine3 fluor; Cy5™, cyanine5 fluor; B, breaker stage; B+5, five days post breaker; B+10, 10 days post breaker; B+15, 15 days post breaker; DAP, days after pollination; MG, mature-green.

Alba et al., 2005 Plant Cell 17: 2954-2965

Nr

Up

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d E

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n-r

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Ex

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WT

Array ElementTim

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Rel

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Figure 3. The effect of the NR lesion on transcriptome dynamics. NR has a global effect on gene expression in developing tomato pericarp and this gene serves as both a positive and negative regulator of expression throughout fruit maturation and ripening. Surprisingly, NR impacts the expression of apparently thousands of genes prior to fruit ripening. These data also indicate that the expression of many ripening-related genes is not influenced by theNr allele in this cultivar. Expression profiles for the Nr pericarp transcriptome (right panel) were plotted in the same order as those for WT pericarp transcriptome (left panel), such that the EST represented at position 1 in the WT plots is the same EST represented at position 1 in the Nr plots. Two different perspectives of each transcriptome are shown, with the upper view in each panel showing relative gene activation and the lower view in each panel showing relative gene repression. Abbreviations: Nr, Never-ripe; WT, wild-type.

Alba et al., 2005 Plant Cell 17: 2954-2965

The effects of exogenous epi-BL and Brz (an inhibitor of BR biosynthesis) on grape berry ripening.

Changes in BR levels during grape berry development.

Brassinosteroids promote ripening

brassinolide

Symons et al., 2006 Plant Physiol 140: 150-158

Brassinosteroids promote ripening

brassinolide

Lisso et al., 2006 Phytochemistry 67: 2232 -2238

FACTORS OTHER THAN ETHYLENE THAT CONTROL RIPENING

ripening inhibitor

(rin)

non-ripening (nor)

Colorless non-ripening

(Cnr)

wild type

Single monogenic mutants that severely inhibit fruit ripening. The biochemical and gene expression changes that occur during normal ripening do not occur. Mutants lack the normal increase in respiration and ethylene production that occurs during ripening. Application of ethylene to these mutants can stimulate the expression of some ripening related genes but this does not lead to ripening. This suggests that other factors in addition to ethylene control ripening.

ripening inhibitor

(rin)

Colorless non-ripening

(Cnr)

RIN encodes a MADS-box gene. Member of a familyof transcription factors that control development. In plants essential for floral and fruit development.Act as multimeric complexes which implies other MADS transcription factors are likely to be involved in regulating fruit ripening.Vrebalov et al., 2002 Science 296: 343-346

CNR encodes an SBP-box protein. Also a class of transcription factor involved in plant development. Have been implicated in regulating the expression of MADS-box genes.Manning et al., 2006 Nat Genetics 38: 948-952

Interesting questions

1) Comparison between dry and fleshy fruit development what genes control this transition – are similar genes involved in the development of both fruit types?

5) What signals other than ethylene promote ripening ?

6) Are genes like RIN , NOR and CNR master regulators of ripening across diverse species ?

4) What will genome / transcriptome sequencing of fleshy fruit bearing species reveal ?

2) What are the regulatory networks that control the development and ripening of fleshy ?

3) How is fleshy fruit development regulated in species that are not true berries ?