G. ZHANG - Weevil-plant associations revealed by multi-locus plant DNA barcoding
1
Guanyang Zhang, Juyan Pourturk & Nico Franz Figure 1. Host specificity. Proportions of generalists (associated with ≥ 2 plant families) = 38% and specialists (1 family) = 62%. Generalist Specialist n = 16 (38%) n = 26 (62%) (weevil species) No. host plant families No. weevil species Figure 2. Host range/breadth. Distribution of the number of plant families associated with generalist weevil species. Figure 3. Weevil phylogeny and plant hosts. Phylogenetic relationships are not strongly correlated with host plant associations in the sampled weevil species. The same plant families are scattered across distant weevil lineages, and sister groups often do not share hosts. • Weevils (Curculionoidea; ~ 70,000 species) are primarily phytophagous insects. • Host plant specificity and range information is traditionally based on field studies or specimen label data (1), and remains unknown for most species. • Previous DNA-based studies of weevil-host plant associations were performed at broad taxonomic scales (2), as opposed to comparing closely related species. School of Life Sciences, Arizona State University, Tempe, Arizona, USA [email protected] taxonbytes.org somanyinsects.org @GYZhang2 1. What host plants do weevils feed on? We identified weevil-host plant associations by sequencing plant DNA extracted from weevil gut contents. 2. Are weevils specialists or generalists? We characterized host plant specificity and range, focusing on a densely sampled group Eustylini + Geonemini (Entiminae) from the Neotropics. 3. Are weevil phylogenetic relationships correlated with host plant associations? We investigated whether host distributions on the sampled weevil phylogeny appear to be clustered (with correlation) or scattered (lack of correlation). References & Acknowledgments (1) Vaurie, P. 1961. A review of the Jamaican species of the genus Exophthalmus (Coleoptera, Curculionidae, Otiorhynchinae). American Museum Novitates 2062: 1–42. (2) Navarro, S.P. et al. 2010. DNA profiling of host-herbivore interactions in tropical forests. Ecological Entomology 35: 18–32. • Support by the National Science Foundation (DEB-1155984) and the United States Department of Agriculture (USDA) (Agreement No. 58-1275-1-335) is kindly acknowledged. • Dr. Robert Andersion (CMNC) contributed specimens to this study, and members of the Franz Lab are thanked for their curatorial contributions and fruitful discussions. Background: Weevil-Plant Associations Research Questions & Objectives • 49 weevil specimens were sampled from the Neotropics. • Two major lineages were represented; i.e., members of the tribes Eustylini and Geonemini, each with numerous closely related species. • The six-loci weevil phylogeny was reconstructed using Maximum Likelihood. • Host plant DNA was extracted from samples of adult weevil gut contents. • Three plant barcoding genes were targeted and sequenced: trnL, rbcL and matK. • Multiple plants in the same individual were isolated by molecular cloning. • Plant sequences were identified to family based on the best hit of BLAST search. Material & Methods Table 1. Plant families and frequencies of associations. 28 families were identified. Fabaceae and Melastomataceae were most common host plants, comprising 41% of all associations. 1. Host plant DNA were successfully extracted and sequenced for 86% of the weevil samples (42/49). 2. In total, 28 host plant families were identified to be fed on by the 42 analyzed weevil species (Table 1). 3. The most common plant hosts were Fabaceae and Melastomataceae, comprising 41% of all associations. 4. 38% of the weevils (n = 16) appeared to be generalists associated with 2+ plant families, and 62% (n = 26) fed on one family and were specialists (Fig. 1). Surprisingly, three generalists fed on 4-5 families (Fig. 2). 5. The phylogenetic relationships were not reliable predictors of weevil/host plant associations (Fig. 3). 6. Overall, our analysis demonstrates the effectiveness of multi-locus DNA-barcoding in characterizing weevil host plant associations. However the gut content analysis approach is limited by spatio-temporally restricted sampling, the lack of plant vouchers for verification, and by incomplete database of sequences in Genbank. Results & Discussion Weevil-plant associations revealed by multi-locus plant DNA barcoding Habitus of representative weevils Phylogeny of weevils Host plant family 33637 33751 33635 33607 33779 33777 33774 33679 33766 33797 33604 33676 33602 33612 33664 33658 33609 33662 33627 33623 33666 33671 33670 33669 33608 33667 33672 33668 33638 33603 33674 33626 33621 33630 33610 33641 33629 33764 33762 33792 33794 33756 WEEVIL Adoxaceae Aquifoliaceae Asteraceae Burseraceae Calophyllaceae Dryopteridaceae Euphorbiaceae Fab. (Caesalpinioideae) Fab. (Papilionoidea) Fagaceae Melastomataceae Meliaceae Moraceae Musaceae Nyctaginaceae Olygonaceae Phyllanthaceae Picramniaceae Pinaceae Piperaceae Poaceae Primulaceae Ranunculaceae Rhamnaceae Rosaceae Rubiaceae Sapotaceae Solanaceae Trebouxiophyceae Generalist Association Total number of weevil-plant associations recovered: 73. 43 (59%) generalist / 30 (41%) specialist species associations. Fabaceae (Papilionoidea) 14 Melastomataceae 9 Fabaceae (Caesalpinioideae) 7 Aquifoliaceae 4 Asteraceae 3 Calophyllaceae 3 Piperaceae 3 Ranunculaceae 3 Rubiaceae 3 Meliaceae 2 Moraceae 2 Poaceae 2 Primulaceae 2 Plant family No. associated weevil species Plant family No. associated weevil species Adoxaceae 1 Burseraceae 1 Dryopteridaceae 1 Euphorbiaceae 1 Fagaceae 1 Musaceae 1 Nyctaginaceae 1 Olygonaceae 1 Phyllanthaceae 1 Picramniaceae 1 Pinaceae 1 Rhamnaceae 1 Rosaceae 1 Sapotaceae 1 Solanaceae 1 Trebouxiophyceae 1
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Transcript of G. ZHANG - Weevil-plant associations revealed by multi-locus plant DNA barcoding
Guanyang Zhang, Juyan Pourturk & Nico Franz
Figure 1. Host specificity. Proportions of generalists (associated with ≥ 2 plant
families) = 38% and specialists (1 family) = 62%.
Generalist Specialist
n = 16 (38%) n = 26 (62%)
(weevil species)
No. host plant families
No. w
eevil
sp
ecie
s
Figure 2. Host range/breadth. Distribution of the number of plant families
associated with generalist weevil species.
Figure 3. Weevil phylogeny and plant hosts. Phylogenetic relationships are not strongly correlated with host plant associations in the sampled weevil species.
The same plant families are scattered across distant weevil lineages, and sister groups often do not share hosts.
• Weevils (Curculionoidea; ~ 70,000 species) are primarily phytophagous insects.
• Host plant specificity and range information is traditionally based on field
studies or specimen label data (1), and remains unknown for most species.
• Previous DNA-based studies of weevil-host plant associations were performed at
broad taxonomic scales (2), as opposed to comparing closely related species.
School of Life Sciences, Arizona State University, Tempe, Arizona, USA [email protected] taxonbytes.org somanyinsects.org @GYZhang2
1. What host plants do weevils feed on?
We identified weevil-host plant associations by sequencing plant DNA
extracted from weevil gut contents.
2. Are weevils specialists or generalists?
We characterized host plant specificity and range, focusing on a densely
sampled group Eustylini + Geonemini (Entiminae) from the Neotropics.
3. Are weevil phylogenetic relationships correlated with host plant associations?
We investigated whether host distributions on the sampled weevil phylogeny
appear to be clustered (with correlation) or scattered (lack of correlation).
References & Acknowledgments (1)Vaurie, P. 1961. A review of the Jamaican species of the
genus Exophthalmus (Coleoptera, Curculionidae,
Otiorhynchinae). American Museum Novitates 2062: 1–42.
(2)Navarro, S.P. et al. 2010. DNA profiling of host-herbivore
interactions in tropical forests. Ecological Entomology 35:
18–32.
• Support by the National Science Foundation (DEB-1155984)
and the United States Department of Agriculture (USDA)
(Agreement No. 58-1275-1-335) is kindly acknowledged.
• Dr. Robert Andersion (CMNC) contributed specimens to this
study, and members of the Franz Lab are thanked for their
curatorial contributions and fruitful discussions.
Background: Weevil-Plant Associations
Research Questions & Objectives
• 49 weevil specimens were sampled from the Neotropics.
• Two major lineages were represented; i.e., members of the tribes Eustylini and
Geonemini, each with numerous closely related species.
• The six-loci weevil phylogeny was reconstructed using Maximum Likelihood.
• Host plant DNA was extracted from samples of adult weevil gut contents.
• Three plant barcoding genes were targeted and sequenced: trnL, rbcL and matK.
• Multiple plants in the same individual were isolated by molecular cloning.
• Plant sequences were identified to family based on the best hit of BLAST search.
Material & Methods
Table 1. Plant families and frequencies of associations. 28 families were identified. Fabaceae and Melastomataceae were
most common host plants, comprising 41% of all associations.
1. Host plant DNA were successfully extracted and sequenced for 86% of the weevil samples (42/49).
2. In total, 28 host plant families were identified to be fed on by the 42 analyzed weevil species (Table 1).
3. The most common plant hosts were Fabaceae and Melastomataceae, comprising 41% of all associations.
4. 38% of the weevils (n = 16) appeared to be generalists associated with 2+ plant families, and 62% (n = 26)
fed on one family and were specialists (Fig. 1). Surprisingly, three generalists fed on 4-5 families (Fig. 2).
5. The phylogenetic relationships were not reliable predictors of weevil/host plant associations (Fig. 3).
6. Overall, our analysis demonstrates the effectiveness of multi-locus DNA-barcoding in characterizing weevil
host plant associations. However the gut content analysis approach is limited by spatio-temporally restricted
sampling, the lack of plant vouchers for verification, and by incomplete database of sequences in Genbank.
Results & Discussion
Weevil-plant associations revealed by multi-locus plant DNA barcoding
Habitus of representative weevils
Phylogeny of weevils
Host plant family
WEEVIL
Ado
xace
ae
Aqu
ifolia
ceae
Aster
acea
e
Bur
sera
ceae
Calop
hylla
ceae
Dry
opte
ridac
eae
Eup
horb
iace
ae
Fab. (
Cae
salpin
ioid
eae)
Fab. (
Papilion
oide
a)
Fagac
eae
Mel
asto
mat
acea
e
Mel
iace
ae
Mor
acea
e
Mus
acea
e
Nyc
tagina
ceae
Olygo
nace
ae
Phy
llant
hace
ae
Picra
mniac
eae
Pinac
eae
Piper
acea
e
Poa
ceae
Prim
ulac
eae
Ran
uncu
lace
ae
Rha
mna
ceae
Ros
acea
e
Rub
iace
ae
Sap
otac
eae
Solan
acea
e
Trebo
uxioph
ycea
e
33637
33751
33635
33607
33779
33777
33774
33679
33766
33797
33604
33676
33602
33612
33664
33658
33609
33662
33627
33623
33666
33671
33670
33669
33608
33667
33672
33668
33638
33603
33674
33626
33621
33630
33610
33641
33629
33764
33762
33792
33794
33756
WEEVIL
Ado
xace
ae
Aqu
ifolia
ceae
Aster
acea
e
Bur
sera
ceae
Calop
hylla
ceae
Dry
opte
ridac
eae
Eup
horb
iace
ae
Fab. (
Cae
salpin
ioid
eae)
Fab. (
Papilion
oide
a)
Fagac
eae
Mel
asto
mat
acea
e
Mel
iace
ae
Mor
acea
e
Mus
acea
e
Nyc
tagina
ceae
Olygo
nace
ae
Phy
llant
hace
ae
Picra
mniac
eae
Pinac
eae
Piper
acea
e
Poa
ceae
Prim
ulac
eae
Ran
uncu
lace
ae
Rha
mna
ceae
Ros
acea
e
Rub
iace
ae
Sap
otac
eae
Solan
acea
e
Trebo
uxioph
ycea
e
Generalist
Association
Total number of weevil-plant associations recovered: 73.
43 (59%) generalist / 30 (41%) specialist species associations.
Fabaceae
(Papilionoidea)14
Melastomataceae 9
Fabaceae
(Caesalpinioideae)7
Aquifoliaceae 4
Asteraceae 3
Calophyllaceae 3
Piperaceae 3
Ranunculaceae 3
Rubiaceae 3
Meliaceae 2
Moraceae 2
Poaceae 2
Primulaceae 2
Plant family
No.
associated
weevil
species
Plant family
No.
associated
weevil
species
Adoxaceae 1
Burseraceae 1
Dryopteridaceae 1
Euphorbiaceae 1
Fagaceae 1
Musaceae 1
Nyctaginaceae 1
Olygonaceae 1
Phyllanthaceae 1
Picramniaceae 1
Pinaceae 1
Rhamnaceae 1
Rosaceae 1
Sapotaceae 1
Solanaceae 1
Trebouxiophyceae 1
