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      Geometric morphometrics reveals shifts in flower shape symmetry and size following gene knockdown of CYCLOIDEA and ANTHOCYANIDIN SYNTHASE

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          Abstract

          Background

          While floral symmetry has traditionally been assessed qualitatively, recent advances in geometric morphometrics have opened up new avenues to specifically quantify flower shape and size using robust multivariate statistical methods. In this study, we examine, for the first time, the ability of geometric morphometrics to detect morphological differences in floral dorsoventral asymmetry following virus-induced gene silencing (VIGS). Using Fedia graciliflora Fisch. & Meyer (Valerianaceae) as a model, corolla shape of untreated flowers was compared using canonical variate analysis to knockdown phenotypes of CYCLOIDEA2A ( FgCYC2A), ANTHOCYANIDIN SYNTHASE ( FgANS), and empty vector controls .

          Results

          Untreated flowers and all VIGS treatments were morphologically distinct from each other, suggesting that VIGS may cause subtle shifts in floral shape. Knockdowns of FgCYC2A were the most dramatic, affecting the position of dorsal petals in relation to lateral petals, thereby resulting in more actinomorphic-like flowers. Additionally, FgANS knockdowns developed larger flowers with wider corolla tube openings.

          Conclusions

          These results provide a method to quantify the role that specific genes play in the developmental pathway affecting the dorsoventral axis of symmetry in zygomorphic flowers. Additionally, they suggest that ANS may have an unintended effect on floral size and shape.

          Electronic supplementary material

          The online version of this article doi: (10.1186/s12870-017-1152-x) contains supplementary material, which is available to authorized users.

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          Most cited references81

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          Virus-induced gene silencing in tomato.

          We have previously demonstrated that a tobacco rattle virus (TRV)-based vector can be used in virus-induced gene silencing (VIGS) to study gene function in Nicotiana benthamiana. Here we show that recombinant TRV infects tomato plants and induces efficient gene silencing. Using this system, we suppressed the PDS, CTR1 and CTR2 genes in tomato. Suppression of CTR1 led to a constitutive ethylene response phenotype and up-regulation of an ethylene response gene, CHITINASE B. This phenotype is similar to Arabidopsis ctr1 mutant plants. We have constructed a modified TRV vector based on the GATEWAY recombination system, allowing restriction- and ligation-free cloning. Our results show that tomato expressed sequence tags (ESTs) can easily be cloned into this modified vector using a single set of primers. Using this vector, we have silenced RbcS and an endogenous gene homologous to the tomato EST cLED3L14. In the future, this modified vector system will facilitate large-scale functional analysis of tomato ESTs.
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            Fluctuating Asymmetry: Measurement, Analysis, Patterns

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              R: a language and environment for statistical computing

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                Author and article information

                Contributors
                +1 (718) 990-6790 , brent.a.berger@gmail.com
                vincent.ricigliano@ars.usda.gov
                yoland.savriama@helsinki.fi
                aedriclim@gmail.com
                vlthompson@ucdavis.edu
                howarthd@stjohns.edu
                Journal
                BMC Plant Biol
                BMC Plant Biol
                BMC Plant Biology
                BioMed Central (London )
                1471-2229
                17 November 2017
                17 November 2017
                2017
                : 17
                : 205
                Affiliations
                [1 ]ISNI 0000 0001 1954 7928, GRID grid.264091.8, Department of Biological Sciences, , St. John’s University, ; 8000 Utopia Parkway, Queens, NY 11439 USA
                [2 ]USDA-ARS, Carl Hayden Bee Research Center, 2000 E. Allen Road, Tucson, AZ 85719 USA
                [3 ]ISNI 0000 0004 0410 2071, GRID grid.7737.4, Institute of Biotechnology, , University of Helsinki, ; PO Box 56 (Viikinkaari 5), FI-00014 Helsinki, Finland
                Article
                1152
                10.1186/s12870-017-1152-x
                5693587
                29149840
                f022cf8f-b62b-4f2f-b0ce-95630df0a4c4
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 3 February 2017
                : 7 November 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: IOS 1121301
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Plant science & Botany
                geometric morphometrics,virus-induced gene silencing,cycloidea,fedia graciliflora,floral symmetry,anthocyanidin synthase

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