Virus-induced gene silencing (VIGS) in Cysticapnos vesicaria, a zygomorphic-flowered Papaveraceae (Ranunculales, basal eudicots).
OBJECTIVE
Studies of evolutionary diversification in the basal eudicot family Papaveraceae, such as the transition from actinomorphy to zygomorphy, are hampered by the lack of comparative functional studies. So far, gene silencing methods are only available in the actinomorphic species Eschscholzia californica and Papaver somniferum. This study addresses the amenability of Cysticapnos vesicaria, a derived fumitory with zygomorphic flowers, to virus-induced gene silencing (VIGS), and describes vegetative and reproductive traits in this species.


METHODS
VIGS-mediated downregulation of the C. vesicaria PHYTOENE DESATURASE gene (CvPDS) and of the FLORICAULA gene CvFLO was carried out using Agrobacterium tumefaciens transfer of Tobacco rattle virus (TRV)-based vectors. Wild-type and vector-treated plants were characterized using reverse transcription-PCR (RT-PCR), in situ hybridization, and macroscopic and scanning electron microscopic imaging.


RESULTS
Cysticapnos vesicaria germinates rapidly, can be grown at high density, has a short life cycle and is self-compatible. Inoculation of C. vesicaria with a CvPDS-VIGS vector resulted in strong photobleaching of green parts and reduction of endogenous CvPDS transcript levels. Gene silencing persisted during inflorescence development until fruit set. Inoculation of plants with CvFLO-VIGS affected floral phyllotaxis, symmetry and floral organ identities.


CONCLUSIONS
The high penetrance, severity and stability of pTRV-mediated silencing, including the induction of meristem-related phenotypes, make C. vesicaria a very promising new focus species for evolutionary-developmental (evo-devo) studies in the Papaveraceae. This now enables comparative studies of flower symmetry, inflorescence determinacy and other traits that diversified in the Papaveraceae.
Department of Environmental and Plant Biology, Ohio University, Athens, OH 45701, USA.
Oriane
O
Hidalgo
Conny
C
Bartholmes
Stefan
S
Gleissberg
D015536:Down-Regulation / Q000235:genetics; D005638:Fruit / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology / Q000821:virology; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D020868:Gene Silencing; D005838:Genotype; D056626:Inflorescence / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology / Q000821:virology; D018519:Meristem / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology / Q000821:virology; D010088:Oxidoreductases / Q000235:genetics; D028101:Papaveraceae / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology* / Q000821:virology; D010641:Phenotype; D018515:Plant Leaves / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology / Q000821:virology; D010940:Plant Proteins / Q000235:genetics*; D010942:Plant Viruses / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D012098:Reproduction / Q000235:genetics
D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D010088:Oxidoreductases; C022398:phytoene dehydrogenase