Virus-induced gene silencing(VIGS)is a natural defense mechanism of plants,which can cause sequence-specific degradation of viral RNA and is often used for studying the functional genome.At present,the VIGS system med...Virus-induced gene silencing(VIGS)is a natural defense mechanism of plants,which can cause sequence-specific degradation of viral RNA and is often used for studying the functional genome.At present,the VIGS system mediated by Tobacco rattle virus(TRV)has been successfully established in many plant species.However,the VIGS system in Chinese narcissus had not been reported yet.In this paper,the first use of the TRV-VIGS system in Chinese narcissus is described in detail.By injecting pTRV2-GFP into narcissus leaves,we confirmed that TRV could infect narcissus and move in narcissus plants.After the inoculation of pTRV2-Nt PDS by two methods,phenomena such as photo-bleaching appeared in narcissus leaves and qRT-PCR results proved that PDS gene of narcissus was successfully silenced.Besides,pTRV2-NtMYB3 was inoculated into basal plates of narcissus to study the function of NtMYB3;the results showed that NtMYB3 is an inhibitor of flavonoids biosynthesis,which can increase the content of proanthocyanidins in basal plates of narcissus by repressing the expression level of NtFLS.These results indicate that the TRV-VIGS system has been successfully established in Chinese narcissus and successfully applied to the functional study of NtMYB3.展开更多
Background:Spodoptera litura(Lepidoptera:Noctuidae),commonly known as tobacco cutworm or cotton leafworm,is a polyphagous pest which causes considerable damage to cotton(Gossypium hirsutum)and other crops.Herbivore-in...Background:Spodoptera litura(Lepidoptera:Noctuidae),commonly known as tobacco cutworm or cotton leafworm,is a polyphagous pest which causes considerable damage to cotton(Gossypium hirsutum)and other crops.Herbivore-induced defence response is activated in plants against chewing pests,in which plant secondary metabolites play an important role.Dhurrinase2(SbDhr2),a cyanogenicβ-glucosidase from Sorghum bicolor,is the key enzyme responsible for the hydrolysis of dhurrin(cyanogenicβ-glucosidic substrate)to phydroxymandelonitrile.Hydroxynitrile lyase(Me HNL)from Mannihot esculanta catalyses the dissociation of cyanohydrins to hydrogen cyanide and corresponding carbonyl compound,both enzymes play a pivotal role in plant defence mechanism.Results:SbDhr2 and Me HNL genes were expressed individually and co-expressed transiently in cotton leaves.We examined the feeding response of S.litura to leaves in the choice assay.The S.litura population used in this study showed better feeding deterrence to leaves co-expressing both genes compared with the expression of an individual gene.Conclusion:Our results suggest that co-expression of SbDhr2 and Me HNL genes in cotton leaves demonstrate feeding deterrence to S.litura.Engineering cyanogenic pathway in aerial parts of cotton would be an additional defence strategy against generalist pests and can be enhanced against specialist pests.展开更多
基金the Science and technology innovation fund of Fujian Agriculture and Forestry University(Grant Nos.KFA17352A,KFA17602A)。
文摘Virus-induced gene silencing(VIGS)is a natural defense mechanism of plants,which can cause sequence-specific degradation of viral RNA and is often used for studying the functional genome.At present,the VIGS system mediated by Tobacco rattle virus(TRV)has been successfully established in many plant species.However,the VIGS system in Chinese narcissus had not been reported yet.In this paper,the first use of the TRV-VIGS system in Chinese narcissus is described in detail.By injecting pTRV2-GFP into narcissus leaves,we confirmed that TRV could infect narcissus and move in narcissus plants.After the inoculation of pTRV2-Nt PDS by two methods,phenomena such as photo-bleaching appeared in narcissus leaves and qRT-PCR results proved that PDS gene of narcissus was successfully silenced.Besides,pTRV2-NtMYB3 was inoculated into basal plates of narcissus to study the function of NtMYB3;the results showed that NtMYB3 is an inhibitor of flavonoids biosynthesis,which can increase the content of proanthocyanidins in basal plates of narcissus by repressing the expression level of NtFLS.These results indicate that the TRV-VIGS system has been successfully established in Chinese narcissus and successfully applied to the functional study of NtMYB3.
基金Project supported by the National Basic Research Program of China(No.2009CB119000)the Genetically Modified Organisms Breeding Major Projects(No.2009ZX08009-044B)+2 种基金the Program for New Century Excellent Talents in University(No.NCET-08-0485)the Program for New Century 151Talents of Zhejiang Provincethe National Natural Science Foundation of China(No.30871608and 30771406)
基金funded by the Dept.of Botany,Maulana Azad College of Arts,Science and Commerce,Aurangabad,Maharashtra,India and Government Institute of Science,Aurangabad,Maharashtra,India。
文摘Background:Spodoptera litura(Lepidoptera:Noctuidae),commonly known as tobacco cutworm or cotton leafworm,is a polyphagous pest which causes considerable damage to cotton(Gossypium hirsutum)and other crops.Herbivore-induced defence response is activated in plants against chewing pests,in which plant secondary metabolites play an important role.Dhurrinase2(SbDhr2),a cyanogenicβ-glucosidase from Sorghum bicolor,is the key enzyme responsible for the hydrolysis of dhurrin(cyanogenicβ-glucosidic substrate)to phydroxymandelonitrile.Hydroxynitrile lyase(Me HNL)from Mannihot esculanta catalyses the dissociation of cyanohydrins to hydrogen cyanide and corresponding carbonyl compound,both enzymes play a pivotal role in plant defence mechanism.Results:SbDhr2 and Me HNL genes were expressed individually and co-expressed transiently in cotton leaves.We examined the feeding response of S.litura to leaves in the choice assay.The S.litura population used in this study showed better feeding deterrence to leaves co-expressing both genes compared with the expression of an individual gene.Conclusion:Our results suggest that co-expression of SbDhr2 and Me HNL genes in cotton leaves demonstrate feeding deterrence to S.litura.Engineering cyanogenic pathway in aerial parts of cotton would be an additional defence strategy against generalist pests and can be enhanced against specialist pests.
