Plant-mediated RNA interference(RNAi)has emerged as a promising technology for insect control.The green peach aphid,Myzus persicae,feeds on over 400 species of host plants.Brassica napus(rape)is the second most import...Plant-mediated RNA interference(RNAi)has emerged as a promising technology for insect control.The green peach aphid,Myzus persicae,feeds on over 400 species of host plants.Brassica napus(rape)is the second most important oilseed crop worldwide.Myzus persicae is highly reproductive and causes severe damage to the rape plants due to its quite flexible life cycle.In this study,we tested the RNAi effects of transgenic rape plants on M.persicae.By in vitro feeding M.persicae with artificial diets containing double-stranded RNAs(dsRNAs)targeting seven aphid genes,we identified a new gene encoding the partitioning-defective protein 6(Par6)as the most potent RNAi target.Tissue-and stage-expression analysis of Par6 suggested this gene is highly expressed in the embryo and adult stage of M.persicae.We next generated transgenic rape plants expressing ds Par6 by Agrobacteriummediated transformation and obtained nine independent transgenic lines.Compared to wild-type control plants,transgenic rape lines expressing ds Par6 showed strong resistance to M.persicae.Feeding assays revealed that feeding transgenic rape plants to M.persicae significantly decreased MpPar6 expression and survival rate and impaired fecundity.Furthermore,we showed that the resistance levels to M.persicae are positively correlated with ds Par6 expression levels in transgenic rape plants.Our study demonstrates that transgenic rape plants expressing ds Par6 are efficiently protected from M.persicae.Interfering with the genes involved in embryo development could be the effective RNAi targets for controlling aphids and potentially other insect pests.展开更多
With the development of tourism at home and abroad,Rapeseed(Brassica napus)has become an important ornamental plant.However,its ornamental value at the inflorescence stage is greatly reduced by Sclerotinia sclerotioru...With the development of tourism at home and abroad,Rapeseed(Brassica napus)has become an important ornamental plant.However,its ornamental value at the inflorescence stage is greatly reduced by Sclerotinia sclerotiorum.Identification of important genes in the defense responses is critical for molecular breeding,which is an important strategy for controlling the disease.In this study,we isolated a B.napus WRKY transcription factor gene,BnaWRKY75.BnaWRKY75 was found to encode a nucleus-localized protein and exhibited relatively high expression in the stems.Arabidopsis thaliana transgenic plants expressing BnaWRKY75 showed enhanced resistance to S.sclerotiorum,and both ProBnaWRKY75:GUS and gene expression analyses showed that BnaWRKY75 was highly responsive to S.sclerotiorum infection,indicating the involvement of BnaWRKY75 in response to this infection.Furthermore,overexpression(OE)of BnaWRKY75 in B.napus significantly enhanced the resistance to S.sclerotiorum,whereas the resistance was reduced in RNAi transgenic B.napus plants.Moreover,the BnaWRKY75-OE B.napus plants exhibited constitutive activation of salicylic acid-,jasmonic acid-,and ethylene-mediated defense responses and the inhibition of both H_(2)O_(2)and O_(2)·^(-)accumulation in response to this pathogen.By contrast,BnaWRKY75-RNAi plants showed a reverse pattern,suggesting that BnaWRKY75 is involved in hormonal signaling pathways and in the control of reactive oxygen species accumulation.In conclusion,these data indicate that BnaWRKY75,a regulator of multiple defense responses,positively regulates resistance against S.sclerotiorum,which may guide the improvement of resistance in rapeseed.展开更多
Five stems of rapeseed with abundant black microsclerotia were collected from Huangyuan County of Qinghai Province,China,and fungal isolates were obtained from the stems.They were identified based on morphology,molecu...Five stems of rapeseed with abundant black microsclerotia were collected from Huangyuan County of Qinghai Province,China,and fungal isolates were obtained from the stems.They were identified based on morphology,molecular features and specific PCR detection.The results showed that the 10 fungal isolates belonged to Verticillium longisporum lineage A1/D3.One of the 10 isolates(HW7-1)was tested for virulence on three species of rapeseed,including B.napus Zhongshuang 9,B.rapa Qingyou 9 and B.juncea Tayou 2 by conidia inoculation of HW7-1 on roots of young seedlings.Control seedlings were inoculated with V.dahliae conidia or water alone.The seedlings of these treatments were transplanted in culture mix and incubated in a growth chamber(20℃).Results suggested that the control seedlings of three cultivars appeared quite healthy,while the seedlings inoculated with HW7-1 turned yellowing leaves,seedling stunting or even death after 22 days post-inoculation.V.longisporum was re-isolated from he yellow leaves,thus fulfilling Koch's postulates.Moreover,compared to the control treatments,inoculation with HW7-1 caused flowering delay and seed yield reduction on Tayou 2 with production of microsclerotia on the stems.To our knowledge,this is the first report of V.longisporum lineage A1/D3 on rapeseed in northwestern China.展开更多
基金supported by the National Natural Science Foundation of China(32102297 and 32272634)。
文摘Plant-mediated RNA interference(RNAi)has emerged as a promising technology for insect control.The green peach aphid,Myzus persicae,feeds on over 400 species of host plants.Brassica napus(rape)is the second most important oilseed crop worldwide.Myzus persicae is highly reproductive and causes severe damage to the rape plants due to its quite flexible life cycle.In this study,we tested the RNAi effects of transgenic rape plants on M.persicae.By in vitro feeding M.persicae with artificial diets containing double-stranded RNAs(dsRNAs)targeting seven aphid genes,we identified a new gene encoding the partitioning-defective protein 6(Par6)as the most potent RNAi target.Tissue-and stage-expression analysis of Par6 suggested this gene is highly expressed in the embryo and adult stage of M.persicae.We next generated transgenic rape plants expressing ds Par6 by Agrobacteriummediated transformation and obtained nine independent transgenic lines.Compared to wild-type control plants,transgenic rape lines expressing ds Par6 showed strong resistance to M.persicae.Feeding assays revealed that feeding transgenic rape plants to M.persicae significantly decreased MpPar6 expression and survival rate and impaired fecundity.Furthermore,we showed that the resistance levels to M.persicae are positively correlated with ds Par6 expression levels in transgenic rape plants.Our study demonstrates that transgenic rape plants expressing ds Par6 are efficiently protected from M.persicae.Interfering with the genes involved in embryo development could be the effective RNAi targets for controlling aphids and potentially other insect pests.
基金supported by the National Natural Science Foundation of China(Grant No.31771836)the National Key Research and Development Program of China(Grant No.2018YFD0201003).
文摘With the development of tourism at home and abroad,Rapeseed(Brassica napus)has become an important ornamental plant.However,its ornamental value at the inflorescence stage is greatly reduced by Sclerotinia sclerotiorum.Identification of important genes in the defense responses is critical for molecular breeding,which is an important strategy for controlling the disease.In this study,we isolated a B.napus WRKY transcription factor gene,BnaWRKY75.BnaWRKY75 was found to encode a nucleus-localized protein and exhibited relatively high expression in the stems.Arabidopsis thaliana transgenic plants expressing BnaWRKY75 showed enhanced resistance to S.sclerotiorum,and both ProBnaWRKY75:GUS and gene expression analyses showed that BnaWRKY75 was highly responsive to S.sclerotiorum infection,indicating the involvement of BnaWRKY75 in response to this infection.Furthermore,overexpression(OE)of BnaWRKY75 in B.napus significantly enhanced the resistance to S.sclerotiorum,whereas the resistance was reduced in RNAi transgenic B.napus plants.Moreover,the BnaWRKY75-OE B.napus plants exhibited constitutive activation of salicylic acid-,jasmonic acid-,and ethylene-mediated defense responses and the inhibition of both H_(2)O_(2)and O_(2)·^(-)accumulation in response to this pathogen.By contrast,BnaWRKY75-RNAi plants showed a reverse pattern,suggesting that BnaWRKY75 is involved in hormonal signaling pathways and in the control of reactive oxygen species accumulation.In conclusion,these data indicate that BnaWRKY75,a regulator of multiple defense responses,positively regulates resistance against S.sclerotiorum,which may guide the improvement of resistance in rapeseed.
基金supported by the Earmarked Fund for CARS-12 from National Modern Agricultural Technology System.
文摘Five stems of rapeseed with abundant black microsclerotia were collected from Huangyuan County of Qinghai Province,China,and fungal isolates were obtained from the stems.They were identified based on morphology,molecular features and specific PCR detection.The results showed that the 10 fungal isolates belonged to Verticillium longisporum lineage A1/D3.One of the 10 isolates(HW7-1)was tested for virulence on three species of rapeseed,including B.napus Zhongshuang 9,B.rapa Qingyou 9 and B.juncea Tayou 2 by conidia inoculation of HW7-1 on roots of young seedlings.Control seedlings were inoculated with V.dahliae conidia or water alone.The seedlings of these treatments were transplanted in culture mix and incubated in a growth chamber(20℃).Results suggested that the control seedlings of three cultivars appeared quite healthy,while the seedlings inoculated with HW7-1 turned yellowing leaves,seedling stunting or even death after 22 days post-inoculation.V.longisporum was re-isolated from he yellow leaves,thus fulfilling Koch's postulates.Moreover,compared to the control treatments,inoculation with HW7-1 caused flowering delay and seed yield reduction on Tayou 2 with production of microsclerotia on the stems.To our knowledge,this is the first report of V.longisporum lineage A1/D3 on rapeseed in northwestern China.
