杨婷婷, 赵立华, 邱润霜, 等. 番茄斑萎病毒2个非结构蛋白VIGS体系的初步构建[J]. 西南林业大学学报（自然科学）, 2022, 42(2): 151–156 . DOI: 10.11929/j.swfu.202009054
引用本文: 杨婷婷, 赵立华, 邱润霜, 等. 番茄斑萎病毒2个非结构蛋白VIGS体系的初步构建[J]. 西南林业大学学报（自然科学）, 2022, 42(2): 151–156 . DOI: 10.11929/j.swfu.202009054
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Yang Tingting, Zhao Lihua, Qiu Runshuang, Chen Si, Zhang Jie, Li Bowen, Zhang Zhongkai, Ma Changle. Construction of 2 Non-structural Proteins VIGS System of TSWV[J]. Journal of Southwest Forestry University, 2022, 42(2): 151-156. DOI: 10.11929/j.swfu.202009054
Citation: Yang Tingting, Zhao Lihua, Qiu Runshuang, Chen Si, Zhang Jie, Li Bowen, Zhang Zhongkai, Ma Changle. Construction of 2 Non-structural Proteins VIGS System of TSWV[J]. Journal of Southwest Forestry University, 2022, 42(2): 151-156. DOI: 10.11929/j.swfu.202009054
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番茄斑萎病毒2个非结构蛋白VIGS体系的初步构建

Construction of 2 Non-structural Proteins VIGS System of TSWV

    摘要
  • 摘要: 将番茄斑萎病毒（TSWV）的NSm、NSs基因200 bp左右构建到pTRV−PTV00沉默载体上，通过农杆菌GV3101浸润本氏烟。注射10 d左右，待注射PDS基因的植株发白，将注射pTRV−PTV00−NSm、pTRV−PTV00−NSs以及pTRV−PTV00载体的植株接种TSWV，显症后采集系统叶（接种叶上一叶）应用实时荧光定量PCR检测NSm、NSs基因表达量。通过测定NSm、NSs基因表达量发现 NSm基因的沉默效率为86.7%、NSs基因的沉默效率为92.00%。结果表明：通过pTRV−PTV00载体构建NSm、NSs 基因沉默载体沉默效率接近90.00%，此载体适用于病毒基因沉默载体的构建，且NSm、NSs基因沉默载体能应用后续基因功能研究。

     

    Abstract: In this study, the sequences of 200 bp of tomato spotted wilt orthotospovirus(TSWV) NSm, NSs genes were constructed to pTRV−PTV00 silencing vector and infiltrated agrobacterium GV3101 in Nicotiana benthamiana. About 10 days after injection, the plants with injecting PDS gene became white, the plants with injecting the vectors of pTRV−PTV00−NSm, pTRV−PTV00−NSs and pTRV−PTV00 were inoculated with TSWV. The expression of NSm and NSs genes was detected by qRT−PCR in the systematic leaves(one leaf above the inoculated leaf) after manifestation, and the gene silencing efficiency was obtained. The silencing efficiency of NSm and NSs gene was 86.7% and 92.00%, respectively. The results showed that silencing efficiency of NSm and NSs gene silencing was close to 90.00% constructed by pTRV−PTV00 vector, which was suitable for the construction of viral gene silencing vectors, and NSm and NSs gene silencing vectors could be used for subsequent gene function studies.

     

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