RNAi介导双生病毒和马铃薯Y病毒的分子调控技术研究
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摘要
本文以由B型烟粉虱传播的中国番茄黄化曲叶病毒(tomato yellow leaf curl China virus, TYLCCNV)和由烟蚜传播的马铃薯Y病毒(potato virus Y, PVY)为研究对象,构建TYLCCNV Y10外壳蛋白dsRNA植物表达载体和B型烟粉虱传毒相关蛋白GroEL基因片段的dsRNA植物表达载体,分别转化烟草和番茄,探索利用RNAi切断病毒传播途径的可能性；构建三个含烟蚜(Myzus persicae) RPS2基因片段的dsRNA原核表达载体,将这三种体外表达的dsRNA加入烟蚜人工饲料中,观察食用该饲料对烟蚜传播PVY的影响。本论文的研究为进一步构建双价基因,从病毒侵染和介体传毒两条途径切断病毒的侵染和流行,建立病毒分子调控技术体系打下了基础。
本文的主要结论是：
1.对TYLCCNV不同氨基酸同源性CP基因片段构建dsRNA转化烟草和番茄及其抗病性进行了研究。
2.对TYLCCNV不同长度CP基因片段构建dsRNA转化烟草和番茄及其抗病性进行了研究。
3.对利用B型烟粉虱GroEL基因片段构建dsRNA转化本氏烟和番茄干扰烟粉虱传毒进行了研究。
4.初步分析了在本氏烟中瞬时表达TYLCCNV CP基因dsRNA的抗病性。
5.构建三个含RPS2基因片段的dsRNA原核表达载体,并将这三种体外表达的dsRNA加入烟蚜人工饲料中,观察食用该饲料对烟蚜传播PVY的影响。
This study focused on tomato yellow leaf curl China virus which transmits by Bemisia tabaci and potato virus Y which transmits by aphids. TYLCCNV Y10coat protein gene and GroEL gene—related to virus transferring are selected to construct dsRNA plant expression vector respectively, then they are transformed into tobacco and tomato to explore the use of RNAi to cut off the transmission of the virus. Besides,we also constructed three dsRNA prokaryotic expression vectors used different length of Myzus persicae RPS2gene. Add these three dsRNAs into the artificial diet of Myzus persicae, and then feed Myzus persicae. Find the effect that PVY transmitted by Myzus persicae. These studies provided a basis for further construction of dual-gene, infection and epidemic are cut off in two ways, which are virus infection and virus-transferring mediator, thus laying the foundation work on establishing the technical system of molecular mechanism for virus.
The results and main conclusions are as follows:
1. Studies on TYLCCNV different homologous in amino acid sequence of CP gene dsRNA vectors being transformed N. benthamiana and tomato, and the resistance of the transgenic plant.
2. Studies on TYLCCNV different length of CP gene dsRNA vectors being transformed N. benthamiana and tomato, and the resistance of the transgenic plant.
3. Studies on RNAi interfere Whitefly transmission capacity by transforming N. benthamiana and tomato with the GroEL gene segments dsRNA.
4. The preliminary analysis of the resistance in thetransient expression of dsRNA TYLCCNV CP gene in N. benthamiana.
5. Constructed three dsRNA prokaryotic expression vectors and add these three dsRNAs into the artificial diet of Myzus persicae, and then feed Myzus persicae.Find the effect that PVY transmitted by Myzus persicae.
本文的主要结论是：
1.对TYLCCNV不同氨基酸同源性CP基因片段构建dsRNA转化烟草和番茄及其抗病性进行了研究。
2.对TYLCCNV不同长度CP基因片段构建dsRNA转化烟草和番茄及其抗病性进行了研究。
3.对利用B型烟粉虱GroEL基因片段构建dsRNA转化本氏烟和番茄干扰烟粉虱传毒进行了研究。
4.初步分析了在本氏烟中瞬时表达TYLCCNV CP基因dsRNA的抗病性。
5.构建三个含RPS2基因片段的dsRNA原核表达载体,并将这三种体外表达的dsRNA加入烟蚜人工饲料中,观察食用该饲料对烟蚜传播PVY的影响。
This study focused on tomato yellow leaf curl China virus which transmits by Bemisia tabaci and potato virus Y which transmits by aphids. TYLCCNV Y10coat protein gene and GroEL gene—related to virus transferring are selected to construct dsRNA plant expression vector respectively, then they are transformed into tobacco and tomato to explore the use of RNAi to cut off the transmission of the virus. Besides,we also constructed three dsRNA prokaryotic expression vectors used different length of Myzus persicae RPS2gene. Add these three dsRNAs into the artificial diet of Myzus persicae, and then feed Myzus persicae. Find the effect that PVY transmitted by Myzus persicae. These studies provided a basis for further construction of dual-gene, infection and epidemic are cut off in two ways, which are virus infection and virus-transferring mediator, thus laying the foundation work on establishing the technical system of molecular mechanism for virus.
The results and main conclusions are as follows:
1. Studies on TYLCCNV different homologous in amino acid sequence of CP gene dsRNA vectors being transformed N. benthamiana and tomato, and the resistance of the transgenic plant.
2. Studies on TYLCCNV different length of CP gene dsRNA vectors being transformed N. benthamiana and tomato, and the resistance of the transgenic plant.
3. Studies on RNAi interfere Whitefly transmission capacity by transforming N. benthamiana and tomato with the GroEL gene segments dsRNA.
4. The preliminary analysis of the resistance in thetransient expression of dsRNA TYLCCNV CP gene in N. benthamiana.
5. Constructed three dsRNA prokaryotic expression vectors and add these three dsRNAs into the artificial diet of Myzus persicae, and then feed Myzus persicae.Find the effect that PVY transmitted by Myzus persicae.
引文
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