| 转CmTreⅡ基因水稻RNAi效应的室内和田间抗性及遗传性鉴定 |
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| 引用本文： | 王爽,杜娟,李尚伟.转CmTreⅡ基因水稻RNAi效应的室内和田间抗性及遗传性鉴定[J].贵州大学学报(农业与生物科学版),2017(2):40-45. |
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| 作者姓名： | 王爽 杜娟 李尚伟 |
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| 作者单位： | 1. 贵州大学昆虫研究所/贵州山地农业病虫害重点实验室,贵州 贵阳,550025;2. 贵州大学昆虫资源开发利用特色重点实验室,贵州 贵阳,550025 |
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| 基金项目： | 国家自然科学基金项目(31360443)，贵州省科技创新人才团队项目(黔科合人才团队[2014]4001号) |
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| 摘 要： | 本研究采用农杆菌介导的遗传转化技术,将CmTreⅡ(膜结合型海藻糖酶基因一个靶位点)导入粳稻中花11号中,利用CmTreⅡ-dsRNA(含有膜结合型海藻糖酶基因一个靶位点的双链RNA)的沉默效应,沉默稻纵卷叶螟的膜结合型海藻糖酶,最终获得一批抗虫性较好的转基因纯合株系。研究中进一步检测了这些株系的室外卷叶率、白叶率以及室内抗虫性。采用实时荧光定量PCR(RT-q PCR)检测转基因水稻室内抗虫性CmTre2基因的相对表达量。通过RT-PCR技术检测了转CmTreⅡ基因水稻的遗传性。结果显示:在室外抗虫性试验中,稻纵卷叶螟对水稻的取食量明显下降,转CmTreⅡ基因水稻的卷叶率和白叶率分别为13.40%和12.47%,相比对照组分别下降了11.32%和10.10%;在室内抗虫性试验中,RT-q PCR检测结果表明,稻纵卷叶螟在取食转CmTreⅡ基因水稻的96 h后,CmTre2基因的相对表达量比对照组下降了29.00%;7 d后,实验组的死亡率为56.67%,相比对照组增加了46.67%。在遗传性试验中,检测发现转CmTreⅡ基因水稻的遗传性良好。因此本研究成功鉴定了转CmTreⅡ基因水稻对稻纵卷叶螟的抗虫性以及遗传性。
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| 关 键 词： | 稻纵卷叶螟 转基因 抗虫性 膜结合型海藻糖酶 RNA干扰 |
Indoor and Field Resistance and Genetic Identification of RNAi in the Transgenic CmTre Ⅱ Gene Rice |
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| WANG Shuang,DU Juan,LI Shang-wei.Indoor and Field Resistance and Genetic Identification of RNAi in the Transgenic CmTre Ⅱ Gene Rice[J].Journal of Mountain Agriculture & Biology,2017(2):40-45. |
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| Authors: | WANG Shuang DU Juan LI Shang-wei |
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| Abstract: | The CmTre2 gene,CmTreⅡ,was introduced into the rice (Chinese Flower 11) by agrobacteriummediated genetic transformation technology.A number of good insect resistance transgenic plants were obtained by using the silencing effect of CmTre Ⅱ-dsRNAn to silent the membrane-bound trehalase of Cnaphalocrocis medinalis.The rate of white leaves and the identification of insect resistance were further investigated.The relative expression of CmTre2 gene in transgenic rice was detected by real-time fluorescence quantitative PCR (RT-qPCR).At the same time,RT-PCR technology was adopted to detect the genetic characteristics of transgenic CmTre Ⅱ gene rice.The results showed that in the outdoor insect resistance experiments,the feeding amount by C.medinalis of transgenic CmTre Ⅱ gene rice decreased significantly.The leaf roll rate and white leaf rate of transgenic CmTre Ⅱ gene rice were 13.40％ and 12.47％,respectively,which were a decrease of 11.32％ and 10.10％,respectively,compared to the control group.In the indoor experiment of insect resistance,RT-qPCR results showed that,after 96 hours of C.medinalis feeding on transgenic CmTre Ⅱ gene rice,the relative expression of CmTre2 was decreased by 29％ of the control group.After 7 days of feeding on transgenic CmTre Ⅱ gene rice,the mortality rate of insects was 56.67％,an increase of 46.67％ compared to the control group.In the genetic test,the genetic characteristics of the transgenic CmTre Ⅱ gene rice was detected.This study successfully identified the pest resistance and genetics of transgenic CmTre Ⅱ gene rice. |
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| Keywords: | Cnaphalocrocis medinalis transgenosis insect resistance the membrane-bound trehalase RNAi |
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