Transgenic Cotton Plants Expressing Double-stranded RNAs Target HMG-CoA Reductase (HMGR) Gene Inhibits the Growth, Development and Survival of Cotton Bollworms.
Transgenic Cotton Plants Expressing Double-stranded RNAs Target HMG-CoA Reductase (HMGR) Gene Inhibits the Growth, Development and Survival of Cotton Bollworms.
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表达靶向 HMG-CoA 还原酶 (HMGR) 基因的双链 RNA 的转基因棉花植物抑制棉铃虫的生长、发育和存活
DOI：
10.7150/ijbs.12463
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发表时间：
2015
影响因子：
9.2
通讯作者：
Jin S
中科院分区：
文献类型：
--
作者：
Tian G;Cheng L;Qi X;Ge Z;Niu C;Zhang X;Jin S
RNA干扰（RNA interference, RNAi）技术已成为功能基因组学研究和植物病虫害防治的有力手段。本文通过农杆菌介导的转化，在棉花中表达了靶向3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）基因的双链rna (dsRNA)，该基因在棉棉虫合成幼激素（JH）的甲戊酸盐途径中催化限速酶反应。PCR和southern分析显示HMGR基因已整合到棉花基因组中。RT-PCR和qRT-PCR证实了dsHMGR基因在转基因棉系中的高转录水平。棉铃虫幼虫取食转基因植株叶片后，HMGR在转录和翻译水平上的表达均显著下调。转基因棉花叶片上饲养的幼虫HMGR基因转录水平比野生型低80.68%。此外，卵黄原蛋白（vitellogenin，卵黄原蛋白是子代胚胎发育的重要营养来源）基因的相对表达量也降低了76.86%。昆虫生物实验结果表明，含有dsHMGR的转基因棉铃虫不仅能抑制棉铃虫的净增重，而且还能延缓棉铃虫幼虫的生长。综上所述，表达dsRNAs的转基因棉花成功下调了HMGR基因，并破坏了目标昆虫的发育和存活，为植物病虫害防治提供了更多的选择。
RNA interference (RNAi) has been developed as a powerful technique in the research of functional genomics as well as plant pest control. In this report, double-stranded RNAs (dsRNA) targeting 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene, which catalyze a rate-limiting enzymatic reaction in the mevalonate pathway of juvenile hormone (JH) synthesis in cotton bollworm, was expressed in cotton plants via Agrobacterium tumefaciens-mediated transformation. PCR and Sothern analysis revealed the integration of HMGR gene into cotton genome. RT-PCR and qRT-PCR confirmed the high transcription level of dsHMGR in transgenic cotton lines. The HMGR expression both in transcription and translation level was significantly downregulated in cotton bollworms (helicoverpa armigera) larvae after feeding on the leaves of HMGR transgenic plants. The transcription level of HMGR gene in larvae reared on transgenic cotton leaves was as much as 80.68% lower than that of wild type. In addition, the relative expression level of vitellogenin (Vg, crucial source of nourishment for offspring embryo development) gene was also reduced by 76.86% when the insect larvae were fed with transgenic leaves. The result of insect bioassays showed that the transgenic plant harboring dsHMGR not only inhibited net weight gain but also delayed the growth of cotton bollworm larvae. Taken together, transgenic cotton plant expressing dsRNAs successfully downregulated HMGR gene and impaired the development and survival of target insect, which provided more option for plant pest control.