The overexpression of insect endogenous microRNA in transgenic rice inhibits the pupation of Chilo suppressalis and Cnaphalocrocis medinalis
The overexpression of insect endogenous microRNA in transgenic rice inhibits the pupation of Chilo suppressalis and Cnaphalocrocis medinalis
复制标题
转基因水稻中昆虫内源microRNA的过表达抑制二化螟和稻纵卷叶螟化蛹
DOI：
10.1002/ps.6422
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发表时间：
2021
影响因子：
4.1
通讯作者：
Han Lanzhi
中科院分区：
文献类型：
--
作者：
Wen Ning;Chen Junjie;Chen Geng;Du Lixiao;Chen Hao;Li Yunhe;Peng Yufa;Yang Xiaowei;Han Lanzhi
背景二化螟和稻纵卷叶蛾是中国稻区主要稻区共同发生的破坏性水稻害虫。基于RNA干扰(RNAi)的抗虫基因工程(IRGE)作物通过抑制基因表达或翻译为害虫管理提供了一种很有前途的方法。通过miRNA(MiRNA)介导的表达二化螟抑制二化螟内源miRNA CSU-NOVICE-260的转基因水稻品系CSU-NOVICE-260对二化螟表现出显著的抗性，这为抑制二化螟提供了一种诱人的控制策略，但该转基因株系是否也表现出对稻纵卷叶蛾的抗性尚不清楚。在食饵实验中，化学合成的CSU-NOVICE-260agomir在200fmol g~(-1)下显著抑制二化螟的盐化。但对幼虫发育、存活率和蛹重无显著影响。此外，转基因品系显著影响稻纵卷叶蛾的化蛹，但不影响幼虫的存活率。QRT-PCR结果表明，当两种害虫取食转基因品系时，Csdiband Cmdib的表达水平受到显著抑制。此外，在组织饲养试验中，转基因株系显著降低了抗Cry1C和敏感Cry1C的二化螟幼虫的存活率，表明抗Cry1C的二化螟对miRNA介导的IRGE水稻中表达的CSU-NOVICE-260没有交叉抗性。结论本研究表明，miRNA介导的IRGE水稻显著抑制二化螟和中枢虫的化蛹。研究结果为RNAi植物的应用提供了一个新的视角，并对环境风险评估提供了启示。
BACKGROUNDChilo suppressalisandCnaphalocrocis medinalisare destructive rice pests co‐occurring in major rice‐growing areas in China. RNA interference (RNAi)‐based insect‐resistant genetically engineered (IRGE) crops provide a promising approach for pest management by suppressing gene expression or translation. A microRNA (miRNA)‐mediated IRGE rice line expressing endogenousChilo suppressalismiRNA Csu‐novel‐260, showing significant resistance againstChilo suppressalis, provides an attractive control strategy forChilo suppressalisby suppressing the expression of thedisembodied(dib) gene expression.However, whether this transgenic line also shows the resistance againstCnaphalocrocis medinalisremains unknown.RESULTSA spatiotemporal expression analysis of Csu‐novel‐260 in the transgenic rice line was performed by quantitative reverse transcription polymerase chain reaction (qRT‐PCR) to determine the paddy field pest exposure dose. In diet feeding assays, a chemically synthesized Csu‐novel‐260 agomir at 200 fmol g–1significantly inhibitedChilo suppressalispupation. However, larval development, survival and pupal weight were not significantly affected. Additionally, the transgenic line significantly affectedCnaphalocrocis medinalispupation but not larval survival. The qRT‐PCR showed thatCsdibandCmdibexpression levels were significantly suppressed when the two pests fed on the transgenic line. Additionally, the transgenic line significantly decreased Cry1C‐resistant and Cry1C‐susceptibleChilo suppressalislarval survival in detached rice tissue feeding assays, indicating that Cry1C‐resistantChilo suppressaliswas not cross‐resistant to Csu‐novel‐260 expressed in miRNA‐mediated IRGE rice.CONCLUSIONOur study demonstrated that miRNA‐mediated IRGE rice significantly inhibitedChilo suppressalisandCnaphalocrocis medinalispupation. The results provide a new viewpoint for the application of RNAi‐based plants and the inspiration for environmental risk assessment.