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 Our group is mainly focusing on exploring the molecular basis of plant-insect interaction. On one hand, to fully understand plant resistance to insects, we have been screening the rice T-DNA insertion library by evaluating rice resistance against different insect pest species, e.g. brown planthopper and white back planthopper. These results will help identify the resistant-related genes responsible for rice resistance to insect pests; the potential targets can be further employed in transgenic plants for crop protection. On the other hand, we are also interested in selecting the key genes related to insect development and reproduction. We lay our emphasis on those gene targets causing a lethal phenotypic effect via RNAi technology and try to understand their mode of action based on the knowledge of insect physiology and biochemistry. And hopefully, our data will provide an insight for some new strategies in developing the dsRNA-based insecticides.
Pricipal Investigator: Dr. Xue-Xia Miao, Professor. Email: xxm@sippe.ac.cn
Permanent Staff: Hai-Chao Li, Research Asisstant; Yu-Bing Wang, Research Intern.
Graduate Students: Hao Zhang; Hui-Min Guo; Ruo-Bing Guan.
Rice endogenous insect resistant-related genes
The plants have constitutive resistance and inducible resistance to insect pests. These two resistance strategies play important roles in self-defense when plants are being attacked by herbivores. Our aim is to study the molecular basis of plant resistance against insect pests, which will be helpful and commercially useful for taking advantage of plant self-raised resistance in pest managements. Thirteen thousands of T-DNA insertion mutants have been screened, and have got 45 high resistant materials against brown planthopper, 285 high resistant materials against white back planthopper and 139 high resistant materials against rice leaf-roller (Figure 1). At present, large-scale gene cloning and potential resistant mechanism are undertaken in the laboratory aiming to identify constitutive and inducible genes for future transgenic engineering against different insect pests.
High-throughput RNAi target gene screening from insects
We adopted second-generation sequencing technology to screen RNAi targets from Ostrinia furnalalis. Total 14690 stage specific genes were obtained which can be considered as potential targets. Ten larval stage specific expression genes were selected for RNAi test. When 50 ng/µl dsRNAs of the genes were directly sprayed on the newly hatched larvae, the mortalities reached 73% to 100% at 5 d after treatment. The qRT-PCR analysis verified the correlation between larval mortality and the down-regulation of the target gene expression. Topically applied fluorescent dsRNA confirmed that dsRNA did penetrate the body wall and circulate in the body cavity. It seems likely that the combination of DGE-tag with RNA-seq is a rapid, high-throughput, cost less and an easy way to select the candidate target genes for RNAi. More importantly, it demonstrated that dsRNAs are able to penetrate the integument and cause larval developmental stunt and/or death in a lepidopteron insect (Figure 2). This finding largely broadens the target selection for RNAi from just gut-specific genes to the targets in whole insects and may lead to new strategies for designing RNAi-based technology against insect damage.
Publications:
Yubing Wang, Hao Zhang, Haichao Li, Xuexia Miao*. 2011. Second-Generation Sequencing Supply an Effective Way to Screen RNAi Targets in Large Scale for Potential Application in Pest Insect Control. PLoS one 6(4): e18644. DOI:10.1371/journal.pone.0018644 Yubing Wang, Haichao Li,Yuan Si, Hao zhang, Xuexia Miao*. 2011. Microarray analysis of broad-spectrum resistance derived from an indica cultivar Rathu Heenati. Planta published online: 15, November 2011. DOI: 10.1007/s00425-011-1546-1 Xuexia Miao, Weihua Li, Muwang Li, Yupo Zhao, Xianru Guo and Yongping Huang. 2008. Inheritance and linkage analysis of co-dominant SSR markers on the Z chromosome of the silkworm (Bombyx mori L.). Genetics Research 90(2): 151-156. Legendre Ariele, Xuexia Miao#. Jean-Luc Da Lage, Claude Wicker-Thomas, 2008. Evolution of a desaturase involved in female pheromonal cuticular hydrocarbon biosynthesis and courtship behavior in Drosophila. Insect Biochemistry and Molecular Biology 38(2): 244-255. (#co-first author). Miao Xuexia, Li Muwang, Dai Fangyin, Lu Cheng, Goldsmith M.R. & Huang Yongping. 2007. Linkage analysis of the visible mutations Sel and Xan of Bombyx mori (Lepidoptera: Bombycidae) using SSR markers. European Journal of Entomology 104(4): 647-652. Miao Xue-Xia, Xu Shi-Jie, Li Ming-Hui, Li Mu-Wang, Huang Jian-Hua, Dai Fang-Yin, Susan W. Marino, David R. Mills, Zheng Pei-Yu, Kazuei Mita, Jia Shi-Hai, Xu An-Ying, Zhang Yong, Liu Wen-Bing, Xiang Hui, Guo Qiu-Hong, Kong Xiang-Yin, Lin Hong-Xuan, Shi Yao-Zhou, Yuji Yasukochi, Toshiyuki Sugasaki, Toru Shimada, Javaregowda Nagaraju, Xiang Zhong-Huai, Wang Sheng-Yue*, Marian R. Goldsmith*, Lu Chen*, Zhao Guo-Ping*, Huang Yong-Ping*. 2005. Simple sequence repeat-based consensus linkage map of Bombyx mori. Proceedings of the National Academy of Sciences USA 102(45): 16303-16308. Xuexia Miao, Yongping Huang, Xiangxiong Zhu and Decheng Ding. 2004. A comparative study on development and reproduction of the parasitoid Lysiphlebus japonicus (Hymenoptera: Aphidiidae) in symbiotic and aposymbiotic host aphids. Applied Entomology and Zoology 39 (2): 243-248. Jianhua Huang, Xuexia Miao*, Weirong Jin, Kasuei Mita, Yong Zhang, Wenbin Liu, Celine Keime, Olivier Gandrillon, Patrick Brouilly, Jerome Briolay, Pierre Cobble, Guoping Zhao, Yongping Huang. 2005. Profile of the genes expressed in silkworm, Bombyx mori through serial analysis of gene expression (SAGE). Genomics 86, 231-245. (*co-first author)
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