增强植物的抗虫能力课件

1、增强植物的抗虫能力dsRNA和遗传防治，抗性系，Bt化学农药为主的害虫治理不可持续强大的杀虫剂严重的污染巨大的灾难农药基因突变疾病靶标药物健康问题害虫抗性加量强抗性再加量经济问题环境污染生物多样性、残留、贸易等生态问题意义和价值意义和价值害虫防治需要新的方法高密度增强植物抗性传统方法辅助方法农药+辅助方法Bt, dsRNA, 抗性系天敌，微生物，其他低密度现代方法种群的遗传调节通过致死基因(经济阈值以下)种类特异的基因调控是未来害虫防治的发展方向意义和价值对环境无污染对目标以外的生物无危害对农作物无残留“三无”要求意义和价值基因组学进展为害虫治理提供了机遇30余种重要昆虫基因组测序功能基因组研

2、究平台逐渐建立RNA干涉从单基因到规模化可供选择的靶标基因数量增加控制翅伸展基因RNA干涉，2005植物介导RNA干涉，2007重要的害虫防治对象意义和价值农林业害虫中70以上是鳞翅目昆虫害虫防治涉及的主体昆虫靶标基因的筛选植物转基因变态发育蜕皮免疫嗅觉害虫的基因调控高密度传统防治化学农药+辅助方法低密度现代方法(经济阈值以下)环保农药杀虫蛋白(Bt等)+生物防治棉铃虫、东亚飞蝗豌豆蚜遗传不育靶标基因水平介导沉默机理环境安全评估垂直介导基因调控致死机理昆虫自体植物、微生物致死基因基因调控的路线图基因水平的害虫控制RNA干涉基因调控+玉米螟棉铃虫小菜蛾果蝇、家蚕、蚊、蜜蜂、赤拟谷盗、豆蚜等变态、

3、发育先天免疫取食靶标基因及其应用害虫靶标基因研究高通量测序基因芯片cDNA文库15000个基因5-10个基因种类特异性高RNA干涉高效？关于SAGESerial Analysis of Gene Expression昆虫靶标基因的大规模筛选方法优点缺点EST sequencing高通量;易于操作不能定量,灵敏度低Microarray高通量; 半数量化; 灵敏度高;已商业运营封闭体系SAGE定量化; 灵敏度高;开放体系不经济基因组水平的表达谱研究基于SAGE方法的研究不同发育时期基因表达谱辐射诱变对基因表达影响从SAGE标签到全长基因SAGEMicroarray结合表达谱与遗传图谱的整合获得SA

4、GE标签的基本程序家蚕转录组研究Huang et al. Genomics, 2005Cumulative curve of the frequencies of unique tag detection derived from the sequencing of 257,964 tags using serial analysis of gene expression (SAGE) libraries of four developmental stages of silkworms EggsLarvaePupamothTotaltagunitag= 2 copies69100177185

5、8156223213242473663966140445171626661522452563154214654017802The percentage distribution of unique tags detected from different developmental stages of the silkworm (egg, larva, pupa, and adult). TagcopiesTotal numberof tagsTotal numberOf unique tagsMatches to silkwormEST database (1050)10010-996-92

6、-51Total115,30575,24514,41227,46824,458257,964383(1%)2935(7.8%)2018(5.3%)9691(24.2%)24458(61.7%)195(50.9%)363(12.4%)96(4.7%)129(1.3%)178(0.7%)961(2.4%)Tag frequency and percentage distribution of unique tags from four silkworm SAGE libraries Gene expression profiles of all four SAGE libraries Expres

7、sion profilesRedGreenBlackGrayhighlowmoderatezeroGene expression of the 27 genes that were differentially expressed (P0.01) in four SAGE libraries RedGreenBlackGrayhighlowmoderatezeroRT-PCR analysis of genes expressed differently from four developmental stages of the silkworm. G3PDH was employed as

8、control. Serine protease geneEggLarvaPupaAdult102101340431416145006Chorion protein geneRibosomal protein S11StageEggLarvaPupaAdultEgg- Larva710- Pupa846663- Adult1106994744-Matrix of differential tag expression between the developmental stagesStatistically significant differences (P1.00.2-1.00.05-0.

9、20.2-1.00.05-0.0050.002Total297613439131931992576641190369100271510558951320338431210021574573562102247329911903177182491813122215610021155313(100%)45(80%)109(51.9%)290(12.9%)124(3.8%)160(1.34%)731(4.1%)normalirradiateNumber of uniqueSAGE tagsnormalirradiateThe abundance classes of these analysed ta

10、gs from the normal and irradiated Bombyx mori embryo SAGE librariesSAGE 标签基因111MM1SAGE标签使用中存在的问题Generation of Longer cDNA fragments from SAGE tags for Gene IdentificationGLGI的主要程序20001000750500250100GLGI resultsSGRS-SAGEG-GLGIR-RACESGR鞣化激素基因RNAi结果对照结果RNAi处理表型Ubx基因RNAi结果SAGE标签获得需要的目的标签通过GLGI得到3cDNA通过SGR得到全长基因序列目标基因的功能研究SAGE-GLGI-Microarray结合StageEggLarvaPupaAdultEgg- Larva710- Pupa846663- Adult1106994744-Matrix of differential tag expression between the developmental stagesStatistically significant differences (P3）CopyEggLarvaPupaAdultTotal216111551178119086851393003289932903788129