【摘 要】
：
MicroRNA (miRNA)是由MIR基因转录后加工成的21 ～ 24nt的非编码RNA,参与精细调控生长发育、应对生物与非生物胁迫等.目前在水稻中已经报道了超过700条miRNA,明确了miRNA在水稻的生长发育中起着重要的精细调控作用.为了明确哪些miRNA参与水稻-稻瘟病菌的相互作用,我们利用稻瘟病普感材料丽江新团黑谷和含Pi-km位点的抗病单基因系IRBLkm-Ts为对象,混合接种20
【机 构】
：
四川农业大学水稻所,成都 611130 四川农业大学农学院,成都 611130
【出 处】
：
中国植物病理学会2014年学术年会
论文部分内容阅读
MicroRNA (miRNA)是由MIR基因转录后加工成的21 ～ 24nt的非编码RNA,参与精细调控生长发育、应对生物与非生物胁迫等.目前在水稻中已经报道了超过700条miRNA,明确了miRNA在水稻的生长发育中起着重要的精细调控作用.为了明确哪些miRNA参与水稻-稻瘟病菌的相互作用,我们利用稻瘟病普感材料丽江新团黑谷和含Pi-km位点的抗病单基因系IRBLkm-Ts为对象,混合接种20个稻瘟病菌菌株,分别于接种前(0h)和接种后12h、24h三个时间点取样,提取小RNA构建了6个小RNA文库,对小RNA文库进行深度测序分析,挑选出读数超过50且在任意两个文库间差异在两倍以上的miRNA共30余个.定量PCR分析验证了部分miRNA的表达量与其靶基因的表达量呈负相关.我们将这30余个miRNA按其读数在三个时间点的变化趋势分为三类,第一类是稻瘟病抗性的正调控因子,表现为在感病材料丽江新团黑谷中稻瘟病菌接种前后表达没有什么变化或接种后下调,但在抗病材料IRBLkm-Ts中上调表达,挑选出这类miRNA 5个；第二类是稻瘟病抗性的负调控因子,表现为在抗病材料IRBLkm-Ts中稻瘟病菌接种前后表达没有什么变化或接种后下调,但在感病材料丽江新团黑谷中接种后上调表达,挑选出这类miRNA 18个；第三类可能是应对稻瘟病菌侵染的基础反应调控因子,表现为在抗病和感病材料中均在稻瘟病菌接种后上调表达,挑选出这类miRNA 16个.从这三类小RNA中分别选取一个进行过量表达转基因分析,结果发现过量表达miR160a和miR398b的转基因植株对稻瘟病的抗性明显增强,而过表达miR444b.2的转基因植株对稻瘟病的抗性明显减弱.这些研究结果说明miRNA通过多种方式调控水稻对稻瘟病菌的防御反应,获得的在抗感材料间及在稻瘟病菌侵染前后差异表达的miRNA为进一步研究稻瘟病抗性的精细调控网络奠定了基础.
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