玉米抗茎腐病微效QTL-qRfg2候选基因功能验证.pdfVIP

内容摘要 玉米茎腐病是危害玉米生产的主要病害之一，在世界各玉米产区普遍发生， 严重的影响玉米的品质和产量。本实验室的前期研究选用玉米抗茎腐病自交系1 145和感病自交系Y331组配群体，对玉米抗茎腐病进行基因定位，在基因组上扫 描到两个QTL ：位于10号染色体的主效QTL-qrfg1和位于1号染色体的微效QTL- qrfg2。对QTL- qrfg2进行精细定位，并通过BAC序列分析，发现其只包括一个候选基因，在抗感 亲本间序列相对保守，命名为ZmARP1 。表达分析发现，ZmARP1在R/S- NIL 之间表达量差异显著，因此构建了过表达载体和RNAi载体，对ZmARP1进行 功能验证。本实验主要是在此基础上，通过设计候选基因的特异引物，检测转基 因植株基因型，并观察田间和苗期的转基因植株表型，从而验证候选基因ZmARP 1的功能。 关键词：玉米 茎腐病 转基因 功能验证 ABSTRACT Stalk rot in maize is one of the most serious diseases worldwide that could limit the maize production and could severely cause yield loss and degrade kernel quality. In our previous study, two QTLs, a major QTL-qRfg1 on bin10.03/04 and a minor QTL-qRfg2 on bin1.08/1.1, have been identified in a segregation population derived from the resistant inbred line ‘1145’ and the susceptible inbred line ‘Y331’. Fine mapping of the QTL-qRfg2，with the analysis of BAC sequence, finds that it includes only one candidate gene named ZmARP1,which has relative conservative sequence between resisit-suscept parents. Expression Analysis finds that ZmARP1 displays significant differencs in expression level between the R-NIL and the S-NIL. So that we have built the overexpression vector and RNAi vector for the functional verification of ZmARP1.This experiment is to verify the function of the candidiate gene ZmARP1 through designing specific primer, detecting the genetype of the transgenetic seedlings and observing the phenotype of the transgenetic seedlings during the field and seedling stages. Key words ：maize stalk rot transgenic functional verification 目录 1．选题背景1 1.1茎腐病 介1 1.2抗病研究进展2 1.3克隆方法3 1.4转基因现状4 1.5本实验研究背景5 1.6研究意义与内容5 2．材料和方法5 2.1实验材料5 2.2实验方法6 3．实验结果11 3.1设计开发鉴定转基因植株的