水稻种子活力相关基因OsIPMS12的功能分析
关键词：水稻；OsIPMS12；种子活力；基因表达
Function Analysis of OsIPMS12 Ralated the Rice Seed Vigor Gene
Abstract：Seed vigor is an important trait of seed quality.It has a direct effects on the speed and uniformity of seed germination, and thereby affects the speed and strength of the seeding growth.In previous study, one gene associated with seed vigor was identified using two-dimensional electrophoresis method in rice. The gene is located on rice chomosome 12, and then named as OsIPMS12. This experiment uses the material of point mutations,excessive expression and transgenic line of promoter analysis,which is built by CRISPR-Cas9 technology.The experimental results show the OsIPMS12 express mainly in spike ,floret,internode and the seeds of development;and expression under salt and drought induced in seeding. The gene effects on the seed vigor and adversity to adapt after mutation. The germination results show that the genetic mutation can reduce rice seed germination rate,and affect seeding root length and shoot length. The study above,OsIPMS12 may be involved in the process of seed germination, and showed improving effect on seed vigor, that also played the role of the preliminary appraisal for subsequent reseach.
Key words: Rice；OsIPMS12；Seed vigor；Gene expression
摘要1
关键词.1
Abstract1
Key words.1
引言.1
1材料与方法.1
1.1材料准备 .1
1.2 RNA的提取 .2
1.3RNA反转录2
1.4 Real-Time PCR.3
1.5 DNA提取3
1.6GUS染色.4
1.7 基因表达分析4
1.8 种子活力鉴定4
2结果与分析4
2.1 基因获得.4
2.2 材料鉴定5
2.2.1点突变植株鉴定5
2.2.2 GUS转基因株系阳性5
2.2.3 过表达转基因植株阳性验证.5
2.3 基因组织表达特点6
2.4 OsIPMS12基因在不同发育阶段及逆境条件下的表达.6
2.5种子活力鉴定.7
2.5.1 正常条件下突变体及转基因材料的种子活力鉴定.7
2.5.2 200mM NaCL胁迫条件下突变体及转基因材料的种子活力鉴定8
3讨论8
致谢9
参考文献9
水稻种子活力相关基因OsIPMS12的功能分析
引言我国是世界上最大的稻米生产和消费国，水稻作为我国第一大粮食作物，它的稳定生产是国家粮食安全的基本保障，具有十分重要的意义。水稻种子活力是一个极其复杂的农艺性状，是由基因控制的数量性状，同时还受到各种非生物胁迫的影响，如盐[1]、低温[2]等。Lin等[3]利用Nona Bokra和Koshihikari构建了F2及F3群体对水稻苗期耐盐性进行了QTL定位，共定位了11个影响8个耐盐相关性状的QTL，其中位于第7染色体中部检测到1个同时影响4个性状的QTL簇，这个QTL簇对地上部Na+含量效应最大，能解释表型变异的48.5%。水稻在受到低温胁迫时，细胞内主要有两类蛋白质分子在发挥作用：一类是直接保护细胞的渗透调节功能蛋白；另一类就是参与胁迫信号传导和表达调控的转录因子[4]，因此，揭示盐胁迫[5]和低温胁迫[6]下水稻种子萌发并形成正常幼苗的分子遗传激励，选育和利用高活力耐盐和耐低温水稻品种是解决水稻盐害和冻害最经济有效的方法，这已经成为水稻育种的一个重要目标。