干旱胁迫下赖草抑制性消减文库的构建与分析  

Construction of suppression subtractive hybridization library of Leymus secalinus exposed to drought stress

作　　者：杨满业[1] 王丽焕[1] 叶煜辉[2] 江明锋[2] 肖冰雪[1] 

机构地区：[1]四川省草原科学研究院,四川成都611731 [2]西南民族大学生命科学与技术学院,四川成都610041

出　　处：《草业科学》2011年第7期1320-1325,共6页Pratacultural Science

基　　金：四川省科技厅应用基础研究项目(2006-J13-134)

摘　　要：用根际水分胁迫对赖草(Leymus secalinus)植株进行干旱胁迫,利用抑制消减技术,以干旱胁迫处理为test-er,正常生长对照组为driver,构建了包含576个克隆的赖草消减cDNA文库。通过斑点杂交筛选,得到16个EST序列,大小居于100～310bp。Blast比较发现,这些序列与大麦(Hordeum vulgare)和小麦(Triticum aesti-vum)干旱诱导下表达的序列、甜菜碱醛脱氢酶核酸序列、植物热激蛋白hsp70核酸序列、1,5-二磷酸核酮糖羧化酶氨基酸序列、植物遍在蛋白缀合酶氨基酸序列、植物50S核糖体蛋白L20基因序列等相关抗旱基因具有较高的同源性。以上结果说明,本研究得到了部分赖草抗旱相关基因片段,可为赖草抗旱机理的研究以及抗逆基因的克隆奠定一定的基础。The drought stress method of root without water was used to construct the subtracted cDNA li- brary of Leymus secalinus containing 576 clone by the method of suppression subtractive hybridization. In this study, the leaves under drought stress were considered as a tester and the leaves under normal condi- tion were considered as a driver. The dot blot hybridization and sequencing was applied to discover the 16 EST with length 100--310 bp. The searching against GenBank with blast algorithm showed that these se- quences were high homologous to the gene sequence of wheat and barley under drought stress, the gene se- quence of LcBADH2, the gene sequence of hsp70 of Oryza sativa, the gene sequence of RubisCO of Hordeurn vulgate, the amino acid sequence of Ubiquitin-conjugating enzyme of Zea mays, the amino acid se- quence of L20 of 50S Ribosomal protein of plant. This study suggested that these known genes and pro- teins play an important role in controlling plant osmotic pressure, heat adjustment, photosynthesis and hy- drolysis, and had established a basis for disclose the mechanism of drought resistance and cloning stress-resistance genes of Leymus secalinus.

关 键 词：赖草 抑制性消减杂交 干旱胁迫 EST 

分 类 号：S543.9[农业科学—作物学]