草原龙胆盐胁迫差减文库的构建及分析

Construction and Analysis of Eustoma grandiflorum Subtracted cDNA Library

为了研究草原龙胆抗盐性机理,初步获得其抗盐性相关基因,分别以400mmol·L-1的NaCl及清水处理的草原龙胆叶片mRNA为tester和driver方,利用抑制差减杂交技术构建了cDNA文库。经过测序和与Gene Ontology数据库序列比对,获得了658条单一序列,其中61条序列分子功能未知,535条序列无同源性,62条序列有显著同源性。此62条序列注释按照功能分为21类。获得了活性氧清除系统基因DHAR、GST、TRX、CAT,转录因子SMT3,调控基因丝氨酸/苏氨酸激酶基因等。结果说明草原龙胆的盐抗性相关基因与数据库中的模式植物基因既有一定的共性,同时绝大部分的序列又无法用已知的基因进行解释。

To research sah tolerance mechanism and get the salt tolerance relative genes ot Eustoma grandiflorum, a cDNA library was made by suppression subtractive hybridization technique. The leave mRNA of Eustorna grandiflorurn treated with 400 mmol· L^-1 NaCl was tester, and that treated with pure water was driver. From the library, we got 658 unique sequences by sequencing and sequence blasting. Sixty-one of the total sequences are functioning unknown, 535 sequences have no homology and 62 sequences have significance homology to the Gene Ontology sequence library. The 62 sequences can be classified into 21 function categories. And we finally got the reactive oxygen species scavenging genes： DHAR, GST, TRX, CAT, transcription factor SMT3, and serine/threonine protein kinase gene. The results indicate that there are general characters between the salt tolerance genes of Eustoma grandiflorum and model plants, and the most of the sequences can not be explained by the known genes.