低温胁迫下花生差异表达基因的分离与分析  

Isolation and Analysis of Differentially Expressed Genes from Peanut under Low-Temperature Stress

作　　者：于树涛 于洪波 苏君伟 赵立仁 史普想 赵艳 唐月异[2] 王秀贞[2] 吴琪[2] 王传堂[2] 

机构地区：[1]辽宁省风沙地改良利用研究所,辽宁阜新123000 [2]山东省花生研究所,山东青岛266100

出　　处：《核农学报》2014年第4期569-576,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家花生产业技术体系建设(CARS-14);辽宁省科技厅科技攻关项目(2011201021);国家科技支撑计划课题(2012BAD36B00)

摘　　要：筛选74个花生种质材料,最后以发芽率在80%以上的9650作为试验材料。对9650分别进行2℃和25℃(对照)处理,分别提取2、6和8h花生样品的总RNA。利用GeneFishing随机引物,克隆测序,获得34个独特的基因序列。经BLAST2GO注释,包括参与细胞合成的组分、应激蛋白反应、代谢过程中的转运与运输、蛋白质的合成、以及耐低温相关的基因序列。进一步利用定量RT-PCR证实了4个基因的相对表达量,选择其中亲环蛋白基因相关序列,通过5'-RACE获得cDNA全长,为通过转基因技术对其进行功能分析奠定了基础。A total of 74 accessions of peanut materials were screened for chilling stress resistance based on a previously established method, resulting in a germplasm line 9650 with 〉 80% germination. Total RNA was extracted from seeds of 9650 imbibed in distilled water at 2 （low temperature treatment） or 25℃ （check） for 2h 6 and 8h, and differentially expressed genes （DEGs） were isolated using Genefishing technology. 34 unique DEGs were obtained. Of them, 5 sequences have not been annotated before, and the rest 29 were found to be highly similar to the sequences in the GeneBank database. BLAST2Go analysis revealed that 34 unigenes belonged to diverse functional categories including metabolism, signal transduetion, stress response, cell defense and transcriptional regulation. Four genes potentially related to low-temperature stress tolerance in peanut were further validated by qRT-PCR The full length eDNA of cyelophilin gene was obtained with 5＇ - RACE, laying foundation for functional analysis of its role in peanut chilling stress tolerance through transgenesis.

关 键 词：花生 低温 GeneFishing 差异表达 亲环蛋白 

分 类 号：S565.2[农业科学—作物学]