陕229干旱复水诱导基因表达及小麦乙烯受体(TaERS)基因特征分析  

Genes Expression in Response to Re-watering after Drought Stress in Shaan 229 and Characterization of Ethylene Receptor Genes(TaERS) in Wheat

作　　者：张宏[1] 陈锐 黄林周[1] 胡银岗[1,3] 

机构地区：[1]旱区作物逆境生物学国家重点实验室,西北农林科技大学农学院,杨凌712100 [2]常州市生态技术应用研究所,常州213131 [3]中国旱区节水农业研究院,西北农林科技大学,杨凌712100

出　　处：《农业生物技术学报》2012年第5期497-505,共9页Journal of Agricultural Biotechnology

基　　金：澳大利亚ACIAR项目(No.CIM/2005/111);基本科研业务费(No.QN2011002);唐仲英育种基金

摘　　要：为了解小麦响应水分胁迫后复水条件下的基因表达特征，以小麦（TriticumaestivumL．1主栽品种陕229的3叶1心期幼苗为材料，采用消减杂交技术，构建了干旱复水条件下的SSH-cDNA表达文库。从消减文库中随机挑选59个插入片段大于400bp的阳性克隆测序，去除冗余序列和嵌合序列后，获得高质量EST序列32条（GenBank登录号为ES466767-ES466798）。序列比对分析表明，小麦干旱后复水的基因表达与植物对其他非生物胁迫的响应具有交叉性；17条EST序列与已知编码蛋白的基因同源性较高，涉及植物的信号传导、能量代谢、转录调控等方面；其他序列为新的EST。以其中一个与乙烯受体基因（ERS）同源性较高的EST序列为基础，采用同源克隆及RACE技术从小麦中分离了4个乙烯受体基因的全长cDNA序列，长度分别为2090、2271、2216和1886bp。4个全长cDNA序列所编码氨基酸序列的同源性极高（99％以上），且均具有ERS典型的GAF、HisKA和HATPase．c跨膜结构，分别命名为TaERSl（HM347272），TaERS2（HM601437），TaERS3（HM601438）和TaERS4（HQl11523）。植物ERS氨基酸序列多重比较表明，小麦与水稻的相似性最高（93％）；TaERS基因的全长cDNA序列间比较共发现SNP位点23个。定量PCR表达分析显示，小麦TaERS家族基因参与了小麦植株响应水分胁迫和复水的调控机制。研究结果有助于进一步认识小麦干旱后复水的基因表达谱和小麦水分高效利用机制，探索小麦乙烯受体在水分高效利用中的作用。To explore more genes for the improvement of drought-tolerance and water use efficiency in crops, the differential gene expression during re-watering after serious drought stress in common wheat （Triticumaestivum L.） was investigated by constructing SSH-cDNA library with the seedlings of Shaan 229 as materials. A total of 59 positive clones with insertions more than 400 bp were randomly picked from the library and sequenced. Then redundancy sequences were removed, finally 32 high quality EST （GenBank accessions of ES466767-ES466798） were identified. Homologous comparison of those EST sequences indicated that there were some similar responses of wheat to re-watering after serious drought stress with that to the other abiotic stresses. Among the 32 EST identified, 17 EST sequences were similar to known genes encoding functional proteins, which involved in the signal transduction, energy metabolism and transcriptional regulation, etc., theothers were new EST in wheat. Based on an EST sequence highly similar to ERS gene in the SSH-cDNA library, four eDNA sequences encoding ethylene receptor in wheat were isolated using RT-PCR and RACE approaches. All the deduced proteins of these four eDNA sequences had the typical domains of ERS （three transmembrane segments, GAF domain, HisKA and HATPase-c domain）. The full-length cDNA sequence of TaERS genes in wheat were 2 090, 2 271, 2 216 and 1 886 bp, and designated as TaERS1 （HM347272）, TaERS2 （HM601437）, TaERS3 （HM601438） and TaERS4 （HQl11523）, respectively. Multiple sequence alignment analysis based on the amino acids encoded by the ERS genes from wheat （T. aestivum L.）, rice （Oryza sabiva）, corn （Zay mays）, sugarcane （Saccharum officinarum）, Arabidopsis （Arabidopsis thaliana）, tobacco （Nicotiana tabacum） and tomato （Lycopersicoum esculentum） indicated that TaERS had the highest similarity with that of rice （93%）. The sequence alignment analysis revealed 23 SNPs among the four eDNA sequences of TaERS genes in wheat

关 键 词：小麦 干旱胁迫后复水 基因表达 乙烯受体(TaERS) 

分 类 号：S667[农业科学—果树学]