棉花S-腺苷蛋氨酸脱羧酶基因(GhSAMDC2/3/4)的克隆及其诱导表达分析  

Cloning and Induced Expression Analysis of Gh SAMDC2/3/4 in Cotton(Gossypium hirsutum L.)

作　　者：王凡龙[1] 朱华国[1] 程文翰[1] 刘永昌[1] 成新琪 孙杰[1] 

机构地区：[1]石河子大学农学院/新疆生产建设兵团绿洲生态农业重点实验室,新疆石河子832003

出　　处：《棉花学报》2015年第2期176-183,共8页Cotton Science

基　　金：国家自然科学基金(31301363);棉花生物学国家重点实验室开放课题(CB2014A12);兵团种质资源创新与功能基因发掘利用专项(2012BB049);国家"十二五"科技支撑计划(2011BAD35B05-3-6)

摘　　要：利用电子克隆结合RT-PCR技术克隆获得陆地棉(Gossypium hirsutum L.)S-腺苷蛋氨酸脱羧酶(S-adenosylmethionine decarboxylase,SAMDC)基因家族3个基因,分别命名为Gh SAMDC2、Gh SAMDC3和GhSAMDC4。序列分析显示,该基因c DNA包含的upstream ORF(u ORF)和main ORF(m ORF)为植物SAMDC基因特征ORF,其中m ORF长度分别为1068 bp、1110 bp和1032 bp,分别编码355、369和343个氨基酸。聚类分析表明,Gh SAMDC2/3蛋白与可可树(Theobroma cacao)SAMDC聚为一类,且Gh SAMDC2与GhSAMDC3蛋白亲缘关系最近;Gh SAMDC4与拟南芥At SAMDC4聚为一类。实时荧光定量PCR分析表明,Gh SAMDC2在茎中表达相对较高,随着纤维发育其表达量不断增加,在纤维发育后期其表达量达到最高;Gh SAMDC2/3/4在不同的胁迫条件下表现出不同的表达模式,Gh SAMDC2受低温和干旱胁迫诱导最强烈,Gh SAMDC3响应盐胁迫显著,Gh SAMDC4受ABA诱导强烈。上述结果为进一步研究棉花SAMDC基因功能奠定了一定基础。S-adenosine methionine decarboxylase（SAMDC） is one of three key enzymes in the polyamine synthesis pathway. Gh- SAMDC2/3/4 （ GhSAMDC2, GhSAMDC3 and GhSAMDC4） were acquired from Gossypium hirsutum L. by electronic cloning and reverse-transcription polymerase chain reaction （RT-PCR） technology. Sequence analysis showed that the gene cDNA con- tained two plant general SAMDC open reading frames （ORFs）, the upstream ORF （uORF） and the main ORF （mORF）. The mORFs of GhSAMDC2/3/4 were 1068, 1110 and 1032 bp, encoding 355, 369 and 343 amino acids, respectively. Cluster analy- sis showed that GhSAMDC2/3 and cacao tree SAMDC clustered within a class and GhSAMDC4 and AtSAMDC4 clustered within a separate category. Real-time PCR analysis showed GhSAMDC2/3/4 were expressed in all testing organization and fiber. GhSAMDC2 was obviously expressed in stem and late developed fiber. GhSAMDC2/3/4 were induced by abiotic stress, but Gh- SAMDC2 was strongly affected by low temperature and drought stress. GhSAMDC3 responded significantly to salt stress and GhSAMDC4 was strongly induced by ABA.

关 键 词：棉花 多胺 S-腺苷蛋氨酸脱羧酶基因 非生物胁迫 

分 类 号：S562[农业科学—作物学]