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当前位置: 首页 > 文章 > 烟草NtSnRK2.1基因的克隆及其在非生物胁迫条件下的表达 中国烟草学报 2014,20 (4) 94-100
Position: Home > Articles > Cloning and expression of SnRK2.1 gene under abiotic stress Acta Tabacaria Sinica 2014,20 (4) 94-100

烟草NtSnRK2.1基因的克隆及其在非生物胁迫条件下的表达

作  者：
张洪映;贾宏昉;张松涛;杨永霞;崔红
单  位：
河南农业大学烟草学院;烟草行业栽培重点实验室
关键词：
普通烟草;NtSnRK2.1;非生物胁迫;克隆;表达分析
摘  要：
从普通烟草腺毛cDNA文库中筛查烟草SnRK2基因的EST序列,以此序列为信息探针,通过电子克隆和RT-PCR的方法从烟草中克隆到一个包含1017 bp开放阅读框、编码338个氨基酸的cDNA序列,命名为NtSnRK2.1。生物信息学分析表明,NtSnRK2.1蛋白同时具有磷酸化丝氨酸/苏氨酸和酪氨酸的活性。氨基酸序列比对发现,NtSnRK2.1与拟南芥、水稻和小麦等植物中受逆境胁迫诱导表达的直系同源基因高度同源。组织表达分析结果显示,烟草NtSnRK2.1基因的组织表达差异较大,在根部的表达量最高,其次是叶片,茎部的表达量最低。胁迫处理下的基因表达分析结果表明,NtSnRK2.1受高盐、高渗、低温胁迫和ABA处理诱导表达,对各胁迫条件的应答模式不同,其对各胁迫条件的敏感度为:高渗>高盐>低温>ABA。
译  名：
Cloning and expression of SnRK2.1 gene under abiotic stress
作  者：
ZHANG Hongying;JIA Hongfang;ZHANG Songtao;YANG Yongxia;CUI Hong;Key Laboratory of Tobacco Cultivation, College of Tobacco Science, Henan Agricultural University;
关键词：
Nicotiana tabacum;;NtSnRK2.1;;abiotic stress;;cloning;;expression analysis
摘  要：
One EST of SnRK2 was screened from glandular trichome cDNA library of tobacco. Based on the EST sequence, NtSnRK2.1 was isolated from tobacco(Nicotiana tabacum L.) by in silico cloning and RT-PCR. NtSnRK2.1 includes an open reading frame(ORF) of 1017 bp and encodes 338 deduced amino acid residues(AAR) with a calculated molecular mass of 43 kDa and a predicted pI of 5.78. Scansite analysis indicated that NtSnRK2.1 contained potential serine/threonine protein kinase activities like other SnRK2 family members. Phylogenetic analysis suggested NtSnRK2.1 was high homologous to its orthologous genes from Arabidopsis, rice and wheat, which were also induced by abiotic stresses. Reverse transcription-polymerase chain reaction(RT-PCR) and quantitative real-time PCR(qRT-PCR) was used to determine expression patterns of NtSnRK2.1 in tobacco. Results revealed that NtSnRK2.1 expressed most strongly in tobacco roots, more in leaves, and marginally in stems. Expression patterns under abiotic stress responses suggested that NtSnRK2.1 was involved in response to NaCl, PEG, cold stresses and ABA treatment, with significant different responsive profiles. The sensitivity degrees of NtSnRK2.1 responding to four treatments was in the order of hyperosmolality > high salinity > low temperature(4℃) > abscisic acid.

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