马铃薯StSnRK2.4基因的序列分析及其在非生物胁迫下的表达分析

Sequence Analysis and Expression Analysis under Abiotic Stress of Potato StSnRK2.4 Gene

SnRK2是植物特有的一种丝氨酸/苏氨酸类蛋白激酶,在ABA信号转导和植物抵御逆境中起着重要作用。为探究马铃薯SnRK2在非生物胁迫下的调控作用,本研究以‘青薯9号’为试验材料,克隆得到一个SnRK2家族基因StSnRK2.4,利用PlantCARE、ExPASy、Cell-PLoc 2.0等软件对其进行序列分析,并通过RT-qPCR技术分析该基因在低温胁迫(4℃)、干旱胁迫(10%PEG-8000)以及NaCl胁迫(NaCl浓度为300 mmol/L)下的表达模式。结果表明:StSnRK2.4基因的开放阅读框(ORF)为1 083 bp,编码360个氨基酸。StSnRK2.4蛋白的相对分子质量为41.491 03 kD,理论等电点(pI)为5.52,脂肪系数为80.69,为不稳定亲水蛋白;亚细胞主要定位于细胞核;与AMP活化蛋白激酶-γ调节亚基、非特异性蛋白-BZIP转录因子、蛋白酪氨酸及PP2C ABI2同源物等互作,可能参与植物MAPK信号通路和植物激素信号转导;与野生番茄、番茄亲缘关系最近。RT-qPCR分析表明,StSnRK2.4基因的相对表达量在低温胁迫(4℃)、干旱胁迫(10%PEG-8000)以及NaCl胁迫(NaCl浓度为300 mmol/L)下均极显著上调,说明该基因受低温、干旱和盐胁迫诱导。StSnRK2.4基因的相对表达量在低温胁迫、干旱胁迫和NaCl胁迫0、3、6 h以及在干旱胁迫和NaCl胁迫0、1、2 d均呈上调-下调模式;StSnRK2.4基因的相对表达量在低温胁迫0、1、2 d呈上调-上调模式,研究认为StSnRK2.4基因可能参与非生物胁迫及相关信号分子应答。

Sn RK2 is a serine/threonine protein kinase unique to plants,which plays an important role in ABA signal transduction and plant resistance to adversity.In order to explore the regulatory role of potato SnRK2 under abiotic stress,in this study,a SnRK2 family gene StSnRK2.4 was cloned from the test material ’Qingshu No.9’.Employed PlantCARE,ExPASy,Cell-PLoc 2.0 and other software for its sequence analysis.The expression patterns of this gene under low temperature stress(4 ℃),drought stress(10% PEG-8000) and salt stress(NaCl concentration of 300 mmol/L) were analyzed by RT-qPCR technology.The results showed that the open reading frame(ORF) of the StSnRK2.4 gene was 1 083 bp,encoding 360 amino acids.The relative molecular mass was 41.491 03 kD;the theoretical isoelectric point(pI) was 5.52;the fat coefficient was 80.69,which is an unstable hydrophilic protein;sub-cells are mainly located in the nucleus;it interacts with AMP-activated protein kinase-γ regulatory subunit,uncharacterized protein-BZIP domain class transcription factor,protein tyrosine and protein phosphatase 2 C ABI2 homolog and other proteins;it may be involved plant MAPK signal pathway and plant hormone signal transduction;it has the closest relationship with Solanum pennellii and Solanum lycopersicum.RT-qPCR analysis showed that the relative expression of StSnRK2.4 gene was significantly up-regulated under low temperature stress(4 ℃),drought stress(10% PEG-8000) and salt stress(NaCl concentration of 300 mmol/L),indicating that the gene is induced by low temperature,drought and salt stress.The relative expression level of the StSnRK2.4 gene under low temperature stress,drought stress and salt stress 0,3,6 h,and drought stress and salt stress 0,1,2 d all showed up-regulation-decreasing pattern;the relative expression of StSnRK2.4 gene showed an up-regulation-upregulation pattern at 0,1 and 2 d under low temperature stress.Research suggests that StSnRK2.4 gene may be involved in abiotic stress and related signal molecular responses.