谷子C2H2型锌指蛋白基因SiZFP182的克隆及表达分析

以‘晋谷45’和‘晋谷42’2个品种作为试验材料,采用聚乙二醇(PEG-6000)模拟干旱胁迫处理谷子幼苗,检测不同处理时间下丙二醛(MDA)的含量,评价其抗旱性。此外,在2个品种中克隆获得SiZFP182基因的cDNA全长,分析比较它们之间的序列差异;通过定量RT-PCR的方法分析SiZFP182基因在受到干旱胁迫的谷子幼苗中的表达特性。结果表明,‘晋谷45’的抗旱性强于‘晋谷42’,2个品种中SiZFP182基因的序列存在5处单碱基差异,使得编码的氨基酸序列不同。此外,PEG胁迫处理后24h内,SiZFP182基因在2个品种中的表达水平均表现出先升高后降低的趋势,处理后6~12h,该基因在‘晋谷45’中表达水平高于‘晋谷42’。初步认为‘晋谷45’的抗旱特性与‘晋谷42’对干旱敏感的特性,可能与干旱胁迫下处于幼苗期的2个品种中SiZFP182基因序列差异及表达差异相关。

Isolation and functional analysis of a C2H2-type zinc finger protein gene SiZFP182 in foxtail millet

Two foxtail millet cultivars,Jingu 45 and Jingu 42,were selected as research materials in this study.Millet seedlings were exposed to PEG-6000 stress treatment.We detected MDA content in seedlings at different treatment time in order to evaluate their drought resistance.In addition, SiZFP182 genes were isolated from cDNAs from these two cultivars and their sequences were blasted and analyzed.The expression pattern of SiZFP182 in millet seedlings after PEG treatment was analyzed by quantitative RT-PCR.The results showed that Jingu 45 had a stronger drought resistance than Jingu 42.Five single nucleotide sequence differences were discovered between these two cultivars,which resulting in encoded protein differences.The expression level of SiZFP182 genes in two cultivars were increased followed by decreasing under drought stress treatment within 24 h.During 6 to 12 h treatment period,the expression level of SiZFP182 in Jingu 45 was higher than that in Jingu 42.The above results indicated that differences in drought resistance between Jingu 45 and Jingu 42 might be related to sequence variation and expression difference of SiZFP182 under drought stress.