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谷子C2H2型锌指蛋白基因SiZFP182的克隆及表达分析
张彬1,2,3, 禾璐1, 候蕊1, 路阳1, 马芳芳1,2, 王兴春2,4, 杨致荣2,5, 韩渊怀1,2,3, 李红英1,2,3
1.山西农业大学 农学院, 山西 太谷 030801;2.山西农业大学 农业生物工程研究所, 山西 太谷 030801;3.农业部黄土高原作物基因资源与种质创制重点实验室, 太原 030001;4.山西农业大学 生命科学学院, 山西 太谷 030801;5.山西农业大学 文理学院, 山西 太谷 030801
摘要:
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6~12 h,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。
关键词:  谷子  C2H2型锌指蛋白  SiZFP182  抗旱性
DOI:10.11841/j.issn.1007-4333.2015.05.02
分类号:
基金项目:教育部2013年高等学校博士学科点专项科研基金联合资助课题(20131403110001);山西特色植物基因资源开发与利用平台项目(2012091004-0103);国家自然科学基金项目(31371693,31471556);山西省青年科技研究基金(2013021024-2)
Isolation and functional analysis of a C2H2-type zinc finger protein gene SiZFP182 in foxtail millet
ZHANG Bin1,2,3, HE Lu1, HOU Rui1, LU Yang1, MA Fang-fang1,2, WANG Xing-chun2,4, YANG Zhi-rong2,5, HAN Yuan-huai1,2,3, LI Hong-ying1,2,3
1.School of Agricultural Science, Shanxi Agricultural University, Taigu 030801, China;2.Institute of Agricultural Bioengineering, Shanxi Agricultural University, Taigu 030801, China;3.Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture, Taiyuan 030031, China;4.College of Life Science, Shanxi Agricultural University, Taigu 030801, China;5.College of Arts and Sciences, Shanxi Agricultural University, Taigu 030801, China
Abstract:
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.
Key words:  foxtail millet  C2H2-type zinc-finger proteins  SiZFP182  drought resistance
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