东农冬麦1号<i>TabZIP2</i>基因的克隆及低温和ABA作用下的表达调控

doi:10.7668/hbnxb.20191127

摘要/Abstract

摘要： 东农冬麦1号（Dn1）是首例能够在黑龙江地区安全过冬的强抗寒冬小麦品种，目前其体内很多相关抗寒基因已被研究。为了探索外源ABA对冬小麦TabZIP2基因在低温胁迫下的影响，以强抗寒冬小麦品种东农冬麦1号（Dn1）为试验材料，克隆了TabZIP2基因的CDS序列全长，并且利用生物信息学手段对其进行了预测分析。之后又以弱抗寒冬小麦品种济麦22（JM22）为对比材料，应用RT-PCR技术对TabZIP2基因进行逆境下不同冬小麦品种的组织表达分析，并且探讨不同温度下外源ABA对Dn1分蘖节和叶片中TabZIP2相对表达量的影响。生物信息学分析结果表明： TabZIP2基因包含759 bp的完整开放阅读框，其编码的蛋白由252个氨基酸组成，是一个分子量为26.595 6 ku的不稳定蛋白；二级结构预测α-螺旋（H）和无规则卷曲（C）占大部分；该蛋白无跨膜区和信号肽，亚细胞定位表示该蛋白在细胞核中，亲水能力较强，并且与二穗短柄草以及水稻亲缘关系较近，而且该蛋白有保守的碱性亮氨酸拉链结构域。表达分析结果显示：寒胁迫下，Dn1叶片要比分蘖节提前应答低温胁迫；Dn1分蘖节和叶片中的TabZIP2表达量明显高于JM22；经过ABA处理后，-10，-25℃下Dn1分蘖节和叶片中TabZIP2的表达增加，即ABA正向调节TabZIP2的表达，初步判断ABA可以提高冬小麦抗寒性。本研究初步探究了TabZIP2在Dn1抗寒方面发挥的功能，也为低温下ABA信号调控机制提供了支持。

关键词: 冬小麦, TabZIP2, 低温, ABA, 生物信息学

Abstract: Winter wheat Dongnongdongmai 1(Dn1) is the first strong cold-resistant variety that can safely overwinter in Heilongjiang Province. At present, many related cold resistance genes have been studied. To explore the effect of exogenous ABA on the winter wheat TabZIP2 gene under low temperature stress, used Dn1 as experimental material, the full length of CDS sequence of TabZIP2 gene was obtained by PCR, and used bioinformatics to predict it. Later, we used the weak cold-resistant winter wheat variety Jimai 22(JM22) as comparative material, the tissue stress analysis of TabZIP2 gene was carried out by qRT-PCR, to investigate the effect of ABA on the relative expression of TabZIP2 in tiller node and leaves of Dn1 at different temperatures. Bioinformatics analysis results showed that:TabZIP2 contained a complete open reading frame of 759 bp, the TabZIP2 gene-encoded protein consisted of 252 amino acids and was an unstable protein with a molecular weight of 26.595 6 ku;Secondary structure prediction α-helix(H) and random coils(C) account for the majority;The protein had no transmembrane region and signal peptide, and was located in the nucleus with strong hydrophilic ability, and TabZIP2 was closely related to Brachypodium serrata and rice, the protein had a conserved basic leucine zipper domain. The results of expression analysis showed that Dn1 leaves responded to low temperature stress earlier than tillers under cold stress;TabZIP2 expression in tiller node of Dn1 and leaves was significantly higher than that in JM22;ABA could promote the increase of TabZIP2 expression in tiller nodes and leaved at-10, -25℃, preliminary judgment that ABA could improve cold resistance of winter wheat. This study preliminarily explored the function of TabZIP2 in the cold resistance of Dn1, and also laid a theoretical foundation for the regulation mechanism of ABA signal at low temperature.

Key words: Winter wheat, TabZIP2, Low temperature, ABA, Bioinformatics

中图分类号:

Q94
S512.03

关荧, 刘楠, 苍晶, 魏铁锁, 吴晶晶, 齐越, 郭富烨, 李沅珊. 东农冬麦1号TabZIP2基因的克隆及低温和ABA作用下的表达调控[J]. 华北农学报, 2021, 36(4): 75-81. doi: 10.7668/hbnxb.20191127.

GUAN Ying, LIU Nan, CANG Jing, WEI Tiesuo, WU Jingjing, QI Yue, GUO Fuye, LI Yuanshan. Cloning of TabZIP2 Gene and Expression Analysis under Low Temperature and ABA in Winter Wheat Dn1[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 75-81. doi: 10.7668/hbnxb.20191127.

http://www.hbnxb.net/CN/Y2021/V36/I4/75

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东农冬麦1号TabZIP2基因的克隆及低温和ABA作用下的表达调控

Cloning of TabZIP2 Gene and Expression Analysis under Low Temperature and ABA in Winter Wheat Dn1

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东农冬麦1号TabZIP2基因的克隆及低温和ABA作用下的表达调控

Cloning of TabZIP2 Gene and Expression Analysis under Low Temperature and ABA in Winter Wheat Dn1

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