沟叶结缕草<i>ZmCOR410</i>基因抗逆功能鉴定

doi:10.7668/hbnxb.20192992

摘要/Abstract

摘要：

为研究结缕草抗逆分子机制,报道了沟叶结缕草一个低温响应基因ZmCOR410的抗逆功能。基于转录组数据克隆获得了ZmCOR410 基因,采用实时定量PCR技术检测了其表达的低温响应模式,将其转化拟南芥中和酵母中鉴定了其抗逆功能。结果表明,ZmCOR410 基因编码区序列长927 bp,编码308个氨基酸,为酸性脱水素。在同源基因的系统进化树中ZmCOR410蛋白与耐旱性强的无芒隐子草和弯叶画眉草的同源基因亲缘关系最近。在基因组中,ZmCOR410基因编码区上游1 700 bp范围内有4个DRE顺式元件核心序列,其mRNA在低温胁迫下显著积累。转化了ZmCOR410基因的拟南芥,叶片抗冻能力增强,植株在干旱和高温胁迫下的存活率提高。转化了ZmCOR410基因的酵母细胞抗高温能力增强。结果说明,ZmCOR410 基因的表达产物在冷冻、高温和干旱胁迫下对细胞起保护作用,将有利于沟叶结缕草度过不良环境。

关键词: 沟叶结缕草, COR基因, 非生物胁迫, 脱水素

Abstract:

In order to explore the molecular mechanism of Zoysia grass in resistance to abiotic stresses,in the present study we reported the function of a cold responding gene from Zoysia matrella designated as ZmCOR410. The sequence information of the gene was obtained on the base of transcriptome and genomic data. The expression profile of ZmCOR410 in response to low temperature was detected by qRT-PCR,and the functions of the gene under abiotic stresses were estimated in Arabidopsis and yeast by genetic transformation.The results showed that the CDS of the gene was 927 bp in length,encoding an acid dehydrin that contains 308 amino acids. In the polygenetic tree of COR410 homologues from gramineous grasses,ZmCOR410 had a close relationship with the homologues of Cleistogenes songorica and Eragrostis curvula,two species that were high tolerant to drought stress. In the genomic DNA,there were four copies of core sequence of DRE-cis element in the 1 700 bp region upstream of the CDS of ZmCOR410,and its mRNA was accumulated in leaves exposed to cold. Compared to WT plants,Arabidopsis plants over expressing ZmCOR410 showed reduced injury in leaves after a freezing-temperature exposure,and showed higher survival rates under drought and high temperature stresses. Yeast cells harboring ZmCOR410 were also more tolerant to high temperature stress than the control cells. The results indicated that the product of ZmCOR410 could enhance cells resistance to freezing cold,high temperature and drought stresses,which would help Zoysia matrella going through adverse environments.

Key words: Zoysia matrella, COR genes, Abiotic stress, Dehydrins

张晓寒, 陈彦良, 马欣, 张珊珊, 韦善君. 沟叶结缕草ZmCOR410基因抗逆功能鉴定[J]. 华北农学报, 2022, 37(4): 53-61. doi: 10.7668/hbnxb.20192992.

ZHANG Xiaohan, CHEN Yanliang, MA Xin, ZHANG Shanshan, WEI Shanjun. Function Identification of ZmCOR410 from Zoysia matrella in Resistance to Abiotic Stresses[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(4): 53-61. doi: 10.7668/hbnxb.20192992.

http://www.hbnxb.net/CN/Y2022/V37/I4/53

图/表 9

图1 ZmCOR410 编码区的PCR扩增

Fig.1 The amplification of the coding sequence of ZmCOR410 by PCR

图2 基因组DNA中ZmCOR410 的结构白色方块为外显子,黑色方块为内含子;(+)和(-)分别代表有义链和无义链;1为翻译起始位点。

Fig.2 The structure of ZmCOR410 in the genome DNA Exon and intron is showed as white block and black block,respectively;(+)and(-)represents sense strand and anti sense strand,respectively;1 indicates the translation initiation site.

图3 ZmCOR410基因序列特征方框内为K-片段;下划线处为S-片段。

Fig.3 Sequence characters of ZmCOR410 The box is K-segment;Underline is S-segment.

图4 ZmCOR410蛋白疏水性分析及二级结构预测 A.COR410蛋白疏水性分析; B.COR410蛋白的二级结构预测。

Fig.4 Hydrophobicity analysis and secondary structure prediction of ZmCOR410 protein A.Hydrophobicity analysis of COR410 protein;B.Predicted secondary structure of COR410 protein.

图5 ZmCOR410蛋白氨基酸序列跨膜分析

Fig.5 Transmembrane analysis of ZmCOR410 protein

图6 ZmCOR410蛋白的系统进化树分析

Fig.6 Phylogenetic tree analysis of ZmCOR410 protein

图7 低温胁迫下的结缕草中ZmCOR410基因的相对表达量不同小写字母表示各处理时间存在显著差异(P<0.05)。图9同。

Fig.7 Relative expression of ZmCOR410 gene in Z. matrella under low temperature stress Different lowercase letters indicate that there are significant differences between processing times at 0.05 level.

图8 转基因植株的耐逆能力不同小写字母表示差异显著(P<0.05)。

Fig.8 Tolerance ability of transgenic plants to abiotic stresses Different lowercase indicate significant difference(P<0.05).

图9 高温胁迫下酵母细胞的存活率 A. 重组酵母表达载体;B. 酵母细胞存活率。

Fig.9 The survival rate of yeast cells exposed to high temperature stress A.The recombinant yeast-expression vector;B.The survival rate of yeast cells.

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沟叶结缕草ZmCOR410基因抗逆功能鉴定

Function Identification of ZmCOR410 from Zoysia matrella in Resistance to Abiotic Stresses

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