Cloning and expression characteristics of ClWRKY44 gene from Cunninghamia lanceolata

LIN Lili, HU Anqi, CHEN Gang, ZHANG Jiyue, CAO Guangqiu, CAO Shijiang

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (1) : 203-209.

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南京林业大学学报（自然科学版）
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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (1) : 203-209. DOI: 10.12302/j.issn.1000-2006.202011005

Cloning and expression characteristics of ClWRKY44 gene from Cunninghamia lanceolata

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Abstract

【Objective】The aim of this study is to clone the Cunninghamia lanceolata ClWRKY44 gene and analyze its expression characteristics, so as to provide a theoretical basis for revealing the molecular mechanism of stress resistance physiology of Cunninghamia lanceolata. 【Method】 We cloned the Chinese fir ClWRKY44 gene by using RT-PCR technology, conducted bioinformatics analysis and subcellular localization study, and used real-time fluorescent quantitative PCR technology to analyze its expression. 【Result】The open reading frame (ORF)of ClWRKY44 gene is 1 776 bp, which encodes 591 amino acids and has a WRKY domain. Phylogenetic tree analysis showed that ClWRKY44 gene was closely related to Arabidopsis thaliana ATWRKY44. It was expressed in different organs (leaves, roots, stems) of Cunninghamia lanceolata. After 6 h of low temperature stress treatment, the relative expression of ClWRKY44 gene was the highest, about 79.8 times that of the control group. After 24 h of drought stress treatment, the relative expression of ClWRKY44 gene was the highest under high concentration drought stress treatment, about 46.6 times that of the control group. 【Conclusion】The protein sequence encoded by Chinese fir ClWRKY44 gene has high homology with Arabidopsis ATWRKY44. It can play a role in regulating plant stress response, and provide a reference for the adaptability of Chinese fir seedling growth and development.

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Cunninghamia lanceolate / gene cloning / gene expression analysis / protein subcellular localization / stresses

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LIN Lili , HU Anqi , CHEN Gang , et al . Cloning and expression characteristics of ClWRKY44 gene from Cunninghamia lanceolata[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2022, 46(1): 203-209 https://doi.org/10.12302/j.issn.1000-2006.202011005

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Abstract
WRKY是植物特有的转录因子基因, 在植物对外界胁迫响应及生长发育的过程中发挥重要作用。本研究克隆了一个新的小麦WRKY转录因子基因TaWRKY44, 获得其全长cDNA, 其中开放阅读框长度为897 bp, 编码298个氨基酸。半定量RT-PCR的结果表明, TaWRKY44在叶片中表达水平较高, 并且受干旱和低温胁迫诱导表达。转基因功能分析结果表明, TaWRKY44的拟南芥超表达株系叶片变小, 叶柄缩短, 并且叶片细胞也明显小于野生型。另外, 转基因系对ABA、干旱和盐等胁迫处理的敏感性也高于野生型, 说明该基因可能作为一个转录抑制子参与逆境胁迫信号转导过程。
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