CDPK20基因表达及其对山药块茎膨大调控作用研究
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引用本文:王金鑫,季 祥,高圆丽,张艳芳,邵 盈,邢丽南,霍秀文.CDPK20基因表达及其对山药块茎膨大调控作用研究[J].西北植物学报,2021,41(2):189~196
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作者单位
王金鑫,季 祥,高圆丽,张艳芳,邵 盈,邢丽南,霍秀文* (内蒙古农业大学 园艺与植保学院 呼和浩特010019) 
基金项目:国家自然科学基金(31860558);
中文摘要:为探讨钙依赖性蛋白激酶(CDPK)在山药块茎淀粉及内源激素合成中的作用,该研究以‘毕克齐’和‘大和长芋’山药为试验材料,测定了块茎的淀粉、糖、内源激素含量等指标并进行相关性分析;采用RT PCR技术克隆了钙依赖性蛋白激酶基因(CDPK20),并进行生物信息学分析,构建CDPK∶GFP融合载体,对CDPK蛋白进行亚细胞定位,实时荧光定量PCR对CDPK20在山药块茎不同发育时期的表达进行分析。结果表明:(1)CDPK20基因开放阅读框长为1 047 bp,共编码348个氨基酸。(2)CDPK20蛋白定位于细胞核和细胞膜。(3)CDPK20的表达量在块茎种植后的105~165 d呈现先升后降的趋势。(4)CDPK酶活性与淀粉含量呈极显著正相关,与可溶性总糖含量和还原糖呈显著负相关,与ABA呈极显著负相关,与ZR呈极显著正相关。研究表明,CDPK参与块茎中的淀粉和糖代谢及植物内源激素ABA、ZR的合成。
中文关键词:山药块茎  钙依赖性蛋白激酶  基因表达分析  相关性分析
 
Regulation of CDPK20 on Tuber Enlargement of Yam
Abstract:To explore the role of calcium dependent protein kinase (CDPK) in the synthesis of yam tuber starch and endogenous hormones, we used Bikeqi yam and Dahechangyu yam as the materials to determine the tuber starch, sugar, endogenous hormone contents and other indicators, as well as did the correlation analysis. The RT PCR (Reverse Transcription Polymerase Chain Reaction) technology has been used to clone the open reading frame of calcium dependent protein kinase gene (CDPK20). The bioinformatics analysis and subcellular location were performed. Real time fluorescent quantitative PCR was applied to analyze the expression of CDPK20 and the correlation analysis of physiological indicators. The results showed that: (1) the open reading frame length of CDPK20 gene is 1 047 bp, encoding a total of 348 amino acids. (2) CDPK20 protein is subcellularly located in the nucleus and cell membrane. (3) The expression of CDPK20 showed a trend of increasing first and then decreasing from 105 d to 165 d after tuber planting. (4) CDPK enzyme activity was extremely significantly positively correlated with starch content, significantly negatively correlated with total soluble sugar and reducing sugar contents, extremely significantly negatively correlated with ABA, and extremely significantly positively correlated with ZR. This result suggests that CDPK is involved in the metabolism of starch and sugar in tubers and the synthesis of plant endogenous hormone ABA and ZR.
keywords:yam tuber  calcium dependent protein kinase  gene expression  correlation analysis
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