黄瓜果实发育相关基因<i>CsIAA29</i>的克隆与表达分析

doi:10.7668/hbnxb.20190916

摘要/Abstract

摘要： 前期通过EMS诱变方法获得黄瓜短果突变体，并对其进行了转录组分析，获得了一个在短果突变体中上调表达的差异表达基因CsIAA29（Csa2G381840），log₂FC=2.09。该基因属于Aux/IAA家族基因，参与植物生长发育早期生长素应答过程。为了研究CsIAA29基因在黄瓜生长发育期的作用，采用PCR技术从黄瓜649中克隆CsIAA29基因的CDS序列。利用Expasy等在线软件对CsIAA29进行蛋白质理化分析，结果表明，CsIAA29蛋白质编码207个氨基酸，分子式为C₁₀₆₂H₁₆₃₄N₃₀₂O₃₁₃S₇，脂肪系数为71.16，总平均亲水性为-0.831。蛋白质结构域预测发现，CsIAA29有4个结构域，符合Aux/IAA家族的结构域特征，初步估计4个结构域的功能应该和Aux/IAA家族Ⅰ、Ⅱ、Ⅲ和Ⅳ是一致的。进化分析表明，黄瓜CsIAA29与南瓜和甜瓜中的IAA29亲缘关系较近。qRT-PCR分析结果表明，CsIAA29基因在黄瓜子房表达量最高，而在雄花、叶片中的表达量相对较低。利用PLANTCAT在线软件对CsIAA29基因启动子元件进行分析，结果表明，CsIAA29基因的启动子包含了多个与调节植物内源生长激素运输相关的调控元件，推测其可能参与黄瓜生长素极性运输等生物途径。上述试验结果证明，CsIAA29基因可能通过调控果实内源生长激素等途径影响黄瓜果实生长发育。

关键词: 黄瓜, 果实发育, 表达分析, CsIAA29

Abstract: Transcriptome analysis was conducted on cucumber short fruit mutants by EMS in the previous experiment,a differentially expressed gene up-regulated in short-fruit mutants was obtained, CsIAA29 (Csa2G381840),log₂FC=2.09. This gene belonged to Aux/IAA family and is involved in auxin response in early plant growth and development. In order to study the role of CsIAA29 gene in the growth and development of cucumber,the CDS sequence of CsIAA29 gene was cloned from cucumber 649 by PCR. Expasy and other online software were used for the physicochemical analysis of CsIAA29 protein. The results showed that CsIAA29 protein encoded 207 amino acids,the molecular formula was C₁₀₆₂H₁₆₃₄N₃₀₂O₃₁₃S₇,the fat coefficient was 71.16,and the total average hydrophilicity was-0.831. Protein structure prediction found four CsIAA29 structure domains,conform to the domain of Aux/IAA family structure characteristics,a preliminary estimate the function of the four structural domain and Aux/IAA family Ⅰ,Ⅱ,Ⅲ and Ⅳ was consistent. Evolutionary analysis showed that CsIAA29 of cucumber was closely related to IAA29 of pumpkin and melon. The results of qRT-PCR showed that the expression of CsIAA29 gene was the highest in cucumber ovary,while the expression of CsIAA29 gene in male flowers and leaves was relatively low. The CsIAA29 gene promoter component was analyzed using the online software PLANTCAT,the results of promoter element analysis showed that the CsIAA29 gene promoter contained several regulatory elements related to the regulation of endogenous growth hormone transport in plants,which were involved in photosynthesis and auxin polarity transport in cucumber. The above experimental results proved that CsIAA29 might affect the growth and development of cucumber fruits by regulating endogenous growth hormone.

Key words: Cucumber, Fruit development, Expression analysis, CsIAA29

中图分类号:

S642.2
X172

于越, 陈海燕, 周龙, 郭冬雪, 宫思宇, 郝宁, 杜亚琳, 武涛. 黄瓜果实发育相关基因CsIAA29的克隆与表达分析[J]. 华北农学报, 2020, 35(S1): 11-17. doi: 10.7668/hbnxb.20190916.

YU Yue, CHEN Haiyan, ZHOU Long, GUO Dongxue, GONG Siyu, HAO Ning, DU Yalin, WU Tao. Cloning and Expression Analysis of CsIAA29 Gene Related to Fruit Development in Cucumber[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 11-17. doi: 10.7668/hbnxb.20190916.

http://www.hbnxb.net/CN/Y2020/V35/IS1/11

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黄瓜果实发育相关基因CsIAA29的克隆与表达分析

Cloning and Expression Analysis of CsIAA29 Gene Related to Fruit Development in Cucumber

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黄瓜果实发育相关基因CsIAA29的克隆与表达分析

Cloning and Expression Analysis of CsIAA29 Gene Related to Fruit Development in Cucumber

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