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节瓜(Benincasa hispida Cogn. var. Chieh-qua How)中WRKY基因同源物的克隆、特征分析及其在镰刀菌酸处理后的表达

Cloning and characterization of WRKY gene homologs in Chieh-qua (Benincasa hispida Cogn. var. Chieh-qua How) and their expression in response to fusaric acid treatment.

作者信息

Mao Yizhou, Jiang Biao, Peng Qingwu, Liu Wenrui, Lin Yue, Xie Dasen, He Xiaoming, Li Shaoshan

机构信息

Vegetable Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, China.

Guangdong Key Lab for New Technology Research of Vegetables, Guangzhou, 510640, China.

出版信息

3 Biotech. 2017 May;7(1):86. doi: 10.1007/s13205-017-0711-z. Epub 2017 May 13.

Abstract

The WRKY transcription factors play an important role in plant resistance for biotic and abiotic stresses. In the present study, we cloned 10 WRKY gene homologs (CqWRKY) in Chieh-qua (Benincasa hispida Cogn. var. Chieh-qua) using the rapid-amplification of cDNA ends (RACE) or homology-based cloning methods. We characterized the structure of these CqWRKY genes. Phylogenetic analysis of these sequences with cucumber homologs suggested possible structural conservation of these genes among cucurbit crops. We examined the expression levels of these genes in response to fusaric acid (FA) treatment between resistant and susceptible Chieh-qua lines with quantitative real-time PCR. All genes could be upregulated upon FA treatment, but four CqWRKY genes exhibited differential expression between resistant and susceptible lines before and after FA application. CqWRKY31 seemed to be a positive regulator while CqWRKY1, CqWRKY23 and CqWRKY53 were negative regulators of fusaric resistance. This is the first report of characterization of WRKY family genes in Chieh-qua. The results may also be useful in breeding Chieh-qua for Fusarium wilt resistance.

摘要

WRKY转录因子在植物对生物和非生物胁迫的抗性中发挥着重要作用。在本研究中,我们使用cDNA末端快速扩增(RACE)或基于同源性的克隆方法,从节瓜(Benincasa hispida Cogn. var. Chieh-qua)中克隆了10个WRKY基因同源物(CqWRKY)。我们对这些CqWRKY基因的结构进行了表征。对这些序列与黄瓜同源物进行系统发育分析,表明这些基因在葫芦科作物中可能具有结构保守性。我们使用定量实时PCR检测了这些基因在抗、感节瓜品系中对镰刀菌酸(FA)处理的表达水平。所有基因在FA处理后均能上调表达,但有4个CqWRKY基因在FA处理前后的抗、感品系间表现出差异表达。CqWRKY31似乎是抗镰刀菌酸的正调控因子,而CqWRKY1、CqWRKY23和CqWRKY53是负调控因子。这是关于节瓜中WRKY家族基因表征的首次报道。这些结果可能也有助于节瓜抗枯萎病的育种。

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