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香蕉几丁质酶基因家族的鉴定及在枯萎病侵染时的表达分析
林秋妹1, 王 霞2, 赵东方2, 贾彩红2, 王静毅2, 刘菊华2, 徐碧玉2, 张玄兵1, 王 卓2
0
(1.海南大学园艺学院;2.中国热带农业科学院热带生物技术研究所/农业部热带生物技术重点开放实验室)
摘要:
几丁质酶(Chitinase, CHi)是一种能够催化真菌细胞壁中几丁质水解的病程相关蛋白,在植物与真菌互作中起着重要的作用。本研究以拟南芥几丁质酶基因的蛋白序列为查询序列,在香蕉A基因组(DH-Pahang)中进行BLASTp比对,鉴定出23个MaCHis基因家族成员。根据其染色体上的位置,分别命名为MaCHi01~MaCHi23。系统进化树分析表明23个MaCHis可分为糖苷水解酶18(glycosyl hydrolase 18, GH18)和糖苷水解酶19(GH19)两个亚家族。转录组数据分析表明接种Foc TR4后MaCHi05、MaCHi06、MaCHi07、MaCHi11、MaCHi18、MaCHi19和MaCHi20基因在感病品种‘巴西蕉’中显著下调或不表达,在抗病品种‘GCTCV-119’中显著上调。利用RT-qPCR技术对MaCHi05、MaCHi06、MaCHi07、MaCHi11和MaCHi19在抗病和感病品种接种Foc TR4中进行表达分析,结果发现在抗病品种这些基因的表达都被激活,而在感病品种中没有激活,且在抗病品种中的相对表达量远大于感病品种,表明这些基因以正调控方式参与香蕉抗枯萎病过程。本研究结果为进一步解析MaCHi基因家族成员在香蕉抗枯萎病过程中的功能提供理论参考。
关键词:  香蕉  几丁质酶基因  表达分析  枯萎病
DOI:10.13430/j.cnki.jpgr.20210622001
投稿时间:2021-06-22修订日期:2021-07-28
基金项目:海南省科技计划三亚崖州湾科技城联合项目资助(320LH020);现代农业产业技术体系(CARS-32)
Identification of Banana Chitinase Gene Family and Analysis of Its Expression in Fusarium Wilt Infection
LIN Qiu-mei1, WANG Xia2, ZHAO Dong-fang2, JIA Cai-hong2, WANG Jing-yi2, LIU Ju-hua2, XU Bi-yu2, ZHANG Xuan-bing1, WANG Zhuo2
(1.College of Horticulture, Hainan University;2.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Tropical Crop Biotechnology of the Ministry of Agriculture and Rural Affairs)
Abstract:
Chitinase (CHi) is a disease-related protein that can catalyze the hydrolysis of chitin in fungal cell walls and plays an important role in the interaction between plants and fungi. In this study, using the protein sequence of the Arabidopsis chitinase gene as the query sequence, BLASTp alignment was performed in banana A genome (DH-Pahang), and 23 MaCHis gene family members were identified. According to their position on the chromosome, they are named MaCHi01 up to MaCHi23, respectively. Phylogenetic tree analysis showed that the 23 MaCHis can be divided into two subfamilies, GH18 (glycosyl hydrolase 18) and GH19 (glycosyl hydrolase 19). Transcriptome data analysis showed that MaCHi05, MaCHi06, MaCHi07, MaCHi11, MaCHi18, MaCHi19 and MaCHi20 genes were significantly down-regulated or not expressed in the susceptible variety ‘Baxijiao’ after inoculation with Foc TR4, and significantly up-regulated in the disease-resistant variety ‘GCTCV-119’. Using RT-qPCR technology to analyze the expression of MaCHi05, MaCHi06, MaCHi07, MaCHi11 and MaCHi19 in the resistant and susceptible varieties inoculated with Foc TR4, it was found that the expression of these genes was activated in the resistant variety, but not in the susceptible variety, and the relative expression in the resistant variety was much greater than that in the susceptible variety, indicating that these genes participated in the process of banana fusarium wilt resistance in a positively regulated manner. The results of this study provides a theoretical reference for further analysis of the functions of the members of the MaCHi gene family in the process of banana fusarium wilt resistance.
Key words:  banana  chitinase gene  expression analysis  fusarium wilt