| 杧果炭疽病菌漆酶基因Cglac3序列特征及其在两个侵染相关基因突变体中的表达分析 |
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| 引用本文： | 钟昌开,肖春丽,张贺,蒲金基,吴秋玉,刘燕莉,刘晓妹.杧果炭疽病菌漆酶基因Cglac3序列特征及其在两个侵染相关基因突变体中的表达分析[J].热带作物学报,2020,41(6):1202-1207. |
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| 作者姓名： | 钟昌开 肖春丽 张贺 蒲金基 吴秋玉 刘燕莉 刘晓妹 |
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| 作者单位： | 1.海南大学植物保护学院/热带农林生物灾害绿色防控教育部重点实验室,海南海口 5702282.中国热带农业科学院环境与植物保护研究所/农业农村部热带作物有害生物综合治理重点实验室/海南省热带农业有害生物监测与控制重点实验室,海南海口 571101 |
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| 基金项目： | 国家自然科学基金项目(31860479);海南省重大科技专项(ZDKJ2017003) |
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| 摘 要： | 本研究运用同源克隆技术克隆了杧果炭疽病菌（Colletotrichum gloeosporioides）的Cglac3漆酶基因,并采用实时荧光定量PCR分析了该基因在侵染过程、漆酶基因LAC1突变体、外切葡聚糖酶基因CgCBH1突变体中的差异表达情况。结果表明,杧果炭疽病菌漆酶Cglac3基因大小为1842 bp,含3个内含子,大小分别为56、51、58 bp。Cglac3开放读码框编码558个氨基酸,蛋白分子量为61.38 kDa,等电点（PI）为4.50。与未侵染对照0 h的表达量相比,Cglac3在6 h孢子萌发形成附着胞时表达量迅速升高,在12 h侵入寄主后达到最高,之后在菌丝扩展、叶片显症过程中出现波动,但均远高于0 h的表达量,显示Cglac3在侵染的不同阶段均发挥着重要的作用,可能是胶孢炭疽菌的重要致病因子之一。与野生型相比,Cglac3表达量在杧果炭疽病菌漆酶基因LAC1敲除突变体中下降了74%,在外切葡聚糖酶基因CgCBH1敲除突变体中下降了56%;在CgCBH1缺失敲除突变体中,LAC1的表达下降了68%,与Cglac3的表现趋势一致,说明Cglac3的表达受LAC1调控,外切葡聚糖酶基因CgCBH1也会影响Cglac3和LAC1的表达。
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| 关 键 词： | 杧果 漆酶 Cglac3基因 qRT-PCR |
| 收稿时间： | 2019-08-01 |
Sequence Characteristics of Laccase Gene Cglac3 and Its Expression in Two Infection-Related Gene Mutants from Colletotrichum gloeosporioides on Mango |
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| ZHONG Changkai,XIAO Chunli,ZHANG He,PU Jinji,WU Qiuyu,LIU Yanli,LIU Xiaomei.Sequence Characteristics of Laccase Gene Cglac3 and Its Expression in Two Infection-Related Gene Mutants from Colletotrichum gloeosporioides on Mango[J].Chinese Journal of Tropical Crops,2020,41(6):1202-1207. |
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| Authors: | ZHONG Changkai XIAO Chunli ZHANG He PU Jinji WU Qiuyu LIU Yanli LIU Xiaomei |
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| Abstract: | In this experiment, a laccase gene Cglac3 was cloned by homologously from Colletotrichum gloeosporioides, which is the fungal pathogen of mango anthracnose, and its expressions during infection process, in laccase gene Cglac1 and exoglucanase gene CgCBH1 deletion knockout mutants, were comparatively analyzed using real-time fluorescent quantitative PCR. Cglac3 was 1842 bp in size with three introns of 56, 51 and 58 bp respectively. The open reading frame of Cglac3 encoded 558 amino acids with a protein molecular weight of 61.38 kDa and an isoelectric point (PI) of 4.50. Cglac3 was highly expressed at 6 h after inoculation when the fungal spores were germing and forming appressoria, increased rapidly at 12 h after inoculation when the fugus were invading. Since then, the expresson of Cglac3 fluctuated during hyphal expansion and leaf necrosis, but both were much higher than the expression level at 0 h. It suggested that Cglac3 played an important role during C. gloeosporioides infection, might be a key virulent factor. Compared with the wild type, the expression of Cglac3 decreased by 74% in the C. gloeosporioides ΔLAC1 mutant, and decreased by 56% in the ΔCgCBH1 mutant. Interestingly, the expression of LAC1 was also decreased by 68% in the ΔCgCBH1 mutant, consistent with the trend of Cglac3 expression, indicating that LAC1 affected the expression of Cglac3, and CgCBH1 affected both the expressions of Cglac3 and LAC1. |
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| Keywords: | mango laccase Cglac3 gene qRT-PCR |
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