玉米大斑病菌转录因子StMR1的结构特征及其表达模式分析

doi:10.3969/j.issn.1674-7968.2019.10.002

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摘要黑色素作为重要的致病因子直接影响植物病原真菌的侵染能力,而黑色素调控因子(melanin regulation factor, MR)在黑色素生物合成中发挥着重要作用。本研究通过比对玉米大斑病菌(Setosphaeria turcica)基因组序列,鉴定并克隆得到调控黑色素合成的转录因子基因Stmr1 (GenBank No. NW007359937)。该基因全长3 282 bp,含有3个内含子和4个外显子,编码1 021个氨基酸,编码蛋白StMR1含有2个Cys2His2锌指基序和1个Zn(Ⅱ)2Cys6锌簇蛋白基序,氨基酸序列分析发现,其与甘蓝链格孢(Alternaria brassicicola)和玉米小斑病菌(Cochliobolus heterostrophus)的MR转录因子相似性最高。自激活活性检测表明,StMR1具有较高的转录激活活性,进一步荧光定位表明,该蛋白在细胞核中表达。qRT-PCR检测Stmr1基因在病菌分生孢子萌发到穿透过程的5个时期的相对表达量,结果显示,其表达水平具有明显差异且在侵入钉时期表达量明显上调(P<0.05),推测其参与病菌侵染及菌丝的扩展。本研究结果为深入探究玉米大斑病菌黑色素合成及StMR1转录因子在病菌侵染中的调控机制提供理论依据。

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	贾慧
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	曹志艳
	董金皋

关键词 ：玉米大斑病菌, 转录因子, 黑色素调控因子, 基因结构, 表达模式

Abstract：As an important pathogenic factor, melanin directly affects the infection ability of plant pathogenic fungi, and the transcription factor MR (melanin regulation factor) plays an important role in melanin biosynthesis. In this study, the transcription factor gene Stmr1 (GenBank No. NW007359937) in Setosphaeria turcica was identified and cloned through genome BLAST. The gene was 3 282 bp in length and consisted of 3 introns and 4 exons, encoding 1 021 amino acids. The homology analysis of amino acid sequence indicated that the encoded protein StMR1 had the highest similarity to the MR transcription factors of Alternaria brassicicola and Cochliobolus heterostrophus. It contained 2 Cys2His2 zinc finger motifs and 1 Zn(Ⅱ)2Cys6 zinc cluster protein. Self-activation assay showed that StMR1 had high transcriptional activation activity and was located in the nucleus by fluorescence localization. Detected by qRT-PCR, the expression of Stmr1 gene was significantly different in the 5 stages from germination of conidia to penetration, and the expression was significantly up-regulated in the invasive nail stage, so it was speculated that it was involved in infection and expansion of the pathogen. The results of this study provide a theoretical basis for further exploration of the mechanism of regulation of melanin synthesis and StMR1 transcription factors in germ infection.

Key words： Setosphaeria turcica Transcription factor Melanin regulation factor Gene structure Expression pattern

收稿日期: 2019-06-21

ZTFLH:

S435.131.4+9

基金资助:国家玉米产业技术体系(CARS-02)和河北省自然科学基金(No. C2018204059)

通讯作者: caoyan208@126.com;dongjingao@126.com

引用本文:

贾慧, 张泽雪, 刘宁, 孟庆江, 曹志艳, 董金皋. 玉米大斑病菌转录因子StMR1的结构特征及其表达模式分析[J]. 农业生物技术学报, 2019, 27(10): 1720-1728.
JIA Hui, ZHANG Ze-Xue, Liu Ning, MENG Qing-Jiang, CAO Zhi-Yan, DONG Jin-Gao. Structural Characteristics and Expression Pattern Analysis of Transcription Factor StMR1 in Setosphaeria turcica. 农业生物技术学报, 2019, 27(10): 1720-1728.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.10.002 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I10/1720

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