华北农学报 ›› 2018, Vol. 33 ›› Issue (3): 56-62. doi: 10.7668/hbnxb.2018.03.010

所属专题: 生物技术

• 论文 • 上一篇    下一篇

谷瘟病菌交配型基因克隆和不同地区交配型基因检测

任世龙1,2, 杨振立3, 白辉1, 王永芳1, 全建章1, 董志平1, 李志勇1, 邢继红2   

  1. 1. 河北省农林科学院 谷子研究所, 河北省杂粮重点实验室, 国家谷子改良中心, 河北 石家庄 050035;
    2. 河北农业大学 生命科学学院, 河北 保定 071001;
    3. 河北省农林科学院, 河北 石家庄 050051
  • 收稿日期:2018-01-20 出版日期:2018-06-28
  • 通讯作者: 李志勇(1976-),男,河北邯郸人,研究员,博士,主要从事谷子病害研究;邢继红(1977-),女,河北东光人,教授,博士,主要从事分子植物病理学研究。
  • 作者简介:任世龙(1993-),男,河北邯郸人,在读硕士,主要从事谷瘟病菌相关研究。
  • 基金资助:
    河北省优秀专家出国培训项目;国家现代农业产业技术体系(CARS-07-13.5-A8)

Cloning of Mating Type Genes from Foxtail Millet Blast and Molecular Detection of Magnaporthe oryzae Mating Type Genes in Different Region

REN Shilong1,2, YANG Zhenli3, BAI Hui1, WANG Yongfang1, QUAN Jianzhang1, DONG Zhiping1, LI Zhiyong1, XING Jihong2   

  1. 1. Millet Institute, Hebei Academy of Agriculture and Forestry Sciences, Minor Cereal Crops Laboratory of Hebei Province, National Foxtail Millet Improvement Center, Shijiazhuang 050035, China;
    2. College of Life Sciences, Hebei Agricultural University, Baoding 071001, China;
    3. Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051, China
  • Received:2018-01-20 Published:2018-06-28

摘要: 为明确我国谷子产区谷瘟病菌交配型基因及其分布情况对谷瘟病菌群体结构分析的重要意义。根据已知稻瘟病菌交配型基因MAT1-1-1(GenBank登录号AB080672.2)和MAT1-2-1(GenBank登录号AB080673.2)的部分序列设计3对特异性引物,利用PCR技术对186株谷瘟病菌交配型基因进行扩增检测,比对分析谷瘟病菌和稻瘟病菌的交配型基因MAT1-1-1的alpha盒子部分氨基酸序列和MAT1-2-1高迁移率蛋白盒子氨基酸序列。结果显示,引物JPX1-1-S3/JPX1-1-A3对谷瘟病菌交配型基因MAT1-1-1的扩增效果较好,而JPX1-2-S1/JPX1-2-A1对谷瘟病菌交配型基因MAT1-2-1的扩增效果较好。序列比对发现谷瘟病菌与稻瘟病菌交配型基因极为相似,但并不完全相同。对2010-2016年采自河北、山东、山西、陕西、吉林、黑龙江、辽宁、内蒙古、新疆和海南等采集地点的共186株谷瘟病菌单胞菌株的交配型基因进行检测,发现夏谷区交配型为MAT1-1的谷瘟病菌占69.15%,而交配型为MAT1-2的菌株仅占26.60%,其余4.25%菌株为双交配型菌株。春谷区交配型为MAT1-1的谷瘟病菌占38.55%,而交配型为MAT1-2的菌株仅占56.63%,其余4.82%菌株为双交配型菌株。建立了谷瘟病菌交配型基因PCR检测体系,通过该方法检测多数谷子种植区存在MAT1-1与MAT1-2 2种交配型的谷瘟病菌菌株,且总体上2种交配型菌株比例接近1:1。但不同地区交配型菌株所占比例有一定差异,并且自然界中存在谷瘟病菌两性菌株。快速检测谷瘟病菌的交配型等位基因和不同区域谷瘟病菌交配型基因分布对研究谷瘟病菌群体结构与遗传分析具有重要意义。

关键词: 谷瘟病菌, 交配型, PCR检测

Abstract: To explore the mating type genes and distribution of foxtail millet blast in different millet production areas in China is important for the analysis of population structure of Magnaporthe oryzae. The three pairs of primers were designed according to the partial sequences of MAT1-1-1 (GenBank accession No.AB080672.2)and MAT1-2-1 (GenBank accession No.AB080673.2) genes of M. oryzae infecting rice,and the mating type genes of 186 strains of M. oryzae of foxtail millet was detected by PCR technique. In addition,amino acid sequence of α-box domain and highly mobility group protein-box of mating type genes were compared between M. oryzae of rice and M. oryzae of foxtail millet. The results showed that the amplification of mating type gene MAT1-1-1 was better by primer JPX1-1-S3/JPX1-1-A3,while the amplification of mating genotype MAT1-2-1 by JPX1-2-S1/JPX1-2-A1 was better. Amino acid sequence of α-box domain and highly mobility group protein-box of two mating type genes between M. oryzae of rice and M. oryzae of foxtail millet were highly similarity,but no identical. Mating type genes of 186 single ascospore isolates of M. oryzae in millet collected from Hebei,Shandong,Shanxi,Shaanxi,Jilin,Heilongjiang,Liaoning,Inner Mongolia,Xinjiang and Hainan Province in 2010-2016 were detected with primer JPX1-1-S3/JPX1-1-A3 and JPX1-2-S1/JPX1-2-A1. In the summer valley,the mating type of MAT1-1 was 69.15%,while the mating type MAT1-2 only accounted for 26.60%,the other 4.25% strains were double mating type strains. In the spring valley,the mating type of MAT1-1 was 38.55%,while the mating type MAT1-2 only accounted for 56.63%,the other 4.82% strains were double mating type strains. Conclusion,established the mating type gene of M. oryzae PCR detection system. Most of the millet growing areas were detected by this method have MAT1-1 and MAT1-2 two mating types,overall,the ratio of two mating type strains was close to 1:1. However,there were some differences in the mating type strains in different areas,and there were two strains of both mating type genes in nature. Rapid detection of mating type alleles and distribution of mating type genes in different regions of M. oryzae from foxtail millet is important for the study of population structure and genetic analysis of M. oryzae.

Key words: Magnaporthe oryzae, Mating type, PCR detection

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引用本文

任世龙, 杨振立, 白辉, 王永芳, 全建章, 董志平, 李志勇, 邢继红. 谷瘟病菌交配型基因克隆和不同地区交配型基因检测[J]. 华北农学报, 2018, 33(3): 56-62. doi: 10.7668/hbnxb.2018.03.010.

REN Shilong, YANG Zhenli, BAI Hui, WANG Yongfang, QUAN Jianzhang, DONG Zhiping, LI Zhiyong, XING Jihong. Cloning of Mating Type Genes from Foxtail Millet Blast and Molecular Detection of Magnaporthe oryzae Mating Type Genes in Different Region[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(3): 56-62. doi: 10.7668/hbnxb.2018.03.010.

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