芸薹生链格孢致病缺陷突变体的筛选及初步分析
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摘要
芸薹生链格孢(Alternaria brassicicola (Schwein.) Wiltshire)引起的黑斑病是十字花科蔬菜生产上的重要病害之一,在全球主产区都有报道。目前关于该病菌的致病机理的研究还较少。因此,查找和鉴定芸薹生链格孢致病相关基因,不仅有助于认识该类病原真菌致病的分子机制,还有助于发现病害防治新靶标。本研究从本实验室已构建的芸薹生链格孢转化子库中筛选得到了7株致病性缺陷的突变菌株。进一步利用Hi TAIL-PCR技术对插入位点的侧翼序列进行了扩增,并通过生物学分析方法进行了初步分析。相关研究结果将有助于对芸薹生链格孢致病机理的认识,为发现病害防治新靶标提供理论基础。
Alternaria brassicicola(Schwein.) Wiltshire is a phytopathogenic fungus that causes one of the most economically important diseases of black rot on cruciferous vegetables, which was reported in global major production area. However, our knowledge of its pathogenic mechanism is still limited. Therefore, identification of pathogenicity-related genes not only helps to increase our understanding of molecular mechanisms, but also helps to identify new targets for disease control. In this research, seven pathogenicity defective mutants were identified from the transformants in our laboratory. The insertion site flanking sequence was amplified by HiTAIL-PCR and the disruption gene was analyzed. These results will be helpful for our understanding on pathogenic mechanism of A. brassicicola and provide theoretical basis of indentifing new targets for disease control.
Alternaria brassicicola(Schwein.) Wiltshire is a phytopathogenic fungus that causes one of the most economically important diseases of black rot on cruciferous vegetables, which was reported in global major production area. However, our knowledge of its pathogenic mechanism is still limited. Therefore, identification of pathogenicity-related genes not only helps to increase our understanding of molecular mechanisms, but also helps to identify new targets for disease control. In this research, seven pathogenicity defective mutants were identified from the transformants in our laboratory. The insertion site flanking sequence was amplified by HiTAIL-PCR and the disruption gene was analyzed. These results will be helpful for our understanding on pathogenic mechanism of A. brassicicola and provide theoretical basis of indentifing new targets for disease control.
引文
Akimitsu K.,Ohtani K.,Shimagami T.,Shimagami T.,Katsumoto M.,Igarashi C.,Tanaka S.,Matsuoka S.,Mochizuki S.,Tsuge T.,Yamamoto M.,Kodama M.,Ichimura K.,and Gomi K.,2016,Citrus as a molecular contact point for co-evolution of Alternaria pathogens,Physiological and Molecular Plant Pathology,95:93-96
Cho Y.,Ohm R.A.,Grigoriev I.V.,and Srivastava1 A.,2013,Fungal-specific transcription factor AbPf2 activates pathogenicity in Alternaria brassicicola.,Plant Journal,75(3):498-514
Kang Y.,Feng H.,Zhang J.,Chen S.,and Valverde B.E.,2017,TeA is a key virulence factor for Alternaria alternata(Fr.)keissler infection of its host,Plant Physiology and Biochemistry,115:73-82
Liu Y.G.,and Chen Y.,2007,High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences,Biotechniques,43(5):649
Saha U.,Mazumder M.,Mukherjee A.,Mukherjee A.,Parveen S.,Mondal B.,Maji S.R.,and Basu D.A.,2016,Critical analysis of phosphatidic acid mediated resistance response in Sinapis alba,against Alternaria brassicicola,Physiological and Molecular Plant Pathology,94:90-99
Srivastava A.,Cho I.K.,and Cho Y.,2013,The bdtf1 gene in Alternaria brassicicola is important in detoxifying brassinin and maintaining virulence on Brassica species,Molecular Plant-microbe Interactions,26(12):1429
Tsuji G.,Kenmochi Y.,Takano Y.,Sweigard J.,Farrall L.,Furusawa I.,Horino O.,and Kubo Y.,2000,Novel fungal transcriptional activators,Cmr1p of Colletotrichum lagenarium and pig1p of magnaporthe grisea,contain Cys2His2 zinc finger and Zn(Ⅱ)2Cys6 binuclear cluster DNA-binding motifs and regulate transcription of melanin biosynthesis genes in a developmentally specific manner,Molecular Microbiology,38(5):940-954
Xiao C.K.,Li Y.,and Li J.Q.,2003,Research in seed borne black sport disease in cruciferous vegetables,Zhongguo Nongye Daxue Xuebao(Journal of China Agricultural University),8(5):61-68(肖长坤,李勇,李健强,2003,十字花科蔬菜种传黑斑病研究进展,中国农业大学学报,8(5):61-68)
Cho Y.,Ohm R.A.,Grigoriev I.V.,and Srivastava1 A.,2013,Fungal-specific transcription factor AbPf2 activates pathogenicity in Alternaria brassicicola.,Plant Journal,75(3):498-514
Kang Y.,Feng H.,Zhang J.,Chen S.,and Valverde B.E.,2017,TeA is a key virulence factor for Alternaria alternata(Fr.)keissler infection of its host,Plant Physiology and Biochemistry,115:73-82
Liu Y.G.,and Chen Y.,2007,High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences,Biotechniques,43(5):649
Saha U.,Mazumder M.,Mukherjee A.,Mukherjee A.,Parveen S.,Mondal B.,Maji S.R.,and Basu D.A.,2016,Critical analysis of phosphatidic acid mediated resistance response in Sinapis alba,against Alternaria brassicicola,Physiological and Molecular Plant Pathology,94:90-99
Srivastava A.,Cho I.K.,and Cho Y.,2013,The bdtf1 gene in Alternaria brassicicola is important in detoxifying brassinin and maintaining virulence on Brassica species,Molecular Plant-microbe Interactions,26(12):1429
Tsuji G.,Kenmochi Y.,Takano Y.,Sweigard J.,Farrall L.,Furusawa I.,Horino O.,and Kubo Y.,2000,Novel fungal transcriptional activators,Cmr1p of Colletotrichum lagenarium and pig1p of magnaporthe grisea,contain Cys2His2 zinc finger and Zn(Ⅱ)2Cys6 binuclear cluster DNA-binding motifs and regulate transcription of melanin biosynthesis genes in a developmentally specific manner,Molecular Microbiology,38(5):940-954
Xiao C.K.,Li Y.,and Li J.Q.,2003,Research in seed borne black sport disease in cruciferous vegetables,Zhongguo Nongye Daxue Xuebao(Journal of China Agricultural University),8(5):61-68(肖长坤,李勇,李健强,2003,十字花科蔬菜种传黑斑病研究进展,中国农业大学学报,8(5):61-68)