苜蓿假盘菌产孢生物学特性和致病力分化及ISSR标记研究
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摘要
本文对苜蓿假盘菌产孢生物学特性和寄主范围进行了初步研究,在对分离的7个不同菌株致病力分化研究的基础上,建立了苜蓿假盘菌生理分化的鉴别体系。通过ISSR分子标记,研究了菌株多态性和致病力分化及地理来源间的关系。
1.通过在不同温度、氮源、碳源、培养基和光照条件下,对苜蓿假盘菌产孢特性进行了研究。苜蓿假盘菌最适宜产生子囊盘的培养基是V-8碳酸钙培养基和苜蓿汁培养基。在不同碳氮源上不宜形成子囊盘。最适宜的温度是25~28℃,随着温度的升高子囊盘成熟率增大,但超过30℃不宜形成子囊盘。最适宜的光照条件是2000lux,黑暗不适合子囊盘的形成。人工诱导可以产生子囊盘但不产生子囊孢子。
2.对豆科牧草的5个属14个种在室内条件下进行苜蓿假盘菌接种,并观察苜蓿假盘菌的寄主范围。试验结果表明:苜蓿假盘菌可以危害苜蓿属的紫花苜蓿、杂花苜蓿、镰荚苜蓿,草木樨属的白花草木樨、黄花草木樨,三叶草属的甜三叶。说明苜蓿假盘菌具有寄主专化性。经室内鉴定,苜蓿假盘菌可以在苜蓿属和黄花草木樨上产生成熟的子实体及子囊孢子,而在白花草木樨和甜三叶上不能形成子实体。
3.从我国苜蓿品种资源中首次成功筛选出9个具有栽培代表性的品种,即宝鸡苜蓿、泾阳苜蓿、沙湾苜蓿、沙河苜蓿、晋南苜蓿、天水苜蓿、龙牧一号、保定苜蓿和伊鲁瑰斯,作为苜蓿假盘菌的生理分化鉴别寄主。初步构建了生理分化的鉴别体系。
4.采用室内接种盘倒扣接菌法,根据在鉴别寄主上的发病等级,将采自我国苜蓿主产区的7个苜蓿褐斑病菌菌株划分成5个致病类型,其中致病类型A为强致病类型,致病类型E为弱致病类型,C、D和G组致病类型介于两者之间。
5.对苜蓿假盘菌的ISSR遗传多态性,致病力分化和地理来源的关系进行了探讨。结果表明:苜蓿褐斑病具有丰富的遗传多样性,不同地区的菌株存在很大的遗传分化现象,ISSR遗传聚类组群的划分与菌株的致病力差异有一定的相关性,但菌株的致病力差异与菌株的来源,遗传聚类组群的划分没有明显的相关性。
The objective of this paper is to study on biology of spores and apothecium formation and host category of pseudopeziza medicaginis; to establish the differentiate hosts system on the base of different pathogen;to study relativity of ISSR polymorphism and pathogenecity by using clustering analysis for issr polymorphic marker of pseudopeziza medicaginis.
1. Effect of temperature, carbon, nitrogen, light and media on sporulation of pseudopeziza medicaginis were studied.The results showed that V8-CaCO3 and juice of alfalfa culture medium were the best medium for apothecium. P. medicaginis could not form apothecium on different types of carbon and nitrogen source culture medium.The optimum range of temperature was 25℃-28℃. The optimum illumination time was 2000 lux. P. medicaginis were not sporulate on the condition of artificial culture.
2.The virulence and infectivity of Pseudopeziza medicaginis to some lequminosae were observed by means of artificial inoculating pathogens of P. medicaginis to fourteen spieces of five category. The results showed the hosts of P. medicaginis were Medicago sativa, Medicago varia, Medicago falcate, Melilotus medic, Melilotus officinalis and Melilotus Mill. P. medicaginis could sporulate on Medicago L and Melilotus Mill .It showed that P. medicaginis is host specialization and take sexual stage in Medicago L. and Melilotus Mill..
3. A set of Alfalfa germplasms were screened as P.m(Pseudopeziza medicaginis) pathogen differential hosts which were currently cultivated in China , such as M.sativa Baoji , M.sativa Jingyang, M.sativa Shawan , M.sativa Shahe, M.sativacv.Jinnan , M.sativacv.Tianshui, M.sativacv. TumuNo.1 , M.sativa Baoding and M.sativacv.Iroquois.
4.Sever P.m strains were classified into five pathogenecity groups through identification at adult stage according to disease severity on differential host. Among pathogenecity types we detected, the pathogenicity type A appeared to be highest virulence to host plants , and pathogenicity type E was weak one , pathogenicity types C, D and G were in middle place.
5.The relationship of genetic diversity,virulence and geographical origin were studied.The analysis showed that there were abundant genetic diversity among Pseudopeziza medicaginis, and the genetic variation was very significant from different area.The ISSR clustering groups had correlation with virulence variation,but virulence variation had no significant correlation with geographical origin and with ISSR groups.
1.通过在不同温度、氮源、碳源、培养基和光照条件下,对苜蓿假盘菌产孢特性进行了研究。苜蓿假盘菌最适宜产生子囊盘的培养基是V-8碳酸钙培养基和苜蓿汁培养基。在不同碳氮源上不宜形成子囊盘。最适宜的温度是25~28℃,随着温度的升高子囊盘成熟率增大,但超过30℃不宜形成子囊盘。最适宜的光照条件是2000lux,黑暗不适合子囊盘的形成。人工诱导可以产生子囊盘但不产生子囊孢子。
2.对豆科牧草的5个属14个种在室内条件下进行苜蓿假盘菌接种,并观察苜蓿假盘菌的寄主范围。试验结果表明:苜蓿假盘菌可以危害苜蓿属的紫花苜蓿、杂花苜蓿、镰荚苜蓿,草木樨属的白花草木樨、黄花草木樨,三叶草属的甜三叶。说明苜蓿假盘菌具有寄主专化性。经室内鉴定,苜蓿假盘菌可以在苜蓿属和黄花草木樨上产生成熟的子实体及子囊孢子,而在白花草木樨和甜三叶上不能形成子实体。
3.从我国苜蓿品种资源中首次成功筛选出9个具有栽培代表性的品种,即宝鸡苜蓿、泾阳苜蓿、沙湾苜蓿、沙河苜蓿、晋南苜蓿、天水苜蓿、龙牧一号、保定苜蓿和伊鲁瑰斯,作为苜蓿假盘菌的生理分化鉴别寄主。初步构建了生理分化的鉴别体系。
4.采用室内接种盘倒扣接菌法,根据在鉴别寄主上的发病等级,将采自我国苜蓿主产区的7个苜蓿褐斑病菌菌株划分成5个致病类型,其中致病类型A为强致病类型,致病类型E为弱致病类型,C、D和G组致病类型介于两者之间。
5.对苜蓿假盘菌的ISSR遗传多态性,致病力分化和地理来源的关系进行了探讨。结果表明:苜蓿褐斑病具有丰富的遗传多样性,不同地区的菌株存在很大的遗传分化现象,ISSR遗传聚类组群的划分与菌株的致病力差异有一定的相关性,但菌株的致病力差异与菌株的来源,遗传聚类组群的划分没有明显的相关性。
The objective of this paper is to study on biology of spores and apothecium formation and host category of pseudopeziza medicaginis; to establish the differentiate hosts system on the base of different pathogen;to study relativity of ISSR polymorphism and pathogenecity by using clustering analysis for issr polymorphic marker of pseudopeziza medicaginis.
1. Effect of temperature, carbon, nitrogen, light and media on sporulation of pseudopeziza medicaginis were studied.The results showed that V8-CaCO3 and juice of alfalfa culture medium were the best medium for apothecium. P. medicaginis could not form apothecium on different types of carbon and nitrogen source culture medium.The optimum range of temperature was 25℃-28℃. The optimum illumination time was 2000 lux. P. medicaginis were not sporulate on the condition of artificial culture.
2.The virulence and infectivity of Pseudopeziza medicaginis to some lequminosae were observed by means of artificial inoculating pathogens of P. medicaginis to fourteen spieces of five category. The results showed the hosts of P. medicaginis were Medicago sativa, Medicago varia, Medicago falcate, Melilotus medic, Melilotus officinalis and Melilotus Mill. P. medicaginis could sporulate on Medicago L and Melilotus Mill .It showed that P. medicaginis is host specialization and take sexual stage in Medicago L. and Melilotus Mill..
3. A set of Alfalfa germplasms were screened as P.m(Pseudopeziza medicaginis) pathogen differential hosts which were currently cultivated in China , such as M.sativa Baoji , M.sativa Jingyang, M.sativa Shawan , M.sativa Shahe, M.sativacv.Jinnan , M.sativacv.Tianshui, M.sativacv. TumuNo.1 , M.sativa Baoding and M.sativacv.Iroquois.
4.Sever P.m strains were classified into five pathogenecity groups through identification at adult stage according to disease severity on differential host. Among pathogenecity types we detected, the pathogenicity type A appeared to be highest virulence to host plants , and pathogenicity type E was weak one , pathogenicity types C, D and G were in middle place.
5.The relationship of genetic diversity,virulence and geographical origin were studied.The analysis showed that there were abundant genetic diversity among Pseudopeziza medicaginis, and the genetic variation was very significant from different area.The ISSR clustering groups had correlation with virulence variation,but virulence variation had no significant correlation with geographical origin and with ISSR groups.
引文
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