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| 土壤样品中长喙壳属真菌诱导分离方法的研究 | |
| 引用本文： | 张治萍,李倩,韩惠兰,周佩,张丹丹,王玉玺,罗来鑫,蒋娜,李健强.土壤样品中长喙壳属真菌诱导分离方法的研究[J].植物病理学报,2019,49(4):560-568. |
| 作者姓名： | 张治萍 李倩 韩惠兰 周佩 张丹丹 王玉玺 罗来鑫 蒋娜 李健强 |
| 作者单位： | 中国农业大学植物保护学院/种子病害检验与防控北京市重点实验室,北京 100193; 先正达(中国)投资有限公司种子保护学院,北京 102206; 北京绿农康地生物工程技术研究有限公司,北京101407 |
| 基金项目： | 国家自然基金(自然和耕作土壤生境中甘薯长喙壳菌群体遗传多样性及致病性研究,31571958) |
| 摘 要： | 长喙壳属真菌(Ceratocystis spp.)是一类重要的土传植物病原真菌,可危害多种作物。由于技术方法的限制,对于土壤样品中该病原菌的分离比较困难。本研究通过比较以胡萝卜、马铃薯、甘薯作为诱导富集载体对土壤中长喙壳属真菌的富集分离效果,确定了胡萝卜为最优诱导富集载体。进一步通过对胡萝卜片厚度、土壤相对湿度、培养温度、光照条件、保湿方式等培养条件的优化,建立了从土壤中分离长喙壳属真菌的有效方法,具体为:以厚度为3~5 mm的胡萝卜片作为诱导富集载体;土壤样品相对含水量为10%~30%;12 h光暗交替或24 h黑暗;在22℃~26℃条件下不保湿进行培养,8 d后可从携带最低数量为10个孢子/g的土壤样品中分离到靶标菌。采用该方法,对采自新疆、河南、湖南、云南等地共计108份自然或耕作土壤样品进行了分离,结果表明从采自云南蒙自市、临沧市的45份土壤样品中均分离获得了长喙壳属真菌。该方法具有操作简便、所需仪器设备简单、灵敏度较高、成本低和高通量等特点,适用于田间调查,可对样本量较大的土壤样品进行快速及时分离,以获得不同土壤生境中的长喙壳属真菌分离物。 |
| 关 键 词： | 长喙壳菌 土传病原菌 分离方法 胡萝卜诱导 |
| 收稿时间： | 2018-10-12 |
Development of a protocol for effective isolation of Ceratocystis spp. from soil by baiting technique |
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| ZHANG Zhi-ping,LI Qian,HAN Hui-lan,ZHOU Pei,ZHANG Dan-dan,WANG Yu-xi,LUO Lai-xin,JIANG Na,LI Jian-qiang.Development of a protocol for effective isolation of Ceratocystis spp. from soil by baiting technique[J].Acta Phytopathologica Sinica,2019,49(4):560-568. | |
| Authors: | ZHANG Zhi-ping LI Qian HAN Hui-lan ZHOU Pei ZHANG Dan-dan WANG Yu-xi LUO Lai-xin JIANG Na LI Jian-qiang |
| Institution: | College of Plant Protection/Beijing Key Laboratory of Seed Disease Testing and Control (BKL-SDTC), China Agricultural University, Beijing 100193, China; The Seedcare Institute, Syngenta (China) Investment Co. Ltd, Beijing 102206, China; Lvnongkangdi Bio-Engineering Technology Research Co. Ltd, Beijing 101407, China |
| Abstract: | Ceratocystis spp. are important soilborne pathogens to many crops. Isolating the pathogen from soil sample has been a challenge due to various technical issues. In this study, carrot was selected as a bait for enriching and isolating Ceratocystis spp. from soil after comparing with potato and sweet potato. A series of experiments were conducted including thickness of carrot slices, relative humidity of soil, temperature of incubation, lighting, and moisture maintenance, which resulted in an optimized protocol as following. Carrot was sliced around 3-5 mm thick, which were placed in soil with water content of 10% to 30% in a container closed. The samples were incubated at 22℃-26℃ for either 24 h with a 12 h light /12 h dark cycle or 24 h in the dark. The pathogen could be recovered from the minimum number of spores 10 per gram soil after 8 days of incubation. This method was successfully applied on 108 soil samples of natural and tillage areas from Xinjiang, Henan, Hunan and Yunnan provinces of China, in which Ceratocystis spp. were isolated for all the 45 samples from Mengzi and Lincang cities of Yunnan. The developed method provide advantages of fast and easy operation, low equipment requirement, and low cost. It can be effectively applied to Ceratocystis isolation from large soil samples. |
| Keywords: | Ceratocystis fimbriata soilborne pathogen isolation method carrot bait |
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