| Biological sciences & biotechnology |
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| Identification of pathotype of Verticillium dahliae isolates on cotton in Zhejiang Province and phenotypic analysis on inhibitory effect by high temperature |
| SUN Xiaoting1, LU Xiuyun2, ZHANG Jingze1*, ZHU Shuijin3* |
| (1. Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China; 2. Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences, Baoding 071000, Hebei, China; 3. Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China) |
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Abstract China is the largest cotton producing country in the world. Zhejiang Province owned cotton growing area of about 17 264 hectares in 2013, was not major cotton producing area in China. Verticillium wilt, caused by Verticillium dahliae, is one of the most important disease of cotton and causes great economic losses in almost all cotton producing areas, but has not been found in fields in Zhejiang region. However, in 2014, the serious verticillium wilt disease of cotton was observed in the experimental field of Zhejiang University. The disease began to appear in the early June and occurred seriously in the early July. The disease caused the serious defoliation on the middle and lower parts of plants and the diseased plants were significantly dwarf. However, the disease became to remit gradually in the early August during high temperature in summer and dwarf phenomenon of the diseased plants was not obvious with new leaf formation. Since then, the diseased plants were still able to bloom and produce bolls. The symptom of the disease was similar to that caused by the defoliating pathotype strains of V. dahliae but in the later period, not identical to that.
Based on the importance of verifying pathotype of pathogen and understanding the phenomenon of high temperature inhibiting disease for disease control, we detected the isolates causing the verticillium wilt disease of cotton by using the specific primers of defoliation and nondefoliation pathotypes of V. dahliae and characteristics of microsclerotia. Meanwhile, the effect of temperature on growth and development of pathogen was determined and the reason for the phenomenon of high temperature inhibiting disease was analyzed as well.
Results of polymerase chain reaction (PCR) amplification using the specific primers showed that the isolates (VD-h1, VD-h2, VD-h6, VD-h3 and VD-h5) obtained all belonged to the defoliating pathotype. The results of observation for characters of microsclerotia formation confirmed that the difference of microsclerotia length-to-width ratio between defoliating and nondefoliating pathotype isolates was remarkable. Temperature had a significant influence on Verticillium isolates. The optimal growth temperature of two isolates (VD-h1 and VD-h6) from Zhejiang was 22-28 ℃ and two (VD-101 and VD-086) from Xinjiang was 22-26 ℃, whereas at 32 ℃, fungal hyphal growth was all inhibited. At the same time, the numbers of produced conidial decreased and conidial germination was delayed. Combining biological properties of fungus and meteorological data, research results indirectly verified that the reason for disappearance of disease symptoms was that the growth and reproduction of pathogen were inhibited under the condition of high temperature in summer.
In sum, this study provides important scientific data for studying the regulation of disease occurrence in future.
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Published: 19 November 2016
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Corresponding Authors:
ZHANG Jingze(http://orcid.org/0000-0001-8604-8280)，E-mail:jzzhang@zju.edu.cn；ZHU Shuijin(http://orcid.org/0000-0001-6209-9630)，E-mail:sjzhu@zju.edu.cn
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浙江棉花黄萎病菌致病型菌株的鉴定及高温抑制病害的表征分析
用大丽轮枝菌(Verticillium dahliae)落叶和非落叶致病型菌株特异性引物扩增并结合微菌核特征观察，鉴定了浙江地区棉花黄萎病菌菌株的致病型，测定了温度对病原菌生长发育的影响，分析了浙江地区夏季高温对棉花黄萎病抑制现象的原因。特异性引物的聚合酶链式反应扩增结果表明，来自于浙江的5个代表性分离菌株（VD-h1、VD-h2、VD-h6、VD-h3和VD-h5）均属于落叶型菌株。对不同类型菌株产生微菌核的观察结果证实，落叶型和非落叶型的微菌核间形态及其长宽比存在差异。温度对病原菌发育的影响结果显示，来自浙江的2个菌株（VD-h1和VD-h61）和来自新疆的2个菌株（VD-101和VD-086）的最适生长温度分别在22~28 ℃和22~26 ℃之间；32 ℃高温显著抑制了菌丝生长和分生孢子产生，并延迟了分生孢子萌发的时间。不同温度试验的结果间接证实了,棉花黄萎病在浙江夏季高温条件下表现的隐退症状是因为高温抑制了病原菌的生长和繁殖，这为病害发生规律研究提供了重要的科学数据。
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