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林业科学 ›› 2019, Vol. 55 ›› Issue (5): 85-94.doi: 10.11707/j.1001-7488.20190510

• 论文与研究报告 • 上一篇    下一篇

油茶苗圃炭疽病原菌鉴定及抗药性

李河, 李司政, 王悦辰, 刘君昂, 徐建平, 周国英   

  1. 森林有害生物防控湖南省重点实验室 南方人工林病虫害防控国家林业和草原局重点实验室、经济林培育与保护省部共建教育部重点实验室 中南林业科技大学 长沙 410004
  • 收稿日期:2018-03-02 修回日期:2019-03-14 出版日期:2019-05-25 发布日期:2019-05-20
  • 基金资助:
    国家自然科学基金项目(31570641)。

Identification of the Pathogens Causing Anthracnose of Camellia oleifera in Nursery and Their Resistence to Fungicides

Li He, Li Sizheng, Wang Yuechen, Liu Jun, Xu Jianping, Zhou Guoying   

  1. Hunan Provincial Key Laboratory for Control of Forest Diseases and Pests, Key Laboratory of National Forestry and Grassland Administration for Control of Diseases and Pests of South Plantation, Key Laboratory for Non-Wood Forest Cultivation and Conservation of Ministry of Education, Central South University of Forestry and Technology Changsha 410004
  • Received:2018-03-02 Revised:2019-03-14 Online:2019-05-25 Published:2019-05-20

摘要: [目的]探讨中国油茶苗圃炭疽病原菌种类,揭示油茶苗圃炭疽病菌抗药性,为其防治提供依据和指导。[方法]分离油茶苗圃炭疽病菌,并对分离菌株的ApMat基因进行PCR扩增和测序,通过对序列系统发育分析鉴定炭疽病原菌种类;采用区分剂量法测定所有菌株对杀菌剂多菌灵、乙霉威、戊唑醇和咪鲜胺的抗药性。[结果]湖南、江西、海南和广东4个省份分离的95株的油茶苗圃炭疽病病原菌主要是果生炭疽菌(Colletotrichum fructicola)、暹罗炭疽菌(C.siamense)、山茶炭疽菌(C.camelliae)和胶胞炭疽菌(C.gloeosporioides)4个种,其中炭疽病菌分布范围最广、分离数量最多(83株,占总分离菌株的87.4%)。抗药性测定结果表明,对多菌灵产生抗性的菌株有31个,占总分离菌株的32.6%;对戊唑醇产生了抗性的菌株有31株,占总分离菌株的32.6%。其中对多菌灵和戊唑醇这2种杀菌剂均具有抗性的菌株有6株,占总分离菌株的6.3%;对多菌灵、乙霉威和戊唑醇3种杀菌剂均有抗性的菌株有2株,占所有分离菌株的2.1%。但所有95个菌株对咪鲜胺敏感,说明目前苗圃油茶炭疽病菌尚未对咪鲜胺产生抗药性。29个高抗多菌灵而对乙霉威敏感菌株第198位氨基酸由谷氨酸(Glu)突变为丙氨酸(Ala),2株对多菌灵和乙霉威具有双重抗药性的菌株第200位氨基酸由苯丙氨酸(Phe)突变为酪氨酸(Tyr)。[结论]我国油茶苗圃炭疽病菌已对多菌灵/乙霉威和戊唑醇产生严重的抗药性,但对咪鲜胺尚未检测到抗药性。

关键词: 油茶, 炭疽病, 病原种类, 抗药性

Abstract: [Objective]The objectives of this study were to identify the pathogens causing anthracnose of Camellia oleifera(oil-tea) in nursery in China and to reveal the resistance of anthracnose pathogens to fungicides, in order to provide targeted strategy for controlling oil-tea tree anthracnose.[Method]The anthracnose pathogen was isolated from C. oleifera in nursery, and its ApMat gene was amplified and sequenced by PCR. The anthracnose pathogen species were identified by phylogenetic analysis. The resistance of all strains to fungicides carbendazim, ethomyl, tebuconazole and prochlorazolam was further determined by differentiated dose method.[Result]Based on phylogenetic analysis of the ApMat locus with the neighbor-joining (NJ) method, the 95 isolates of anthracnose pathogens, collected from oil-tea trees in nurseries in Hunan, Jiangxi, Hainan, and Guangdong provinces (the important oil-tea trees production regions in China), were found belonging to four species of Colletotrichum genus:C. fructicola,C. siamense,C. camelliae and C. gloeosporioides. Among them, C. fructicola was the most abundant, accounted for 87.4%. Among these 95 strains, 31 were respectively resistant to either carbendazim or tebuconazole, with 6 of the 56 showing resistance to both carbendazim and tebuconazole. In contrast, all 95 strains were very sensitive to prochloraz. Two strains were resistant to three fungicides, carbendazim, carbendazim and tebuconazole. Comparison of the β-tubulin amino acid sequences between carbendazim-resistant and susceptible strains of Colletotrichum spp. revealed that 29 resistant strains had a mutation leading to an amino acid substitution at the position 198, from glutamicacid in the susceptible strain to alanine in the resistant strains.Furthermore, the 200th amino acid was also mutated from phenylalanine (Phe) to tyrosine (Tyr) of β-tubulin in the two strains showing resistance to both carbendazim and carbendazim.[Conclusion]The results suggest that the Colletotrichum spp. collected from oil-tea tree nurseries in China have developed serious resistance to carbendazim, diethofencarb and tebuconazole but not to prochloraz.

Key words: Camellia oleifera, anthracnose, Colletotrichum spp, fungicide resistance

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