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猕猴桃果实贮藏期主要真菌病害的rDNA-ITS鉴定及序列分析
引用本文:段爱莉,雷玉山,孙翔宇,高贵田,赵金梅,谷留杰.猕猴桃果实贮藏期主要真菌病害的rDNA-ITS鉴定及序列分析[J].中国农业科学,2013,46(4):810-818.
作者姓名:段爱莉  雷玉山  孙翔宇  高贵田  赵金梅  谷留杰
作者单位:1. 陕西师范大学食品工程与营养科学学院,西安710062;甘肃省武威市工业企业人才服务中心,甘肃武威733000
2. 陕西省农村科技开发中心,西安,710054
3. 陕西师范大学食品工程与营养科学学院,西安,710062
基金项目:陕西省“13115”科技创新工程重大科技专项(2009ZDKG-05)、陕西省科技统筹创新工程计划项目(2012KTJD03-05)、猕猴桃质量安全技术研究与示范(771845)、西安市科技项目:果树等品种选育、快繁、贮藏及病虫害防治技术研究与应用--猕猴桃贮藏期灰霉病、青霉病防治关键技术研究(NC1116(2))
摘    要:【目的】对陕西省周至县猕猴桃主栽品种‘华优’、‘海沃德’及‘秦美’贮藏期霉烂果实中的病原菌进行分离与鉴定,分析比较病原菌rDNA-ITS序列的差异和种内与种间群体分化状况,确定优势菌,为病害防治提供依据。【方法】采用病原菌形态鉴定与rDNA-ITS鉴定相结合的方法,确定引起猕猴桃霉烂的病原菌种类;通过构建系统发育树,分析病原菌种之间的亲缘关系。【结果】从贮藏期霉烂的猕猴桃果实中共分离出30株病原菌,主要为青霉属(Penicillium)、木霉属(Trichoderma)、交链孢霉属(Alternaria) 、毛霉属(Mucor)、拟青霉属(Simplicillium)等5个属。其中青霉属(Penicillium)19株占63.3%,为优势菌,包括Penicillium sp. 、Penicillium chrysogenum、Penicillium paneum、Penicillium purpurogenum和Penicillium commune;木霉属(Trichoderma)8株占26.7%较次之,包括Trichoderma longibrachiatum和Trichoderma sp;交链孢霉属(Alternaria) 、毛霉属(Mucor)、拟青霉属(Simplicillium)各1株,各占3.3%;聚类分析表明,30株病原聚为五大类群。【结论】引起陕西省周至县‘华优’、‘海沃德’和 ‘秦美‘猕猴桃贮藏期果实霉烂优势病原菌为Penicillium sp.、Penicillium chrysogenum、Penicillium paneum、Penicillium purpurogenum和Penicillium commune;rDNA-ITS区序列在青霉菌属内种间差异鉴别具有较高的参考价值。

关 键 词:猕猴桃    真菌    鉴定    rDNA-ITS序列分析
收稿时间:2012-02-07

Molecular Identification and rDNA-ITS Sequence Analysis of Fungal Pathogens in Kiwifruit During Storage
DUAN Ai-Li,LEI Yu-Shan,SUN Xiang-Yu,GAO Gui-Tian,ZHAO Jin-Mei,GU Liu-Jie.Molecular Identification and rDNA-ITS Sequence Analysis of Fungal Pathogens in Kiwifruit During Storage[J].Scientia Agricultura Sinica,2013,46(4):810-818.
Authors:DUAN Ai-Li  LEI Yu-Shan  SUN Xiang-Yu  GAO Gui-Tian  ZHAO Jin-Mei  GU Liu-Jie
Affiliation:1.College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an 710062; 2.Rural Science and Technology Development Center of Shaanxi Province, Xi’an 710054; 3. Wuwei Industrial Enterprises Talent Service Center of Gansu Province, Wuwei 733000, Gansu
Abstract:【Objective】The fungal pathogens of ‘China optimal’, ‘Haiwode’ and ‘Qinmei’ kiwi fruit that were harvested in Zhouzhi County, Shaanxi Province, were isolated during storage and identified by rDNA-ITS sequences analysis.【Method】With rDNA-ITS analysis and morphological characterization, the species of Fungal pathogens were differentiated and identified. By means of the genetic relationship analysis and multiple comparison analysis of ITS sequences of fungal pathogens, the diversity of intra-species in the pathogens were investigated.【Result】Five strains were identified, e.g. Penicillium, Trichoderma, Alternaria, Mucor and Simplicillium. Penicillium was the dominant species, accounting for 63.3% of the tested strains. And Trichoderma was the second dominant species, accounting for 26.7%. There were little differences in ITS sequences among Trichoderm, approval ratings were 99%. But the differences of ITS sequences between the Penicillium were significant .Such as Penicillium sp., Penicillium purpurogenum, Penicillium chrysogenum, Penicillium paneum and Penicillium commune.【Conclusion】Penicillium are mainly fungal pathogens of ‘China optimal’, ‘Haiwode’ and ‘Qinmei’ kiwifruit that were harvested in Zhouzhi County, Shaanxi Province. The differences of rDNA-ITS sequences between Penicillium are obvious. So, rDNA-ITS sequence between ourtra-species and intra-species have a certain reference value.
Keywords:kiwifruit  fungal  identification  rDNA-ITS sequence analysis
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