库尔勒香梨果萼黑斑病病原鉴定及其ITS、GPD和EF-1α序列分析.pdfVIP

园艺学报，2016 ，43 (2) ：329 –336. Acta Horticulturae Sinica doi ：10.16420/j.issn.0513-353x.2015-0879；http ：//www. ahs. ac. cn 329 库尔勒香梨果萼黑斑病病原鉴定及其 ITS 、GPD 和 EF-1α序列分析 宋 博，朱晓锋，徐兵强，阿布都克尤木·卡德尔，杨 森* （新疆农业科学院植物保护研究所，特色林果产业国家地方联合工程研究中心；农业部西北荒漠作物有害生物综合 治理重点实验室，乌鲁木齐 830091 ） 摘 要：采用常规组织分离法对库尔勒香梨果萼黑斑病病样进行病原菌分离，依据科赫氏法则进行 病原菌田间致病性测定，并利用分子生物学手段结合形态学鉴定确定病原菌及其分类学地位。田间致病 性测定结果表明链格孢属菌株为致病菌株，利用真菌通用引物进行致病菌株保守基因（ITS 、GPD 和 EF-1α） PCR 扩增及测序，获得序列 GenBank 数据库登录号为 KU145269 、KU145271 、KU145273 。根据田间致 病性、形态学观察以及保守基因遗传进化分析结果确定库尔勒香梨果萼黑斑病病原为链格孢属链格孢 （Alternaria alternata ）。 关键词：梨；黑斑病；鉴定；链格孢；保守基因 中图分类号：S 661.2 文献标志码：A 文章编号：0513-353X （2016 ）02-0329-08 Identification of Kolra Fragrant Pear Calyx-end Black Spot Pathogen and Its Sequence Analysis of ITS ，GPD and EF-1α * SONG Bo，ZHU Xiao-feng ，XU Bing-qiang ，Abudu Keyimu · Kader ，and YANG Sen （Institute of Plant Protection ，Xinjiang Academy of Agricultural Sciences ，National Local Joint Engineering Research Center of Special Forestry and Fruit Industry；Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis Ministry of Agriculture，Urumqi 830091 ，China ） Abstract ：A study was conducted to definitude the disease pathogen and its taxonomic status. The pathogen was isolated from disease tissue samples of Kolra Fragrant pear calyx-end black spot with conventional tissue isolation. The pathogenicity test was conducted and confirmed by Koch’s postulates. The pathogen was identified based on morphology and molecular identification. Determination of pathogenic field confirmed that the Alternaria sp. as the pathogen ，and meanwhile，the general primer of fungal conserved genes （ITS ，GPD and EF-1α）was taken advantage of for PCR amplification. After that ， it w