| 李吉二, 温思思, 张羽加, 金洛稼, 赵伟春*.浙贝母灰霉病病原真菌的分子鉴定[J].植物保护,2022,48(2):151-156. |
| 浙贝母灰霉病病原真菌的分子鉴定 |
| Identification of pathogenic fungi species of gray mold disease of Fritillaria thunbergii |
| 投稿时间：2021-02-15 修订日期：2021-04-21 |
| DOI：10.16688/j.zwbh.2021091 |
| 中文关键词: 浙贝母 灰霉病 病原真菌 灰葡萄孢 分子鉴定 |
| 英文关键词:Fritillaria thunbergii grey mold disease pathogenic fungus Botrytis cinerea molecular identification |
| 基金项目:浙江省基础公益研究计划(LGN21C140004, 2015C32076); 浙江省中医药科技计划(2020B060) |
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| 中文摘要: |
| 为了鉴定引起浙贝母Fritillaria thunbergii Miq. 灰霉病的病原真菌, 于2015年-2019年收集浙贝母灰霉病样品, 采用常规组织分离法在PDA培养基上分离纯化获得10株葡萄孢属Botrytis真菌?进一步以内转录间隔区(internal transcribed spacer, ITS)?RNA聚合酶亚基Ⅱ(the second largest subunit of the nuclear RNA polymerase enzyme Ⅱ, RPB2)?热激蛋白60(heat shock protein 60, HSP60)和甘油醛-3-磷酸脱氢酶(glyceraldehyde 3-phosphate dehydrogenase, G3PDH)为DNA条形码, 用特异性引物进行PCR扩增并测序, 运用CodonCode Aligner拼接序列及BLAST分析, 这10个菌株序列相同, 为同一菌株, 在GenBank中进行同源性比对, 用MEGA 10.1构建系统发育树, 确定此菌株与灰葡萄孢Botrytis cinerea相应序列的一致性为100%?盆栽接种法检测表明此菌株可引起浙贝母灰霉病?据此认为浙贝母灰霉病的病原真菌是灰葡萄孢而非普遍认为的椭圆葡萄孢Botrytis elliptica? |
| 英文摘要: |
| In order to identify the pathogenic fungi causing grey mold on Fritillaria thunbergii Miq., the disease samples were collected from 2015 to 2019, conventional tissue isolate method was used to isolate and purify the pathogen from samples on PDA medium and the ten isolates belonging to Botrytis were obtained. The specific sequence of ITS (internal transcribed spacer), RPB2 (the second largest subunit of the nuclear RNA polymerase enzymeⅡ), HSP60 (heat shock protein 60) and G3PDH (glyceraldehyde 3-phosphate dehydrogenase) were used as DNA barcode. They were amplified by PCR and sequenced, respectively. The sequences were spliced using CodonCode Aligner software and then BLAST analysis was carried out. These ten isolates are same strain with same sequence. The sequences were aligned in GenBank, and the phylogenetic trees were constructed using MEGA 10.1 software. The results showed that this strain shared 100% identity with Botrytis cinerea. The pathogenicity test by in vivo pot inoculation revealed that this strain can cause gray mold disease on F.thunbergii. Therefore, the pathogenic fungus causing mold disease of F.thunbergii is B.cinerea rather than B.elliptica, which was generally believed. |
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