江西省抚州烟区青枯雷尔氏菌的分离鉴定与遗传多样性分析

Identification and genetic diversity analysis of Ralstonia solanacearum from Fuzhou tobacco-planting areas in Jiangxi Province

为揭示青枯雷尔氏菌的遗传多样性,进而有效防治烟草青枯病,从江西省抚州市宜黄、黎川、乐安、广昌、资溪和南丰等烟叶产区采集青枯病发病烟株进行病菌分离,并以青枯雷尔氏菌种特异引物759/760进行PCR鉴定,获得了青枯雷尔氏菌33株。同时采用RAPD熔解曲线分析法(McRAPD)及内切葡聚糖酶基因(egl)测序法进行遗传多样性分析,确定其序列变种。McRAPD结果显示,多数青枯雷尔氏菌菌株遗传分化与地理来源相关性不强。egl基因测序结果表明,33株菌属于亚洲分支(演化型Ⅰ)的序列变种13、15、17和34,序列变种和地理来源具有较强相关性。序列变种15菌株占比最高,为57.6%,19株分离自广昌、黎川、乐安、南丰、宜黄和资溪等6县10个乡镇,是优势序列变种。序列变种17占比最少,为9.1%,3株全部分离自宜黄县黄陂镇。而4株序列变种34主要分离自乐安县增田镇,黎川县潭溪乡和资溪县高阜镇,占比12.1%。广昌县尖峰乡和头陂镇,南丰县傅坊乡和太源乡等产区在云烟87上分离到序列变种13,占分离菌株的21.2%,在江西省抚州市为首次发现。

To investigate the genetic diversity of Ralstonia solanacearum and effectively control tobacco bacterial wilt, 33 strains were isolated from Yihuang, Lichuan, Le’an, Guangchang, Zixi and Nanfeng in Fuzhou tobacco-planting areas and identified by PCR with primer 759/760. The genetic diversity of the 33 stains was analyzed by McRAPD and sequence of endoglucanase gene(egl) and the sequevar type of those strains were confirmed. McRAPD results showed that geographical origin influenced the genetic diversity of most strains slightly. Sequencing results indicated that the 33 strains belonged to the sequevars(13, 15, 17 and 34) from Asian phylotype I and the sequevar closely corrected to geographical origin. Sequevar 15(including 19 strains)was most abundant, accounting for 57.6%, which was isolated from samples from 10 towns in 6 counties including Guangchang, Lichuan, Le’an, Nanfeng, Yihuang, Zixi, etc., therefore sequevar 15 was the dominant sequevar. The proportion of sequevar 17(including 3 strains) was the lowest, accounting for 9.1%, which was isolated from Huangpi Town in Yihuang County. Sequevar 34(including 4 strains) accounted for 12.1%, which was isolated from Zengtian Town in Le’an County, Tanxi Town in Lichuan County and Gaofu Town in Zixi County. Sequevar 13 accounted for 21.2%, which was isolated from Yunyan 87 planted in Jianfeng Town and Toupi Town in Guangchang County and Fufang Town and Taiyuan Town in Nanfeng County. It was the first time that sequevar 13 was found in Fuzhou.