园艺学报 ›› 2009, Vol. 36 ›› Issue (3): 341-346.

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桃黄化病病原植原体的鉴定及16S rDNA序列分析

黄 云1*;王 靖1;赵艳琴2;何 苗1   

  1. (1四川农业大学植物病理系, 四川雅安625014; 2内蒙古民族大学植保系, 内蒙古通辽028043)
  • 收稿日期:2008-08-26 修回日期:2008-12-15 出版日期:2009-03-25 发布日期:2009-03-25
  • 通讯作者: 黄 云

The Identif ica tion and Sequence Analysis of 16S Ribosomal DNA of the Phytoplasma sp. Associated with Peach Yellow Disease

HUANG Yun1*, WANG Jing1,ZHAO Yan-qin2,and HE Miao1   

  1. (1Departm ent of Plant Pathology, S ichuan AgriculturalUniversity, Yapan, Sichuan 625014, China; 2Departm ent of Plant Protection, Inner Mongolia University for the N ationalities, Tongliao, InnerM ongolia 028043, China )
  • Received:2008-08-26 Revised:2008-12-15 Online:2009-03-25 Published:2009-03-25
  • Contact: HUANG Yun

摘要: 对四川省成都市龙泉驿区桃植原体黄化病病原进行了研究。经DAP I荧光染色, 在荧光显微镜下观察到病叶叶脉韧皮部存在荧光点; 在透射电镜下病叶韧皮部存在植原体菌体, 其大小为40~650 nm, 平均大小350 nm, 单位膜厚度10~16 nm。利用植原体16S rDNA通用引物对桃植原体黄化病患病植株提取的DNA进行nested2PCR扩增, 获得115 kb的特异片段, 同时构建了16S rDNA系统进化树。桃黄化病植原体序列与油桃黄化NecY2In1 (nectarine yellows) 病原同源性最高。从而探明了桃黄化病病原为植原体及其分类地位。

关键词: 桃, 桃黄化病, 植原体, 病原形态, 系统发育, 16S rDNA

Abstract: The phytop lasma associated with peach yellow disease occurred in Longquanyi District in Sichuan Province was studied. The specific fluorescence dots for the pathogen were observed in phloem cells of leaf veins under a fluorescence microscope by staining with 4, 62diamidino222phenylindole diacetate. Phytoplasma-like particles were also found in the same tissues under a transmission electronic microscope. Its diameter was 40-650 nm, 350 nm on average and the thickness of the unitmembrane of the pathogen was 10 - 16 nm. The 16S rDNA about 115 kb was amplified by nested-PCR with universal p rimers for Phytoplasm a using the total DNA extracted from diseased peach as the temp lates. A phylogenetic tree was constructed based on the sequences of 16S rDNA. The result showed that Phytoplasm a associated with peach yellow disease was closely related to nectarine yellows (NecY2In1).

Key words: peach, peach yellow disease, Phytoplasma, morphology, phylogenetics, 16S rDNA