开发高效引物检测槟榔黄化病叶片及种果内植原体  

Developing Efficient Primers to Detect Phytoplasmas in Areca Palms Infected with Yellow Leaf Disease

作　　者：孟秀利 唐庆华[1] 林兆威 牛晓庆[1] 刘博 宋薇薇[1] Meng Xiuli;Tang Qinghua;Lin Zhaowei;Niu Xiaoqing;Liu Bo;Song Weiwei(The Innovation Plaorm for Academicians of Hainan Province,Coconut Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wenchang,571339)

机构地区：[1]中国热带农业科学院椰子研究所,海南省院士创新平台,文昌571339

出　　处：《分子植物育种》2022年第14期4624-4633,共10页Molecular Plant Breeding

基　　金：中国热带农业科学院基本科研业务费专项(No.1630152021006);海南省重大科技计划项目(No.zdkj201817);海南省院士创新平台科研专项共同资助。

摘　　要：槟榔黄化病是由植原体引起的致死侵染性病害,严重制约海南省槟榔产业发展。为了提高槟榔黄化植原体的检测效率,本研究设计了新的巢氏PCR引物:F4/R1和F2/R2,并利用该引物检测海南省300份槟榔黄化叶部样品,其中184份呈阳性,比植原体通用引物(R16mF2/R16mR1和R16mF2n/R16mR2)检测率提高了58.00%。利用新引物首次在槟榔种果中检测到植原体。将本研究扩增的约1.2 kb的特异性片段测序,并进行序列比对和系统进化树分析。结果显示,该槟榔黄化植原体属于16SrI组,且与16SrI-M序列完全一致。此外,本研究所测序的槟榔黄化植原体(GenBank登录号:MZ971180)与已知的海南省槟榔黄化植原体(GenBank登录号:FJ694685;FJ998269)分别存在两个碱基差异,序列一致性为99.80%。本研究为槟榔黄化病诊断及防控提供理论依据。Yellow leaf disease(YLD) of areca palm caused by phytoplasmas is a lethal infectious disease, which has severely restricted the healthy development of areca-nut industry in Hainan, China. In order to improve the detection rate of YLD phytoplasmas, two pairs of nested-PCR primers named F4/R1 and F2/R2 were designed and used to detect 300 leaf samples collected in Hainan. 184 out of 300 samples were positive and the detection rate is 58.00%higher than the rate of universal primers(R16mF2/R16mR1 and R16mF2n/R16mR2). Phytoplasmas were detected in the areca-nut old fruits by new primers for the first time. The approximate 1.2 kb amplified fragments were sequenced. Sequence alignment and phylogenetic tree construction results indicated that the YLD phytoplasmas in this paper were belong to16SrI group and completely consistent with 16SrI-M subgroup. In addition, the YLD phy toplasmas in this study(GenBank accession number: MZ971180) shared the sequence identity of 99.80% with known YLD phytoplasmas in Hainan(GenBank accession number: FJ694685;FJ998269). This study provides theoretical basis for the diagnosis and control of the disease.

关 键 词：槟榔黄化病 植原体 巢氏PCR 序列比对 系统进化树 

分 类 号：S763.7[农业科学—森林保护学]