叶下珠黄化植原体和丛枝植原体的分子鉴定  

Molecular Identification of Phyllanthus urinaria Yellows Phytoplasma and Witches’-broom Phytoplasma

作　　者：林兆威 牛晓庆[1] 唐庆华[1] 王晔楠 孟秀利 宋薇薇[1] LIN Zhaowei;NIU Xiaoqing;TANG Qinghua;WANG Yenan;MENG Xiuli;SONG Weiwei(Coconut Research Institute,Chinese Academy of Tropical Agricultural Sciences/Hainan Innovation Center of Academician Team,Wenchang,Hainan 571339,China)

机构地区：[1]中国热带农业科学院椰子研究所/海南省院士团队创新中心,海南文昌571339

出　　处：《热带作物学报》2024年第4期864-871,共8页Chinese Journal of Tropical Crops

基　　金：海南省院士创新平台科研专项(No.YSPTZX202138,No.YSPTZX202151);海南省自然科学基金项目(No.321QN345)。

摘　　要：2022年8月在海南省文昌市一个槟榔黄化病园内,分别发现叶片黄化和丛枝症状的叶下珠,前期检测均为植原体感染。为了明确叶下珠黄化植原体和丛枝植原体的分类鉴定,本研究通过克隆16S rDNA基因和核糖体蛋白(rp)基因,并进行基因序列一致性、系统发育树及虚拟RFLP等分析。结果显示:克隆获得叶下珠黄化植原体16S rDNA片段1246 bp,rp基因1212 bp;叶下珠丛枝植原体16S rDNA片段1827 bp,rp基因1240 bp。基因序列一致性显示,叶下珠黄化植原体与丛枝植原体的16S rDNA基因序列均与16SrⅠ组的植原体一致性高达98%以上,与槟榔黄化植原体海南株系的16S rDNA序列一致性达100%;而二者的rp基因序列均与rpⅠ组的植原体一致性高达99%以上。系统发育树分析显示,叶下珠黄化植原体和丛枝植原体16S rDNA基因均与翠菊黄化组(16SrⅠ)植原体聚于一个大分支,且与16SrⅠ-B亚组的槟榔黄化植原体聚于同一个小分支,亲缘关系接近;而rp基因均与翠菊黄化组(rpⅠ)植原体集聚于一个大分支,且与rpⅠ-B亚组的翠菊黄化植原体聚于同一个小分支,亲缘关系接近。虚拟RFLP分析显示,叶下珠黄化植原体和丛枝植原体的16S rDNA基因序列虚拟RFLP图谱与16SrⅠ-B的洋葱黄化植原体的参考图谱相同,且相似系数为1.00。综上表明,叶下珠黄化植原体和丛枝植原体均属于16SrⅠ-B亚组成员。研究结果可对采用铲除槟榔黄化病中间寄主的防控手段提供理论依据。In August 2022,we found Phyllanthus urinaria with yellows and witches’-broom symptoms,respectively,in areca yellows disease garden in Wenchang city,Hainan province,which were phytoplasma infection.In order to clarify the taxonomic status of the phytoplasma strain in the yellows leaves and witches’-broom of P.urinaria.In this study,we cloned the 16S rDNA gene and ribosomal protein(rp)gene,and analyzed the gene sequence consistency,phylogenetic tree and virtual RFLP.The results showed that the 16S rDNA fragment 1246 bp and rp gene 1212 bp were cloned of P.urinaria yellows phytoplasma,the 16S rDNA fragment 1827 bp and rp gene 1240 bp were cloned of P.urinaria witches’-broom phytoplasma.The gene sequence consistency showed that the 16S rDNA gene sequence of the P.urinaria yellows phytoplasma and the P.urinaria witches’-broom phytoplasma was more than 98%consistent with the phytoplasma of 16SrⅠgroup,and 100%consistent with the 16S rDNA sequence of areca yellows phytoplasma Hainan strain;the consistency of them in rp gene sequences was more than 99%to those of the rpⅠgroup.The phylogenetic tree analysis showed that the 16S rDNA genes of P.urinaria yellows phytoplasma and P.urinaria witches’-broom phytoplasma were clustered in a large branch with those of aster yellows phytoplasma group(16SrⅠ),and clustered in the same small branch with those of areca yellows phytoplasma of 16SrⅠ-B subgroup,with the close genetic relationship;in the phylogenetic tree of rp gene,they were clustered in a large branch with the aster yellows phytoplasma group(rpⅠ),and clustered in the same small branch with those of aster yellows phytoplasma of rpⅠ-B subgroup,with the close genetic relationship.The virtual RFLP analysis showed that the RFLP map obtained by the 16S rDNA gene of the P.urinaria yellows phytoplasma and P.urinaria witches’-broom phytoplasma was the same as the reference map of the onion yellows phytoplasma of 16SrⅠ-B,and the similarity coefficient was 1.00.In summary,the P.urinaria yellows phytoplasma a

关 键 词：叶下珠 黄化 丛枝 植原体 分子鉴定 

分 类 号：S763.7[农业科学—森林保护学]