| 李正男1, 马强1, 张磊1, 孙平平1, 赵文军2, 周洪友1*.宁夏马铃薯僵顶植原体的分子鉴定[J].植物保护,2021,47(2):56-61. |
| 宁夏马铃薯僵顶植原体的分子鉴定 |
| Detection and identification of potato stolbur-associated phytoplasmas in Ningxia |
| 投稿时间：2019-12-09 修订日期：2020-03-08 |
| DOI：10.16688/j.zwbh.2019679 |
| 中文关键词: 马铃薯僵顶植原体 16S rRNA基因 rp基因 |
| 英文关键词:potato stolbur phytoplasmas 16S rRNA gene rp gene |
| 基金项目:国家重点研发计划（2017YFD0201101）；国家自然科学基金（31670128）；内蒙古自治区科技攻关项目（2019GG180）；内蒙古自然科学基金（2019MS03021） |
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| 中文摘要: |
| 通过透射电子显微镜, 在从宁夏回族自治区固原市彭阳县红河镇采集的表现叶片上卷、红叶、气生薯症状的马铃薯样品叶脉韧皮部筛管细胞内观察到大量直径为500~700 nm的球形植原体粒子。以提取的感病和健康马铃薯叶片总DNA为模板, 应用植原体16S rRNA基因和rp基因通用引物进行PCR扩增, 从感病样品中扩增得到了长度均约为1.2 kb的片段。对获得基因核酸一致性比较分析表明, 马铃薯僵顶植原体宁夏株系16S rRNA基因与‘Candidatus Phytoplasma fragariae’槭树株系(MK501642)16S rRNA基因核酸一致性最高, 为99.7%, rp基因与‘Ca. P. fragariae’云南马铃薯YN-2G株系(KJ144889)rp基因核酸一致性最高,为100%; 基于16S rRNA基因和rp基因构建系统进化树发现, 马铃薯僵顶植原体宁夏株系与16SrⅫ-E亚组成员聚在一起。基于透射电镜观察和基因序列比较分析, 证明宁夏发生的马铃薯僵顶病与植原体侵染相关, 该植原体在分类地位上属于植原体16SrⅫ-E亚组。 |
| 英文摘要: |
| Potato plants showing the symptoms of leaf rolling, reddening and air tuber were collected in Honghe town, Pengyang county, Guyuan city, Ningxia Hui autonomous region, China. The phytoplasma-like particles of 500700 nm in diameter were observed in the phloem sieve elements of the leaves of the collected samples under a transmission electron microscope (TEM). Total DNAs were extracted from the symptomatic and healthy samples, respectively, as templates, and PCR products of around 1.2 kb were amplified from the symptomatic samples using phytoplasma universal primers for 16S rRNA and rp genes. Analyses of the nucleotide sequences indicated that the 16S rRNA of potato stolbur phytoplasma shared the highest identity of 99.7% with that of ‘Candidatus Phytoplasma fragariae’ strain Acer sp. (MK501642); the rp gene of potato stolbur phytoplasma had the highest identity of 100% with that of Ca. P. fragariae-related strain YN-2G (KJ144889). On the phylogenetic trees constructed based on 16S rRNA and rp gene sequences, the potato stolbur phytoplasma from Ningxia clustered with the members of 16SrⅫ-E subgroup. The results of TEM examination and molecular detection suggested that the potato stolbur in Ningxia is associated with the phytoplasma of the subgroup 16SrⅫ-E. |
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