| 何梅,朱淑珠,王雅丽,王林,李燏,宋常美.玛瑙红樱桃植原体鉴定及qPCR检测体系建立[J].中国南方果树,2025,54(3): |
| 玛瑙红樱桃植原体鉴定及qPCR检测体系建立 |
| Identification of phytoplasma and establishment of qPCR detection system for Prunus Pseudocerasus ‘Manaohong’ |
| 投稿时间：2024-09-11 修订日期：2024-10-13 |
| DOI： |
| 中文关键词: 玛瑙红樱桃 花变绿植原体 病原鉴定 实时荧光PCR 检测 |
| 英文关键词:Prunus pseudocerasus ‘Manaohong’ floral greenery phytoplasma identification real-time fluorescence PCR detection |
| 基金项目:国家自然科学基金地区基金 |
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| 中文摘要: |
| 为确定贵州玛瑙红樱桃上发生的花变绿病原分类地位及构建快速有效的检测体系，该研究利用植原体16S rRNA基因引物R16m F2/R16m R1对感病玛瑙红樱桃总DNA进行常规PCR扩增、克隆和测序，通过同源性比较、系统进化分析和在线虚拟RFLP分析，确定其属于植原体16SrV-B亚组。然后根据16S rRNA基因设计并合成引物对MN16SF-2/MN16SR-2，用重组质粒pEASY-T1-MN16S为模板建立标准曲线，构建玛瑙红樱桃花变绿植原体实时荧光定量PCR 检测体系；运用该体系检测喷施了外源茉莉酸甲酯、褪黑素、四环素和未施外源物质的患病苗木体内植原体含量，施加外源物质组体内植原体含量与未施组相比均大幅下降。该研究明确了玛瑙红樱桃花变绿植原体的分类地位并成功建立了qPCR检测体系，为玛瑙红樱桃植原体检测提供了技术支持。 |
| 英文摘要: |
| In order to determine the taxonomic status of the phytoplasma of Virescence on Guizhou Prunus pseudocerasus ‘Manaohong’ and to construct a rapid and effective detection system, the phytoplasma 16S rRNA gene primer R16m F2/R16m R1 was used to routinely PCR amplify, clonate and sequence the total DNA of susceptible Prunus pseudocerasus ‘Manaohong’, and it was determined that it belonged to the phytoplasma 16SrV-B subgroup through homology comparison, phylogenetic analysis and online virtual RFLP analysis. Then, according to the 16S rRNA gene, primer pairs MN16SF-2/MN16SR-2 were designed and synthesized, and the standard curve was established with the recombinant plasmid pEASY-T1-MN16S as template, and a real-time fluorescence quantitative PCR detection system for Prunus pseudocerasus ‘Manaohong’ Virescence phytoplasma was constructed. In this study, the taxonomic status of Prunus pseudocerasus ‘Manaohong’ Virescence phytoplasma was clarified, and a qPCR detection system was successfully established, which provided technical support for the detection of Prunus pseudocerasus ‘Manaohong’ phytoplasma. |
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