| 刘建密1, 纪翠红2, 陈细红3, 林积秀2, 黄培枝2, 严叔平1, 陈文乐1, 陈彩霞1, 张宇翔2, 牟海青4*.福建永安市辣椒丛枝病植原体的分子检测及鉴定[J].植物保护,2015,41(3):115-118. |
| 福建永安市辣椒丛枝病植原体的分子检测及鉴定 |
| Molecular detection and identification of the phytoplasma associated with pepper witches’ broom in Yong’an |
| 投稿时间：2014-03-13 修订日期：2014-04-17 |
| DOI： |
| 中文关键词: 辣椒 植原体 16S rDNA 巢式-PCR 系统进化分析 |
| 英文关键词:pepper phytoplasma 16S rDNA nested-PCR phylogenetic analysis |
| 基金项目:国家科技重大专项（2013ZX08003-001）; 云南省科技计划项目（2012CH007） |
| 作者 | 单位 | | 刘建密1, 纪翠红2, 陈细红3, 林积秀2, 黄培枝2, 严叔平1, 陈文乐1, 陈彩霞1, 张宇翔2, 牟海青4* | 1. 福建三明市植保植检站, 三明365000
2. 福建永安市植保植检站, 永安366000
3. 福建厦门出入境检验检疫局, 厦门361012
4. 中国检验检疫科学研究院, 北京100029 |
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| 中文摘要: |
| 永安地区发病辣椒植株表现小叶、黄化、丛枝、簇芽等症状。利用植原体16S rDNA基因的通用引物R16 mF2/R16 mR2和R16F2n/R16R2, 对发病辣椒植株总DNA进行巢式PCR检测, 获得约1.2 kb的特异性DNA片段。经测序并在GenBank数据库进行比对分析, 共获得4条植原体特定的16S rDNA基因序列(CHY-C4-1、CHY-Y1-1、CHY-Y7-1、CHY-G1-1)。将测得的4条序列与已报道的植原体序列进行同源性比对, 并构建系统进化树, 结果显示获得的4条植原体序列均聚类到16SrI组, 其中CHY-Y1-1、CHY-Y7-1、CHY-G1-1与16SrI-B亚组植原体聚类到同一支, 而CHY-C4-1与已报道的16SrI组内的6个亚组均未聚类到一支, 因此建议将CHY-C4-1命名为新的亚组。利用iPhyClassifier 在线分析软件对获得的4条植原体序列进行虚拟RFLP分析, 结果与进化树获得的结果一致。 |
| 英文摘要: |
| Symptoms of little leaf, leaf yellowing, witches’ broom and bunching of small leaves were observed in pepper fields in Yong’an City. By use of nested-PCR, the 16S rRNA gene of phytoplasma associated with infected pepper was amplified with universal primer pairs R16 mF2/R16 mR2 and R16F2n/R16R2. The specific DNA fragments of ca. 1.2 kb were obtained from the total DNA of diseased samples. After nucleotide sequencing and BLAST analysis in GenBank, four sequences of phytoplasma 16S rRNA gene (CHY-C4-1, CHY-Y1-1, CHY-Y7-1 and CHY-G1-1) were obtained. The similarity and phylogenetic analysis showed that the four sequences were closely related to 16SrI group of phytoplasma. In the phylogenetic trees constructed with 16S rDNA, CHY-Y1-1, CHY-Y7-1 and CHY-G1-1 were clustered together with subgroup B of aster yellows group (16SrI-B), and it was suggested that CHY-C4-1 should belong to a new subgroup because CHY-C4-1 was not clustered together with any other subgroups of 16SrI. The results of four 16S rDNA sequences were analyzed by virtual RFLP online using iPhyClassifier software, and the result was correspondent with phylogenetic tree. |
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