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Chinaberry tree samples showing witches' broom were collected in Danzhou, Hainan Province. Using universal primers Rl6mF2/Rl6Mr1 for phytoplasma 16 S rDNA, a fragment of about 1.4 kb was amplified, cloned and sequenced from the DNA samples extracted from the diseased Chinaberry tree.
利用植原体16S rDNA通用引物对Rl6mF2/Rl6mR1,通过PCR技术从表现典型丛枝症状的苦楝植株总DNA扩增到约1.4 kb的特异片段,将此片段克隆后进行序列测定、分析及系统关系树构建。
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The phytoplasmas diseases were investigated in 30 counties and cities of 10 areas in Yunnan Province. There are 12 phytoplasma diseases, including paulownia witches-broom,chinaberry witches-broom, cactus witches-broom, christmas cactus witches-broom, orange little leaf, petunia flat stem and so on were indentified using PCR with universal primers, which amplified phytoplasma 16SrRNA gene and/or ribosomal protein genes. So the kind of phytoplasma diseases in Yunnnan Province were understood. Identified the characteristic of 16SrRNA and rp gene of phytoplasma and the kind, classification and genitic relativity of these strains using RFLP, clonging and sequence analysis (nearly 20 full-length sequences of phytoplasma 16SrRNA and/or rp gene were submitted in GenBank ); The phytoplasma strains isolated from herbage plants were conserved in storeroom. 9 articles were published in the key journals of China and 1 technique invention patent was applied. The net-page about phytoplasma disease was established. 4 graduate student were bringed up.
对云南省10个地区的30余个县市地区进行了植原体病害的调查采集工作;采用植原体16SrRNA基因通用引物(R16mF2/R16mR1及R16F2/R16R2)和/或植原体核糖体蛋白基因引物(rpF1/R1)对所采集到的病害标样总DNA进行PCR扩增,根据植原体特异扩增条带的出现,共鉴定出泡桐丛枝、苦楝丛枝、仙人掌丛枝、蟹爪兰丛枝、柑桔小叶、矮牵牛扁茎等12种植原体病害,初步明确了云南省植原体病害的种类;通过RFLP技术、克隆及测序技术获得其16SrRNA及部分核糖体蛋白基因的近全长序列(已在GenBank中共登录近20余个相关序列),通过序列比较分析,明确了上述植原体株系中这两个基因的特征并确定了云南省植原体株系的种类、分类地位及遗传相关性;对一些发生在草本植物上的植原体进行了株系保存;在国内核心期刊及一般期刊发表研究论文 9 篇,申请技术发明专利1项;初步建立了有关植原体病害的网页;培养研究生4名。
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Using universal primers (R16mF2/Rl and R16F2/R2) for 16S rRNA and primers (rpF1/rpR1) for ribosomal protein gene to detect phytoplasmas associated with of chinaberry tree witches'-broom from Yunnan and Guizhou Province by polymerase chain reaction.
根据植原体16S rRNA基因通用引物序列(R16mF2/R16mRl、R16F2/R16R2)及核糖体蛋白基因通用引物序列(rpF1/rpR1),应用PCR技术对采自不同省份的苦楝丛枝病植原体进行检测,结果从两个植物上分别获得苦楝丛枝病植原体云南株系和贵州株系。
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