Phylogenetic relationships of coconut phytoplasmas and the development of specific oligonucleotide PCR primers.
Phylogenetic relationships of coconut phytoplasmas and the development of specific oligonucleotide PCR primers.
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椰子植原体的系统发育关系和特异性寡核苷酸 PCR 引物的开发。
DOI：
10.1111/j.1744-7348.1998.tb05220.x
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发表时间：
1998
影响因子：
2.6
通讯作者：
N. Harrison
中科院分区：
文献类型：
--
作者：
A. Tymon;P. Jones;N. Harrison
利用聚合酶链式反应技术，从佛罗里达和墨西哥尤卡坦地区以及东非和西非的感染植株中扩增出与椰子致死性衰退病相关的植质16S rRNA基因和16S-23S间隔区。在对rDNA产物进行测序后，系统发育分析证实，这些椰子植原体在植原体分支中形成了一个单独的簇，而在西非导致疾病的病原体在这个簇中形成了一个新的亚分支。对16S-23S基因间隔区的分析证实了该区域的序列多样性，并能够设计出两个针对东非和西非发现的疾病的引物。当与通用引物配对时，这些特异引物都不能从健康的椰子DNA、来自加勒比海的受感染的椰子DNA或各种长春花(长春花属)维持的植质体的DNA中产生PCR扩增产物。这些特异的引物可以作为鉴定田间样品中特定椰子植原体的有效工具。
Using the polymerase chain reaction the 16S rRNA genes and the 16S-23S spacer regions of phytoplasmas associated with lethal decline diseases of coconut palm (Cocos nucifera), were amplified from infected plants from Florida and the Yucatan region in Mexico and from east and west Africa. Following sequencing of the rDNA products, phylogenetic analysis confirmed that these coconut phytoplasmas form a separate cluster within the phytoplasma clade and that the pathogen causing diseases in west Africa formed a new sub-clade within this cluster. Analysis of the 16S-23S intergenic spacer regions confirmed the sequence diversity of this region and enabled two primers to be designed which were specific for the diseases found in east and west Africa. None of these specific primers, when paired with a universal primer, produced PCR amplification products from healthy coconut DNA, infected coconut DNA from the Caribbean or DNA from a variety of periwinkle (Catharanthus roseus)-maintained phytoplasmas. These specific primers can serve as effective tools for identifying particular coconut phytoplasmas in field samples.