Kaplan D T, Vanderspool M C, Garrett C, Chang S, Opperman C H
USDA-ARS, U.S. Horticultural Research Laboratory, Orlando, FL 32803, USA.
Mol Plant Microbe Interact. 1996 Jan;9(1):32-8. doi: 10.1094/mpmi-9-0032.
Six polymorphic bands of DNA were amplified from purified Radopholus citrophilus genomic DNA from one strain of each of the sibling species R. citrophilus and R. similis in random amplified polymorphic DNA analyses involving 380 single 10-base primers. Four of these polymorphic DNA fragments were successfully cloned and amplified through subsequent use of primers designed to complement the terminal sequences of the polymorphic DNA. Results of ensuing studies using mini-prepped DNA from 14 burrowing nematode strains collected from Florida, Hawaii, and Central America, characterized for their ability to parasitize citrus, indicated that a 2.4-kb fragment appeared to be associated with citrus parasitism in burrowing nematode populations from Florida. However, a fragment of comparable size was also detected in R. citrophilus from Hawaii and from burrowing nematode populations collected from Belize and Puerto Rico. Overall, findings suggest that the genome organization of the burrowing nematode sibling species R. citrophilus and R. similis is highly conserved. This remarkable genetic similarity should facilitate identification of genetic sequence related to important phenotypes such as citrus parasitism. Detection of R. citrophilus-specific DNA fragments in burrowing nematodes collected from Belize and Puerto Rico suggests that R. citrophilus is resident in some Central American countries.
在涉及380种单链10碱基引物的随机扩增多态性DNA分析中，从嗜柠檬酸穿孔线虫（Radopholus citrophilus）和相似穿孔线虫（R. similis）各一个菌株的纯化基因组DNA中扩增出6条DNA多态性条带。其中4条多态性DNA片段通过随后使用设计用于互补多态性DNA末端序列的引物成功克隆并扩增。对从佛罗里达州、夏威夷州和中美洲收集的14个穿孔线虫菌株的小量制备DNA进行后续研究，这些菌株以其寄生柑橘的能力为特征，结果表明，一个2.4kb的片段似乎与佛罗里达州穿孔线虫种群中的柑橘寄生现象有关。然而，在来自夏威夷的嗜柠檬酸穿孔线虫以及从伯利兹和波多黎各收集的穿孔线虫种群中也检测到了大小相当的片段。总体而言，研究结果表明，穿孔线虫近缘种嗜柠檬酸穿孔线虫和相似穿孔线虫的基因组组织高度保守。这种显著的遗传相似性应有助于鉴定与重要表型（如柑橘寄生）相关的基因序列。在从伯利兹和波多黎各收集的穿孔线虫中检测到嗜柠檬酸穿孔线虫特异性DNA片段，表明嗜柠檬酸穿孔线虫存在于一些中美洲国家。
