Pooler M R, Hartung J S
Fruit Laboratory, United States Department of Agriculture, Beltsville Agricultural Research Center, MD 20705, USA.
Curr Microbiol. 1995 Dec;31(6):377-81. doi: 10.1007/BF00294703.
By cloning and sequencing specific randomly amplified polymorphic DNA (RAPD) products, we have developed pairs of PCR primers that can be used to detect Xylella fastidiosa in general, and X. fastidiosa that cause citrus variegated chlorosis (CVC) specifically. We also identified a CVC-specific region of the X. fastidiosa genome that contains a 28-nucleotide insertion, and single base changes that distinguish CVC and grape X. fastidiosa strains. When using RAPD products to develop specific PCR primers, we found it most efficient to screen for size differences among RAPD products rather than presence/absence of a specific RAPD band.
通过对特定随机扩增多态性DNA（RAPD）产物进行克隆和测序，我们开发出了几对PCR引物，这些引物可用于一般情况下检测木质部难养菌，特别是检测引起柑橘杂色黄化病（CVC）的木质部难养菌。我们还鉴定出了木质部难养菌基因组中一个CVC特异性区域，该区域包含一个28个核苷酸的插入片段以及区分CVC和葡萄木质部难养菌菌株的单碱基变化。在使用RAPD产物开发特异性PCR引物时，我们发现筛选RAPD产物之间的大小差异而非特定RAPD条带的有无最为有效。
