Molecular Genotyping of Colletotrichum Species Based on Arbitrarily Primed PCR, A + T-Rich DNA, and Nuclear DNA Analyses

S. Freeman, M. Pham, R.J. Rodriguez
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引用次数: 120

Abstract

Freeman, S., Pham, M., and Rodriguez, R. J. 1993. Molecular genotyping of Colletotrichum species based on arbitrarily primed PCR, A + T-rich DNA, and nuclear DNA analyses. Experimental Mycology 17, 309-322. Isolates of Colletotrichum were grouped into 10 separate species based on arbitrarily primed PCR (ap-PCR), A + T-rich DNA (AT-DNA) and nuclear DNA banding patterns. In general, the grouping of Colletotrichum isolates by these molecular approaches corresponded to that done by classical taxonomic identification, however, some exceptions were observed. PCR amplification of genomic DNA using four different primers allowed for reliable differentiation between isolates of the 10 species. HaeIII digestion patterns of AT-DNA also distinguished between species of Colletotrichum by generating species-specific band patterns. In addition, hybridization of the repetitive DNA element (GcpR1) to genomic DNA identified a unique set of Pst 1-digested nuclear DNA fragments in each of the 10 species of Colletotrichum tested. Multiple isolates of C. acutatum, C. coccodes, C. fragariae, C. lindemuthianum, C. magna, C. orbiculare, C. graminicola from maize, and C. graminicola from sorghum showed 86-100% intraspecies similarity based on ap-PCR and AT-DNA analyses. Interspecies similarity determined by ap-PCR and AT-DNA analyses varied between 0 and 33%. Three distinct banding patterns were detected in isolates of C. gloeosporioides from strawberry. Similarly, three different banding patterns were observed among isolates of C. musae from diseased banana.

基于任意引物PCR、富A + t DNA和核DNA分析的炭疽菌分子基因分型
弗里曼,S.,范,M.和罗德里格斯,R. J. 1993。基于任意引物PCR、富A + t DNA和核DNA分析的炭疽菌分子基因分型。真菌学学报,17,309-322。根据任意引物PCR (ap-PCR)、富含A + t的DNA (AT-DNA)和核DNA带带模式将炭疽菌分离菌株分为10个独立的种。总的来说,这些分子方法对炭疽菌分离物的分类与经典的分类鉴定一致,但也有例外。使用四种不同引物对基因组DNA进行PCR扩增,可以可靠地区分10种菌株。HaeIII的AT-DNA消化模式也可以通过产生种特异性带图来区分炭黑菌的种间差异。此外,将重复DNA元件(GcpR1)与基因组DNA杂交,在10种炭疽菌中鉴定出一组独特的Pst - 1酶切核DNA片段。基于ap-PCR和AT-DNA分析,玉米和高粱中不同分离株的acutatum、C. cocodes、C. fragariae、C. lindemuthianum、C. magna、C. orbiculare、C. graminicola的种内相似性为86-100%。通过ap-PCR和AT-DNA分析确定的种间相似性在0 - 33%之间。从草莓中分离到三种不同的带状带。同样,在患病香蕉分离株中观察到三种不同的条带模式。
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