Balali G R, Neate S M, Kasalkheh A M, Stodart B J, Melanson D L, Scott E S
Department of Biology, The University of Isfahan, Isfahan, Iran.
Mycopathologia. 2007 Feb;163(2):105-15. doi: 10.1007/s11046-006-0089-7. Epub 2007 Jan 23.
Pectic zymogram, RFLP and PCR analyses were used to characterize Rhizoctonia solani AG 3 isolates collected from diseased potatoes in South Australia. The pectic zymogram data were compared with those obtained for isolates collected from central Iran. Analyses of bands corresponding to pectin esterase and polygalacturonase revealed three zymogram subgroups (ZG) in AG 3. In addition to the previously reported ZG7 (here renamed ZG7-1), two new zymogram subgroups, ZG7-2 and ZG7-3, were identified. Of the 446 isolates tested, 50% of the South Australian and 46% of the Iranian isolates were ZG7-1. The majority of the isolates originating from stem and root cankers were ZG7-1, whereas most of the isolates designated ZG7-2 and ZG7-3 originated from tuber-borne sclerotia. Pathogenicity tests revealed that ZG7-1 generally produced fewer sclerotia and more severe cankers of underground parts of the potato plants than the other two ZGs. Two random DNA clones, one originating from an AG 3 isolate and the other from an AG 4 isolate, were used as probes for RFLP analyses of Australian isolates. The AG 3 probe, previously identified to be specific to this group, detected a high level of genetic diversity, with 11 genotypes identified amongst 50 isolates analysed. The low-copy AG 4 probe resolved three genotypes amongst 24 isolates. For 23 isolates analysed with both markers, the combined data distinguished a total of six genotypes and similarity analysis resolved the isolates into two main groups with 50% homology. PCR, using primers for the plant intron splice junction region (R1), also revealed variation. No obvious relationship among pectic zymogram groups, RFLP and PCR genotypes was observed.
采用果胶酶谱分析、限制性片段长度多态性(RFLP)分析和聚合酶链反应(PCR)分析,对从南澳大利亚患病马铃薯中分离得到的立枯丝核菌AG 3菌株进行特征鉴定。将果胶酶谱数据与从伊朗中部分离得到的菌株的数据进行比较。对与果胶酯酶和多聚半乳糖醛酸酶相对应的条带分析显示,AG 3中有三个酶谱亚组(ZG)。除了先前报道的ZG7(此处重新命名为ZG7-1),还鉴定出两个新的酶谱亚组ZG7-2和ZG7-3。在测试的446个菌株中,50%的南澳大利亚菌株和46%的伊朗菌株为ZG7-1。大多数源自茎和根溃疡的菌株为ZG7-1,而大多数被指定为ZG7-2和ZG7-3的菌株源自块茎上的菌核。致病性测试表明,与其他两个ZG相比,ZG7-1通常产生的菌核较少,马铃薯植株地下部分的溃疡更严重。两个随机DNA克隆,一个源自AG 3菌株,另一个源自AG 4菌株,用作澳大利亚菌株RFLP分析的探针。先前鉴定为该组特异性的AG 3探针检测到高度的遗传多样性,在分析的50个菌株中鉴定出11种基因型。低拷贝的AG 4探针在24个菌株中解析出三种基因型。对于用两种标记分析的23个菌株,综合数据共区分出六种基因型,相似性分析将这些菌株分为两个主要组,同源性为50%。使用针对植物内含子剪接连接区(R1)的引物进行的PCR也显示出变异。未观察到果胶酶谱组、RFLP和PCR基因型之间存在明显关系。