Datta J, Lal N
Department of Life Sciences, C.S.J.M. University, Kanpur 208 024, India.
Cell Mol Biol (Noisy-le-grand). 2012 Dec 22;58(1):55-65.
(foc) and Fusarium udum (Fud) collected from major pulse growing regions of India. Out of 247 bands produced by 24 Randomly Amplified Polymorphic DNA (RAPD) primers in Foc isolates, 210 (85%) were polymorphic. A maximum of 14 amplicons were generated by primer OPF 05 whereas minimum 7 amplicons were generated by primer K7. A total of 24 alleles were produced by twelve Simple Sequence Repeats (SSR) primers with an average of two alleles per marker in foc isolates. The maximum number of 4 alleles was obtained with primer SSR 12. SSR amplicon size ranged from 100 to 400 bp. The Unweighted Pair Group Method with Arithmetic average (UPGMA) cluster analysis based on RAPD and SSR profiles grouped the fourteen foc isolates into four major clusters. The universal Inter Transcribed Spacer (ITS) primer pair amplified 630 bp bands in all fourteen foc isolates while significant length polymorphism was obtained only when analysed by restriction digestion with EcoRI and MspI enzymes. The cluster analysis of ITS—RFLP grouped all 14 Foc isolates into three major clusters. Twenty four RAPD primers generated a total of 226 bands (ranging 0.3 to 3.0 kb) in Fusarium udum with an average of 9.4 bands per primer and a total of 27 alleles were produced by twelve SSR primers with an average of 2.25 alleles per marker. All isolates amplified a single band ranging from 100 to 450 bp. The universal ITS primer pair amplified 650 bp bands in all fourteen fud isolates while significant length polymorphism was obtained only when analysed by restriction digestion with EcoRI and Hind III enzymes. The cluster analysis of ITS—RFLP grouped all 14 Fud isolates into three major clusters. The cluster analysis using various markers show the grouping of Fusarium isolates strictly according to their cultural characteristics and degree of pathogenicity and not the geographical origin. This information will be helpful for pathologists and plant breeders to design effective resistance breeding programs in chickpea and pigeonpea taking into account the diversity in wilt pathogen.
从印度主要豆类种植区采集的尖镰孢菌（Foc）和木贼镰孢菌（Fud）。在Foc分离株中，24个随机扩增多态性DNA（RAPD）引物产生了247条带，其中210条（85%）具有多态性。引物OPF 05产生的扩增子最多，为14个，而引物K7产生的扩增子最少，为7个。12个简单序列重复（SSR）引物在Foc分离株中共产生24个等位基因，每个标记平均有两个等位基因。引物SSR 12获得的等位基因数量最多，为4个。SSR扩增子大小在100至400 bp之间。基于RAPD和SSR图谱的算术平均非加权配对组法（UPGMA）聚类分析将14个Foc分离株分为4个主要类群。通用的转录间隔区（ITS）引物对在所有14个Foc分离株中扩增出630 bp的条带，而只有在用EcoRI和MspI酶进行限制性消化分析时才获得显著的长度多态性。ITS-RFLP聚类分析将所有14个Foc分离株分为3个主要类群。24个RAPD引物在木贼镰孢菌中总共产生了226条带（范围为0.3至3.0 kb），每个引物平均产生9.4条带，12个SSR引物共产生27个等位基因，每个标记平均有2.25个等位基因。所有分离株均扩增出一条100至450 bp的单带。通用的ITS引物对在所有14个Fud分离株中扩增出650 bp的条带，而只有在用EcoRI和Hind III酶进行限制性消化分析时才获得显著的长度多态性。ITS-RFLP聚类分析将所有14个Fud分离株分为3个主要类群。使用各种标记的聚类分析表明，镰孢菌分离株严格按照其培养特征和致病程度进行分组，而不是根据地理来源。这些信息将有助于病理学家和植物育种者在考虑枯萎病病原菌多样性的情况下，设计有效的鹰嘴豆和木豆抗性育种计划。
