Sheikholeslami M, Seidel M, Okhovvat S M, Hedjaroude G, Javan-Nikkhah M
Department of Plant Protection, College of Agriculture, University of Tehran, Karaj, Iran.
Commun Agric Appl Biol Sci. 2005;70(3):327-32.
107 samples of E. betae were collected on infected leaves from all over Iranian beet cultivation areas. Their choosing were based on geographical and host origin(sugar beet, red beet, fodder beet and wild beet). 30 isolates were single colonized and grown on sugar beet susceptible genotype 7233. 107 specimens were analyzed by restriction fragment length polymorphism (RFLP) of the ribosomal internal transcribed spacers (ITS) and 5.8s DNA which previously amplified by the polymerase chain reaction (PCR) with 2 universal primers, ITS1 and ITS4. PCR product was affected by 9 different restriction enzymes. PCR product was a 645 bp band for all of the isolates. 3 restriction enzymes; CfoI, MspI and HaeIII could cut this fragment into smaller bands, but electrophoretic patterns were identical for all of the isolates. 30 single colonized isolates were used in RAPD experiments. In RAPD-PCR experiment genetic diversity was investigated with 30 isolates from different parts of the country. 59 random primers were used and then 21 primers that displayed good consistency and reproducibility were selected. Most of the primers revealed identical patterns between 3 to 14 bands. 5 primers that showed more polymorphism were selected to analyze 30 isolates. For these 5 primers 61 distinct bands were obtained which 62% of these bands were polymorphic. Results indicated that there is no relationship between cluster grouping and geographical origin and the isolates showed a high similarity.
从伊朗各地的甜菜种植区感染叶片上采集了107份甜菜尾孢菌样本。样本的选取基于地理和寄主来源(甜菜、红甜菜、饲料甜菜和野生甜菜)。30个分离株在甜菜易感基因型7233上单独定殖并生长。通过核糖体内部转录间隔区(ITS)和5.8s DNA的限制性片段长度多态性(RFLP)对107个样本进行分析,此前用两种通用引物ITS1和ITS4通过聚合酶链反应(PCR)扩增了这些片段。PCR产物受到9种不同限制性内切酶的作用。所有分离株的PCR产物均为645 bp条带。3种限制性内切酶(CfoI、MspI和HaeIII)可将该片段切割成较小的条带,但所有分离株的电泳图谱相同。30个单独定殖的分离株用于随机扩增多态性DNA(RAPD)实验。在RAPD-PCR实验中,用来自该国不同地区的30个分离株研究遗传多样性。使用了59个随机引物,然后选择了21个显示出良好一致性和可重复性的引物。大多数引物在3至14条带之间显示出相同的图谱。选择了5个显示出更多多态性的引物来分析30个分离株。对于这5个引物,获得了61条不同的条带,其中62%的条带具有多态性。结果表明,聚类分组与地理来源之间没有关系,分离株显示出高度相似性。