利用R.A.P.D.技术对三叶烟叶根瘤中抗重金属根瘤菌的遗传特征及多样性分析

Q4 Environmental Science Environment and Ecology Research Pub Date : 2023-09-01 DOI:10.60151/envec/nvud9012
Pallaval Veera Bramhachari, G. Kranthi Kumar, D. Shilpa, Ekamber Kariali
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引用次数: 0

摘要

本研究对从印度安得拉邦不同地区采集的三叶甘油三酯(Vigna trilobata)品种根瘤中分离的21株根瘤菌进行了研究。采用选择性培养基酵母提取物甘露醇琼脂培养基(Y.E.M.A.)分离根杆菌。根据菌落形态和生化特征对21株菌株进行了鉴定,并通过16S rDNA测序分析达到了种水平。不同种类的细菌存在于三叶藤蔓的根瘤中。短随机序列引物扩增基因组dna的过程称为随机扩增多态性dna (R.A.P.D.),用于评估分离菌株的遗传变异。采用随机引物OPA-4、OPA-9、OPA-11、OPA-12和OPA-15对分离株基因组dna进行rap - pd分析,获得条带模式。分别比较了21株根细菌的rappd谱,通过多态dna片段的出现来确定它们之间的差异。对21株分离细菌的dna进行PCR扩增,得到70个扩增产物,其中68个为多态产物,2个为单态产物。调查发现,R.A.P.D.可以非常精确地区分这两种菌株。利用分子标记研究根瘤菌的遗传多样性有助于根瘤菌新品种的鉴定和培育。
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Genetic characterization and diversity analysis of heavy metal-resistant rhizobacterial strains isolated from the root nodules of Vigna trilobata determined by the R.A.P.D. technique
The present study characterizes twenty-one rhizobacterial strains isolated from root nodules of Vigna trilobata cultivars collected from various Andhra Pradesh, India districts. Rhizobacteria was isolated using selective medium Yeast Extract Mannitol Agar medium (Y.E.M.A.). All 21 strains were characterized based on colony morphology and biochemical characterization and identified up to species level by 16S rDNA sequencing analysis. Different genera of the bacteria are present in the root nodules of Vigna trilobata. Short random sequence primers amplify genomic D.N.A. in a process called random amplified polymorphic D.N.A. (R.A.P.D.), which were employed to assess genetic variation across isolated bacterial strains. Banding patterns were obtained via R.A.P.D. analysis of genomic D.N.A. from the isolates using five random primers, including OPA-4, OPA-9, OPA-11, OPA-12, and OPA-15. The R.A.P.D. profiles of twenty-one rhizobacterial isolates were compared separately to determine their differences by the occurrence of polymorphic D.N.A. fragments. PCR amplified of the D.N.A. isolated from twenty-one bacterial isolates yielded seventy amplified products, of which sixty-eight were polymorphic, and two were monomorphic. The investigation found that R.A.P.D. could differentiate between the strains very precisely. Characterizing and creating novel Rhizobium species can be aided by using molecular markers to investigate genetic diversity.
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Environment and Ecology Research
Environment and Ecology Research Environmental Science-Pollution
CiteScore
1.30
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59
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