ABSTRACT
Lentil is one of the most important legumes cultivated in various provinces of Iran. However, there is limited information about the symbiotic rhizobia of lentils in this country. In this study, molecular identification of lentil-nodulating rhizobia was performed based on 16S-23S rRNA intergenic spacer (IGS) and recA, atpD, glnII, and nodC gene sequencing. Using PCR-RFLP analysis of 16S-23S rRNA IGS, a total of 116 rhizobia isolates were classified into 20 groups, leaving seven strains unclustered. Phylogenetic analysis of representative isolates revealed that the rhizobia strains belonged to Rhizobium leguminosarum and Rhizobium laguerreae, and the distribution of the species is partially related to geographical location. Rhizobium leguminosarum was the dominant species in North Khorasan and Zanjan, while R. laguerreae prevailed in Ardabil and East Azerbaijan. The distribution of the species was also influenced by agroecological climates; R. leguminosarum thrived in cold semiarid climates, whereas R. laguerreae adapted to humid continental climates. Both species exhibited equal dominance in the Mediterranean climate, characterized by warm, dry summers and mild, wet winters, in Lorestan and Kohgiluyeh-Boyer Ahmad provinces.
Subject(s)
DNA, Bacterial , Lens Plant , Phylogeny , Rhizobium , Lens Plant/microbiology , Iran , Rhizobium/genetics , Rhizobium/classification , Rhizobium/isolation & purification , DNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Climate , DNA, Ribosomal Spacer/genetics , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA , RNA, Ribosomal, 23S/genetics , Rhizobium leguminosarum/genetics , Rhizobium leguminosarum/classification , Rhizobium leguminosarum/isolation & purification , Symbiosis , Bacterial Proteins/genetics , Polymerase Chain ReactionABSTRACT
Abstract The effect of different fungicides on mycorrhizal fungi should be investigated in different plants and environmental conditions. Thus, the purpose of this study was to appraise the effect of simultaneous fungicides application (including benomyl, rovral TS, mancozeb, and tilt) on the efficiency of Rhizophagus irregularis in cultivations of maize and wheat. This study was conducted in two separate experiments in the laboratory and greenhouse. The results of the laboratory stage showed that the use of all four fungicides significantly reduced the spore number compared to the conditions of non-use of the fungicide, although only rovral TS and mancozeb led to a significant reduction in root colonization percentage of R. irregularis. In the greenhouse, the benomyl significantly increased root dry weight in maize although tilt significantly reduced root colonization of maize with R. irregularis. The tilt and rovral TS had a positive effect and benomyl had a negative effect on wheat growth traits, but the root colonization of wheat with R. irregularis was not affected by fungicides. Generally, benomyl (2 g L-1) in maize and tilt (2 mL L-1) in wheat and rovral TS in both plants could be recommended with the combined application of R. irregularis inoculants. Therefore, depending on the type of fungicide and the host plant, the effect of the fungicide on colonization and association of mycorrhiza varies.
ABSTRACT
Azotobacter chroococcum and A. salinestris do not possess significant and distinct morphological and physiological differences and are often mistaken with each other in microbiological research. In this study, 12 isolates of Azotobacter isolated by standard protocol from soils were identified morphologically and physiologically as A. chroococcum. The isolates were more closely investigated for the molecular differentiation and diversity of A. chroococcum and A. salinestris. For this purpose, the ARDRA technique including HpaII, RsaI, and AluI restriction enzymes, and REP, ERIC, and BOX markers were used. The nifD and nifH genes were also utilized to evaluate the molecular identification of these two species. The 16S rDNA evaluation showed that only four out of the 12 isolates were identified as A. chroococcum and the rest were A. salinestris. The results revealed that HpaII was able to differentiate A. chroococcum from A. salinestris whereas RsaI and AluI were not able to separate them. Moreover, BOX and REP markers were able to differentiate between A. chroococcum and A. salinestris. However, ERIC marker and nifD and nifH genes were unable to separate these species. According to the results, HpaII restriction enzyme is suggested to save time and cost. BOX and REP markers are recommended for differentiation and clear discrimination not only between A. chroococcum and A. salinestris but also among their strains.
