Rhizobial Isolates Research Articles

Survival of white clover rhizobium isolate S11N9 in different delivery systems: assessment of potential for commercialisation

Survival of white clover rhizobium isolate S11N9 in different delivery systems: assessment of potential for commercialisation

Potential of novel Rhizobium strains as biofertilizer for improved groundnut (Arachis hypogaea L.) cultivation in Eastern India

ABSTRACT Comprehensive assessments were conducted on eight fast growing, novel Rhizobium isolates, short listed from a preliminary selection of 68 isolates obtained from root nodules of groundnut plants grown in diverse agro-climatic regions of eastern India. Additionally, two conventional rhizobia strains, viz. IGR6 and GN2, (collected from the Nodule Research Center, BCKV), were utilized for comparative analysis. Cluster analysis, based on carbon utilization and biochemical parameters, grouped all the test isolates with an average similarity level of 73%. The ability of the isolates to utilize a wide range of carbon sources, including hexoses, pentoses, and disaccharides, confirms their fast-growing nature. Further analysis through 16S rDNA extraction and nucleotide sequencing revealed that the promising fresh isolates exhibited 95–99% similarity to known Rhizobium species in public databases. When evaluated in pot culture experiments using new alluvial soil from the Gangetic Plains in eastern India, the native fresh isolate NRA1(PP355674) outperformed other isolates and conventional strains in terms of nodulation efficiency, plant dry matter production, and plant nitrogen uptake. This study highlights the significant potential of native Rhizobium isolates to surpass conventionally used strains in biological nitrogen fixation, supporting the development of sustainable, environmentally-friendly, and cost-effective agricultural practices.

Potential Nitrogen Fixing Rhizobia Isolated from Some Wild Legumes of Nagaland Based on RAPD with Nif-directed Primer and Their Biochemical Activities

Wild legumes are widely dispersed and can survive in challenging environments as bacteria dwell in their nodules and help each other. Although Nagaland is home to many wild legume varieties, research on the microbial diversity that goes along with them is still in its infancy. This work aimed to characterize several wild legume root nodules and distinguish possible rhizobial isolates using RAPD and nif-directed RPO1 primer. Nodule bacteria were isolated in Yeast extract culture media. Based on their colony morphology, 150 isolates were selected for performing RAPD with nif-directed RPO1 primer. Eighty-four isolates were bonded with RPO1 primer, and a few biochemical tests were conducted on RPO1-positive isolates. Activities that promoted plant development were also investigated for these isolates. Of all the isolates, 18 exhibited phosphate solubilization capacity, while 38 isolates were found to produce IAA. This study entails a large variety of rhizobia in the nodules, which were able to promote growth. Hence, these isolates promise to be bio-fertilizers that could improve agricultural operations.

Study of Plant Growth Promoting Abilities of Rhizobium Strains Isolated from Mungbean Root Nodules

Background: Plant Growth Promoting Rhizobacteria (PGPR) are a type of bacteria that use biofertilizers and other plant growth-promoting agents to increase plant output and growth. In the last several decades, there has been a global surge in the usage of beneficial soil microorganisms like PGPR for safe and sustainable agriculture due to the detrimental effects of artificial fertilizers on the environment and their rising prices. Plant-beneficial rhizobacteria have the potential to reduce the world's reliance on dangerous agricultural chemicals that upset agro-ecosystems. Today a large part of yield is lost due to the various stress mechanisms of plants. It could be classified into biotic and abiotic stress. These are generally a group of microorganisms that is found either in the rhizosphere or in the root nodules of plants. PGPR has beneficial impact, they colonize in the plant roots and improve crop yields and agricultural productivity and also produce some useful growth promoting growth hormones viz., IAA, Auxins, Cytokinin and Gibberellins. Methods: The bacterial isolates were screened for plant growth promoting traits such as phosphate solubilization, IAA production and siderophore production was tested qualitatively by using pikovskayas media, TSB (Tryptone Soy Broth) agar and Chrome Azurol Sulphate (CAS) blue agar respectively. Results: In present study out of the eight rhizobial isolates, five exhibited the phosphate solubilization zones, three shows negative result. However out of eight isolates five isolates produced IAA and three showed negative result. Among all isolates seven isolates were able to produce siderophore, while one showed negative result.

Unearthing Optimal Symbiotic Rhizobia Partners from the Main Production Area of Phaseolus vulgaris in Yunnan.

Phaseolus vulgaris is a globally important legume cash crop, which can carry out symbiotic nitrogen fixation with rhizobia. The presence of suitable rhizobia in cultivating soils is crucial for legume cropping, especially in areas beyond the plant-host native range, where soils may lack efficient symbiotic partners. We analyzed the distribution patterns and traits of native rhizobia associated with P. vulgaris in soils of Yunnan, where the common bean experienced a recent expansion. A total of 608 rhizobial isolates were tracked from soils of fifteen sampling sites using two local varieties of P. vulgaris. The isolates were discriminated into 43 genotypes as defined by IGS PCR-RFLP. Multiple locus sequence analysis based on recA, atpD and rpoB of representative strains placed them into 11 rhizobial species of Rhizobium involving Rhizobium sophorae, Rhizobium acidisoli, Rhizobium ecuadorense, Rhizobium hidalgonense, Rhizobium vallis, Rhizobium sophoriradicis, Rhizobium croatiense, Rhizobium anhuiense, Rhizobium phaseoli, Rhizobium chutanense and Rhizobium etli, and five unknown Rhizobium species; Rhizobium genosp. I~V. R. phaseoli and R. anhuiense were the dominant species (28.0% and 28.8%) most widely distributed, followed by R. croatiense (14.8%). The other rhizobial species were less numerous or site-specific. Phylogenies of nodC and nifH markers, were divided into two specific symbiovars, sv. phaseoli regardless of the species affiliation and sv. viciae associated with R. vallis. Through symbiotic effect assessment, all the tested strains nodulated both P. vulgaris varieties, often resulting with a significant greenness index (91-98%). However, about half of them exhibited better plant biomass performance, at least on one common bean variety, and two isolates (CYAH-6 and BLYH-15) showed a better symbiotic efficiency score. Representative strains revealed diverse abiotic stress tolerance to NaCl, acidity, alkalinity, temperature, drought and glyphosate. One strain efficient on both varieties and exhibiting stress abiotic tolerance (BLYH-15) belonged to R. genosp. IV sv. phaseoli, a species first found as a legume symbiont.

Selection of effective Rhizobium isolates of chickpea (Cicer arietinum L.) from Bilaspur and Mungeli districts of Chhattisgarh Plain region

Selection of effective Rhizobium isolates of chickpea (Cicer arietinum L.) from Bilaspur and Mungeli districts of Chhattisgarh Plain region

Genetic Characterization of Rhizobium spp. Strains in an Organic Field Pea (Pisum sativum L.) Field in Lithuania.

Biological nitrogen fixation in legume plants depends on the diversity of rhizobia present in the soil. Rhizobial strains exhibit specificity towards host plants and vary in their capacity to fix nitrogen. The increasing interest in rhizobia diversity has prompted studies of their phylogenetic relations. Molecular identification of Rhizobium is quite complex, requiring multiple gene markers to be analysed to distinguish strains at the species level or to predict their host plant. In this research, 50 rhizobia isolates were obtained from the root nodules of five different Pisum sativum L. genotypes ("Bagoo", "Respect", "Astronaute", "Lina DS", and "Egle DS"). All genotypes were growing in the same field, where ecological farming practices were applied, and no commercial rhizobia inoculants were used. The influence of rhizobial isolates on pea root nodulation and dry biomass accumulation was determined. 16S rRNA gene, two housekeeping genes recA and atpD, and symbiotic gene nodC were analysed to characterize rhizobia population. The phylogenetic analysis of 16S rRNA gene sequences showed that 46 isolates were linked to Rhizobium leguminosarum; species complex 1 isolate was identified as Rhizobium nepotum, and the remaining 3 isolates belonged to Rahnella spp., Paenarthrobacter spp., and Peribacillus spp. genera. RecA and atpD gene analysis showed that the 46 isolates identified as R. leguminosarum clustered into three genospecies groups (B), (E) and (K). Isolates that had the highest influence on plant dry biomass accumulation clustered into the (B) group. NodC gene phylogenetic analysis clustered 46 R. leguminosarum isolates into 10 groups, and all isolates were assigned to the R. leguminosarum sv. viciae.

Molecular identification and characterizations of rhizobacterial isolates collected from lentil rhizosphere of Indo-gangetic plains.

Plant growth promoting rhizobacteria (PGPR) are also known to colonize in the soil rhizosphere and prevent the development of other soil borne pathogens residing in the root surface. These microorganisms play a vital role in growth and development of the plant and also enhances the soil fertility by enriching the soil with different beneficial nutrients. This study was aimed at isolation of different rhizobacteria and their molecular characterization in search of efficient bacterial strains with multiple growth regulating activities. A total 36 bacteria were isolated from lentil root nodule as well as soil from different lentil growing fields with a view to screen/evaluate their plant growth promoting potential. Morphological characterization of isolated rhizobacterial candidates were done by observing the colonies on YEMA and nutrient agar media. Determination of CFU, Congo red test and gram staining tests were done to further screen them according to their morphology. All the isolates were then undergone molecular phylogenetic analysis using the partial sequences of the 16S rDNA. Based upon the Gram staining test, all the isolates were negative in gram reaction except six Bacillus isolates, PSB2 and AB3. Results of Ribosomal Database Project (RDP) and Basic Local Alignment Search Tool for Nucleotide Sequences (BLASTn) from 16S rDNA gene sequences showed that these isolates are genetically diverse. A total of 15 isolates of Rhizobium, 6 isolates of Bacillus, 3 isolates of Pseudomonas, 2 isolates of Phosphate Solubilizing Bacteria, 4 isolates of actinomycetes were identified by molecular sequencing of their 16S rDNA region and comparing them with the other isolates enlisted in the database of NCBI for the similarity percentage, query coverage. The purpose of the present study was to select native rhizosphere bacteria from the lentil nodule and soil of Lentil field and to evaluate their plant growth promoting potential as an alternative of chemical fertilizer for sustainable, environment friendly agriculture and assessment of their phylogenetic characterization.

Characterization and Evaluation of Native Rhizobium of Groundnut (Arachis hypogaea L.) and Soybean (Glycine max L.)

The present investigation was conducted to characterize and evaluate native Rhizobium isolates of groundnut and soybean to enhance the Legume-Rhizobium symbiosis as well as increase sustainable crop production. In this connection, all the native isolates under study were characterized biochemically and tested with N-free sand culture in the glasshouse to select effective native Rhizobium isolates through proper screening. 10 isolates of each crop were collected from the culture collection bank of the Department of Agricultural Microbiology, Indira Gandhi Krishi Vishwavidyalaya Raipur (Chhattisgarh). Out of 10 isolates, 3 isolates of each crop were selected on the basis of their growth performance and the purity of the culture. The biochemical studies of selected isolates include starch hydrolysis, catalase, urease, citrate utilization, indole production, gelatine iron, and methyl red tests, which were further evaluated for nodulation and biological nitrogen fixation in groundnut and soybean under controlled conditions with four treatments for each crop and five replications. The experiment result revealed that the evaluation of native Rhizobium isolate no. 1185 of groundnut and isolate no. 1076 of soybean was found to be superior among all the rhizobial isolates taken for study. These isolates accumulated 1.24, 1.44 mg/plant extra amount of atmospheric nitrogen over un-inoculated control plants, respectively. Further, it can be concluded that these native isolates may be the most effective nitrogen fixers for groundnut and soybean over the control.

Phenotypic and genotypic diversity of root nodule bacteria from wild Lathyrus and Vicia species in Gaziantep, Turkey.

This study identified the phenotypic and genotypic characteristics of the bacteria that nodulate wild Lathyrus and Vicia species natural distribution in the Gaziantep province of Turkey. Principle component analysis of phenotypic features revealed that rhizobial isolates were highly resistant to stress factors such as high salt, pH and temperature. They were found to be highly sensitive to the concentrations (mg/mL) of the antibiotics neomycin 10, kanamycin, and tetracycline 5, as well as the heavy metals Ni 10, and Cu 10, and 5. As a result of REP-PCR analysis, it was determined that the rhizobial isolates were quite diverse, and 5 main groups and many subgroups being found. All of the isolates nodulating wild Vicia species were found to be related to Rhizobium sp., and these isolates were found to be in Clades II, III, IV, and V of the phylogenetic tree based on 16S rRNA. The isolates that nodulated wild Lathyrus species were in Clades I, II, IV, V, VI, VII, and VIII, and they were closely related to Rhizobium leguminasorum, Rhizobium sp., Phyllobacterium sp., Serratia sp., and Pseudomonas sp. According to the genetic analyses, the isolates could not be classified at the species level, the similarity ratio was low, they formed a distinct group that was supported by strong bootstrap values in the phylogenetic tree, and the differences discovered in the network analysis revealed the diversity among the isolates and gave important findings that these isolates may be new species.

SeqCode facilitates naming of South African rhizobia left in limbo

South Africa is well-known for the diversity of its legumes and their nitrogen-fixing bacterial symbionts. However, in contrast to their plant partners, remarkably few of these microbes (collectively referred to as rhizobia) from South Africa have been characterised and formally described. This is because the rules of the International Code of Nomenclature of Prokaryotes (ICNP) are at odds with South Africa's National Environmental Management: Biodiversity Act and its associated regulations. The ICNP requires that a culture of the proposed type strain for a novel bacterial species be deposited in two international culture collections and be made available upon request without restrictions, which is not possible under South Africa’s current national regulations. Here, we describe seven new Mesorhizobium species obtained from root nodules of Vachellia karroo, an iconic tree legume distributed across various biomes in southern Africa. For this purpose, 18 rhizobial isolates were delineated into putative species using genealogical concordance, after which their plausibility was explored with phenotypic characters and average genome relatedness. For naming these new species, we employed the rules of the recently published Code of Nomenclature of Prokaryotes described from Sequence Data (SeqCode), which utilizes genome sequences as nomenclatural types. The work presented in this study thus provides an illustrative example of how the SeqCode allows for a standardised approach for naming cultivated organisms for which the deposition of a type strain in international culture collections is currently problematic.

Selection of multi-purpose native rhizobia from a new red clover cultivar developed in Brazil

Red clover is an important forage legume in the livestock sector in southern Brazil, but regional climatic conditions compromise its management. Therefore, UFRGS and EMBRAPA have developed the cultivar URS BRS Mesclador adapted to the southern region of Brazil, but efficient rhizobia for it is still missing. This study aimed to select symbiotic rhizobia for the URS BRS Mesclador and evaluate their efficiency in biological nitrogen fixation in red clover and growth promotion in the rye. Seven rhizobia isolates from red clover nodules of the new cultivar and nine strains from the SEMIA Rhizobia Culture Collection were evaluated for their ability to promote plant growth. Certain strains exhibited notable abilities among the studied rhizobia, such as Rhizobium changzhiense Tp46 producing indole compounds (5.33 μg ml−1), Semia 2076 with a phosphate solubilization index of 2.4, and R. leguminosarum Tp57 and Tp34, SEMIA 2038, and SEMIA 276 with high siderophore production index. Isolates Tp46 and Tp34 demonstrated potential by displaying high biological nitrogen fixation efficiency in red clover and promoting rye growth. These promising results warrant further investigation for field studies with a long-term goal of developing inoculants for red clover cultivation in Brazil.

Rhizobium acaciae and R. anhuiense are the dominant rhizobial symbionts of Pisum sativum L. from Yunnan-Guizhou Plateau.

The aim of this study is to investigate the diversity and geographic distribution of pea-nodulating rhizobia in the subtropical region of Yunnan Province from Yunnan-Guizhou Plateau. A total of 615 rhizobial isolates were obtained from root nodules of the trapping plants and characterized genetically and symbiotically. The isolates discriminated into 43 genotypes by PCR-RFLP of IGS DNA. Multiple locus sequence analysis based on 16S rRNA, recA, atpD, dnaK, and rpoB genes placed them into eight clusters corresponding to species R. acaciae, R. anhuiense, R. binae, R. bangladeshense, R. hidalgonense, and three suspected novel populations of Rhizobium genosp. I-III. R. acaciae was the dominant group (52.5%) followed by R. anhuiense (30.7%). The other species were minor groups. Based on nodC phylogeny, all of them were the symbiovar viciae. All the tested strains showed efficient symbiotic N2 fixation on pea plants, in which WLB27, WCB18, and WNY29 presented the best PGP effects. Some of the tested strains had better IAA production, with WCB18 as the best producer (64.556 mg/L). Their distribution was mainly affected by soil available phosphorus, available potassium, and effective nitrogen. According to the results of symbiotic effect and resistance tests, strains of WLB27, WCB18, and WNY29 were selected as candidates for creating inoculants. This suggests that the pea-nodulating rhizobia in Yunnan Province form a unique community. The results gave some novel information about the diversity, diversification, and biogeography of pea-nodulating rhizobia.

Bio-prospecting the Efficacy of Carrier Based Inoculant of Rhizobium Strains as Plant Growth Promoter for the Development of Albizia procera Seedling

The carrier-based formulation of rhizobial cultures in general, excels to register their effects on various plant growth parameters of Albizia procera seedlings. Carrier based bio-inoculant prepared with rhizobial isolate FA6 of Uttrakhand performed better for all plant growth characteristics. Charcoal based application of rhizobial isolates also increased the N, P, K content of soil. In case of seedlings treated with charcoal-based formulation of rhizobial isolate from Himachal Pradesh maximum available N (323.30 kg/ha), P (38.97 kg/ha) and K (395.9 kg/ha) content was recorded in T2 (Charcoal based formulation of BA2 strain+20kg/ha N) which was significantly superior to T3 and uninoculated control. Among the charcoal-based formulation of rhizobial isolate from Uttrakhand seedlings under treatment T2 (Charcoal based formulation of FA6 strain+20kg/ha N) registered maximum available N (327.7kg/ha), P (39.01 kg/ha) and K (397.4 kg/ha) content in soil. Charcoal based application of rhizobial isolates significantly influenced the activity of soil enzymes (Dehydrogenase and Phosphatase). Application of Carrier based formulation of rhizobial isolate BA2 from Himachal Pradesh showed maximum dehydrogenase (11.20 μg TPF g-1h-1) and phosphatase (536.2 μmolL-1g-1h-1) activity as compare to uninoculated control. Whereas, among the rhizobial isolates from Uttrakhand maximum dehydrogenase (11.39 μg TPFg-1h-1) and phosphatase (545.4 μmol L-1g-1h-1) activity was also recorded with T2 (charcoal-based formulation of strain FA6+20kg/ha N) which was significantly superior to T3 and uninoculated control.

Effect of glyphosate on the growth and survival of rhizobia isolated from root nodules of grass pea (Lathyrus sativus L.)

Grass pea (L. sativus L.) is a widely cultivated crop worldwide, forming a symbiotic relationship with nitrogen-fixing rhizobia. Glyphosate is commonly used by farmers for weed control during agricultural processes. However, the application of this chemical herbicide negatively impacts soil fertility by affecting the nitrogen-fixing rhizobia. This study aimed to assess the effects of glyphosate on rhizobia isolated from healthy and robust Grass pea plants. Specifically, Grass pea plants exhibiting vigorous growth and a healthy appearance were intentionally selected to isolate rhizobia from their root nodules. The isolated rhizobia were then characterized based on their morphological features, biochemical properties, and resistance to abiotic traits. Rhizobial isolates from grass peas exhibited Gram-negative, rod-shaped morphology, milky colony color, and variable colony sizes. Additionally, the majority displayed smooth colony surfaces on yeast extract mannitol agar medium. Based on morphological and biochemical characteristics, the isolates could be grouped under the genus Rhizobium. Optimum growth conditions for these isolates were observed at temperatures between 28 and 38 °C, pH levels ranging from 5 to 8, and salt (NaCl) concentrations of 0.5% and 1%. At a concentration of 20 mL L−1, glyphosate inhibited 5.52–47% of the Rhizobium population. The inhibition percentage increased to 17.1–53.38% at a concentration of 40 mL L−1. However, when exposed to a higher concentration (60 mL/L) of glyphosate, 87% of the isolates were inhibited. The number of colonies after glyphosate exposure was significantly dependent on concentration, and there were notable differences between treatments with varying glyphosate concentrations (p < 0.05). Glyphosate negatively impacted the survival of grass pea rhizobia, leading to a reduction in the Rhizobium population (CFU). However, the effect varied between Rhizobium isolated from grass pea root nodules.

Diverse Bradyrhizobium spp. with Similar Symbiosis Genes Nodulate Peanut in Different Regions of China: Characterization of Symbiovar sv. Arachis.

A total of 219 rhizobial strains isolated from peanut grown in soils from six peanut croplands in Zhengyang county, Henan Province, were typed by PCR-RFLP of IGS sequences. Their phylogenetic relationships were refined on representative strains using sequence analyses of 16S rRNA genes, housekeeping genes (atpD, recA, glnII) and symbiosis genes (nodA, nodC and nifH). The 219 rhizobial isolates were classified into 13 IGS types, and twenty representatives were defined within eight Bradyrhizobium genospecies: B. guangdongense covering 5 IGS types (75.2% of total isolates), B. guangzhouense (2 IGS types, 2.7% total isolates), B. zhengyangense (1 IGS type, 11.3% total isolates) and five novel genospecies (5 IGS types, 0.9 to 3.2% total isolates). All representative strains had identical nodA, nodC and nifH sequences except for one nifH sequence. With this one exception, these sequences were identical to those of the type strains of Bradyrhizobium species and several Bradyrhizobium genospecies isolated from peanut in different regions of China. The nodC sequences of all strains showed < 67% similarity to the closest strains on the Genbank database indicating that they are representative of a novel Bradyrhiobium symbiovar. This study has shown that (1) diverse Bradyrhizobium spp. with similar symbiosis genes nodulate peanut in different regions of China. (2) Horizontal transfer of genes involved in nodulating peanut is common between Bradyrhizobium species in soils used to grow the crop in China. (3) The strains studied here are representative of a novel Bradyrhizobium symbiovar that nodulates peanut in China. We propose the name sv. arachis for this novel symbiovar indicating that the strains were isolated from Arachis hypogaea. Results here have practical implications in relation to the selection of rhizobial inoculants for peanut in China.

Comparative efficacy of different salt tolerant rhizobial inoculants in improving growth and productivity of Vigna radiata L. under salt stress

Worldwide, salinity severely affects agricultural production of crops such as mung bean in arid and semi-arid regions. In saline conditions, various species of Rhizobium can be used to enhance nodulation and induce salinity tolerance in maize. The present study conducted a pot experiment to determine the efficiency of three rhizobial isolates under different salinity conditions, such as 1.41, 4 and 6 dS m−1, on mung bean growth parameters, antioxidant status and yield. Results revealed that salt stress imparted adverse effects on the growth, antioxidants, yield and nodulation of mung bean. Under high salt stress conditions, fresh weights were reduced for roots (78.24%), shoots (64.52%), pods (58.26%) and height (32.33%) as compared to un-inoculated control plants. However, an increase in proline content (46.14%) was observed in high salt stressed plants. Three Rhizobium isolates (Mg1, Mg2, and Mg3), on the other hand, mitigated the negative effects of salt stress after inoculation. However, effects of Mg3 inoculation were prominent at 6 dS m−1 and it enhanced the plant height (45.10%), fresh weight of shoot (58.68%), root (63.64%), pods fresh weight (34.10%), pods number per plant (92.04%), and grain nitrogen concentration (21%) than un-inoculated control. Rhizobium strains Mg1, and Mg2 expressed splendid results at 1.41 and 4 dS m−1 salinity stress. The growth promotion effects might be due to improvement in mineral uptake and ionic balance that minimized the inhibitory effects caused by salinity stress. Thus, inoculating with these strains may boost mung bean growth and yield under salinity stress.

Evaluation Effect of Inoculum Prepared from Local Bacterial Isolates of Rhizobium and Nitrogen Fertilization on some Growth Traits of Mung Bean Plant in Gypsiferous Soil

The root nodule bacteria are utilized in the production of natural biological fertilizers to achieve clean agriculture by reducing chemical fertilizers. Therefore, the bacterial inoculum was prepared from local isolates of Rhizobium leguminosarum and Bradyrhizobium japonicum, which were morphologically and molecularly characterized. Subsequently, the efficacy of the prepared inoculum was tested in the formation of root nodules and some growth traits of mung bean plants in gypsiferous soil at three levels of nitrogen fertilization (0, 40, 80) kg N ha-1 The results showed a significant increase in all plant growth traits during the flowering stage for the treatments inoculated with Rhizobium leguminosarum and Bradyrhizobium japonicum bacteria in comparison to the treatment without inoculation. The inoculated treatments outperformed the non-inoculated treatment significantly. Additionally, the treatment inoculated with Bradyrhizobium japonicum showed a significant superiority over the treatment inoculated with Rhizobium leguminosarum in all studied growth traits, the recorded values for the count of root nodules were (0.00, 8.33, 35.11) nodule. plant-1, The wet mass of nodules was (0.00, 0.091, 1.802) g. plant-1, and dry mass of nodules was (0.00, 0.010, 0.337) g. plant-1. The plant height was (52.11, 56.22, 58.05) cm. plant-1. The dry mass of the vegetative part, was (14.03, 20.10, 23.05) g. plant-1, and, the dry mass of the root part was (1.37, 2.57, 2.67) g. plant-1. The nitrogen content was (1.547, 2.765, 3.028) % phosphorus (0.121, 0.236, 0.377) %, and potassium (1.168, 2.342, 3.562) % in the vegetative part. The absorbed nitrogen in the vegetative part was (217.41, 541.10,718.02) mg. plant-1. These values were recorded for the non-inoculated treatment, Rhizobium leguminosarum inoculated treatment, and Bradyrhizobium japonicum inoculated treatment, respectively. The results showed a significant superiority for the 40 kg N ha-1 fertilized treatment in comparison to both the unfertilized treatment and the 80 kg N/ha-1 fertilized treatment in all studied traits. The interaction effect was significant, as the treatment that was inoculated with Bradyrhizobium japonicum and 40 kg N ha-1 fertilized treatment exhibited significant superiority over the other treatments in all studied traits.

Nodulation pattern in different legumes by Rhizobia isolated from rajma

Rhizobium is an important symbiotic nitrogen fixing bacteria. The bacteria colonize leguminous plant cells within root nodules, where they convert atmospheric nitrogen into ammonia using the enzyme nitrogenase and then provide organic nitrogenous compounds such as glutamine or ureides to the plant. The plant, in turn, provides the bacteria with organic compounds made by photosynthesis. In this study, rhizosphere soil samples and nodules were collected from rajma plants of hilly Paderu region of Andhra Pradesh. Rhizobia were isolated from these samples using Yeast Extract Manitol Agar (YEMA) medium. There were 5 Rhizobium isolates obtained from the rhizosphere soil and nodules of rajma. These isolates were purified and inoculated to different legumes viz, black gram, green gram, red gram, Bengal gram and groundnut through seed treatment, planted in glass jars and maintained in green house. Nodulation pattern was studied 45 days after sowing. Rhizobium isolates 1 and 5 produced nodules in black gram and green gram, and Rhizobium isolate 2 produced nodules in black gram and red gram. Rhizobium isolate 3 produced nodules in black gram, green gram and red gram, and Rhizobium isolate 4 produced nodules in blackgram, greengram and groundnut. This study clearly showed that there is a possibility for Rhizobium isolate obtained from one legume to form nodules in other legumes but with variation in nodulation pattern.

Effect of Chemical Mutagenesis on Salt Tolerance of Common Beans (Phaseolus vulgaris L.) Nodulating Rhizobium

The chemical mutation is one of the important factors in the enrichment of the salt resistance capacity of the rhizobium isolates. Therefore, this study focused on 10 isolates of rhizobium collected from soil samples of Babile, Hararghe region, Ethiopia. All the collected isolates turned into a moderately yellow, yellow, and deep yellow color in yeast extract mannitol agar medium (YEMA) containing bromothymol blue after 48 h of incubation. It indicated that all the isolates were acid-producing Rhizobia. Moreover, based on colony morphology and diameter, 70% of the isolates displayed large mucoid colonies, and 30% of the isolates showed large watery colonies in YEMA media. Among the 10 isolates, 9(90%), 9(90%), 6(60%), 6(60%), 2(20%), and 1(10%), were grown at different salinity levels such as 2%, 4%, 6%, 8%, 9% and 10% of NaCl, correspondingly. The most salt-resistant wild isolate was HUCR 6 collected from Babile soil grown at 10% NaCl salinity level. A total of six mutants were considered after chemical mutagenesis based on their capacity to survive in the extreme salinity levels (11 to 14%). Mutant isolates such as HUCRM 4, HUCRM 5, and HUCRM 6 were the most tolerant Rhizobium that grew at the salinity level of 11 to 14% NaCl. The most sensitive mutant isolate was HUCRM 10 followed by the isolates HUCRM 8 and HUCRM 9 was the next sensitive mutant Rhizobia that grew only at 11% of NaCl concentration. Compared to the Rhizobium wild isolates, the mutant isolates were observed to be more tolerant to a medium containing higher concentration of NaCl, as high as 11% to 14%. Besides, 80% of the mutant isolates demonstrated effective nodulation with the common beans. The mutant isolate (HUCRM 4) showed better performance in relation to root nodule performance of Rhizobium species and increased the plant biomass production. In this study, mutant isolates HUCRM 6, which is tolerated to 14% NaCl, and HUCRM 4, HUCRM 5, and HUCRM 6 isolates tolerated at 12% salinity level. Finally, based on their symbiotic efficiency and tolerance to extreme salt levels, these mutant isolates (HUCRM 4, HUCRM 5 and HUCRM 6) were encouraged to be used for the development of Rhizobium inoculants of common beans grown under extreme saline conditions. J. Bio-Sci. 30(2): 13-22, 2022