Identification of Rhizobia Isolated from Nodules of Mexican Commercial Soybean Varieties

Rhizobia, crucial for nitrogen fixation in leguminous plants, play a vital role in soybean cultivation. This study, conducted in Mexico, a major soybean importer, aimed to identify bacteria from nodules of five soybean varieties in high-production regions. Multilocus sequence analysis (MLSA) was employed for enhanced species resolution. The study identified six Bradyrhizobium species: Bradyrhizobium japonicum USDA 110, Bradyrhizobium japonicum USDA 6, Bradyrhizobium elkanii USDA 76, Bradyrhizobium neotropicale, Bradyrhizobium lablabi, and Bradyrhizobium icense. Bradyrhizobium japonicum USDA 110 predominated in the soils, displaying symbiotic preference for the Huasteca 400 variety. However, phylogenetic analysis didn't reveal a clear association between strains, soil, and soybean variety. This research sheds light on the diversity of rhizobia in Mexican soybean cultivation, contributing to the understanding of symbiotic relationships in soybean production systems.

Morpho-Physiological and Biochemical Characterization of Soybean-Associated Rhizobia and Effect of Their Liquid Inoculant Formulation on Nodulation of Host Plants in the Cameroon Cotton Fields Zone

The aim of this research was to assess the diversity of the Cameroon cotton zone in soybean associated rhizobia in order to formulate the most efficient elite inoculant to boost both the cotton and soybean production. Therefore, soybean associated rhizobia were isolated and characterized morphologically, physiologically and biochemically on YEMA culture media. For each of the two soybean varieties (Houla1 and TGX1910 14F) used, the trials were laid out in two IRAD-fields of North Cameroon (Sanguere-Paul) and Far-North (Soukoundou) respectively, under a complete randomized complete block design, the isolate formulations representing the treatments. The six isolated strains (IS1, IS2, IS3, IS4, IS5, IS6) from which seven liquid inoculant were formulated were revealed to belong to the same slow growing group of rhizobia, with a high level of tolerance to temperature, pH, and salinity, with optimum growth at respectively 28˚C, pH (7 - 9), salt (1% - 5%). Not surprisingly, root nodules were formed by both inoculated and uninoculated soybean plants. However, the most efficient soybean-rhizobia symbiosis for nodulations were isolate IS6 associated to TGX1910 14F variety, and isolate IS5 associated to Houla1variety at Sanguere-Paul. Whereas isolate M was associated to TGX1910 14F variety, Houla 1 variety had affinity with native rhizobia isolates at Soukoundou. The present results suggest the adaptability of rhizobia isolates to a particular soybean variety at a particular cotton fields zone. These findings should be taken into consideration for commercial inoculant formulation.

Isolation and Authentication of Local Rhizobia Nodulating Common Bean (Phaseolus vulgaris L.) in Different Agro-Ecological Zones of Côte d’Ivoire

Legumes such as common bean (Phaseolus vulgaris L.) have been introduced into cropping systems for sustainable soil management. Consequently, the loss of fertility of the latter remains a major constraint to bean production because this legume is rarely fertilized, yet it is considered to be a poor nitrogen fixer in the absence of inoculation. To overcome this, this study was undertaken with the objective of seeking efficient local rhizobia in order to propose a bean inoculum formulation. To do this, soil samples taken from twelve localities in the Centre, North and West areas of C?te d’Ivoire were used to trap bean nodulating rhizobia. The ROBA1 bean accession used was sown in pots containing the sampled soils. Seedlings were uprooted at the start of flowering and nodulation was assessed. The isolates obtained were purified and then characterized phenotypically. The infectivity and symbiotic efficacy of these isolates were determined in vitro by the authentication test in which the purified isolates were reinoculated to their original host plant. A total of 24 rhizobium isolates were obtained from the soils of six localities. During morphological characterization, the isolates showed typical characteristics of Rhizobium. With the exception of RPC501, RPC505 and RPC522, all isolates were authenticated and able to nodulate the host plant in controlled culture. Isolates RPC502, RPC507, and RPC508 were effective and significantly increased (P < 0.05) nodule number and weight, height, and plant biomass. This study has, therefore, revealed the presence of effective local rhizobia in Ivorian soils and capable of nodulating common beans. A genetic characterization of efficient rhizobia identified after experimentation in different environmental conditions should be considered before being recommended as bean rhizobia inoculant.

Dynamic Transportation of Endophytic Rhizobia in Plants during Growth Development of Alfalfa

[ Objective] The aim of this study was to investigate the dynamic distribution of endophytic rhizobia in different plant parts during seed development of alfalfa and the carrying mechanism of endophytic rhizobia from alfalfa seeds. [ Method] Root, stem, leaf, flower, pod and seed from two varieties of alfalfa in different development stages were used for surface disinfection, and then the present site and quantity change of rhizobia in alfalfa were studied by the dilution - plate method. [ Result] The number of rhizobia from underground plants in the same stage was as follows： main root 〈 lateral root 〈 hair root. The number of rhizobia from main root and hair root in different stages was as follows： bearing pod stage 〉 branching stage 〉 florescence, while that from lateral root was as follows： branching stage 〉 florescence 〉 bearing pod stage. The distribution of endophytic rhizobia from aerial plants was as follows： few rhizobia was found in stem during branching stage and bearing pod stage, which accounted for 155 cfu/g and 125 cfu/g respectively. Rhizobia mainly distributed in flower bud during branching stage accounting for 9 400 cfu/g, while only in ovary wall during squaring stage accounting for 18 cfu/5 flowers, and few in pollen （26 cfu/5 flowers）, fertilized ovules （26 cfu/50 grains）, receptacle （15 cfu/5 flowers） and ovary wall （95 cfu/5 flowers） during florescence. Rhizobia existed in pod peel （3 550 cfu/5 pods） and immature seed （1 135 cfu/50 grains） during bearing pod stage, and greatly distributed in seed coat of mature seeds （1 435 cfu/50 grains）, but only few in embryo （32 cfu/50 grains） and cotyledon （22 cfu/50 grains）. [ Cenclusion] There is a significant difference in the distribution of endophytic rhizobia from different parts of aerial alfalfa, which only exists in the parts directly related to seed development.

Effects of N,P and K Fertilizers on Growth of Clover Nitrogen-fixing Rhizobia and Soil Fertility after Plantation

In order to develop organic rice and increase paddy soil fertility by cloverorganic rice rotation, the effects of N, P and K fertilizers on growth of clover nitrogen-fixing rhizobia and soil fertility after plantation were investigated, thereby providing certain reference for reasonable cultivation of clover and improvement of soil fertility. A two-year experiment was conducted from 2012 to 2013. The clover was cultivated after rice every year, and different levels of N, P and K fertilizers were applied before winter. In the treatment group, no fertilizer was applied. The effects of different fertilizers and different application amounts on clover yield, nitrogen-fixing rhizobia quantity, nitrogen-fixing rhizobia weight and soil fertility after plantation were analyzed. The results showed that the application of N, P and K fertilizers in the early stage had significantly effect on the growth of clover. When the application amount of N fertilizer was 75 kg/hm^2（N 46%）, the clover yield, nitrogen-fixing rhizobia quantity and nitrogen-fixing rhizobia weight were highest. The soil nitrogen content after plantation increased with the increase of the application amount of N fertilizer, while the P and K content decreased and then increased with the increased application amounts. Soil available P content increased with the increased application amounts of N and P fertilizers, but it did not change significantly with the increased application amount of K fertilizer. Soil available K content increased first and then decreased with the increased application amounts of N and P fertilizers.When the application amounts of N and P fertilizers were 150（N 46%） and 300（P_2O_5 12%） kg/hm^2, soil available K content reached the maximum. Soil organic matter content increased with the increased application amounts of N, P and K fertilizers. Therefore, in the cultivation of clover, appropriate application of N, P and K fertilizers in the early stage can improve clover yield and soil fertility.

Adsorption of Cadmium by Soil Colloids and Minerals in Presence of Rhizobia

Experiments were conducted to study the adsorption of Cd on two soil colloids (red soil and yellow- brown soil) and three variable-charge minerals (goethite, noncrystalline Fe oxide and kaolin) in the absence and presence of rhizobia. The tested strain Rhizobium fredii C6, tolerant to 0.8 mmol L-1 Cd, was selected from 30 rhizobial strains. Results showed that the isotherms for the adsorption of Cd by examined soil colloids and minerals in the presence of rhizobia could be described by Langmuir equation. Within the range of the numbers of rhizobial cells studied, the amount of Cd adsorbed by each system increased with increasing rhizobial cells. Greater increases for the adsorption of Cd were found in red soil and kaolin systems. Rhizobia influence on the adsorption of Cd by examined soil colloids and minerals was different from that on the adsorption of Cu. The presence of rhizobia increased the adsorption sanity of soil colloids and minerals for Cd, particularly for the goethite and kaolin systems. The discrepancies in the influence of rhizobia on the adsorbability and affinity of selected soil colloids and minerals for Cd suggested the different interactions of rhizobia with various soil components. It is assumed that bacterial biomass plays an important role in controlling the mobility and bioavailability of Cd in soils with kaolinite and goethite as the major colloidal components, such as in variable-charge soil.

Phylogenetically clustering of rhizobia by genome structure:application to unclassified Rhizobium

Previous research reveals that the genome structures of rhizobial type strains and reference strains can reflect their phylogenetic relationships. In order to further explore the potential application of genome structure as a phylogenetic marker in rhizobial natural taxonomy, this study analyzed the genome structures of 29 unclassified nodule bacteria isolated from the root nodules of leguminous trees, Robinia sp., Dalbergia spp., and A lbizia spp. and 7 rhizobial reference strains by I-CeuI cleavage, then clustered these bacteria phylogenetically based on their genome structures and compared these clusters with those based on numerical taxonomy and 16S rDNA PCR-RFLP. Eleven phylogenetic clusters were obtained, The clusters were in large part consistent with those based on numerical taxonomy and 16S rDNA PCR-RFLP. Also there are inconsistent clusters based on the above three methods. But results are completely consistent with 16S rRNA clusters. This suggested that the genome structure clustering method can be used to lastly identify root nodule isolates and detect their phylogenetic relationships. The credibility and repeatability of the results, together with the simplicity and possibility to analyze a large number of strains in a short time of the method, indicates the broad potential application of genome structure as phylogenetic marker to categorize rhizobial isolates and should in the future facilitate biodiversity studies.

Isolation and Characterization of Nitrogen Fixing Rhizobia from Cultivated and Uncultivated Soils of Northern Tanzania

Soil bacteria1 called rhizobia are gram-negative capable to colonize the soil immediately surrounding roots under the influence of the plant “rhizosphere” and reduce atmospheric nitrogen into the form available to plants through nitrogen fixation process. Nitrogen is the most limiting and supplied nutrient to most plants, and the determinant of plant growth. Legumes differ with most plants because they have access to nitrogen from both mineral and symbiotic sources. Small-scale farmers who are the major legume producers in Africa rarely apply fertilizers during legume production. Hence, the crop is largely dependent on fixed nitrogen from native nitrogen fixers. Isolation of rhizobia for legume production has been given a little attention in Africa due to inadequate research or negligence of researchers and unawareness of its potential in legume production as well as lack of an intention from skilled personnel to popularize the technology. Evaluation of effectiveness of isolated rhizobia is essential for inoculants preparation, host specificity recommendation and symbiotic effectiveness. The isolation, determination of their population in the soil and assessing factors affecting their population and testing the effectiveness of native nitrogen fixers with respect to right trap host crop are given a special attention in this review.

Symbiotic and Phenotypic Characteristics of Rhizobia Nodulaing Cowpea （Vigna Unguiculata L. Walp） Grown in Arid Region of Libya （Fezzan）

Symbiotic and phenotypic characteristics of thirty rhiobial isolates obtained from root nodules of two cowpea （Vigna unguiculata L. Walp） cultivars that grown in different sites of Fezzan （Southern part of Libya） were studied. Cultural characteristics and cross-nodulation with Arachis hypogega and Faidherbia albida showed that they were slow-growing rhizobia. Each isolate was found to coexist with non-symbiotic bacteria similar in their cultural characteristics to fast-growing rhizobia. All isolates formed symbiosis with the test plants, but different in their nitrogen-fixation efficiency. Numerical analysis of phenotypic characteristics showed that at boundary level of 70% average similarity, the isolates formed four distinguished groups and two isolates remained separate. Most isolates exhibited wide tolerance to acidity, alkalinity and extreme temperatures. They also resistant to some heavy metals such as mercury, copper, zinc, lead, cadmium and aluminum at low concentrations and antibiotics like polymyxin, colistin, bacitracin and nalidixic acid. Isolates displayed different response to salinity ranging from sensitive, which unable to grow in 1% NaCI to resistant and grow at 2% NaCl or above. Urea was hydrolyzed by most of them and carbohydrates utilizations were different. Sucrose and maltose were metabolized by most of the test isolates, whereas, monosaccharide and sugar alcohols were poorly utilized.

Physiological Characteristics of Rhizobia Isolated from Retama raetam （Forsk） and Lupinus various （L.） Indigenous to Libyan Desert

A total of 20 rhizobial isolates obtained from root-nodules of Retama raetam （Forsk） and Lupinus various （L.） were studied for their ability to tolerate local environmental and chemical stresses. Six fast and one slow growing reference strains were included for comparison. Numerical analysis showed that the reference strains were separated from test isolates. The majority of isolates displayed a high tolerance to extreme temperatures; some even grew at 48 ℃ and grew at alkaline and acid pH. Regarding tolerance to salinity on agar medium, the majority of the isolates grew at 6% NaCl, but some isolates from Lupinus various were more resistant, grew even at 8% NaCl. Most isolates were resistant to heavy metals, but were sensitive to most antibiotics tested.

Phenotypic, Stress Tolerance and Plant Growth Promoting Characteristics of Rhizobial Isolates from Selected Wild Legumes of Semiarid Region, Tirupati, India

Rhizobia are vital for nitrogen input, fertility of soil and legume plant growth. Knowledge on rhizobial diversity from arid and semiarid areas is important for dry land agriculture in the context of climatic change and for economic utilization. This study provides morphological, biochemical, stress tolerance and plant growth promoting characteristics of fifteen rhizobial isolates from the nodules of same number of wild legumes and one isolate from cultivated Arachis hypogea from semi-arid region, Tirupati. The bacterial isolates were confirmed as rhizobia based on colony morphology and biochemical tests. Based on the colour change of YMA-BTB medium, eight isolates were identified as slow growers and six were fast growers. The isolates differed in growth pattern, colony morphology, antibiotic resistance at higher concentrations and uniformity in utilization of carbon and nitrogen sources. The isolates are tolerant to NaCl up to one percent, displayed normal growth at temperatures 28℃ - 30℃, at neutral pH and poor growth at pH 5and 9. The isolates varied in the production of EPS and IAA, positive for phosphate solubilization and siderophore formation. This functional diversity displayed by the isolates can be utilised for the legume crop production by cross inoculation.

Comparative analysis of symbiotic phenotypes of soybean ’’Tianlong 1’’ with two different rhizobia and co-inoculation at different developmental stages

Symbiotic nitrogen fixation between soybean and rhizobia, which directly catalyze reduction of atmospheric nitrogen into ammonia, has critical importance in agricultural economy and environment. However, soybean symbiotic nitrogen efficiency, which changes with growth stage, largely relies on symbiotic host-specificity and nodule development. In this report, symbiotic phenotypes of soybean ‘Tianlong 1’ with different inoculation (inoculated with Bradyrhizobium japonicum strain 113-2, Sinorhizobium fredii USDA205 and co-inoculated with 113-2 and USDA205) were comparatively studied at different developmental stages. Without additional nitrogen, chlorophyll content, above ground fresh weight,plant height, nodule number and nodule dry weight changed with different rhizobia at various developmental stages. 113-2 had more advantages than USDA205 in promoting plant growth and nodulation. Nodulation morphology of co-inoculation was more similar to 113-2 than USDA205. Competitive nodulation between two rhizobia promoted plant growth, which had significant agricultural application prospect. For the first time, this study provided evidence for co-inoculation with different rhizobia having positive effect on plant growth, and shed new light on the competitive relationship between different rhizobia.

Phenotypic and Genetic Characterization of Trifolium Rhizobia Isolated from Temperate and Subtropical Regions of China

The clovers that comprise Trifolium genus are naturalized in the subtropical and tropical zones in China. They are valuable bioresources as important green manures and pasture grass, which contribute biologically fixed dinitrogen （N2） and provide nutrition to farming systems. However, there are very few effective strains available for inoculant production and there is little information available about symbiotic rhizobia in Chinese legume clover root nodules. In this study, 139 root nodule bacteria were isolated from two clover species （Trifolium repens and Trifolium pretense） growing in the subtropical and temperate regions of China, 16S rRNA gene sequence analysis, BOX-PCR, whole cell protein SDS-PAGE, and nodulation tests were performed to characterize these strains. The results showed that phenotypic and genetic diversities among 139 isolates were large, with 83 protein patterns and 66 BOX- AIR profiles, respectively. The rhizobial strains were first divided into two large phenotypic protein groups. The sequencing strains representing the two groups were related to Rhizobium leguminosarum USDA2370T and R. sophorae CCBAU03386T and had 99.6%-100% similarities. The phylogeny specificity of the rhizobia from clover was elucidated, while showed a large variation in the fingerprints of the phenotypes and genotypes and genetic diversity was high （revealed by Shannon diversity index, H＇）. The rhizobial isolates from subtropical regions, such as Anhui Province, Yunnan Province and Hubei Province, had higher diversities than those from temperate areas, such as Hebei Province and Shanxi Province, which could be used to identify rhizobial strains from clover and screen efficient inoculum strains. A number of diverse rhizobial strains had been identified and a pool of currently available clover rhizobial strains were increased. This would ultimately increase the likelihood of identifying more efficient strains suited for developing a successful inoculation strategy for the production of white clover.

Phosphorus Fertilizer Rating and Rhizobia Inoculation for Improved Productivity of Cowpea in Northern Uganda

Cowpea (Vigna unguiculata) is an important legume crop in the tropics and subtropical regions of the world. It is mainly grown for its leaves and grains, and to a lesser extent as a fodder crop. Cowpea is considered as the most important food grain legume in the dry savannas of tropical Africa. This study compared the yield of local (Agondire) and improved (SECOW 2W) cowpea varieties grown on an Oxisol. Inorganic P at levels of 0, 10, 20, and 40 kg·ha-1 was tested on each variety with or without rhizobia inoculation. The experiments were set up in a randomized complete block design and replicated thrice during the short and long rains of the 2015/2016 seasons on fifteen fields in Arua district, northern Uganda. Agondire responded significantly (P < 0.05) better than SECOW 2W when high rates of inorganic phosphorus (40 kg P ha-1) were applied. A significant increase of 26.4% and 28.4% in grain yield of Agondire and SECOW 2W, respectively was obtained after inoculation with rhizobia. We concluded that inoculation and P fertilizer application increased the yield of both varieties, but with inoculation, SECOW 2W performs much better at lower P fertilizer rates than Agondire. Therefore, we recommend growing of SECOW 2W under inoculation with 20 kg P ha-1 and an application of 40 kg P ha-1 for Agondire local cowpea variety in northern Uganda.

Evidence of Fungicides Degradation by Rhizobia

Fungicides which are not easily degradable have the greatest adverse effects on soil microbes. These pesticides negatively affect the growth and multiplication of fungi and bacteria and consequently cause the disturbance of the natural soil microbial balance. In this study two fungicide tolerant isolates of rhizobia;clover isolate (TA1) and peanut isolate (8) were assessed in their capacity to degrade Vitavax and Rizolex. The performance of these isolates in fungicides degradation was tested using the colorimetric assay for Rizolex and the HPLC analysis for Vitavax to detect the degradation products. Using HPLC analyses, the control sample showed specific peak indicating the Vitavax presence in the medium. The specific peak did not change in the control samples throughout the experiment.With the strainTA1 the specific peak of the Vitavax fungicides started to reduce as the incubation time goes on. The Vitavax fungicide did not degrade completely after 240 hours of incubation with rhizobial isolate. The Rizolex used in this study contained blend of Thiram (active ingredient of Rizolex) and Tolcofs methyl fungicides in 1:1 ratio. The biodegradation of Rizolex in the liquid media showed the formation of two new intermediates which were released into the medium indicating the degradation of the tested fungicide by peanut rhizobial isolate No. 8 in 48 hrs of incubation 45% of this compound was degraded. This work shows that the selection of fungicides tolerant rhizobial strains is important to protect the rhizobial inoculants from the toxic effect of the pesticides.

Effect of Pesticides on Growth of Rhizobia and Their Host Plants During Symbiosis

Effect of various pesticides (insecticides, fungicides and herbicides) has been studied on growth and efficiency of symbiotic properties of 3 fast growing Rhizobium sp. under green house conditions. The results revealed adverse effects on plant growth and nitrogen fixing capacity as measured by dry weight and total nitrogen content of plants infected with pesticide treated Rhizobium. Of the pesticides tested, herbicides were found to be more effective on the above parameters than the insecticides and fungicides.

Resistance to Abiotic Stress and Effectiveness of Native Rhizobia on Bambara Groundnut [Vigna subterranea (L.) Verdc.] in Benin

Bambara groundnut [Vigna subterranea (L.) Verdc.], as a legume, can establish relationships with nitrogen-fixing bacteria such as Rhizobium. However, Rhizobium efficacy is not always optimal due to the lack or poor efficient strains in the soil. This study aimed to evaluate symbiotic efficiency of endogenous Rhizobia nodulating Bambara groundnut and their resistance to abiotic conditions. Root nodules were randomly sampled from three agroecological zones across the country, surface sterilized, ground and paste plated on YEMA media. After 24 hours, the bacterial colonies were purified. The pure cultures were further characterized using morphological and biochemical methods and their resistance to antibiotics and heavy metals was evaluated. Lastly, the symbiotic efficiency of the isolates was assessed through a greenhouse experiment. A total of eighty-five presumptive strains were isolated from Bambara groundnut roots nodules obtained from the farms. The physiological characterization of the isolated showed a decrease in isolates growth when NaCl concentration was more than 7%. In addition, 47% of the isolates were tolerant to a temperature of 40°C. Most of the isolates were highly resistant to Erythromycin in all its concentration levels and to Kanamycin, Spectinomycin, Neomycin and Ampicillin at 10 μg⋅mL−1. Most of them showed resistance to Cu and Zn at 10 μg⋅mL−1. Results of the effectiveness test on two Bambara groundnut varieties yielded dry shoot matter varying from 3.33 g⋅plant−1 to 7.21 g⋅plant−1 for variety 1 and from 4.38 g⋅plant−1 to 8.38 g⋅plant−1 on variety 2. N uptake ranged between 0.09 g⋅plant−1 and 0.29 g⋅plant−1 for variety 1 and between 0.12 and 0.29 g⋅plant−1 for variety 2. The isolates yielding higher shoot dry weight and N uptake were LMSEM312, LMSEM338, LMSEM307, LMSEM351 for variety 1 and LMSEM338, LMSEM309, LMSEM307 for variety 2. The isolates showing better performance can be used to develop bio-fertilizer for sustainable Bambara groundnut production in Benin.

Response of Indigenous Rhizobia to the Inoculation of Soybean[Glycine max(L.)Merrill]Varieties Cultivated under Controlled Conditions in Cote d’Ivoire

Soybean [Glycine max (L.) Merrill] is an important crop known to improve population nutritional status and increase soil fertility and its productivity through biological nitrogen fixation. In C?te d’Ivoire, the introduced Bradyrhizobium japonicum used as inoculum had slight compatibility to several soybean varieties compromising their vulgarization. Therefore, the present study was conducted to examine the infectiveness and the effectiveness of indigenous rhizobial isolates on three soybean varieties (Canarana, Doko and Piramana) cultivated in C?te d’Ivoire. The experiment was conducted with potted plant filled with sterilized sand and was statistically laid in Completely Randomized Design (CRD) with sixteen (16) natives rhizobia, one (01) reference strain and uninoculated control (with or without nitrogen) with three replications. The results showed that inoculation significantly improved nodule number, nodule dry weight, plant height and total dry matter of soybean over the negative control treatment. Among the inoculated treatments, five indigenous rhizobia RSC119, RSC324, RSC502, RSC504 and RSC508 significantly (P -1) compared to IRAT FA3 strain (95 mg·plant-1) on Doko. The higher effectiveness was recorded with RSC119, RSC504 and RSC502 with 206.73%;201.79% and 200.45% respectively compared to TN (100%). The correlation analysis indicated significant association of nodule number and total biomass indicating the importance of symbiotic nitrogen fixation. Based on their infectiveness and their effectiveness, the isolates RSC119, RSC324, RSC502, RSC504 and RSC508 could be used as elite local rhizobia and tested in field conditions to establish their potential contribution on soybean productivity.

Selection of rhizobial strains differing in their nodulation kinetics under low temperature in four temperate legume species

Background:Winter climate change including frequent freeze-thaw episodes and shallow snow cover will have major impacts on the spring regrowth of perennial crops.Non-bloating perennial forage legume species including sainfoin,birdsfoot trefoil,red clover,and alsike clover have been bred for their adaptation to harsh winter conditions.In parallel,the selection of cold-tolerant rhizobial strains could allow earlier symbiotic nitrogen(N)fixation to hasten spring regrowth of legumes.Methods:To identify strains forming nodules rapidly and showing high N-fixing potential,60 rhizobial strains in association with four temperate legume species were evaluated over 11 weeks under spring soil temperatures for kinetics of nodule formation,nitrogenase activity,and host yield.Results:Strains differed in their capacity to form efficient nodules on legume hosts over time.Strains showing higher nitrogenase activity were arctic strain N10 with sainfoin and strain L2 with birdsfoot trefoil.For clovers,nitrogenase activity was similar for control and inoculated plants,likely due to formation of effective nodules in controls by endophyte rhizobia present in seeds.Conclusions:Selection based on nodulation kinetics at low temperature,nitrogenase activity,and yield was effective to identify performant rhizobial strains for legume crops.The use of cold-tolerant strains could help mitigate winter climatic changes.

达乌里胡枝子根瘤菌的促生作用及高效根瘤菌的鉴定

本研究选择达乌里胡枝子(Lespedeza daurica)根瘤分离且结瘤效果较好的7株根瘤菌(TG64,TG97,TG98,TG100,TG101,TG105和TG115),通过测定达乌里胡枝子生长表型和主要营养成分,结合结瘤和光合特性初步分析促生的生理生化机制,研究不同根瘤菌的促生作用;通过分子生物学、形态学和生理生化特性鉴定高效根瘤菌。结果表明:除TG64,其余菌株显著提高了株高、根长、生物量和蛋白质含量,改善了根系构建,TG101促生最明显。所有处理单株结瘤5~33个,根瘤干重2.55~35.87 mg;TG98,TG100,TG101和TG115显著提高了叶绿素含量、光合和氮吸收能力,是促生的主要生理生化机制。经鉴定,TG101为格木慢生根瘤菌(Bradyrhizobium erythrophlei)。综上,根瘤菌通过改善植株氮营养促进叶绿素合成、增强光系统活性,提高光合能力,进而促进干物质积累调控生长。其中TG101为高效根瘤菌,为后续研发适用黄土高原的达乌里胡枝子根瘤菌剂提供了菌种资源和理论依据。