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Selection of Competitive and Efficient Rhizobia Strains for White Clover.

ABSTRACT: The practice of inoculating forage legumes with rhizobia strains is widespread. It is assumed that the inoculated strain determines the performance of the symbiosis and nitrogen fixation rates. However, native-naturalized strains can be competitive, and actual nodule occupancy is often scarcely investigated. In consequence, failures in establishment, and low productivity attributed to poor performance of the inoculant may merely reflect the absence of the inoculated strain in the nodules. This study lays out a strategy followed for selecting a Rhizobium leguminosarum sv. trifolii strain for white clover (Trifolium repens) with competitive nodule occupancy. First, the competitiveness of native-naturalized rhizobia strains selected for their efficiency to fix N₂ in clover and tagged with gusA was evaluated in controlled conditions with different soils. Second, three of these experimental strains with superior nodule occupancy plus the currently recommended commercial inoculant, an introduced strain, were tested in the field in 2 years and at two sites. Plant establishment, herbage productivity, fixation of atmospheric N₂ (¹⁵N natural abundance), and nodule occupancy (ERIC-PCR genomic fingerprinting) were measured. In both years and sites, nodule occupancy of the native-naturalized experimental strains was either higher or similar to that of the commercial inoculant in both primary and secondary roots. The difference was even greater in stolon roots nodules, where nodule occupancy of the native-naturalized experimental strains was at least five times greater. The amount of N fixed per unit plant mass was consistently higher with native-naturalized experimental strains, although the proportion of N derived from atmospheric fixation was similar for all strains. Plant establishment and herbage production, as well as clover contribution in oversown native grasslands, were either similar or higher in white clover inoculated with the native-naturalized experimental strains. These results support the use of our implemented strategy for developing a competitive inoculant from native-naturalized strains.

SUBMITTER: Irisarri P

PROVIDER: S-EPMC6489563 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Selection of Competitive and Efficient Rhizobia Strains for White Clover.

Irisarri Pilar P Cardozo Gerónimo G Tartaglia Carolina C Reyno Rafael R Gutiérrez Pamela P Lattanzi Fernando A FA Rebuffo Mónica M Monza Jorge J

Frontiers in microbiology 20190423

The practice of inoculating forage legumes with rhizobia strains is widespread. It is assumed that the inoculated strain determines the performance of the symbiosis and nitrogen fixation rates. However, native-naturalized strains can be competitive, and actual nodule occupancy is often scarcely investigated. In consequence, failures in establishment, and low productivity attributed to poor performance of the inoculant may merely reflect the absence of the inoculated strain in the nodules. This s ...[more]

PMID: 31065250

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Project description:Biological nitrogen fixation is the most important eco-friendly approach to nitrogenous fertilizer management in the rhizosphere. Rhizobium is considered the most important symbiotic N-fixing microorganism. Native strains of Rhizobium perform better than the non-native strains by getting ambient conditions for growth and proliferation. Native strains enhance the soil fertility and productivity of pulses. The study was carried out in three phases, i.e., pot experiment, field experiment, and farmers' field demonstrations. In a pot experiment, two isolated rhizobia were inoculated to seeds of French bean (Phaseolus vulgaris) and applied with and without lime to evaluate crop growth, photosynthetic activity, and nodule characteristics of the target crop. In the field, strains were inoculated to seeds of French bean, which received different combinations of inputs- inorganic fertilizers, lime, and boron- to study the influence of native stains on crop productivity and agronomic efficiency. In comparison to non-limed packaging, the amounts of chlorophyll a, chlorophyll b, total chlorophyll, and chlorophyll a:b were, respectively, 13% to 30%, 1% to 15%, 10% to 27%, and 1% to 20% greater in limed packages. In limed packages compared to non-limed packages, the root length, biomass, density, and growth rate were increased by 16% to 17%, 36% to 52%, 38% to 49%, and 36% to 52%, respectively. In contrast to non-limed packages, limed packages had nodule attributes like the number of nodules per plant and nodular weight, which were 28% to 41% and 33% to 37% greater, respectively. Inoculation of native rhizobia strains with liming to acid soil increased 46% to 72% of leaf nitrogen content over non-limed rhizobia inoculated packages. In a field experiment, the adoption of soil test-based fertilizer application had an advantage of 25% in pod yield over farmers' practice. Acid soil amelioration with lime improved pod yield from 14% to 39% over non-limed packages. Farmers' field demonstration recorded the highest pod yield in the package where seeds were inoculated with S2 (RBHR-21) strain added with soil test-based fertilizers (STD) followed by STD + S1 (RBHR-15) with 98% and 84% increase over farmers' practice. However, experimental evidence favored using both strains for bio-inoculation of the French bean crop.

Project description:Establishment of the symbiotic relationship that develops between rhizobia and their legume hosts is contingent upon an interkingdom signal exchange. In response to host legume flavonoids, NodD proteins from compatible rhizobia activate expression of nodulation genes that produce lipochitin oligosaccharide signaling molecules known as Nod factors. Root nodule formation commences upon legume recognition of compatible Nod factor. Rhizobium leguminosarum was previously considered to contain one copy of nodD; here, we show that some strains of the Trifolium (clover) microsymbiont R. leguminosarum bv. trifolii contain a second copy designated nodD2. nodD2 genes were present in 8 out of 13 strains of R. leguminosarum bv. trifolii, but were absent from the genomes of 16 R. leguminosarum bv. viciae strains. Analysis of single and double nodD1 and nodD2 mutants in R. leguminosarum bv. trifolii strain TA1 revealed that NodD2 was functional and enhanced nodule colonization competitiveness. However, NodD1 showed significantly greater capacity to induce nod gene expression and infection thread formation. Clover species are either annual or perennial and this phenological distinction is rarely crossed by individual R. leguminosarum bv. trifolii microsymbionts for effective symbiosis. Of 13 strains with genome sequences available, 7 of the 8 effective microsymbionts of perennial hosts contained nodD2, whereas the 3 microsymbionts of annual hosts did not. We hypothesize that NodD2 inducer recognition differs from NodD1, and NodD2 functions to enhance competition and effective symbiosis, which may discriminate in favor of perennial hosts.IMPORTANCE Establishment of the rhizobium-legume symbiosis requires a highly specific and complex signal exchange between both participants. Rhizobia perceive legume flavonoid compounds through LysR-type NodD regulators. Often, rhizobia encode multiple copies of nodD, which is one determinant of host specificity. In some species of rhizobia, the presence of multiple copies of NodD extends their symbiotic host-range. Here, we identified and characterized a second copy of nodD present in some strains of the clover microsymbiont Rhizobium leguminosarum bv. trifolii. The second nodD gene contributed to the competitive ability of the strain on white clover, an important forage legume. A screen for strains containing nodD2 could be utilized as one criterion to select strains with enhanced competitive ability for use as inoculants for pasture production.

Dataset Information

Selection of Competitive and Efficient Rhizobia Strains for White Clover.

Publications

Selection of Competitive and Efficient Rhizobia Strains for White Clover.

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