Symbiotic effectiveness of Bradyrhizobium strains isolated from Parasponia and tropical legumes on Parasponia host species

The symbiotic effectiveness of Bradyrhizobium strains isolated from three species of Parasponia and from legumes were compared on Parasponia grown in Leonard-jars. Effectiveness of each symbiotic association was estimated from dry weight and total nitrogen of shoots and nodules of plants grown on medium free of combined nitrogen. Twenty strains isolated from three species of Parasponia were found to vary in their effectiveness on P. andersonii, the least effective fixing one fifth of the nitrogen of the most effective strains. The outcome of the symbiosis was not associated with the host source of the test strain. P. andersonii, P. rugosa and P. rigida responded differently to a selection of seven strains of Parasponia Bradyrhizobium; some strains were either ineffective or fully effective on each host, while others varied in their symbiotic performance. P. andersonii fixed significantly (P < 0.001) larger quantities of nitrogen than either P. rugosa or P. rigida with p. rigida being the least effective. In contrast to Bradyrhizobium strains from Parasponia spp. which formed nodules rapidly (within 11–20 days), nine strains isolated from legumes required between 25 and 74 days to form partially effective nodules. The thre Parasponia species formed relatively large quantities of nodule tissue relative to the amount of nitrogen fixed and shoot dry matter produced. The Bradyrhizobium isolated from Parasponia plants growing in Papua New Guinea soils could be grouped together on the basis of their infection characteristics on Parasponia and legumes.     

Chemotaxis,induced gene expression and competitiveness in the rhizosphere

Rhizobia are soil bacteria which symbiotically infect legume roots and generate nodules in which they fix atmospheric nitrogen for the plant in exchange for photosynthetically fixed carbon. A crucial aspect of signal exchange between these symbionts is the secretion of phenolic compounds by the host root which induce nodulation gene expression in the bacteria. Stimulation of nod gene expression by host phenolics is required for nodule formation, is biochemically specific at 10^-6 M, and is mediated by nodD. We and others have shown that rhizobia display chemotaxis to 10^-9 M of the same phenolic compounds. Chemotaxis to inducer phenolics is selectively reduced or abolished by mutations in certain nod genes governing nodulation efficiency or host specificity. Conversely, mutations in rhizobia that affect general motility or chemotaxis have substantial effects on nodulation efficiency and competitiveness. These findings suggest that microbes entering the rhizosphere environment may utilize minor, non-nutrient components in root exudates as signals to guide their movement towards the root surface and elicit changes in gene expression appropriate to this environment.     

大豆结瘤突变体成分的初步分析及其对硝酸还原酶活性和结瘤的影响

超结瘤大豆（Ｇｌｙｃｉｎｅ ｍ ａｘ （Ｌ．） Ｍｅｒｒ．） ｎｔｓ ３８２ 和不结瘤大豆Ｎｏｄ ４９ 的叶和根组织水提取物经Ｓｅｐｈａｄｅｘ Ｇ２５ 过滤、洗脱,再根据洗脱物对硝酸还原酶（ＮＲ）活性的影响可划分为４ 个组分（ｆｒａｃｔｉｏｎ）样品,即ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ１、ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ２、ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ３ 和ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ４。其中, ｎｔｓ３８２ Ｆ２ 和Ｆ４ 抑制ＮＲ 活性作用在接种ＵＳＤＡ１１０ 后明显下降, 但接种的ｎｔｓ ３８２ Ｆ２ 却能提高大豆Ｂｒａｇｇ 的结瘤数达一倍, 而接种的ｎｔｓ ３８２ Ｆ３ 和Ｆ４ 的作用不明显。ＮＲ 活性抑制因子不是刺激结瘤的因子, 刺激结瘤的因子主要分布在接种的ｎｔｓ３８２ Ｆ２ 部分中。与这一现象相反, Ｎｏｄ ４９ Ｆ２ 和Ｆ４ 抑制ＮＲ活性的作用在接种后更强, 且也抑制大豆ｎｔｓ ３８２ 的结瘤, 其中Ｎｏｄ ４９ Ｆ４ 抑制结瘤的作用基本不能逆转。抑制结瘤因子主要分布在接过种的Ｎｏｄ ４９ Ｆ４ 部分中     

Presence of unique repeated insertion sequences in nodulation genes of Rhizobium ‘hedysari’

Cloning and sequencing of DNA from a symbiotic large plasmid in Rhizobium hedysari strain IS 123 required for its nodulation of the mediterranean legume crop Hedysarum coronarium (sulla) and complementation studies of nod^- mutant derivatives led to the characterization of a 30-kb region containing common and host-specific nod genes. This DNA region also contained at least six copies of a novel insertion sequence-like structure, some of which appeared to have suffered deletions. This 0.8 kb novel element carries two 17-bp flanking inverted repeats and an open reading frame showing homology with a transposase from Staphylococcus aureus. Hybridization studies revealed that several strains of Rhizobium hedysari carry this element in various copy number. The six copies in strain IS 123 appear clustered specifically within the pSym nod region.The significance of this IS element in rhizobia and its possible use as a probe for taxonomic and phylogenetic studies of Rhizobiaceae is addressed.     

Alnus rubra nodulation capacity of soil under five species from harvested forest sites in coastal British Columbia

Nodulation of Alnus rubra seedlings after inoculation with soil from under A. rubra, Betula papyrifera. Rubus lacianutus, R. spectabilis, and R.ursinus on 2 recently harvested sites was compared. Nodulation capacity was low compared to other published reports, ranging from 0 to 18.9 infective units cm^-3 of soil and was significantly affected by the site and plant species. Nodulation capacity of soil under alder was significantly higher than under all other species except R. spectabilis, regardless of site. The lowest nodulation capacity was found in soil under B. papyrifera.Joint appointment with Dept. of Soil Science, Faculty of Agricultural Sciences     

Influence of drought on competition between selected Rhizobium meliloti strains and naturalized soil rhizobia in alfalfa

Drought is an important environmental factor that can affect rhizobial competition and N₂ fixation. Three alfalfa (Medicago sativa L. and M. falcata L.) accessions were grown in pots containing soil from an irrigated (Soil 1) and a dryland (Soil 2) alfalfa field in northern Utah, USA. Mutants of three strains of Rhizobium meliloti Dang. from Pakistan (UL 136, UL 210, and UL 222) and a commercial rhizobial strain 102F51a were developed with various levels of resistance to streptomycin. Seeds inoculated with these individual streptomycin-resistant mutants were sown in the two soils containing naturalized rhizobial populations. Soils in the pots were maintained at −0.03, −0.5, and −1.0 MPa. After 10 weeks, plants were harvested and nodule isolates were cultured on agar medium with and without streptomycin to determine nodule occupancy (proportion of the nodules occupied by introduced rhizobial strains). Number of nodules, nodule occupancy, total plant dry weight, and shoot N were higher for Soil 1 than Soil 2. Number of nodules, plant dry weight, and shoot N decreased as drought increased from −0.03 to −1.0 MPa in the three alfalfa accessions. Rhizobial strains UL 136 and UL 222 were competitive with naturalized alfalfa rhizobia and were effective at symbiotic N₂ fixation under drought. These results suggest that nodulation, growth, and N₂ fixation in alfalfa can be improved by inoculation with competitive and drought-tolerant rhizobia and may be one economically feasible way to increase alfalfa production in water-limited environments. Joint contribution from USDA-ARS and the Utah Agric. Exp. Sta., Utah State Univ., Logan, UT 84322-4810, USA. Journal Paper No. 4931. Joint contribution from USDA-ARS and the Utah Agric. Exp. Sta., Utah State Univ., Logan, UT 84322-4810, USA. Journal Paper No. 4931.     

Direct shoot formation in spontaneously occurring root pseudonodules of alfalfa (Medicago sativa L.)

Summary A simple and rapid procedure for direct organogenesis from root nodulelike structures of alfalfa (Medicago sativa L.) line SGg, spontaneously induced on growth regulator-free Gamborg (B₅) medium, was developed. Prolific adventitious shoot initiation was obtained using a combination of 1.0 mg/liter TIBA and 0.5 mg/liter 2iP. Transfer of shoots to a medium containing 0.5 mg/liter ABA and reduced concentration of TIBA (0.5 mg/liter) before rooting markedly stimulated shoot development. Regenerated shoots rooted easily and revealed the early appearance of nodulelike structures on basal medium (B₅) lacking growth regulators. Analysis of endogenous growth regulator levels of SGg roots maintained on growth regulators free media, showed that spontaneous shoot appearances was correlated with high cytokinin-to-auxin ratios.     

Spatial distribution of root activity and nitrogen fixation in sorghum/pigeonpea intercropping on an Indian Alfisol

A medium-duration pigeonpea cultivar (ICP 1–6) and a hybrid sorghum (CSH 5) were grown on a shallow Alfisol in monocropping and intercropping systems. Using a monolith method, spatial distribution of nodulation, acetylene reduction activity (ARA) and root respiration were measured.The number, mass and ARA of nodules decreased exponentially with distance from the plant base except at the late reproductive stage. Nodulation and ARA tended to be higher in the intercrop than in the monocrop.Respiration rate of roots increased with distance from the plant base and reached a maximum value at about 20–30 cm. The rate was higher in pigeonpea than in sorghum and also higher in intercrop than in monocrop.This study suggests that pigeonpea roots are physiologically more active than sorghum roots, implying that pigeonpea may become a strong competitor for nutrients in the soil when intercropped. The nitrogen-fixing ability of pigeonpea may be enhanced by intercropping because the sorghum rapidly absorbed inorganic N which would otherwise inhibit N₂ fixation.     

Metabolite effectors for short-term nitrogen-dependent enhancement of phosphoenolpyruvate carboxylase activity and decrease of net sucrose synthesis in wheat leaves

It has been demonstrated previously that field pea (Pisum sativum L. cv. Express) grown in hydroponic culture on a complete nutrient solution with low NH₄⁺ concentrations (<0.5 mM) will produce a larger than normal proliferation of nodules. Peas grown in the absence of mineral N in hydroponic culture have been shown to rapidly autoregulate nodulation, forming a static nodule number by 14 to 21 days after planting. The present study further characterizes the effect of NH₄⁺ concentration in hydroponic culture on nodulation and nodule growth. Peas were grown continually for 4 weeks at NH₄⁺ concentrations that were autoregulatory (0.0 mM), stimulatory (0.2 mM) or inhibitory (1.0 mM), or peas were transferred between autoregulatory or NH₄⁺ inhibited and stimulatory solutions after 2 weeks. The peas nodulated as expected when grown under constant autoregulatory, stimulatory or inhibitory concentrations of NH₄⁺. When peas were transferred from the inhibitory (1.0 mM) to the stimulatory solution (0.2 mM) a massive proliferation of nodule primordia over the entire root system was observed within 3 days of the transfer. When they were transferred from the autoregulatory (0.0 mM) to the stimulatory (0.2 mM) solution a 10-day delay occurred before a proliferation in nodule primordia occurred at distal regions of the root system. These findings support our hypothesis that low concentrations (<1.0 mM) of NH₄⁺ in hydroponic culture cause a suppression of autoregulation in pea. In addition, the temporal and spatial differences in nodule proliferation between transfer treatments demonstrate at a whole plant level that autoregulation and NH₄⁺ inhibition suppress early nodule development via different mechanisms.     

Soybean response to nodulation by rhizobitoxine-producing bradyrhizobia as influenced by nitrate application

Foliar chlorosis of soybean (Glycine max [L.] Merr.) resulting from nodulation by rhizobitoxine-producing (RT⁺) strains of Bradyrhizobium japonicum is commonly less severe in the field than under greenhouse conditions. Differences in nutritional conditions between the field and greenhouse may contribute to this phenomenon. In particular, field-grown plants obtain some N from soil sources, whereas in the greenhouse soybean is often grown in low-N rooting media to emphasize symbiotic responses. Therefore, we examined the effect of NO₃ ^- on the expression of RT-induced symptoms. Soybean plants inoculated with RT⁺ bradyrhizobia were grown for 42 days in horticultural vermiculite receiving nutrient solution amended with 0.0, 2.5, or 7.5 mM KNO₃. Foliar chlorosis decreased with increasing NO₃ ^- application whereas nodule mass per plant was generally increased by NO₃ ^- application. Total amounts of nodular RT remained constant or increased with NO₃ ^- application, but nodular concentrations of RT decreased. Chlorosis severity was negatively correlated with shoot dry weight, chlorophyll concentration, and total shoot N content. It was concluded that application of NO₃ ^- can reduce the negative effects of RT production on the host plant. This suggests that any NO₃ ^- present in field soils may serve to limit chlorosis development in soybeans.Abbreviations RT rhizobitoxine - RT⁺ rhizobitoxine-producing - Lb leghemoglobin Published as Miscellaneous Paper No. 1429 of the Delaware Agricultural Experiment Station.