Genotypic variation of nodules’ enzymatic activities in symbiotic nitrogen fixation among common bean (Phaseolus vulgaris L.) genotypes grown under salinity constraint

The effect of salt stress, under glasshouse conditions, was studied on plant biomass, nodulation, and activities of acid phosphatases (APase, EC 3.1.3.2) and trehalose 6-phosphate phosphatase (TPP, EC 3.1.3.12) in the symbiosis common bean (Phaseolus vulgaris L.)-rhizobia nodules. Four common bean recombinant inbred lines (147, 115, 104 and 83) were separately inoculated, with CIAT 899 or RhM11 strains and grown in hydroaeroponic culture. Two NaCl levels (0 and 25 mM NaCl plant^?1 week^?1 corresponding, respectively, to the control and the salt treatment) were applied and the culture was assessed during 42 days after their transplantation. The results showed that the nodulation of these lines was not affected by salinity except for the line 83 inoculated with CIAT 899, whose nodule dry weight decreased by 48.24 % compared with the corresponding controls. For the other symbiotic combinations, shoot and root biomasses were not significantly affected by salt constraint. Salinity stress generally reduced acid phosphatise and trehalose phosphate phosphatase activities in nodules that were less affected in plants inoculated with RhM11. Based on our data, it appears that nodule phosphatase activity may be involved in salinity tolerance in common beans and the levels of salt tolerance depend principally on specific combination of the rhizobial strain and the host cultivar.     

Changes in ascorbate peroxidase, catalase, guaiacol peroxidase and superoxide dismutase activities in common bean (Phaseolus vulgaris) nodules under salt stress

To analyse nodular antioxidant enzyme expression in response to salt stress, Phaseolus vulgaris genotype BAT477 was inoculated with reference strain CIAT899, and treated with 50 mM NaCl. Plant growth, nodulation and nitrogen fixing activity were analysed. Results showed that: (1) all parameters, particularly in nodules, were affected by salt treatments, and (2) confirmed preferential growth allocation to roots. The ARA was significantly decreased by salt treatments. Protein dosage confirmed that nodules were more affected by salt treatment than were roots. We analysed superoxide dismutase, catalase, ascorbate peroxidase and peroxidase in nodules, roots and a free rhizobial strain. Our results indicated that SOD and CAT nodular isozymes had bacterial and root origins. The SOD expressed the same CuZn, Fe and Mn SOD isoforms in nodules and roots, whereas in free rhizobia we found only one Fe and Mn SOD. APX and POX nodule and root profiles had only root origins, as no rhizobial band was detected. Under salt stress, plant growth, nitrogen fixation and activities of antioxidant defense enzymes in nodules were affected. Thus, these enzymes appear to preserve symbiosis from stress turned out that NaCl salinity lead to a differential regulation of distinct SOD and POX isoenzyme. So their levels in nodules appeared to be consistent with a symbiotic nitrogen fixing efficiency hypothesis, and they seem to function as the molecular mechanisms underlying the nodule response to salinity.     

Symbiotic performance of common bean and soybean co-inoculated with rhizobia and <Emphasis Type="Italic">Chryseobacterium balustinum</Emphasis> Aur9 under moderate saline conditions

The effect of co-inoculating beans and soybeans with rhizobia and Chryseobacterium, a plant growth promoting bacteria (PGPR), was studied under conditions of mild saline stress. Chryseobacterium balustinum Aur9 was used with Rhizobium tropici CIAT899 or R. etli ISP42 to inoculate common bean (Phaseolus vulgaris L.), or jointly with Ensifer (Sinorhizobium) fredii SMH12 and HH103 to inoculate soybean (Glycine max (L.) Merrill). The effect of co-inoculation was studied by following nodule primordia initiation, nodulation kinetics and symbiotic performance in plants grown under moderate saline conditions (25 mM NaCl). In common bean, co-inoculation improved nodule primordia formation when compared with single inoculation (R. tropici CIAT899). However, co-inoculation did not provide benefits in the development of nodule primordia in soybean with E. fredii SMH12. The kinetic of nodulation in bean was also favored by double inocula resulting in a higher number of nodules. Long-term effects of co-inoculation on beans and soybeans depended on the rhizobial species used. In both, control and saline conditions, co-inoculation of R. tropici CIAT899 and C. balustinum Aur9 improved bean growth when compared with the single inoculation (CIAT899). However, the positive effect of double inocula on plant growth did not occur when using R. etli ISP42. Soybean plants receiving double inoculation (E. fredii SMH12 and C. balustinum Aur9) showed better symbiotic performance, mostly under saline stress, than with a single inoculation. The results indicate that co-inoculation with C. balustinum and rhizobia under mild saline conditions partially relieves the salt-stress effects, although do not always result advantageous for symbiotic N₂ fixation in legume plants.     

Modulation of symbiotic efficiency and nodular antioxidant enzyme activities in two <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> genotypes under salinity

To analyse nodular expression of antioxidant enzymes depending on plant genotype and salinity, two Phaseolus vulgaris genotypes, tolerant BAT477 and sensitive COCOT, were inoculated with the reference strain Rhizobium tropici CIAT899 and grown under 25 and 50 mM NaCl. Plant growth, nodulation and nitrogen fixing activity measured by the acetylene reducing activity (ARA) as an indicator of nitrogenase (E.C. 1.7.9.92) activity were more affected by salt concentrations in COCOT than in BAT477, particularly with 50 mM NaCl. Electrophoresis analysis of antioxidant enzymes in nodules, roots and free-living rhizobia showed that only catalase (CAT E.C. 1.11.1.6) isoenzymes varied with genotype. The sensitive genotype showed lower antioxidant enzyme activities than tolerant genotype and it was more affected by salinity. In the tolerant genotype catalase and ascorbate peroxidase (APX, E.C. 1.11.1.11) were inhibited by salt stress, whereas superoxide dismutase (SOD, E.C. 1.15.1.1) and peroxidase (POX, E.C. 1.11.1.7) were activated by salinity. Statistical analysis allowed suggesting that tolerance to salinity is associated with a differential regulation of distinct superoxide dismutase and peroxidase activities.     

Nitrogenase and antioxidant enzyme activities in Phaseolus vulgaris nodules formed by Rhizobium tropici isogenic strains with varying tolerance to salt stress

Common bean plants inoculated with salt-tolerant Rhizobium tropici wild-type strain CIAT899 formed a more active symbiosis than did its decreased salt-tolerance (DST) mutant derivatives (HB8, HB10, HB12 and HB13). The mutants formed partially effective (HB10, HB12) or almost ineffective (HB8, HB13) nodules (Fix(d)) under non-saline conditions. The DST mutant formed nodules that accumulated more proline than did the wild-type nodules, while soluble sugars were accumulated mainly in ineffective nodules. Under salt stress, plant growth, nitrogen fixation, and the activities of the antioxidant defense enzymes of nodules were affected in all symbioses tested. Overall, mutant nodules showed lower antioxidant enzyme activities than wild-type nodules. Levels of nodule catalase appeared to correlate with symbiotic nitrogen-fixing efficiency. Superoxide dismutase and dehydroascorbate reductase seem to function in the molecular mechanisms underlying the tolerance of nodules to salinity.     

The Physiological Mechanisms Underlying N₂-Fixing Common Bean (<i>Phaseolus vulgaris</i> L.) Tolerance to Iron Deficiency

Iron is an essential element for plants as well as all living organisms, functioning in various physiological and biochemical processes such as photosynthesis, respiration, DNA synthesis, and N₂ fixation. In the soil, Fe bioavailability is extremely low, especially under aerobic conditions and at high pH ranges. In contrast, plants with nodules on their roots that fix atmospheric nitrogen need much more iron. To highlight the physiological traits underlying the tolerance of N₂-fixing common bean to iron deficiency, two genotypes were hydroponically cultivated in a greenhouse: Coco nain (CN) and Coco blanc (CB). Plants were inoculated with an efficient strain of Rhizobium tropici, CIAT899, and received a nutrient solution added with 0 µM Fe (severe Fe deficiency, SFeD), 5 µM Fe (moderate Fe deficiency, MFeD) or 45 µM Fe (control, C). Several physiological parameters related to photosynthesis and symbiotic nitrogen fixation were then analyzed. Iron deficiency significantly reduced whole plant and nodule growth, chlorophyll biosynthesis, photosynthesis, leghemoglobin (LgHb), nitrogenase (N₂ase) activity, nitrogen, and Fe nutrition, with some genotypic differences. As compared to CB, CN maintained better Fe allocation to shoots and nodules, allowing it to preserve the integrity of its photosynthetic and symbiotic apparatus, thus maintaining the key functional traits of the plant metabolism (chlorophyll biosynthesis and photosynthesis in shoots, leghemoglobin accumulation, and nitrogenase activity in root nodules). Plant growth depends on photosynthesis, which needs to be supplied with sufficient iron and nitrogen. Fe deficiency stress index (FeD-SI) and Fe use efficiency (FeUE) are two physiological traits of tolerance that discriminated the studied genotypes.     

Growth, nitrogen fixation and ammonium assimilation in common bean (Phaseolus vulgaris): effect of phosphorus

The impact of phosphorous nutrition on plant growth, symbiotic N₂ fixation, ammonium assimilation, carbohydrate and amino-acid accumulation, as well as on nitrogen, phosphorus and ATP content in tissues in common bean ( Phaseolus vulgaris ) plants was investigated. Plants inoculated with Rhizobium tropici CIAT899 were grown in Leonard jars under controlled conditions, with P-deficient (0 and 0.1 m M ), P-medium (0.5, 1 and 1.5 m M ) and P-high (2 m M ) conditions in a N-free nutrient solution. The P application, increased leaf area, whole plant DW (67%), nodule biomass (4-fold), and shoot and root P content (4- and 6-fold, respectively) in plant harvested at the onset of flowering (28-days-old). However, P treatments decreased the total soluble sugar and amino acid content in vegetative organs (leaf, root and nodules). The root growth proved less sensitive to P deficiency than did shoot growth, and the leaf area was significantly reduced at low P-application. The absence of a relationship between shoot N content, and P levels in the growth medium could indicate that nitrogen fixation requires more P than does plant growth. The optimal amount for the P. vulgaris – R. tropici CIAT899 symbiosis was 1.5 m M P, this treatment augmented nodule-ARA 20-fold, and ARA per plant 70-fold compared with plants without P application.     

Osmotic stress affects water relations, growth, and nitrogen fixation in Phaseolus vulgaris plants

The effects of mannitol-mediated osmotic stress on water relations, plant growth and symbiotic N₂-fixation in four common bean (Phaseolus vulgaris) lines (Coco Blanc, BAT 477, BRB 77 and Flamingo) were studied. After germination, seedlings were inoculated with a reference strain (Rhizobium tropici CIAT 899) and aerohydroponically grown in a glasshouse. Osmotic stress was applied by 50 mM mannitol. Plants were harvested 4 weeks after osmotic stress application. Measured parameters were plant water relations, growth, nodule development, and symbiotic N₂-fixation (SNF) as well as leghemoglobin contents. Osmotic stress induced significant changes in water relations, growth and symbiotic N₂-fixation in stressed plants compared to control ones in all lines studied. A noticeable different behaviour was observed in the end of the treatment: Flamingo was the most tolerant line, whereas Coco blanc was the most sensitive, the two other lines exhibited an intermediate behaviour. The four bean lines displayed significant differences in their responses to osmotic stress. This study indicated that the relative tolerance of Flamingo line seems to be due to its ability to maintain higher leaf water potential, adequate leaf area and abundant and efficient nodular system, which in turn determines an important rate of SNF.     

Response of common bean lines to inoculation: comparison between the <Emphasis Type="Italic">Rhizobium tropici</Emphasis> CIAT899 and the native <Emphasis Type="Italic">Rhizobium etli</Emphasis> 12a3 and their persistence in Tunisian soils

Since Phaseolus vulgaris (L) is poorly nodulated in all regions of Tunisia where this crop is grown, the response of common-bean lines CocoT and Flamingo to inoculation with reference Rhizobium tropici CIAT 899 or native rhizobia, namely Sinorhizobium fredii 1a6, Rhizobium etli 12a3, and Rhizobium gallicum 8a3, was studied in a field station. Since R. etli 12a3 was found to be the most effective native rhizobium, it was subsequently compared with R. tropici CIAT 899 in a broader study in two stations over 3 years. A significant interaction between bean and rhizobia was observed for nodule number, shoot dry weight, grain yield, and contents of nitrogen and chlorophyll. The native rhizobia was more efficient than CIAT899 for Flamingo, though not for CocoT. The Enzyme-linked immunosorbent assay technique was used with polyclonal antibody to assess the occupancy in nodule and persistence in soil of the inoculated rhizobia. For both stations the nodule occupancy was 100% during the first year for each rhizobium, but during the next 2 years, between 7 and 15% of nodules were formed by the rhizobia inoculated in the neighboring plot. It is concluded that the first-year inoculation is sufficient to maintain an adequate rate of nodulation during three growth cycles, and that the native R etli can be recommended for the common-bean inoculation in similar soils of Tunisia.     

Selection of Competitive Strains of Rhizobium Nodulating Phaseolus Vulgaris and Adapted to Environmental Conditions in Egypt,Using the Gus-Reporter Gene Technique

Two Rhizobium etli strains, EBRI 2 and EBRI 26, isolated from Egypt were tested for nodulation competitiveness on beans using Rhizobium tropici CIAT 899G as the competing strain. The insertion of the gus-reporter transposon mTn5ssgusA30 did not alter the nodulation or nitrogen fixation capacity of mutant strain CIAT 899G compared to the wild type. At neutral pH, R. etli strains EBRI 2 and EBRI 26 were more competitive than CIAT 899G with the bean cultivar Saxa. These two strains gave nodule occupancies of 52.1 and 61.1% competing with equal cell numbers of CIAT 899G. Nodule occupancies from these two native strains increased with the bean cultivar Giza 6 from Egypt to 66 and 67.5%. Based on these results, cultivar Giza 6 was used to select the most competitive strains under stress of salinity or alkalinity as a major problem for a large part of Egyptian soils. Under stress of salinity (0.2% NaCl or 34.2 mM NaCl), the salt-sensitive strain EBRI 2 was more competitive than the salt-resistant strain EBRI 26. Strain EBRI 2 gave 87.4% but strain EBRI 26 gave 63.7% nodule occupancy against CIAT 899G. The same trend of results was observed under stress of alkalinity (pH 8). Strain EBRI 2 occupied 83% while Strain EBRI 26 occupied 53.2%.     

Genotypic variation of N2-fixing common bean (Phaseolus vulgaris L.) in response to iron deficiency

In calcareous soils, the yield of grain legumes is often limited by the lower availability of iron (Fe), especially when they depend upon symbiosis with root nodule bacteria for their N nutrition. In order to explore the variability of responses of N(2)-fixing common bean to Fe deficiency the common bean white-seeded lines Striker and Coco blanc, and coloured-seeded lines SVM-29-21 and ARA14 were inoculated with Rhizobium tropici (CIAT 899) and cultivated hydroaeroponically with a N-free nutrient solution supplied or not with 45microM Fe. Differences among lines were observed: Fe-deficiency-induced-chlorosis on young leaves was earlier and more severe in some lines than others. Nodule development and N(2)-fixing capacity was less affected in line ARA14 which preferentially allocated Fe towards nodules. Results suggest that Fe use efficiency for symbiotic nitrogen fixation (FeUE SNF) could be used to screen tolerant bean lines to Fe deficiency in condition of symbiotic nitrogen fixation.     

Combined inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases phosphorus use efficiency for symbiotic nitrogen fixation in common bean (Phaseolus vulgaris L.)

This study compared the response of common bean (Phaseolus vulgaris L.) to arbuscular mycorrhizal fungi (AMF) and rhizobia strain inoculation. Two common bean genotypes i.e. CocoT and Flamingo varying in their effectiveness for nitrogen fixation were inoculated with Glomus intraradices and Rhizobium tropici CIAT899, and grown for 50 days in soil–sand substrate in glasshouse conditions. Inoculation of common bean plants with the AM fungi resulted in a significant increase in nodulation compared to plants without inoculation. The combined inoculation of AM fungi and rhizobia significantly increased various plant growth parameters compared to simple inoculated plants. In addition, the combined inoculation of AM fungi and rhizobia resulted in significantly higher nitrogen and phosphorus accumulation in the shoots of common bean plants and improved phosphorus use efficiency compared with their controls, which were not dually inoculated. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve the efficiency in phosphorus use for symbiotic nitrogen fixation especially under phosphorus deficiency.     

Co-inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases P use efficiency for N2 fixation in the common bean (Phaseolus vulgaris L.) under P deficiency in hydroaeroponic culture

Common bean (Phaseolus vulgaris L.) genotypes CocoT and Flamingo were inoculated with Rhizobium tropici CIAT899 and Glomus intraradices (Schenck & Smith) and grown under sufficient versus deficient phosphorus supply for comparing the effects of double inoculation on growth, nodulation, mycorrhization of the roots, phosphorus use efficiency and total nitrogen. Although the double inoculation induced a significant increase in all parameters whatever the phosphorus supply in comparison to control, significant differences were found among genotypes and treatments. Nevertheless, the highest phosphorus use efficiency and plant total nitrogen were found under P deficiency in combination with arbuscular mycorrhizal fungi. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve symbiotic nitrogen fixation even under phosphorus deficiency.     

Soil chemical properties and dieback of Quercus robur in Atlantic wet forests after a weather extreme

Background and aims

Burkholderia phymatum strain GR01 is a recently reported common bean (Phaseolus vulgaris) symbiont isolated from nodules of plants grown in semi-arid soils in Morocco. The osmotolerance of B. phymatum GR01N under free-living and in symbiotic association with P. vulgaris was investigated in this study.

Methods

The osmotolerance of B. phymatum GR01N was checked by growing cells in the presence of varying concentrations of NaCl or sucrose, and the cellular solutes were analyzed in cell extracts by ¹³C-nuclear magnetic resonance (NMR) spectroscopy. Nodule occupancy was checked in P. vulgaris grown in the presence of 25, 35 or 50 mM NaCl and inoculated with a mixture of B. phymatum GR01N and R. tropici CIAT899R cells. The effect of salt stress on nodule biomass, plant dry weight, plant nitrogen content and leghaemoglobin content of nodules was also analyzed in plants inoculated with either B. phymatum GR01N or R. tropici CIAT899R and grown in the presence of 25 or 35 mM NaCl.

Results

Burkholderia phymatum strain GR01N showed increased tolerance to osmotic stress under free-living conditions as compared to the reference strain R. tropici CIAT899R. Strain GR01N accumulated trehalose, mannitol and alanine in response to saline stress, suggesting their role in the observed osmoloterance. Under conditions of saline stress, P. vulgaris plants nodulated by B. phymatum GR01N showed increased plant dry weight and nitrogen fixation, when compared to those inoculated with R. tropici CIAT899R. Nodule competition assays revealed that B. phymatum GR01N had higher levels of nodule occupancy than R. tropici CIAT899R in P. vulgaris plants grown under saline conditions.

Conclusions

Burkholderia phymatum strain GR01N displays a remarkable osmotolerance under free-living and symbiotic conditions.     

Influence of salinity and abscisic acid on the O2 uptake by N2-fixing nodules of common bean

The effects of NaCl and ABA on the respiration of N₂-fixing nodules were analysed in common bean (Phaseolus vulgaris) inoculated with Rhizobium tropici the reference strain CIAT899. Shoot and nodule growth was more inhibited by NaCl than root growth. The O₂ uptake by nodulated roots at 21 kPa O₂ was significantly inhibited by salinity. Raising pO₂ stimulated nodule respiration more under NaCl treatment than for the control, although it did not compensate totally for the inhibitory effect of NaCl. Short NaCl application was less destructive than long term application. Also, the external application of ABA inhibited nodule respiration, and this inhibition was partly compensated by raising pO₂.     

Quantitative trait locus analysis of symbiotic nitrogen fixation activity in the model legume <Emphasis Type="Italic">Lotus japonicus</Emphasis>

Many legumes form nitrogen-fixing root nodules. An elevation of nitrogen fixation in such legumes would have significant implications for plant growth and biomass production in agriculture. To identify the genetic basis for the regulation of nitrogen fixation, quantitative trait locus (QTL) analysis was conducted with recombinant inbred lines derived from the cross Miyakojima MG-20 × Gifu B-129 in the model legume Lotus japonicus. This population was inoculated with Mesorhizobium loti MAFF303099 and grown for 14 days in pods containing vermiculite. Phenotypic data were collected for acetylene reduction activity (ARA) per plant (ARA/P), ARA per nodule weight (ARA/NW), ARA per nodule number (ARA/NN), NN per plant, NW per plant, stem length (SL), SL without inoculation (SLbac−), shoot dry weight without inoculation (SWbac−), root length without inoculation (RLbac−), and root dry weight (RWbac−), and finally 34 QTLs were identified. ARA/P, ARA/NN, NW, and SL showed strong correlations and QTL co-localization, suggesting that several plant characteristics important for symbiotic nitrogen fixation are controlled by the same locus. QTLs for ARA/P, ARA/NN, NW, and SL, co-localized around marker TM0832 on chromosome 4, were also co-localized with previously reported QTLs for seed mass. This is the first report of QTL analysis for symbiotic nitrogen fixation activity traits.     

Inoculation and production of container-grown red alder seedlings

Summary The inoculation ofAlnus rubra (red alder) withFrankia sp. can lead to a highly efficient symbiosis. Several factors contribute to the successful establishment of nitrogenfixing nodules: (1) quantity and quality ofFrankia inoculant; (2) time and method of inoculation; (3) nutritional status of the host plant.Frankia isolates were screened for their ability to nodulate and promote plant growth of container-grown red alder. Inoculations were performed on seedlings and seeds. Apparent differences in symbiotic performance could be seen when seeds or seedlings were inoculated. Plants inoculated at planting performed significantly better than those inoculated four weeks later in terms of shoot height, nodule number and shoot dry weight. If inoculation was delayed further, reduction in shoot height, nodule number and shoot dry weight resulted. The effect of fertilizer was also investigated with regard to providing optimal plant growth after inoculation. Plants receiving 1/5 Hoagland's solution minus nitrogen showed maximal plant growth with abundant nodulation. Plants receiving 1/5 Hoagland's solution with nitrogen showed excellent plant growth with significantly reduced nodulation.     

Is genotypic variation of H⁺ efflux under P deficiency linked with nodulated-root respiration of N₂ fixing common-bean (Phaseolus vulgaris L.)?

To examine genotypic variation of common bean in growth, phosphorus uptake, nodulated-root proton release, and nodule gas permeability, seven common bean recombinant inbred lines (RIL) from the cross of BAT477×DOR364 were inoculated by Rhizobium tropici CIAT 899 and grown in hydroaeroponic culture under glasshouse conditions. A positive correlation was observed between shoot and nodule biomass for most of the studied RILs. Under P deficiency, the tolerant common bean RILs acidified more of their rhizosphere than the sensitive ones. The proton release of the RILs 147, 124, 104, 75 and RIL34 was positively correlated with nodule O? permeability. We conclude that nodulated common bean plants release a substantial amount of H? into the rhizosphere that is linked to the symbiotic N? fixation. It depends upon the nodule permeability to O? diffusion, and varies with genotype.     

A ClC chloride channel homolog and ornithine-containing membrane lipids of Rhizobium tropici CIAT899 are involved in symbiotic efficiency and acid tolerance

Rhizobium tropici CIAT899 is highly tolerant to several environmental stresses and is a good competitor for nodule occupancy of common bean plants in acid soils. Random transposon mutagenesis was performed to identify novel genes of this strain involved in symbiosis and stress tolerance. Here, we present a genetic analysis of the locus disrupted by the Tn5 insertion in mutant 899-PV9, which lead to the discovery of sycA, a homolog of the ClC family of chloride channels and Cl-/H+ exchange transporters. A nonpolar deletion in this gene caused serious deficiencies in nodule development, nodulation competitiveness, and N2 fixation on Phaseolus vulgaris plants, probably due to its reduced ability to invade plant cells and to form stable symbiosomes, as judged by electron transmission microscopy. A second gene (olsC), found downstream of sycA, is homologous to aspartyl/asparaginyl beta-hydroxylases and modifies two species of ornithine-containing lipids in vivo, presumably by hydroxylation at a still-unknown position. A mutant carrying a nonpolar deletion in olsC is symbiotically defective, whereas overexpressed OlsC in the complemented strain provokes an acid-sensitive phenotype. This is the first report of a ClC homolog being essential for the establishment of a fully developed N2-fixing root nodule symbiosis and of a putative beta-hydroxylase that modifies ornithine-containing membrane lipids of R. tropici CIAT899, which, in turn, are contributing to symbiotic performance and acid tolerance.     

Growth, nitrogen fixation and ion distribution in Medicago truncatula subjected to salt stress

This study compared the growth, nodulation, N₂ fixation, and ion distribution in three Medicago truncatula lines, in response to salt in nutrient solution. Two local lines (TN8.20 and TN6.18) and a reference line (Jemalong 6) were inoculated with a reference strain Sinorhizobium meliloti 2011, a very tolerant strain to salinity (700 mM NaCl) and grown in a controlled glasshouse with or without 75 mM NaCl. A genotypic variation in tolerance to salt was found: TN6.18 was the most sensitive line whereas TN8.20 was the most tolerant. The relative tolerance of TN8.20 was concomitant with the lowest leaf Na⁺ concentration and the highest nodule biomass production. However, nodule efficiency (amount of nitrogen fixed per g dry weight nodule) decreased in all lines. Results suggest that the tolerance to salt seems to depend on the host plant ability to protect its leaves against an excessive Na⁺ (and Cl^?) accumulation, and its ability to maintain the development of an abundant nodular system, which in turn determines an important rate of nitrogen fixation and allows the plants to conserve their growth potentialities. The loss of the nodular efficiency under salt stress seems to be compensated by a large nodule biomass.