Gabriel Favelukes

Adsorption of Rhizobium meliloti to alfalfa roots: Dependence on divalent cations and pH

Adsorption ofRhizobium meliloti L5-30 in low numbers to alfalfa (Medicago sativa L.) roots was dependent on the presence of divalent cations, and required neutral pH. Adsorption was proportional to Ca and/or Mg concentrations up to 1.5 mM. Ca was not substituted by Sr, Ba or Mn. Adsorption was abolished and viability decreased at pH≤6. When lowering pH, higher Ca concentrations were required to attain similar adsorption levels, indicating a marked interactive effect between Ca and H ions. Pretreatment of the roots with Ca and low pH did not affect subsequent adsorption of the bacteria. However, Ca pretreatment ofR. meliloti sustained further adsorption at low Ca levels and low pH substantially affected their ability to adsorb. Low pH appears to affect the stability of binding causing desorption of the previously bound bacteria. The presence of saturating concentrations of heterologousR. leguminosarum bv.trifolii A118, did not prevent the expression of divalent cations and pH requirements, as well as their interaction. Our results suggest that rhizobial binding to the root surface already shows the Ca and pH dependence of alfalfa nodulation, which was generally associated to some event prior to rhizobial penetration of root hairs.

Comparison of drought tolerance in nitrogen-fixing and inorganic nitrogen-grown common beans.

In this work, we evaluated how the use of alternative N sources affects drought-stress tolerance in common beans. To this end, plants were cultivated employing either N(2) fixation or two levels of inorganic nitrogen: 1 mM NH(4)NO(3) (limiting) or 10 mM NH(4)NO(3) (sufficient). Drought was imposed by withholding watering at 30 days after planting (DAP) - coinciding with flowering. At 20 DAP, growth and N content were significantly higher in NH(4)NO(3)-sufficient plants than in N(2)-fixing and NH(4)NO(3)-limited beans. At later times, only N(2)-fixing and NH(4)NO(3)-sufficient plants continued assimilating N and growing, with the NH(4)NO(3)-sufficient plants being consistently bigger. After 10 days of stress (40 DAP), desiccation was evident, but only NH(4)NO(3)-sufficient plants suffered drought-induced senescence. After 20 days of stress (50 DAP), N content increased in NH(4)NO(3)-sufficient but not in N(2)-fixing beans, despite the latters lesser state of wilt. Pod dry weight dropped 43% in NH(4)NO(3)-sufficient beans with respect to well-watered plants, while remaining constant in N(2)-fixing beans. Under drought conditions, the number of pods limited pod yield regardless of the nitrogen source used; nevertheless, the translocation of soluble matter to pods continued in both NH(4)NO(3)-sufficient and N(2)-fixing beans. We conclude that common beans grown under conditions of N(2) fixation were more drought tolerant than those provided with sufficient levels of NH(4)NO(3). The most stress-sensitive traits in these plants were the incorporation of N into their shoots and the number of pods remaining on them.

Improved Soybean Root Association of N-Starved Bradyrhizobium japonicum

In this study, we addressed the effects of N limitation in Bradyrhizobium japonicum for its association with soybean roots. The wild-type strain LP 3001 grew for six generations with a growth rate of 1.2 day(-1) in a minimal medium with 28 mM mannitol as the carbon source and with the N source [(NH(4))(2)SO(4)] limited to only 20 microM. Under these conditions, the glutamine synthetase (GS) activity was five to six times higher than in similar cultures grown with 1 or 0.1 mM (NH(4))(2)SO(4). The NtrBC-inducible GSII form of this enzyme accounted for 60% of the specific activity in N-starved rhizobia, being negligible in the other two cultures. The exopolysaccharide (EPS) and capsular polysaccharide (CPS) contents relative to cell protein were significantly higher in the N-starved cultures, but on the other hand, the poly-3-hydroxybutyrate level did not rise in comparison with N-sufficient cultures. In agreement with the accumulation of CPS in N-starved cultures, soybean lectin (SBL) binding as well as stimulation of rhizobial adsorption to soybean roots by SBL pretreatment were higher. The last effect was evident only in cultures that had not entered stationary phase. We also studied nodC gene induction in relation to N starvation. In the chromosomal nodC::lacZ fusion Bj110-573, nodC gene expression was induced by genistein 2.7-fold more in N-starved young cultures than in nonstarved ones. In stationary-phase cultures, nodC gene expression was similarly induced in N-limited cultures, but induction was negligible in cultures limited by another nutrient. Nodulation profiles obtained with strain LP 3001 grown under N starvation indicated that these cultures nodulated faster. In addition, as culture age increased, the nodulation efficiency decreased for two reasons: fewer nodules were formed, and nodulation was delayed. However, their relative importance was different according to the nutrient condition: in older cultures the overall decrease in the number of nodules was the main effect in N-starved cultures, whereas a delay in nodulation was more responsible for a loss in efficiency of N-sufficient cultures. Competition for nodulation was studied with young cultures of two wild-type strains differing only in their antibiotic resistance, the N-starved cultures being the most competitive.

Effects of Drought Stress on Growth and Nitrogen Assimilation by Common Beans

Drought stress is one of the most important limitations for common bean (Phaseolus vulgaris) crops in the Argentinean North West. Frequently, the combined effects of high temperature and rainfall scarcity cause rapid increments in atmospheric water demand that lead to sudden, severe drought stress situations during the crop season. Approaches to prevent crop damages will derive from optimization of both genetic resources and crop management options. To succeed, these approaches must take into account the physiological plant strategies to face with the stress situation. Nitrogen nutrition in common bean is dual: atmospheric nitrogen may be fixed by the root nodules and/or inorganic nitrogen may be assimilated from soil solution. Therefore, common beans might have developed different strategies according with each N-nutrition way, and in particular, different levels of tolerance might be reached. In a first attempt to identify the best strategy of plants to hold a severe drought stress, we compared the plant tolerance to drought in relation with its growth, as conditioned by each N-nutrition way.

INTERACTION BETWEEN Rhizobium meliloti AND ALFALFA AGGLUTININ

Rhizobium meliloti (Rme) is specifically agglutinated at pH 4.0 by alfalfa agglutinin, and this protein has been proposed as the root lectin hypothetically involved in the mutual recognition of both symbionts (Paau et al., 1981). However, early events in this association which precede infection and nodulation are completely inhibited at pH 4 (Munns, 1968). In this study we have explored whether agglutinin can interact with Rme in conditions which are permissive for preinfection.