Strigolactones in the Rhizobium-legume symbiosis: Stimulatory effect on bacterial surface motility and down-regulation of their levels in nodulated plants.

Strigolactones (SLs) are multifunctional molecules acting as modulators of plant responses under nutrient deficient conditions. One of the roles of SLs is to promote beneficial association with arbuscular mycorrhizal (AM) fungi belowground under such stress conditions, mainly phosphorus shortage. Recently, a role of SLs in the Rhizobium-legume symbiosis has been also described. While SLs' function in AM symbiosis is well established, their role in the Rhizobium-legume interaction is still emerging. Recently, SLs have been suggested to stimulate surface motility of rhizobia, opening the possibility that they could also act as molecular cues. The possible effect of SLs in the motility in the alfalfa symbiont Sinorhizobium meliloti was investigated, showing that the synthetic SL analogue GR24 stimulates swarming motility in S. meliloti in a dose-dependent manner. On the other hand, it is known that SL production is regulated by nutrient deficient conditions and by AM symbiosis. Using the model alfalfa-S. meliloti, the impact of phosphorus and nitrogen deficiency, as well as of nodulation on SL production was also assessed. The results showed that phosphorus starvation promoted SL biosynthesis, which was abolished by nitrogen deficiency. In addition, a negative effect of nodulation on SL levels was detected, suggesting a conserved mechanism of SL regulation upon symbiosis establishment.

24-epibrassinolide modulates growth, nodulation, antioxidant system, and osmolyte in tolerant and sensitive varieties of Vigna radiata under different levels of nickel: a shotgun approach.

The objective of this study was to explore the response of 24-epibrassinolide to improve the biological yield of Ni-tolerant and Ni-sensitive varieties of Vigna radiata and also to test the propositions that 24-epibrassinolide induced up-regulation of antioxidant system protects the efficiency of V. radiata, grown under Ni-stress. Surface sterilized seeds of var. T-44 (Ni-tolerant) and PDM-139 (Ni-sensitive) were soaked in DDW (control), 10(-10), 10(-8), or 10(-6) M of 24-epibrassinolide for 8 h (shotgun approach). These treated seeds were then inoculated with specific Rhizobium grown in sandy loam soil supplemented with different levels of Ni 0, 50, 100, or 150 mg Ni kg(-1) of soil and were allowed to grow for 45-days. At this stage of growth, plants were sampled to assess the various growths and nodule related traits as well as selected biochemical characteristics. The remaining plants were allowed to grow to maturity to study the yield characteristics. The results indicated that plant-fresh and dry mass, number of nodules, their fresh and dry mass, leghemoglobin content, nitrogen and carbohydrate content in the nodules, leaf chlorophyll content, activities of nitrate reductase and carbonic anhydrase decreased proportionately with the increasing concentrations of soil nickel. However, the application of 24-epibrassinolide as shotgun approach (pre-sowing seed soaking) to the nickel-stressed or non-stressed plants improved growth, nodulation and enhanced the activity of various antioxidant enzymes (viz. catalase, peroxidase and superoxide dismutase) and also the content of proline. The up-regulation of antioxidant enzymes as well as proline (osmolyte) triggered by 24-epibrassinolide could have conferred tolerance to the Ni-stressed plants resulting in improved growth, nodulation and yield attributes.

Use of potato extract broth for culturing root-nodule bacteria.

Liquid media containing potato extract and 1% of glucose or sucrose were used to culture root-nodule bacteria (rhizobia) in shaken Erlenmeyer flasks. For comparison, these bacteria were also cultured in yeast extract-mannitol broth (YEMB) as a standard medium. Proliferation of rhizobia was monitored by measuring optical densities (OD550) of the cultures and by plate counting of the viable cells (c.f.u) of the bacteria. In general, multiplication of the rhizobia in potato extract-glucose broth (PEGB) and potato extract-sucrose broth (PESB) was markedly faster, as indicated by higher values of OD550, than in YEMB. The numbers of R. leguminosarum by. vicae GGL and S. meliloti 330 in PEGB and PEGB were high and ranged from 1.2 x 10(10) to 4.9 x 10(10) mL(-1) after 48 h of incubation at 28 degrees C. B. japonicum B3S culture in PEGB contained 6.4 x 10(9) c.f.u. ml(-1) after 72 h of incubation. PEGB and YEMB cultures of the rhizobia were similar with respect to their beneficial effects on nodulation of the host-plants of these bacteria.