Evaluation of N2‐fixation measured by the 15N‐dilution and N‐difference methods in Nicaraguan and Ecuadorian Phaseolus vulgaris L. plants inoculated with Rhizobium leguminosarum biovar

Pot and field experiments were performed to assess N₂ fixation in Nicaraguan (R79 and R84) and Ecuadorian (Imba) common bean (Phaseolus vulgaris L.) cultivars, with the aim of improving their productivity by inoculating them with commercially produced Rhizobium phaseoli. With maize (Zea mays L.) as the non‐N₂‐fixing control, the percentage of N₂ fixed predicted by the ¹⁵N‐dilution method was significantly (P ≤ 0.05) higher than that predicted by the N‐difference method. However, the N₂ amounts predicted by the two methods were not significantly different. The correlation between the two methods was significant and positive (P ≤ 0.0001, n = 36). Compared with the native rhizobial strain, symbiotic associations of the bean cultivars with UMR1073, UMR1077 and UMR1899 rhizobial inoculants did not significantly (P ≤ 0.05) influence plant dry matter (DM) and N yields, the extent of N₂ fixation and uptake of soil and fertilizer N. Nevertheless, the UMR1077 and UMR1899 strains markedly increased the uptake of soil N by R84 plants, while decreasing N2 fixation. In contrast, the Imba‐UMR1899 association enhanced positive effects on all variables. About 60–70% of the total N taken up by the Imba plants was fixed N_2. The R79 and R84 plants fixed about 50% of their total N uptake. N₂ fixation rates were positively correlated with DM and total N yields, while being negatively correlated with soil N uptake (P ≤ 0.001, n = 36). Future research in Nicaragua should focus on selecting rhizobial strains suitable for indigenous common bean cultivars.     

Assessment of the floral origin of honey by SDS-page immunoblot techniques

We report on the development of a novel alternative method for the assessment of floral origin in honey samples based on the study of honey proteins using immunoblot assays. The main goal of our work was to evaluate the use of honey proteins as chemical markers of the floral origin of honey. Considering that honeybee proteins should be common to all types of honey, we decided to verify the usefulness of pollen proteins as floral origin markers in honey. We used polyclonal anti-pollen antibodies raised in rabbits by repeated immunization of Sunflower (Elianthus annuus) and Eucalyptus (Eucalyptus sp.) pollen extracts. The IgG fraction was purified by immunoaffinity. These antibodies were verified with nitrocellulose blotted pollen and unifloral honey protein extracts. The antibodies anti-Sunflower pollen, bound to the 36 and 33 kDa proteins of Sunflower unifloral honey and to honey containing Sunflower pollen; and the antibodies anti-Eucalyptus sp. pollen bound to the 38 kDa proteins of Eucalyptus sp. unifloral honey in immunoblot assays. Satisfactory results were obtained in differentiating between the types of pollen analyzed and between Sunflower honey and Eucalyptus honey with less cross reactivity with other types of honey from different origin and also with good sensitivity in the detection. This immunoblot method opens an interesting field for the development of new antibodies from different plants, which could serve as an alternative or complementary method to the usual melissopalynological analysis to assess honey floral origin.     

Hybridization between Haliotis rufescens and Haliotis discus hannai: evaluation of fertilization,larval development,growth and thermal tolerance

Two introduced abalone species are currently produced in Chile, red abalone Haliotis rufescens and Japanese abalone Haliotis discus hannai. However, red abalone accounts for 99% of total production, while the Japanese abalone has not adapted well to Chilean coastal waters. This study reports the hatching, growth and thermal tolerance performance in interspecific hybrids produced between red (R) and Japanese (J) abalone. Our results show that egg age and sperm concentration were critical factors to produce hybrids. The cross R♀ × J♂ showed a fertilization rate of 55.3 ± 3.5% using 20‐min‐old eggs and sperm concentrations of 14 × 10⁶ cells mL^?1, while the reciprocal cross (J♀ × R♂) was not successful. Further, larval development stages were similar in RR, JJ and RJ hybrid abalones. Among the experimental trials, settlement rate varied from 12.3% to 18.6% and final survival from 20.1% to 31.7%, being the RJ hybrid rates intermediate between parental species. The final shell lengths were similar between RR and RJ hybrids, but significantly higher in JJ abalones. In addition, thermal tolerance was ascertained due its pivotal role for the abalone physiology. Thus, RJ hybrids showed the highest HSP70 gene expression and offers new possibilities to expand Chilean abalone production in warm waters zones.     

Alpha-tocopherol protects against oxidative damage to lipids of the rod outer segments of the equine retina

Oxidative stress is a possible risk factor for eye diseases. Lipid peroxidation is one of the major events induced by oxidative stress and is particularly active in polyunsaturated fatty acid (PUFA)-rich biomembranes. This work evaluated endogenous lipid antioxidants, in vitro non-enzymatic lipid peroxidation of rod outer segment membranes (ROS), the fatty acid composition during oxidative damage of total lipids from equine retina and ROS, and the protective action of alpha-tocopherol (alpha-Toc). The major lipid soluble antioxidant was alpha-Toc followed by retinoids and carotenoids. The retina contained a high percentage of PUFAs, mainly docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6). Lipid peroxidation of the equine ROS, induced by Fe(2+)-ascorbate, was monitored using chemiluminescence (CL) with or without pre-treatment with alpha-Toc. With alpha-Toc pre-treatment, CL values were significantly decreased. The most abundant fatty acid was 22:6n-3. After 3h incubation, 95% of total PUFAs were destroyed by peroxidation, whereas in alpha-Toc pre-treated ROS the percentage was significantly decreased. The results show that the retina has an endogenous lipid soluble antioxidant system. ROS were highly sensitive to oxidative damage, since their fatty acid composition was markedly modified during the lipid peroxidation process. The protective role of alpha-Toc as an antioxidant was evident and it could be used in the treatment of equine ocular diseases in which free radicals are involved.