In vitro regeneration of Salix nigra from adventitious shoots

Black willow (Salix nigra Marsh.) is the largest and only commercially important willow species in North America. It is a candidate for phytoremediation of polluted soils because it is fast-growing and thrives on floodplains throughout eastern USA. Our objective was to develop a protocol for the in vitro regeneration of black willow plants that could serve as target material for gene transformation. Unexpanded inflorescence explants were excised from dormant buds collected from three source trees and cultured on woody plant medium (WPM) supplemented with one of: (1) 0.1 mg l(-1) thidiazuron (TDZ); (2) 0.5 mg l(-1) 6-benzoaminopurine (BAP); or (3) 1 mg l(-1) BAP. All plant growth regulator (PGR) treatments induced direct adventitious bud formation from the genotypes. The percentage of explants producing buds ranged from 20 to 92%, depending on genotype and treatment. Although most of the TDZ-treated inflorescences produced buds, these buds failed to elongate into shoots. Buds on explants treated with BAP elongated into shoots that were easily rooted in vitro and further established in potting mix in high humidity. The PGR treatments significantly affected shoot regeneration frequency (P < 0.01). The highest shoot regeneration frequency (36%) was achieved with Genotype 3 cultured on 0.5 mg l(-1) BAP. Mean number of shoots per explant varied from one to five. The ability of black willow inflorescences to produce adventitious shoots makes them potential targets for Agrobacterium-mediated transformation with heavy-metal-resistant genes for phytoremediation.     

Latex proteins from the plant Calotropis procera are partially digested upon in vitro enzymatic action and are not immunologically detected in fecal material

Soluble proteins from the latex of Calotropis procera (LP) were investigated in vitro and in vivo for digestibility as the latex has previously been shown to produce considerable toxic effects on animals. The latex is also an important biologically active compound that displays antiinflammatory and antidiarrhea properties. The proteins were digested by the action of trypsin, pepsin or chemotrypsin as revealed by gel filtration and SDS-PAGE analysis. Furthermore, the full LP digestion was easily achieved by protease treatment. Rabbit polyclonal antibodies raised against LP failed to detect cross-reactive molecules in fecal material of experimental rats following 35 consecutive days of LP consumption in water. Similar patterns of electrophoresis were observed for the negligible amounts of protein observed in the fecal extracts of control and test animals. No death or toxic effects were observed among animals. Taken together these results suggest that harmful and toxic effects on animals of the latex from C. procera are present in its rubber and low molecular weight fractions rather than its protein content.     

The effect of aluminum on the germination of wheat seeds 1

The effect of aluminum (Al) ions on the germination of wheat (Triticum aestivum L.) seeds has been investigated using two varieties, one sensitive to Al (Robin) and the other relatively Al‐tolerant (Carazinho). High concentrations of Al (i.e. greater than 1 mM) were required to inhibit the growth of the emerging roots and shoots of germinating seeds of both varieties. Moreover, Al at concentrations up to 10 mM did not affect the mobilization of carbohydrate reserves in the germinating seeds. It was concluded that germinating seeds of both wheat varieties were much less sensitive to the effects of Al than were established seedlings.     

Growth attributes,xylem sap composition,and photosynthesis in common bean as affected by nitrogen and phosphorus deficiency

The effects of nitrogen (N‐) and phosphorus (P‐) deficiency, isolatedly or in combination, on growth, nitrogenous fraction, and inorganic phosphate in xylem exudade, and photosynthesis of common bean (Phaseolus vulgaris L. cv. Negrito) were investigated. Plants were grown in nutrient solution adjusted daily to pH 5.5 and aerated continuously. Ten days after emergence mineral deficiency was imposed. Plants were then supplied with high N (7.5 mol m^‐3) or low N (0.5 mol m^‐3), and also with high P (0.5 mol m^‐3) or low P (0.005 mol m^‐3). All sampling and measurements were made 28 days after emergence. N‐ or P‐deprivation brought about large decreases in total leaf area by inhibiting the emergence of new leaves and primarily the expansion of the leaves. The specific leaf area did not change under N‐ but decreased under P‐limitation. The decreased shoot to root ratio in all deficiency treatments was a consequence of a lowering mass of above‐ground organs, especially of leaves.

The content of chlorophylls declined significantly only under N‐deficiency alone; carotenoids declined under both N‐ and combined N‐ and P‐limitation. No alteration in amino acid concentration in xylem exudate occurred in plants experiencing N‐starvation, while ureides increased by 79%, and nitrate and inorganic phosphate decreased greatly. Under P‐deprivation, amino acids and nitrate in xylem sap dropped by about half; ureides were held relatively constant, and phosphate was severely depressed. Total upward translocation of N through xylem was estimated to be about 16% higher in N‐deficient plants than in plants without mineral limitation, but leaf N levels in the former were lower as compared to control plants. The net carbon (C) assimilation decreased similarly regardless of the imposed deficiency treatment. Such a decrease was mainly determined by non‐stomatal factors. In general, no additive effect between N‐ and P‐limitation on any of measured parameters was observed.     

Macronutrient accumulation in wheat crop (Triticum aestivum L.) with Azospirillum brasilense associated with nitrogen doses and sources

Abstract

New studies are needed to optimize the nitrogen (N) amount that can be applied to utilize the Azospirillum brasilense benefits. In addition, information regarding the interaction between the urease inhibitor and biological nitrogen fixation (BNF) and how they affect the macronutrients accumulation are also needed. We evaluate the effect of N sources and doses associated with A. brasilense regarding the macronutrients accumulation in straw and grains and wheat grain yield in tropical conditions. A randomized block experimental design was used with four replications in a 2?×?5?×?2 factorial arrangement as follows: two N sources (urea and urea with urease enzyme inhibitor NBPT; five N doses (0, 50, 100, 150, and 200?kg ha^?1) applied in topdressing; with and without A. brasilense inoculation. We found that an increase in N doses positively influenced the accumulation of macronutrients in straw and grains and the wheat grain yield. N sources have similar effects. Inoculation with A. brasilense increased accumulation of Mg and S in straw and P, Ca, and Mg in grains, regardless of the N dose. The inoculation with A. brasilense associated with 140?kg ha^?1 of N increased wheat grain yield. The inoculation can contribute in a more sustainable way to wheat nutrition and optimizing N fertilization.     

Gene expression and immunolocalization of fibroblast growth factor 2 in the ovary and its effect on the in vitro culture of caprine preantral ovarian follicles

This study quantified Fibroblast growth factor 2 (FGF-2) mRNA and localized FGF-2 protein in different categories of follicles isolated from goat ovaries. In addition, we verified the effects of this factor on the in vitro culture of preantral follicles isolated from goats. For mRNA quantification, we performed real-time PCR using primordial, primary and secondary follicles, as well as cumulus-oocyte complexes (COCs) and mural granulosa and theca cells of small and large antral follicles. For FGF-2 protein localization, the ovaries were subjected to conventional immunohistochemical procedures. Preantral follicles were isolated and cultured in vitro for 12 days in either control (basic) or supplemented with FGF-2 medium. The expression of FGF-2 mRNA was detected in all categories of follicles and there was no difference in preantral follicles and COCs or granulosa/theca cells from small and large antral follicles. However, in large antral follicles, COCs showed expression levels significantly lower than in granulosa/theca cells (p < 0.05). We observed moderate expression of FGF-2 protein in preantral follicles but not in granulosa cells of primordial follicles and theca cells of secondary follicles. In both small and large antral follicles, strong, moderate and weak staining was observed in oocytes, granulosa and theca cells, respectively. The addition of FGF-2 caused a significant increase in the daily follicular growth rate compared to the control group. We conclude that FGF-2 mRNA is expressed throughout follicular development and that its protein can be found in different patterns in preantral and antral follicles. Furthermore, FGF-2 increases the follicular growth rate in vitro.     

Effect of cropping systems and crop successions on fumonisin levels in corn from Northern Paraná State,Brazil

In this study the effect of different cropping systems and crop successions was evaluated on natural Fusarium sp. contamination and fumonisin levels in corn. The cropping systems consisted of a conventional and no-tillage area cultivated with corn in the summer following either oats or fallow in the winter (2006 and 2007 growing seasons). In addition, the effect of applying nitrogen fertilizer (0, 22.5, 45.0, 90.0 and 90.0 kg ha⁻¹ nitrogen supplemented with potassium oxide) on fumonisin contamination was evaluated in the 2006 growing season. Fusarium sp. was detected in 90% samples in 2006 and in 100% samples in 2007. In both growing seasons, no-till corn following oats showed the highest mean fumonisin levels and differed significantly (P < 0.05) from all the others (2006) and from conventional till corn following either oats or fallow in the winter (2007). Fumonisin levels ranged from 0.13 to 19.52 μg g⁻¹ (mean 6.97 μg g⁻¹) and from 3.70 to 7.75 μg g⁻¹ (mean 6.29 μg g⁻¹) in no-till corn following oats from the 2006 and 2007 growing seasons, respectively. Plots treated with 0 kg ha⁻¹ and 22.5 kg ha⁻¹ nitrogen showed the highest mean fumonisin levels and differed significantly from those with 45.0 and 90 kg ha⁻¹ nitrogen. Fumonisin levels correlated negatively (P < 0.05) with the nitrogen fertilization rates. Although no-till is advantageous from a soil conservation standpoint, it may enhance the potential for fumonisin contamination in corn.     

Detection of <Emphasis Type="Italic">Brucella</Emphasis> sp. infection through serological,microbiological, and molecular methods applied to buffaloes in Maranhão State,Brazil

The aim of the current study is to diagnose Brucella spp. infection using methods such as serology, bacterial isolation, and molecular analysis in buffaloes bred in Maranhão State. In order to do so, 390 samples of buffalo serum were subjected to serological tests, to Rose Bengal Plate Test (RBPT) and to 2-mercaptoethanol (2-ME) combined with slow agglutination test (SAT). Vaginal swabs were collected from seropositive animals and subjected to bacterial isolation and to generic PCR. According to the serological test, 16 animals had a positive reaction to the confirmatory test (2-ME/SAT). As for bacterial isolation, three samples resulted in the isolation of Brucella spp.-characteristic colonies, which were confirmed through PCR. These results confirmed Brucella spp. infection in the buffalo herd from Maranhão State.     

Chitosan associated with whole raw soybean in diets for Murrah buffaloes on ruminal fermentation,apparent digestibility and nutrients metabolism

This study aimed to investigate the effects of chitosan and whole raw soybean on nutrient intake, apparent digestibility, nitrogen utilization, microbial protein synthesis, blood metabolites, feeding behavior, ruminal fermentation, digesta kinetics, and reticular flow of nutrients of buffaloes. Four ruminally‐cannulated Murrah buffaloes (351 ± 15 kg of initial BW) were randomly assigned according to a 4 × 4 Latin square design. Treatments were arranged as 2 × 2 factorial arrangement: the first factor was whole raw soybean (WRS), and the second factor was chitosan (CHI) with or without their inclusion in diets. Intake and apparent digestibility of ether extract (p < .01; p = .04, respectively), non‐fiber carbohydrates intake (p = .03) and apparent ruminal digestibility of dry matter (p = .01) were affected by diets. An interaction effect or tendency was observed for microbial nitrogen (p = .09), concentrations, ruminal ammonia nitrogen (p = .05), total volatile fatty acid (p = .03). Association of chitosan with whole raw soybean has potential effects as a modulator of rumen fermentation; therefore, chitosan can be applied as an alternative non‐ionophore for Murrah buffaloes.