Eletroforese de isoenzimas de plântulas na identificação de espécies de Brachiaria

Lotes de sementes de braquiária comercializados no Brasil vem apresentando contaminações com sementes de outras espécies pertencentes ao mesmo gênero. Deste modo, uma das espécies de braquiária atuaria como planta infestante da outra no agroecossistema e a erradicação da espécie infestante seria dificultada pela agressividade característica do gênero e pela falta de seletividade dos herbicidas disponíveis no mercado. Esses fatores ressaltam a importância da comercialização e utilização de lotes de sementes isento de sementes de outras espécies e a utilização de metodologias precisas de identificação das principais espécies de braquiária no controle de qualidade das empresas produtoras de sementes. Neste trabalho, buscou-se avaliar o potencial discriminante da técnica de eletroforese, utilizando quatro sistemas enzimáticos presentes em plântulas de quatro espécies do gênero Brachiaria, quer sejam B. brizantha cv. Marandu, B. decumbens cv. Basilisk, B. humidicola cv. comercial e B. plantaginea. Foram realizadas análises de eletroforese de isoenzimas testando-se 50 indivíduos de cada espécie por tratamento, utilizando-se coleóptilos de plântulas obtidas a partir de sementes germinadas a 30°C, no escuro. Para a eletroforese foi utilizado como meio suporte géis de poliacrilamida, nas concentrações de 7,0 e 7,5%. As isoenzimas Glutamato desidrogenase e Glucose-6-fosfato desidrogenase, embora eficientes na diferenciação entre B. plantaginea e B. humidicola e entre as sementes dessas espécies e as de B. brizantha ou B. decumbens, não se mostraram capazes de diferenciar as sementes destas duas últimas espécies. Entretanto, as izoenzimas α- e β-esterase possibilitaram uma nítida diferenciação das quatro espécies de Brachiaria estudadas.Undesirable physical seed mixture in commercial seed lots of several Brachiaria species is a cause of growing concern in Brazil. Such a mixtures often result in the contamination of cultivated areas by species which are difficult to erradicate in conseqüence of their aggressivity and of the low selectivity of available herbicides. The obtention of contamination-free seed lots is, therefore, of great interest and so are the techniques for the precise identification of Brachiaria species, which would allow the improvement of quality control by commercial enterprises. This paper reports an attempt of evaluating the discriminatory potential of the eletrophoretic technique applied to four enzymatic systems present in seedlings of four Brachiaria species, namely, B. brizantha cv. Marandu, B. decumbens cv. Basilisk, B. humidicola cv. common , and B. plantaginea. Fifty coleoptiles of each species, obtained from seeds germinated in the dark at 30°C, were used in each treatment. Polyacrilylamide gels, at 7.0% and 7.5%, were used as supporting gels. The banding pattern resulted from the electrophoresis of the isozymes Glutamate dehydrogenase and Glucose-6-phosphate dehydrogenase allowed the differentiation of B. plantaginea and B. humidicola between themselves and these from B. brizantha and B decumbens, but not the differention between the later species. However, when the isozymes α- and β-esterase were used, a clear distinction of the four species studied was possible

High accumulation of soluble sugars in deep supercooling Japanese white birch xylem parenchyma cells.

• Seasonal changes in the accumulation of soluble sugars in extracellular freezing cortical parenchyma cells and deep supercooling xylem parenchyma cells in Japanese white birch (Betula platyphylla var. japonica) were compared to identify the effects of soluble sugars on the mechanism of deep supercooling, which keeps the liquid state of water in cells under extremely low temperatures for long periods. • Soluble sugars in both tissues were analyzed by high-performance liquid chromatography (HPLC), and the concentrations of sugars in cells were estimated by histological observation of occupancy rates of parenchyma cells in each tissue. Relative and equilibrium melting points of parenchyma cells were measured by differential thermal analysis and cryoscanning electron microscopy, respectively. • In both xylem and cortical parenchyma cells, amounts of sucrose, raffinose and stachyose increased in winter, but amounts of fructose and glucose exhibited little change throughout the entire year. In addition, no sugars were found to be specific for either tissue. Combined results of HPLC analyses, histological observation and melting point analyses confirmed that the concentration of sugars was much higher in xylem cells than in cortical cells. • It is thought that the higher concentration of soluble sugars in xylem cells may contribute to facilitation of deep supercooling in xylem cells by depressing the nucleation temperature