Haiko Enok Sawazaki

Diversidade genética em palmeiras através de isoenzimas e RAPD

Genetic diversity of palms was studied by means of enzymatic polymorphism using polyacrilamide gel electrophoresis and a DNA polymorphism assay based on the amplification of random DNA segments, denominated RAPD. Species and ecotypes from genus Euterpe, Bactris, Elaeis and Syagrus were utilized. Palm leaf extracts were utilized for isozymes of malate dehydrogenase (MDH), leucine aminopeptidase (LAP), glutamate oxaloacetate transaminase (GOT), phosphoglucose isomerase (PGI), phosphoglucose mutase (PGM), acid phosphatase (ACP), peroxidase (PRX), esterase (EST), and the RAPD markers technique, using the A and B kits from Operon Technologies. Interespecific differences were observed, as well as distinctive patterns for palm hybrids. Some bands were observed in addition to those cited in the literature where starch gel electrophoresis was used for enzymatic polymorphism. The conclusions with regard to polymorphic patterns obtained using the RAPD technique confirmed those observed with isozymes, with the advantages of its greater efficiency and ability to process a large number of samples.

CARACTERIZAÇÃO DA VARIABILIDADE GENÉTICA EM COUVE-MANTEIGA UTILIZANDO ISOENZIMAS E RAPD

Genetic variability of kale plants (Brassica oleracea L. var. acephala D.C.) was studied by means of enzymatic polymorphism using polyacrilamide gel electrophoresis and a DNA polymorphism assay based on RAPD. Fifteen clones of kale var. acephala from IAC germplasm collection were studied. Leaf extracts were analysed for isozymes and RAPD markers using A and B kits of primers. Isozyme polymorphism was observed for phosphoglucose mutase (PGM), peroxidase (PRX) and esterase (EST) and was higher for PGM. Differences among clones were observed by isozymes and RAPD, however, the dendrograms obtained from both kinds of markers were dissimilar, suggesting that the isozymes provided less information than RAPD about the genome. The superior efficiency of the RAPD was due to its ability to process a larger number of samples, making details about genome more evident.

Biochemical and physical modifications of bean seeds during storage

Seeds of dry beans cvs Rico 23 and Carioca, were chemical and physically characterized three times during eleven months of storage. Electrophoresis in polyacrylamide gel with SDS of the globulin fraction showed an increase in the number of bands at the third sampling, probably due to a more efficient protein extraction. However, molecular weight of the G1 and G2 subunits were constant, about 50,000 -43,000 and 18,000, respectively. Total protein, soluble sugars and water contents did not change during storage, while fiber increased and lipids decreased. Starch content, as in case of globulin fraction, also increased due to greater extractability. Both free fatty acids content and peroxide value increased, according to the increase of the acidity value. Water absorption capability increased initially and leveled off thereafter. The percentage of seeds with hard coat was not affected by storage. The behaviour of the two cultivars was very similar.

Identificação de espécies de citros mediante polimorfismo enzimático

Estudou-se, mediante polimorfismo enzimatico em gel de poliacrilamida, a variabilidade genetica das especies de laranja-doce (Citrus sinensis); laranja-azeda (C. aurantium); tangerinas clementina (C. clementina), sunki (C. sunki), cleopatra (C. reshni) e ponca (C. rsticulata); lima-da-persia (C. limettioides); limao-galego (C. aurantifolia); limao-cravo (C. limonia) e trifoliata (Poncirus trifoliata). Extratos de folhas foram analisados para as isoenzimas de malato deidrogenase (MDH), enzima malica (ME), leucino amino peptidase (LAP), glutamato oxaloacetato transaminase (GOT), fosfoglucoisomerase (PGI), fosfoglucomutase (PGM) e isocitrato deidrogenase (IDH). Verificou-se grande variabilidade genetica interespecifica, porem nenhuma entre os cultivares de laranja-doce. Foram encontradas algumas aloenzimas, alem das referidas pela literatura em gel de amido, como aquelas de uma regiao proxima ao loco conhecido por Pgm-1, responsavel por proteinas monomericas. Este sistema, denominado PGM, revelou a maior diferenciacao entre as especies, tendo apresentado duas regioes distintas com 9 alelos. No sistema MDH, foram considerados dois locas codificando para proteinas dimericas com 7 alelos; no ME, um loco com 3 alelos; no LAP, possivelmente dois locos responsaveis por proteinas monomericas com 4 alelos; no GOT, dois focos com 7 alelos; no PGI, um loco com 3 alelos e no IDH, um loco com 4 alelos.

Estresse de água no crescimento, produtividade e acúmulo de prolina em feijão

The objective of this paper was to verify the behavior of dry bean cultivars (Phaseolus vulgaris L.) related to the development, yield and free proline accumulation when subjected to waters stress. Ten dry bean cultivars were grown in greenhouse with two treatments (a) irrigated, and (b) without irrigation for some periods. Leaf area, dry weight, height, leaf proline content were determined several times during the growing season and grain yield at the end of plants cycle. The results showed that the cultivars Moruna and Aroana showed the highest rate of development and the lowest accumulation of free proline when the two treatments were compared. It can be concluded from the data that the cultivars Moruna and Aroana presented the highest drought resistance and the cultivars Jalo and Roseli the lowest.

Avaliação da atividade da lipoxigenase em linhagens de soja

Seed of soybeans, Glycine max L. (Merrill), cultivar IAC-8 and lines A-5, low linolenic acid content, PI 408251, lacking L1 mutant, PI 86023, lacking L2 mutant, Tohoko no 74, lacking L3 mutant, and F2 population from the crosses between IAC-8 and mutant lines (lacking L1, L2 and L3) were analysed for linolenic acid content, lipoxygenase activity and identification of mutant lines lacking L1, L2 or L3 isoenzymes. The purpose was to evaluate methods to identify mutants lines of soybean for lipoxygenase and to verify a possible correlation between enzimatic activity and linolenic acid content. The enzimatic activity was estimated quantitatively by a colorimetric method and lipoxygenase isoenzymes detection by isoelectric focusing. The oil seed content was determined by gravimetry and the fatty acids content by gas liquid chromatography of their methyl esters. The lipoxygenase activity data was good to indicate mutant lines for the final identification using isoelectric focusing. It permits the use of this method for choosing lines in breeding program to obtain a good soybean oil stability. It was not observed correlation, in the material studied, between enzymatic activity and linolenic acid content.

Transporte de compostos nitrogenados em soja cultivada com diferentes fontes de nitrogênio

Plantas de soja inoculadas com Rhizobium japonicum foram cultivadas em vermiculita com solucao nutritiva sem nitrogenio, em casa de vegetacao, no Centro Experimental de Campinas, do Instituto Agronomico. Foi estudado o efeito quanto aos niveis de ureideos (alantoina e acido alantoico), aminoacidos (total e qualitativo por analisador de aminoacidos), NO3- e NH4+, encontrados na seiva do xilema. Para esse fim, as plantas (noduladas) foram tratadas durante sete dias com solucoes nutritivas contendo os ions NO3- (15mM) ou NH4+ (10mM) aplicadas proximo a epoca de floracao, ou sem nitrogenio (controle). O acido alantoico predominou em relacao a alantoina, e essa proporcao tendeu a aumentar com a exposicao da planta a N-mineral. O transporte de nitrogenio total pelo xilema foi maior para o processo de fixacao de N2, decrescendo para o de absorcao de NH4+ e para o de absorcao de NO3-, sugerindo que o transporte de nitrogenio tenha dependido da interacao entre presenca de N-mineral no solo e disponibilidade de energia. A fixacao de N2, processo mais dispendioso de energia, possibilita maior producao de ureideos, que exportam mais nitrogenio que os aminoacidos. A asparagina foi o aminoacido encontrado em maior quantidade no xilema, independente do tratamento. O tratamento NH4+ nao alterou o teor de N-NH4+, mas aumentou o nivel de glutamina e asparagina em comparacao com o tratamento NO3-, que, por sua vez, aumentou o teor de acido aspartico e o de N-NO3-. Esses resultados sugerem que a diferenciacao na formacao de aminoacidos decorreu dos processos especificos de absorcao de nitrogenio.

Caracterização isoenzimática de clones e somaclones de cana-de-açúcar

Extracts from leaves and roots of three sugarcane varieties and their somaclones: NA56-79 (at seven and nine month old plants), IAC68-12 and IAC68-144 (both at nine month old plants) were obtained. The peroxidase and esterase isoenzyme contents of the extracts were determined to verify the genetic variability of tissue culture somaclones. The peroxidase specific activity was studied in relation to the time and isoenzymatic pattern. The peroxidase and esterase zymograms showed distinct differences among varieties, and the esterase presented less bands but easier to characterize than the peroxidase bands. The root zymograms exhibited great enzymatic variation due to the presence of old roots thus not allowing to characterize the varieties. No differences in peroxidase or esterase zymograms among somaclones of the same variety were noticed. The NA56-79 presented lower specific activity of peroxidase and number of isoenzymes than IAC68-144, suggesting that the decrease of isoenzyme bands is associated to the reduction of enzymatic activity. The specific activity of peroxidase changed with maturity and was higher at nine than at seven month old plants.

O pH e a temperatura na produção de biogás a partir de casca de arroz

Rica hulls were studied in order to examine their potential for biogas production. The material was previously treated with 5, 10 and 15% NaOH W/W and incubated at temperatures of 40 and 60o C. Results showed that rice hulls were not good for methane production, but the total production of gases increased with temperatura and alcali concentration up to 10%.

Temperatura, umidade relativa e disponibilidade de água no acumulo de prolina em cultivares de feijoeiro

O estudo de plantas de feijao (Phaseolus vulgaris L.) submetidas a deficiencia hidrica e temperaturas de 35-40o C umidade relativa (UR) 40-100%; 5-10oC, UR 20-30%; 2-5oC, UR 10-20%, realizado em casa de vegetacao e câmara de crescimento, no Instituto Agronomico, em 1982, mostrou haver correlacao negativa entre o acumulo de prolina e o potencial de pressao de agua nas folhas (yp). Os cultivares Curitibano e P-511, este ultimo considerado resistente a seca e baixa temperatura, apresentaram o menor acumulo de prolina e o maior yp, confirmando a hipotese de que e possivel o reconhecimento de material geneticamente resistente a escassez de agua atraves da selecao de plantas que acumulam menor quantidade de prolina.