Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / O Ãcido abscÃsico (ABA) à um fitohormÃnio que desempenha papÃis crÃticos na regulaÃÃo das respostas das plantas ao crescimento e desenvolvimento. O vacÃolo de plantas superiores à uma organela que ocupa a maior parte da cÃlula. A condiÃÃo de acidez à mantida por duas bombas de prÃtons distintas, V-ATPase e V-PPase. Sabe-se que estas bombas de prÃtons desempenham um papel essencial nas respostas das plantas Ãs mudanÃas ambientais. O gradiente eletroquÃmico promovido por essas enzimas à a forÃa motriz para o acÃmulo de Ãons e outros solutos no vacÃolo, sendo importante para manter a homeostase citosÃlica de Ãons e o metabolismo celular. As enzimas antioxidantes constituem um sistema de defesa contra as espÃcies reativas de oxigÃnio, que podem provocar danos ao desenvolvimento das plantas. O objetivo deste trabalho foi estudar o efeito do Ãcido abscÃsico nos parÃmetros de crescimento e fisiolÃgicos, bem como o efeito na atividade hidrolÃtica e na expressÃo de transcritos das bombas de prÃtons vacuolares e na atividade de enzimas antioxidantes de plantas de Vigna unguiculata. As sementes de V. unguiculata foram germinadas em areia, onde cresceram por 15 dias, com aplicaÃÃo de soluÃÃo nutritiva de Hoagland na ausÃncia (controle) ou presenÃa de ABA 0,1 ÂM no terceiro, no sÃtimo e no dÃcimo dias apÃs a germinaÃÃo. Os efeitos do Ãcido abscÃsico (ABA) sobre o crescimento, a condutÃncia estomÃtica, a transpiraÃÃo, a fotossÃntese, a concentraÃÃo interna de CO2, a atividade e a expressÃo da subunidade A da V-ATPase e da V-PPase e a atividade de enzimas antioxidantes de Vigna unguiculata foram analisados. ABA aumentou o crescimento das plantas, mas nÃo afetou os parÃmetros fisiolÃgicos; induziu um aumento na atividade hidrolÃtica da V-ATPase em folhas e da V- PPase em raÃzes; aumentou os transcritos de VuVHA-A e de VuVHP em folhas e diminuiu os transcritos de VuVHA-A em raÃzes e, por fim, causou aumento na atividade da catalase de folhas e de raÃzes. Esses resultados sugerem que o Ãcido abscÃsico regula a atividade e a expressÃo dos genes das bombas de prÃtons vacuolares, bem como as atividades de enzimas antioxidantes, sendo portanto importantes efetores que regulam o desenvolvimento de plantas de V. unguiculata. / Abscisic acid (ABA) is a phytohormone that plays critical roles in regulating plant responses to growth and development. The vacuole of higher plants is an organelle that occupies a larger part of the cell. The acidic condition is maintained by two distinct proton pumps, VATPase and V-PPase. It is known these proton pumps play essential roles in plant responses to environmental changes. The electrochemical gradient promoted by these enzymes is the driving force for the accumulation of ions and other solutes in the vacuole, being important to maintain cytosolic ion homeostasis and cellular metabolism. Antioxidant enzymes constitute a defense system against reactive oxygen species, which can cause damage to plant development. The aim of this study was to study the effect of the abscisic acid in the growth and physiological parameters, as well as the effect on vacuolar proton pumps and antioxidant enzymes (SOD, CAT and APX) from Vigna unguiculata cv. Pitiuba. The seeds of V. unguiculata were germinated in sand and grown for 15 days, with application of Hoagland solution in the absence (control) or presence of 0.1 ÂM ABA in the third, seventh and tenth days after germination.The effects of the abscisic acid (ABA) on the growth, stomatal conductance, transpiration, photosynthesis, internal CO2 concentration, V-ATPase subunit A and V-PPase activities and expression and antioxidant enzymes activities of Vigna unguiculata were analyzed. ABA increased the plants growth but did not affect the physiological parameters; induced an increase on V-ATPase hidrolytyc activity in leaves and on V-PPase in roots; ABA increased the transcripts of VuVHA-A and VuVHP in leaves and decreased the VuVHA-A transcripts in roots; caused increase in the leaves and roots catalase activity. These results suggest that the abscisic acid regulate the activity and the genes expression of the vacuolar proton pumps, as well as the antioxidant enzymes activity, being thus important effectors that regulate the development of V. unguiculata plants.

This study is focused on study of changes of enzyme and low-molecular weight antioxidants in different fruits during long-term storage. In theoretical part individual low-molecular weight antioxidants and enzymes are described. The main causes of fruit decay and some possibilities of fruit preservation and storage are summarized. As biological material some common fruits were chosen - green and red apples, peaches, plums and white grapes. The fruits were stored in laboratory, cellar, in refrigerator and in freezer. In freezing experiments some ways of fruit preparation and processing were tested and their influence on fruit antioxidant status was compared. Shortened storage experiment was applied on blueberries, cranberries, raspberries and strawberries too. In fruits some group parameters – total antioxidant status, dry mass content, ascorbate level, total flavonoids and total phenolics were analyzed spectrophotometrically. Individual flavonoids and phenolics were determined by RP-HPLC/UV-VIS and on-line LC/PDA/ESI-MS. Antioxidant enzyme activities (superoxide dismutase SOD, catalase CAT, polyphenol oxidase PPO and lipoxygenase LOX) were measured by spectrophotometry. The surface microscopy and cultivation of moulds from fruit surface were performed too. Influence of storage conditions on biological activities is dependent on fruit sort. Freezing is the most suitable procedure for long-term storage without significant changes of active substance content. Long-term storage in controlled temperature conditions and/or atmosphere is usable for fruits with longer storage period. In these fruits stabile levels of antioxidant enzymes are stored for relatively long time. Some of enzymes act synergistically. Enzyme activities differed according to storage phase; at the beginning mainly high SOD and LOX activities were observed. CAT and PPO are probably activated as defence systems in rippened and/or damaged fruits. Levels of total as well as individual low molecular weight antioxidants varied during storage in all sorts, generally, increased course with longer storage period can be observed.