Estudo do potencial fitotóxico de extratos de Bauhinia ungulata L. sobre a divisão celular e atividade enzimática em plântulas de alface / Study of phytotoxic potential of extracts of Bauhinia ungulata L. on cell division and enzyme activity in lettuce seedlings

RESUMO : Este trabalho visou avaliar a ação fitotóxica de extrato e frações obtidas das folhas de Bauhinia ungulata L sobre a divisão celular e atividade das enzimas α-amilase, catalase, peroxidase e polifenoloxidase em plântulas de alface (Lactuca sativa, var Grand rapis). Sementes de alface foram expostas às concentrações de 250, 500 e 1000 µg.mL-1 do extrato de Bauhinia ungulata e frações do mesmo, em condições ambientais controladas. Após três dias de protusão radicular, as raízes das plântulas foram cortadas e submetidas a preparo específico para visualização e contagem do número de células em cada fase mitótica. No sétimo dia, as amostras foram submetidas à avaliação da atividade enzimática utilizando técnica específica para cada enzima e leitura espectrofotométrica. Na análise da divisão celular observou-se que houve redução no número de células em mitose nas raízes das plântulas testadas, e a fração acetato de etila apresentou maior efeito inibitório sobre a divisão celular. O efeito sobre as enzimas que demonstram alterações no sistema de defesa antioxidante de L. sativa mostram que o extrato bruto e frações alteraram a produção das enzimas α- amilase, peroxidase, catalase e polifenol oxidase em, ao menos, uma das concentrações testadas, sendo um indicativo de estresse que interfere na divisão celular. Os resultados obtidos apontam para a presença de compostos com atividade inibidora ou estimulatória nas folhas de B. ungulata revelando potencial fitotóxico sobre as plântulas de alface

ABSTRACT Study of phytotoxic potential of extracts of Bauhinia ungulata L. on cell division and enzyme activity in lettuce seedlings. Theaim of this studywas to evaluate thephytotoxic actionofextractand fractionsobtainedfrom the leaves ofBauhiniaungulata L. inbioassaylaboratory, assessing their interferenceon cell division andactivity of the enzymesalpha-amylase, catalase, peroxidase andpolyphenoloxidaseofLactucasativa(lettu ce). The lettuce seedswere exposedto concentrationsof 250,500and 1000µg.mL-1of the extracts and fractions of Bauhinia ungulataunder controlled environmental conditions. After three days ofrootradicleprotrusion, the plants` rootswere cutand subjected tospecific preparationfor visualization andcounting of the number ofcells in eachmitotic stage. On the seventh daythe sampleswere subjected toenzymatic activityusingspecific techniquefor eachenzyme anda spectrophotometer measure. In the analysisof the mitotic indexit was observedthatthere was reduction in the number of cells in mitosis in the roots of the plants tested and thattheethyl acetate fractionsignificantly affected thecell division. The effecton the enzymesthat showed changesinthe antioxidant defensesystemof Lactucasativaindicate thatthe crude extractand the fractionsaltered theproductionofα-amylase, peroxides, catalaseand polyphenol enzymesin at least oneof the testedconcentrations, representingan indicationstress,which interferesin the celldivision.Theobtained results point out the presence ofcompoundswithinhibitoryorstimulatory activityon plant leaves of B. ungulata,revealingphytotoxic potentialtolettuce seedlings

Isoperoxidases show differing sensitivity to salicylic acid.

The effects of salicylic acid (SA) on the activity of total peroxidase and the patterns of isoperoxidases of cultured tobacco cells were investigated. The total peroxidase activity of tobacco cells was inhibited by 70% when the cells were treated with 5 mM SA for one week. The peroxidase activity of tobacco cells is declined by 90% in the presence of 30 mM SA. Moreover, the activity of isoperoxidases C3, A1, and A3 decreased dramatically with increasing SA concentration, while, one of the anodic isoperoxidases, A2, was somewhat resistant to SA treatment. When isoperoxidase C3 was isolated, SA inhibited the activity of purified C3 in a concentration-dependent manner. The IC50 of isoperoxidase C3 was approximately 0.45 mM. However, the inhibition of isoperoxidase C3 activity was removed by the addition of Fe2+ ion. The possible mechanism of inhibition of peroxidase by SA is discussed.

Effects of rogar and endosulfan on the metabolism of fresh water sponge (Spongilla lacustris).

Spongilla lacustris were exposed to sub lethal concentrations of pesticides, rogar and endosulfan for one month period. Metabolites like carbohydrates, protein and enzymes like those that peroxidase and carbonic anhydrase were estimated in the experimental and control animals. The results show, depletion of carbohydrates while protein elevated as the days progressed. Similarly an enzyme activity found to be decreased in exposed Spongilla lacustris.

Environmental stress-induced extracellular isoperoxidase RC3 from rice.

Effects of various environmental stresses such as heavy metals, salts and low (high) temperature on the secretion of peroxidase isozyme into the medium were examined in rice (Oryza sativa cv. Nak-Dong) suspension culture. The major extracellular peroxidases secreted into the medium by various stresses were cationic isoperoxidases. A far migrating cationic isoperoxidase RC3 was isolated from the medium after application of CaCl2, the effective stimulator for peroxidase secretion. Isolation of extracellular isoperoxidase RC3 was accomplished by ammonium sulfate fractionation, CM-cellulose cation-exchange chromatography, and Sephacryl S-100 gel filtration. The enzyme was a glycoprotein having molecular weight of approximately 34 KDa as determined by SDS-PAGE and 38 KDa by Sephacryl S-100 gel filtration. The pI value of the enzyme was 8.9. Kinetic studies revealed that the optimum pH of the enzyme was 6.0 for guaiacol and H2O2, and the Km values for guaiacol and H2O2 were 10.5 mM and 3.2 mM, respectively.

Cadmium toxicity induced changes in plant water relations and oxidative metabolism of Brassica juncea L. plants.

Excess of cadmium (Cd) induced changes in oxidative scenario and water status of plants viz.., total water content, specific water content, water saturation deficit (WSD) and transpiration of Brassica juncea plants grown in soil pot culture. Although lower and marginal levels of excess cadmium (100 and 250 ppm) improved growth but higher levels (500 ppm) caused significant suppression. Significant accumulation of proline, an indicator of water stress, occurred at higher level of Cd. Gradual increases in activities of certain antioxidant enzymes such as catalase and peroxidase along with increased lipid peroxidation are suggestive of disturbed oxidative metabolism. Taking together, the deleterious effects of Cd and its effects on oxidative metabolism clearly indicate enhanced generation of reactive oxygen species (ROS) to be instrumental in producing toxic effects of Cd. The excess levels of Cd also decreased the concentrations of soluble protein and chlorophylls and increased the ratio of chlorophyll a/b.