The relationships between changes in energy metabolism and the antioxidant defense system in the weed species Ipomoea triloba L. during seed germination and early seedling growth were investigated. The effects of some common allelochemicals on these parameters also were studied. Respiratory activity and the activities of alcohol dehydrogenase, superoxide dismutase, catalase, guaicol peroxidase, ascorbate peroxidase, glutathione reductase, and lipoxygenase were measured. Mitochondrial oxidative phosphorylation resumed shortly after the seed imbibition period, as indicated by considerable KCN-sensitive respiratory activity in embryos of I. triloba. The occurrence of superoxide dismutase, catalase, guaicol peroxidase, and lipoxygenase activities in the embryos, along with significant KCN-insensitive respiration, suggest that production of reactive oxygen species (ROS) is initiated as soon as mitochondrial respiration is resumed. All assayed antioxidant enzymes were present in the embryos except ascorbate peroxidase, which appeared only in primary roots. The activities of antioxidant enzymes increased after completion of germination, especially in primary roots. Superoxide dismutase, catalase, and guaicol peroxidase probably were the crucial enzymes involved in the neutralization of ROS, since they had higher levels of activity compared with other enzymes, such as ascorbate peroxidase and glutathione reductase. When seeds were grown in the presence of α-pinene, coumarin, quercetin, and ferulic acid, there was an additional increase in activities of antioxidant enzymes, as well as increases in lipoxygenase activity and KCN-insensitive respiration, suggesting a further increase in ROS generation. The antioxidant defense system of I. triloba was not effective in preventing lipid peroxidation caused by α-pinene. The data indicate that during seed germination and initial growth of I. triloba, a period when antioxidant enzyme activity increases to counteract the harmful ROS effects produced during mitochondrial metabolism resumption, the presence of allelochemicals, which cause further oxidative stress, may leave the seeds/seedlings more vulnerable to cellular dysfunction and cell death.
Nine natural plant compounds were screened for phytotoxicity to Bidens pilosa L. a troublesome weed in field and plantation crops. The sensitivity of three other weed species to coumarin, the most active identified compound, was also evaluated. Coumarin, at a concentration of 500 microM, had little effect on germination and growth of Senna obtusifolia L., Euphorbia heterophylla L., and Ipomoea grandifolia L. when compared with its effects on B. pilosa L. In a concentration range of 10-100 microM, coumarin caused a dose-dependent inhibition of germination and growth of B. pilosa L. The measurements of some parameters of energy metabolism revealed that coumarin-treated root tissues exhibited characteristics of seedlings in an earlier stage of growth, including higher respiratory activity and higher activities of alcohol dehydrogenase and lipoxygenase. These results suggest that coumarin inhibition of germination and growth of B. pilosa L. was not a consequence of an impairment of energy metabolism. Rather, it seems to act as a cytostatic agent, retarding germination. At concentrations above 50 microM, coumarin increased lipoxygenase activity and the level of conjugated dienes of root extracts, suggesting that it may induce oxidative stress in seedling roots.
O emprego do microscópio eletrônico de transmissão (MET) é bastante difundido no estudo de materiais biológicos, pois ele permite definição de imagens intracelulares, permitindo estudos de morfologia celular e de aspectos gerais das organelas. As imagens são produzidas por meio da interação inicial de um feixe de elétrons com a amostra, que é mantida sob condições de alto vácuo. Tanto o ambiente de vácuo como a própria irradiação dos elétrons durante a observação, representam condições adversar para a amostra, especialmente no caso de materiais orgânicos ou biológicos, que são por natureza hidratados, frágeis e elétricamente isolantes. Considerando que existem várias técnicas de preparo para MET, mas pouca descrição detalhada destas técnicas na literatura, principalmente para o preparo de raízes de plântulas, assim, esse trabalho teve como objetivo aplicar e adaptar essas técnicas para o estudo de boa qualidade de raízes de plântulas. As amostras primeiramente foram lavadas em tampão fosfato salina, passando pela fixação primária com glutaraldeído, depois pós-fixação com tetróxido de Ósmio, onde posteriormente foram desidratadas com acetona e infiltradas com resina Epon, finalizando com a polimerização, ultramicrotomia e contrastação da amostra. Contudo, essa descrição detalhada da metodologia aplicada para raiz de plântulas, pode ser utilizada com segurança e eficácia por outras pessoas, pois quase não existem descrições adaptadas de metodologia de MET na literatura, principalmente para raiz de plântulas.
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