No presente trabalho foi avaliada a capacidade da planta de soja de responder ao ferimento mecânico através da via das lipoxigenases. Para isso, foram analisados o efeito do ferimento mecânico em folhas de soja sobre a atividade do “pool” de lipoxigenases e os possíveis produtos finais da via das lipoxigenases. Foram utilizados dois genótipos: um normal (variedade IAC-100) e outro com ausência completa de lipoxigenases em suas sementes (linhagem IAC-100 TN). As plantas dos dois genótipos foram submetidas a ferimento mecânico e coletadas 0, 6, 12, 24, 48 e 168 horas após o ferimento. Foram analisadas a resposta local (folíolos da primeira folha trifoliolar) e a resposta sistêmica (folíolos da segunda folha trifoliolar) de plantas dos dois genótipos.
Os resultados do estudo bioquímico e cinético das lipoxigenases de folhas para os dois genótipos analisados, utilizando ácido linoléico como substrato, revelaram:
- Dois picos mais acentuados de atividade de lipoxigenases a pH 4,5 e 6,0 nos testes e nos respectivos controles, tanto na resposta local quanto na sistêmica. Nos testes, houve aumento de atividade em resposta ao ferimento mecânico.
- Aumento da atividade de lipoxigenase tanto na resposta local quanto na sistêmica, nos dois genótipos estudados, a pH 4,5 e 6,0.
- O valor da temperatura ótima das isoenzimas de lipoxigenases a pH 6,0, em respostas local e sistêmica, foi de 25 oC.
- Os valores de KM aparente das isoenzimas de lipoxigenases foliares, em relação aos controles, aumentaram consideravelmente seis horas após o ferimento e diminuiu a partir de 12 horas, tendo em 168 horas o menor valor tanto na resposta local quanto na sistêmica, mostrando, assim, respostas local e sistêmica nos dois genótipos estudados.
- O menor valor de KMapp em 168 horas após ferimento nas respostas local e sistêmica de ambos os genótipos sugerem uma alteração do “pool” de lipoxigenases em resposta a ferimento.
- Os resultados dos dados de cinética enzimática são semelhantes para os dois genótipos estudados, indicando, possivelmente, que a remoção genética das lipoxigenases de semente não está interferindo na expressão dos genes que codificam para as lipoxigenases que atuam no mecanismo de defesa da planta de soja contra danos causados por ferimento.
- Nos dois genótipos analisados, ocorreu aumento de inibição tríptica após ferimento, tanto na resposta local quanto na sistêmica. Os valores em mg de tripsina inibida/grama de proteína foram maiores nos testes do que nos controles em 6, 12, 24, 48 e 168 horas após ferimento mecânico.
- Ocorreu resposta da planta de soja a ferimento, através do aumento de inibidores de proteases, tanto na variedade quanto no genótipo IAC-100 TN. Assim, pode-se também verificar que a remoção de lipoxigenases de semente de soja não alterou a produção de inibidores de proteases nas folhas.
- Os resultados apresentados sugerem ainda um possível mecanismo de defesa da planta de soja contra ferimento, que passa pela via das lipoxigenases e tem como produto final inibidores de proteases.
- Ocorreu ligeiro aumento dos níveis de hexanal e de aldeídos totais 48 e 168 horas após tratamento, nos dois genótipos estudados. Esses resultados sugerem que o mecanismo de defesa de plantas de soja contra ferimento, através da via das lipoxigenases, não passa pelo caminho que tem como produto final aldeídos (Figura 2).
- É possível que a planta, em resposta ao ferimento mecânico, ative a “via das lipoxigenases” e, após a formação de hidroperóxidos, ocorra a ação enzimática de uma hidroperóxido ciclase, seguindo a via de produção de ácido jasmônico e subseqüente ativação e expressão de genes que expressam inibidores de proteases, uma vez que ferimento mecânico é uma simulação da
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