摘要
Succinate dehydrogenase(SDH) is one of the key enzymes of the tricarboxylic acid cycle(TCA cycle) and the only multi-subunit one embedded in the membrane.To research the roles of the SDHA gene in Phytophthora sojae,we first cloned the conservative PsSDHA gene to construct the PsSDHA silenced expression vector pHAM34-PsSDHA,and then utilized PEG to mediate the P.sojae protoplast transformation experiment.Through transformation screening,we obtained the silenced mutants A1 and A3 identified by PCR and RT-qPCR,which have good suppression effects.Further study showed that the silenced mutant strains were shorter and more bifurcated;the growth of the silenced mutants was clearly inhibited in 10%V8 culture medium containing 0.7mol/L sodium chloride(NaCl),0.01%sodium dodecyl sulfate(SDS),3mmol/L hydrogen peroxide(H_2O_2) and 200ng/mL Congo Red.The pathogenicity of the silenced mutants was significandy reduced compared with the wild-type strain and the control strain.Through studying this gene,we can better understand the position and function of SDH in P.sojae,and we can provide theoretical basis for further research.
Succinate dehydrogenase(SDH) is one of the key enzymes of the tricarboxylic acid cycle(TCA cycle) and the only multi-subunit one embedded in the membrane.To research the roles of the SDHA gene in Phytophthora sojae,we first cloned the conservative PsSDHA gene to construct the PsSDHA silenced expression vector pHAM34-PsSDHA,and then utilized PEG to mediate the P.sojae protoplast transformation experiment.Through transformation screening,we obtained the silenced mutants A1 and A3 identified by PCR and RT-qPCR,which have good suppression effects.Further study showed that the silenced mutant strains were shorter and more bifurcated;the growth of the silenced mutants was clearly inhibited in 10%V8 culture medium containing 0.7mol/L sodium chloride(NaCl),0.01%sodium dodecyl sulfate(SDS),3mmol/L hydrogen peroxide(H_2O_2) and 200ng/mL Congo Red.The pathogenicity of the silenced mutants was significandy reduced compared with the wild-type strain and the control strain.Through studying this gene,we can better understand the position and function of SDH in P.sojae,and we can provide theoretical basis for further research.
引文