Nitric oxide synthase (NOS) associated protein in pea (Pisum sativum L.)
2011
Mui-Yun Wong | Huang, J., Universiti Putra Malaysia, 43400 Serdang (Malaysia). Inst. of Tropical Agriculture and Dept. of Plant Protection | Davis, E.L., North Carolina State Univ., NC27695-7616 Raleigh (USA). Dept. of Plant Pathology
Nitric oxide synthase (NOS) activity was detected in pea (Pisum sativum L.) leaf extracts using a citrulline formation assay that is used in mammalian system. Total protein extraction method was modified from that used in mammalian system based on biochemical activities such as the use of protease inhibitors, pH and precipitation with salts and organic solvents. Psychological aspects such as the effects of various chemicals that induce systematic resistance in plants on NOS activity and immunodetection of a NOS associated protein were also studied. The NOS associated protein was partially isolated using liquid chromatography and characterized based on mammalian NOS inhibitor and co-factor requirement. Co-relation of NOS activity and NOS associated gene expression during incompatible and compatible pea-bacterial interactions were investigated using interactions of Ralstonia solanacearum and Pseudomonas syringae pv pisi,respectively, with pea. NOS activity was detected using citrulline formation assay. NOS associated gene expression was measured using real-time reverse transcription-polymerase chain reactions (RT-PCR) and a probe designed from a cloned cDNA fragment of pea that was homologous to NOS of snail and AtNOS1/ AtNOA1 of Arabidopsis.
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