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Effective Screening Methods for Lipoxygenase Isozymes in Soybean Seeds  

Kim, Young Jin (Honam Agricultural Research Institute, NICS, RDA)
Park, Tae Il (Honam Agricultural Research Institute, NICS, RDA)
Cho, Sang Kyun (Honam Agricultural Research Institute, NICS, RDA)
Oh, Young Jin (Honam Agricultural Research Institute, NICS, RDA)
Kim, Tae Soo (Honam Agricultural Research Institute, NICS, RDA)
Kim, Jung Gon (Honam Agricultural Research Institute, NICS, RDA)
Publication Information
Korean Journal of Breeding Science / v.40, no.1, 2008 , pp. 26-30 More about this Journal
Abstract
Normal soybean seed contains three lipoxygenase isozymes called L-1, L-2, and L-3, respectively, which are responsible for the generation of undesirable grassy-beany flavors. Simple and effective methods for the detection of lipoxygenase isozymes were developed in soybean seeds. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) has been tried in separating these isozymes. It was done effectively on 7.5% separating gel and 4.5% stacking gel. However, no reliable method has been developed specifically for separating L-3, L-13 and L-23. Visual judging methods were based on the bleaching activities of lipoxygenase in contact with methylene blue and ${\beta}$-carotene. Sodium linoleate bleaching method was adopted to determine L-1 and L-2. Carotene bleaching and spectrophotometric methods were used to determine L-3. These systems were very rapid within one minute, furthermore only required a small piece of cotyledon (below 10 mg) and the other part could be used for generation advance after analysis. It was demonstrated that 200 seed samples could be analyzed per day by one laboratory assistant. The combination of visual judging methods and electrophoresis is suitable for breeding programs. It took 6.5 hours for analysis of 100 seed samples by one person.
Keywords
soybean; lipoxygenase isozymes; SDS-PAGE; visual judging method;
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