利用凝胶扩散法测定刺萼龙葵种子中β-甘露聚糖酶的活性

β-甘露聚糖酶是种子萌发过程中降解胚乳细胞壁的关键酶,明确其活性的动态变化可为揭示杂草种子的休眠萌发机制提供重要依据。以外来杂草刺萼龙葵Solanum rostratum Dunal种子为材料,建立了种子中β-甘露聚糖酶活性的检测方法—凝胶扩散法。利用凝胶扩散法对不同贮存时间及贮存条件下刺萼龙葵种子中β-甘露聚糖酶的活性进行了检测,发现贮存3年以上的种子中该酶的活性为0.03 nmol/(min·mg),显著低于贮存3年以下的种子中的酶活性0.15 nmol/(min·mg),而湿润冷藏的种子中β-甘露聚糖酶的活性为0.12 nmol/(min·mg),显著高于干燥冷藏的种子的酶活性0.02 nmol/(min·mg)。实际应用结果表明,凝胶扩散法综合了传统方法的优势,检测特异性强、灵敏度高、重复性好,可同时检测大量种子样品,具有良好的实用性。

Determination of endo-β-mannanase activity in seeds of Solanum rostratum by gel diffusion assay

Endo-β-mannanase(EBM)is a key enzyme in degradation of endosperm cell walls during seed germination process.Understanding the dynamics of EBM activities will provide important guidance for elucidating the mechanisms of weed seed dormancy and germination.For determination of EBM activity in seeds of an invasive plant,Solanum rostratum Dunal(buffalobur),a gel diffusion assay was established.The EBM activities in buffalobur seeds with different storage time and under different storage conditions were detected by this method.The results showed that the EBM activity in seeds stored longer than 3 years0.03 nmol/(min·mg)]was significantly lower than that of seeds stored less than 3 years0.15 nmol/(min·mg)].Whereas for seeds stored wet in the refrigerator0.12 nmol/(min·mg)],the EBM activity was significantly higher than that of seeds stored dry in the refrigerator0.02 nmol/(min·mg)].The practical application verified that the gel diffusion assay,which took advantages of traditional methods,was a specific,highly sensitive and reproducible method,and could analyze a large number of seed samples simultaneously.