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首页> 外文期刊>Acta Physiologiae Plantarum >A Bowman–Birk protease inhibitor with antifeedant and antifungal activity from Dolichos biflorus
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A Bowman–Birk protease inhibitor with antifeedant and antifungal activity from Dolichos biflorus

机译：一种来自Dolichos biflorus的具有拒食和抗真菌活性的Bowman–Birk蛋白酶抑制剂

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摘要

In the present study, 11 varieties of Dolichos biflorus exhibited both protease inhibitor activities as well as in vitro inhibitory activity against Helicoverpa armigera gut protease. A Bowman–Birk protease inhibitor showing activity against trypsinand α-chymotrypsin has been purified from D. biflorus seeds using multi-step strategy. The purified inhibitor revealed a single band on SDS-PAGE corresponding to molecular mass of 16 kDa. The inhibitory constants for the interaction of purified PI withtrypsin and α-chymotrypsin were 0.04 and 0.48 μM, respectively. The purified inhibitor was stable over a pH range of 2–12 and up to a temperature of 100 °C for 20 min. The results of insect bioassay against H. armigera revealed 68 % decline in larvalweight after 7 days of feeding on artificial diet containing the inhibitor. The larval growth and % leaf area eaten were drastically reduced in the presence of inhibitor. The observed cumulative mortality from larval to adult was 51.21 %. The inhibitordisplayed antifungal activity against Alternaria alternata, Fusarium oxysporum, and Aspergillus niger with minimum inhibitory concentration as 0.4, 0.6, and 1.2 mu g mL~(-1), respectively. This is the first report of anti-feedant and anti-fungal activities of D. biflorus protease inhibitor on a single protein, which might be important for developing transgenic plants resistant to insect pests and fungal pathogens.
机译：在目前的研究中，11个种类的Dolichos biflorus既显示蛋白酶抑制剂活性，又显示对棉铃虫肠道蛋白酶的体外抑制活性。一种鲍曼-伯克蛋白酶抑制剂，对胰蛋白酶和α-胰凝乳蛋白酶具有活性，已使用多步策略从双花木seeds种子中纯化出来。纯化的抑制剂在SDS-PAGE上显示出一条条带，对应于16 kDa的分子量。纯化的PI与胰蛋白酶和α-胰凝乳蛋白酶相互作用的抑制常数分别为0.04和0.48μM。纯化的抑制剂在2至12的pH范围内以及在100°C的温度下可稳定20分钟。昆虫对棉铃虫的生物分析结果表明，喂食含有该抑制剂的人工饲料7天后，幼虫体重下降了68％。在抑制剂的存在下，幼虫的生长和所吃的叶面积的百分比大大降低。观察到的从幼虫到成年的累积死亡率为51.21％。该抑制剂对链格孢菌，尖孢镰刀菌和黑曲霉均显示出抗真菌活性，其最低抑菌浓度分别为0.4、0.6和1.2μg mL〜（-1）。这是D. biflorus蛋白酶抑制剂对单一蛋白质的抗饲料和抗真菌活性的首次报道，这对于开发对害虫和真菌病原体具有抗性的转基因植物可能很重要。

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