抗番茄溃疡病菌菱角壳活性物质提取条件优化及抑菌活性研究

Extraction Optimization of Active Substances from Trapa bispinosa Roxb Shell and Antibacterial Activity against Clavibacter michiganensis subsp. michiganensis

在单因素试验基础上,通过L_(27)(3^(13))正交试验对抗番茄溃疡病菌的菱角壳活性物质的提取条件进行优化。结果表明:提取温度和提取时间对活性物质抑菌效果的影响极显著,提取时间与提取温度之间及提取时间与料液比之间存在显著交互作用;优化后提取条件为提取温度60℃,提取时间9 h,料液比1︰5 g/mL。通过检测番茄溃疡病菌(Cmm)被菱角壳活性物质处理后细胞外AKP活性、细胞膜通透性及细胞膜电位,分析菱角壳活性物质抑菌活性。结果表明:菱角壳活性物质破坏Cmm的细胞壁,导致AKP外漏;改变细胞膜电位,损伤Cmm细胞膜的通透性,影响正常生命活动。

Based on single factor experiments, L_(27)(3^(13)) orthogonal experiment was used to enhance the extraction conditions of active compounds in Trapa bispinosa Roxb(TbR) shell against Clavibacter michiganensis subsp. michiganensis(Cmm). The results showed that the effects of extraction temperature and extraction time on the inhibitory effect of the active substances were extremely significant, and there were significant interactions between extraction time and extraction temperature and between extraction time and material-to-liquid ratio. The optimal extraction condition was determined as temperature 60 ℃, time 9 h and material-to-liquid ratio 1︰5 g/mL. The antibacterial activity was analyzed by detecting the changes in extracellular AKP activity, cell membrane permeability and cell membrane potential after treating with active substances of TbR shell. The results showed that the active substances of TbR shell destroyed the cell wall and leaded to AKP leakage;The active substances of TbR shell altered cell membrane potential, and impaired the permeability of the Cmm cell membrane, which interfered with normal life activities.