哈茨木霉β-葡聚糖酶诱导、纯化及对黄瓜幼苗的促生防病作用  

Induction and Purification ofβ-Glucanase from Trichoderma harzianum and Its Growth Promoting and Disease Prevention Effects on Cucumber Seedlings

作　　者：杨春林[1,2] 李洪浩 胡强[3] 席亚东 YANG Chunlin;LI Honghao;HU Qiang;XI Yadong(The Institute of Plant Protection,Sichuan Academy of Agricultural Sciences,Vegetable Germplasm Innovation and Variety Improvement Key Laboratory of Sichuan Province,Chengdu 610066,Sichuan,China;Leshan Plant Protection and Quarantine Station of Sichuan Province,Leshan 614000,Sichuan,China;Bamboo Diseases and Pests Control and Resources Development Key Laboratory of Sichuan Province,Leshan Normal University,Leshan 614000,Sichuan,China)

机构地区：[1]四川省农业科学院植物保护研究所,蔬菜品种改良与种质创新四川省重点实验室,四川成都610066 [2]四川省乐山市植保植检站,四川乐山614000 [3]乐山师范学院,竹类病虫防控与资源开发四川省重点实验室,四川乐山614000

出　　处：《中国蔬菜》2024年第4期121-128,共8页China Vegetables

基　　金：国家现代农业产业体系四川创新团队(川农函〔2019〕472号);四川省科技计划项目(2021YFYZ0021);四川蔬菜花卉种植创新与产业提升关键技术研发专项(2016CYTS-018)。

摘　　要：在实验室条件下分别以β-葡聚糖、麦麸和黄瓜枯萎病菌(Fusarium oxysporum)细胞壁作为唯一碳源诱导哈茨木霉菌(Trichoderma harzianum)Th-30发酵产生β-葡聚糖酶,采用硫酸铵盐析和琼脂糖凝胶色谱柱层析法对β-葡聚糖酶进行分离纯化,以津研4号黄瓜为试验材料,探究β-葡聚糖酶对黄瓜幼苗抗性生理指标、形态指标的影响及黄瓜常见土传病害的防控作用。结果表明:不同诱导条件下木霉Th-30发酵产β-葡聚糖酶活性差异显著,经β-葡聚糖诱导发酵的粗酶液酶活性最高,发酵72 h时达75.63 U·mL^(-1);纯化的β-葡聚糖酶比活力为783.56 U·mg^(-1),是粗酶液的45.32倍;5倍液和10倍液β-葡聚糖酶可显著提高黄瓜幼苗的根系活力、游离脯氨酸含量、苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)活性以及根冠比和壮苗指数;β-葡聚糖酶10倍液处理对黄瓜立枯病、黄瓜枯萎病、黄瓜疫病、黄瓜猝倒病、黄瓜菌核病的防效为50.36%~80.63%。Under laboratory condition,this experiment tookβ-glucan,wheat bran and cell wall of Fusarium oxysporum as the only carbon source to induce Trichoderma harzianum Th-30 fermentation andβ-glucanase production,and isolate and purifyβ-glucanase by ammonium sulfate salting out and agarose gel column chromatography.Taking‘Jinyan 4’cucumber as test material,this experiment also explored the effects ofβ-glucanase on physiological and morphological indexes of resistance,and prevention and control function of cucumber seedling on common soil-borne diseases of cucumber.The results showed that the activities ofβ-glucanase from Trichoderma Th-30 fermentation induced under different conditions were significantly different.The enzyme activity of crude enzyme liquid induced byβ-glucan was the highest,reaching 75.63 U·mL^(-1) at 72 h of fermentation.The specific activity of purifiedβ-glucanase was 783.56 U·mg^(-1),being 45.32 times of the crude enzyme.The treatments of 5-fold solution and 10-fold solution ofβ-glucanase could increase the root activity and free proline content,PAL,POD,and PPO activities of cucumber seedling,as well as the root shoot ratio and healthy seedling strength index,and the control effects of 10-fold solution ofβ-glucanase on cucumber Rhizoctonia root rot,cucumber Fusarium wilt,cucumber blight,cucumber damping-off,cucumber stem rot were 50.36%-80.63%.

关 键 词：哈茨木霉 Β-葡聚糖酶 分离纯化 黄瓜幼苗 生防作用 

分 类 号：S642.2[农业科学—蔬菜学]