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生防因子螺旋毛壳(Chaetomium spirale)ND35生长于含立枯丝核菌(Rhizoctonia solani)菌丝细胞壁制备物的SM培养基中,从培养滤液提取的粗酶液强烈抑制Valsa sordida、Valsa mali、Glomerella cingulata和Curvulavia lunata病原真菌的菌丝生长和孢子萌发。经DNS法检测,粗酶液同时具有β-1,3-葡聚糖酶(包括内切和外切酶)和几丁质酶活性,分别为0.19 U.mg-1和0.09 U.mg-1。生长于3%玉米粉浸渍液的螺旋毛壳ND35的培养滤液也能抑制上述病原菌的菌丝生长和孢子萌发。离体条件下检测了ND35产生的一纯化的具有分子量为73 kD的内切β-1,3-葡聚糖酶(GLUC73)和一纯化的抗生素对苹果炭疽病菌的分生孢子萌发和芽管延长的抑制效果。当内切β-1,3-葡聚糖酶使用浓度为180μg.mL-1时,抑制了测试真菌的孢子萌发,造成细胞壁的改变,导致了菌丝顶端的破裂;抗生素的有效抑菌中浓度ED50为1.75μg.mL-1。单独应用时,1.0μg.mL-1的抗生素或40μg.mL-1的内切β-1,3-葡聚糖酶没有或仅有很低的抑菌效果;两者组合导致孢子萌发约78%受抑制。甚至10μg.mL-1的内切β-1,3-葡聚糖酶也使1.5μg.mL-1抗生素的抑菌作用从低于25%增加到超过50%的抑菌效果。此外,80μg.mL-1的内切β-1,3-葡聚糖酶使1.0μg.mL-1抗生素的抑菌活性从0%提高到90%。抗生素和β-1,3-葡聚糖酶的协同抗菌活性可能在内生菌螺旋毛壳的植病生防中起重要作用。
Biocontrol Factor Chaetomium spirale ND35 is grown in SM medium containing Rhizoctonia solani hyphal cell wall preparations and the crude enzyme solution extracted from the culture filtrate strongly inhibits Valsa sordida, Valsa mali, Mycelial growth and spore germination of Glomerella cingulata and Curvulavia lunata pathogenic fungi. The crude enzyme solution had both β-1,3-glucanase (including endo- and exo-enzyme) and chitinase activity by the DNS method, which were 0.19 U.mg-1 and 0.09 U.mg- 1. The culture filtrates of Sphaerotheca brasiliensis ND35 grown in 3% maize meal dipping solution also inhibited mycelial growth and spore germination of the above pathogenic bacteria. In vitro, a purified ND35-producing endosymbiotic β-1,3-glucanase (GLUC73) with a molecular weight of 73 kD and a purified antibiotic were tested for conidiospore germination and germ tube of apple anthracnose Prolonged inhibitory effect. When the concentration of endo-β-1,3-glucanase was 180μg.mL-1, the spore germination of the tested fungi was inhibited and the cell wall was changed, leading to the rupture of the top of the mycelium. The effective antibacterial activity of antibiotics The concentration of ED50 was 1.75 μg.mL-1. On application alone, either 1.0 μg.mL-1 of the antibiotic or 40 μg.mL-1 of the endo-β-1,3-glucanase had no or very low inhibitory effect; the combination of the two resulted in a spore germination of about 78 % Inhibited. Even 10 μg.mL-1 of endo-β-1,3-glucanase also increased the bacteriostatic effect of 1.5 μg.mL-1 antibiotics from less than 25% to over 50% of the antibacterial effect. In addition, 80μg.mL-1 endo-β-1,3-glucanase increased the antibacterial activity of 1.0μg.mL-1 antibiotics from 0% to 90%. The synergistic antibacterial activity of antibiotics and β-1,3-glucanase may play an important role in the biocontrol of endophyte Helicottifolia.