拮抗菌XY1培养条件优化研究
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摘要
为了研究分离自杏叶上的拮抗菌XY1的最佳培养条件,采用梨黑斑病菌和梨枯萎病菌抑菌活性为靶标,通过单因素试验和3指标5因素4水平的正交试验,筛选出拮抗菌株XY1发酵的最优培养基配方和发酵条件。结果表明,XY1发酵的最优培养基配方为:胰蛋白胨8 g,酵母膏5 g,蔗糖9 g,蒸馏水1 000 mL,初始pH值7.0;最优培养条件为:温度25℃,初始pH值7.0,接种量1%,每250 mL三解瓶装液量40 mL,发酵48 h。在此条件下,与优化前相比,拮抗菌XY1菌体生长量提高6百分点,梨黑斑病抑菌率提高9百分点,梨枯萎病抑菌率提高10.1百分点。
The study was undertaken to investigate the best culture conditions of Pseudomonas aeruginosa XY1 isolated from apricot leaf. Optimal culture media formula for strain XY1 was screened with Alternaria. alternata and Fusarium. oxysporum of gold pear tracer analysis. Fermentation condition for strain XY1 was screened with single factor experiment and three-indicator, five-factor and four-level orthogonal experiment. The results showed that the optimal culture media formula for strain XY1 fermentation was 8 g tryptone,5 g yeast extract, 9 g sucrose, 1 000 mL distilled water and 7.0 initial pH value. The optimal fermentation condition for strain XY1 was 25 ℃ temperature, 40 mL/250 mL loading volume, 1% inoculation volume and 48 h fermentation time. After optimization,the growing mass of XY1 increased for 6% , the inhibition rate causing pear black spot increased for 9% and the inhibition rate causing pear blight increased for 10.1% more than before.
The study was undertaken to investigate the best culture conditions of Pseudomonas aeruginosa XY1 isolated from apricot leaf. Optimal culture media formula for strain XY1 was screened with Alternaria. alternata and Fusarium. oxysporum of gold pear tracer analysis. Fermentation condition for strain XY1 was screened with single factor experiment and three-indicator, five-factor and four-level orthogonal experiment. The results showed that the optimal culture media formula for strain XY1 fermentation was 8 g tryptone,5 g yeast extract, 9 g sucrose, 1 000 mL distilled water and 7.0 initial pH value. The optimal fermentation condition for strain XY1 was 25 ℃ temperature, 40 mL/250 mL loading volume, 1% inoculation volume and 48 h fermentation time. After optimization,the growing mass of XY1 increased for 6% , the inhibition rate causing pear black spot increased for 9% and the inhibition rate causing pear blight increased for 10.1% more than before.
引文
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[19]闫建芳,刘秋,赵柏霞,等.真菌广谱拮抗菌GQ-17的鉴定、发酵液性质研究及发酵条件优化[J].沈阳农业大学学报,2012,43(5):534-540.
[20]邓振山,侯改成,孙志宏,等.番茄灰霉病菌拮抗菌D6和D10发酵条件的优化及其抑菌效果[J].西北农林科技大学学报(自然科学版),213,41(9):73-77.
[21]李红亚,李术娜,朱宝成.棉花黄萎病拮抗细菌B.velezensis6-61拮抗蛋白发酵条件的优化[J].中国农学通报,2011,27(21):93-99.
[22]姜云,尹望,陈长卿,等.人参内生拮抗细菌NJ13的鉴定及发酵条件[J].农药,2013,52(2):97-101.
[23]石炳兴,赵红,刘春朋,等.抗生素AGPM摇瓶发酵条件的正交试验[J].过程工程学报,2001(4):442-444.
[2]吕增芳.花椒树蚜虫危害严重的原因及防治建议[J].山西果树,2010(1):39-40.
[3]张小平.化学农药对农业生态环境的污染及防治[J].生态经济,2003(10):166-168.
[4]周文强,樊慧梅.拮抗微生物在生物防治中的研究进展[J].辽宁农业科学,2005(5):32-34.
[5]范永玲,任璐,刘秀英,等.番茄早疫病菌对3种杀菌剂的抗药性监测[J].安徽农业科学,2008,36(26):11431-11433.
[6]吴永官,陆少峰,黄思良,等.华南地区瓜类疫霉对甲霜灵的田间抗药性[J].微生物学报,2011(8):1078-1086.
[7]张荣意,谭志琼,简日明.芒果炭疽病菌生防细菌的筛选、鉴定及生防潜能的初步研究[J].热带作物学报,1998(3):21-27.
[8]BRAVO O N.In vitro effect of Bacillus sp.on growth,sporulation and germination of conidia and spores of new fungi[J].Fitopatologia Colombiana,1993(17):62-72.
[9]詹喜.枯草芽孢杆菌对柑橘采后病害的生物防治及其机理研究[D].杭州:浙江大学,2006.
[10]陈丽锋.柑橘采后蒂腐病菌生物学特性及其拮抗菌研究[D].武汉:华中农业大学,2007.
[11]郝卫宁,李辉,胡美英,等.柑桔绿霉病拮抗细菌的筛选、鉴定及其抑制效果[J].中国生物防治学报,2011,27(2):284-288.
[12]PUSEY P L,WILSON C L.Postharvest biological control of stone fruit brown rot by Bacillus subtilis[J].Plant Diseases,1984,68(2):753-756.
[13]UTKHEDE R S,SHOLBERG P L.In vitro inhibition of plant pathogens by Bacillus subtilis and Enterobacter aerogenes and in vivo control of two postharvest cherry diseases[J].Canadian Journal of Microbiology,1986,32(4):963-967.
[14]VERO S,MONDINO P,BURGUENO J,et al.Characterization of biocontrol activity of two yeast strains from Uruguay against blue mold of apple[J].Postharvest Biology and Technology,2002,26(1):91-98.
[15]汪腾,段雅婕,刘兵团,等.两株香蕉枯萎病拮抗菌在香蕉体内的定殖[J].基因组学与应用生物学,2011,30(3):342-350.
[16]陈志谊,许志刚,高泰东,等.水稻纹枯病拮抗细菌的评价与利用[J].中国水稻科学,2000,14(2):98-102.
[17]鹿秀云,李社增,栗秋生,等.玉米叶斑病拮抗细菌的筛选及其发酵培养基优化[J].中国生物防治,2006,22(Z):47-53.
[18]朱宏建,欧阳小燕,周倩,等.一株辣椒尖孢炭疽病菌拮抗菌株的分离鉴定与发酵条件优化[J].植物病理学报,2012,42(4):418-424.
[19]闫建芳,刘秋,赵柏霞,等.真菌广谱拮抗菌GQ-17的鉴定、发酵液性质研究及发酵条件优化[J].沈阳农业大学学报,2012,43(5):534-540.
[20]邓振山,侯改成,孙志宏,等.番茄灰霉病菌拮抗菌D6和D10发酵条件的优化及其抑菌效果[J].西北农林科技大学学报(自然科学版),213,41(9):73-77.
[21]李红亚,李术娜,朱宝成.棉花黄萎病拮抗细菌B.velezensis6-61拮抗蛋白发酵条件的优化[J].中国农学通报,2011,27(21):93-99.
[22]姜云,尹望,陈长卿,等.人参内生拮抗细菌NJ13的鉴定及发酵条件[J].农药,2013,52(2):97-101.
[23]石炳兴,赵红,刘春朋,等.抗生素AGPM摇瓶发酵条件的正交试验[J].过程工程学报,2001(4):442-444.