核桃内生细菌HT-6对致病疫霉的持续抑制作用及其抑菌特性研究
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摘要
为进一步了解核桃内生细菌HT-6菌株在防治马铃薯晚疫病中的利用潜力,采取延长对峙培养时间以及致病疫霉预先培养后再接种HT-6的方法,对其抑菌特性及持续抑菌能力进行了评价。结果显示,延长对峙培养时间到第15天时,致病疫霉菌落半径为11.0 mm,与对照(40.0 mm)仍有极显著差异;以可溶性淀粉和葡萄糖代替黑麦培养基中的蔗糖,对于抑菌作用无明显影响;HT-6菌液浓度与抑菌效果关系密切,最适浓度分别为OD值1.088和1.928的菌液,其抑菌率分别为95.33%和91.78%,且随OD值升高其抑菌率降低;预先培养致病疫霉10 d后再接种HT-6菌液继续对峙培养21 d,仍可强效抑制致病疫霉生长;镜检发现受抑制的致病疫霉孢子囊数量显著低于对照,菌丝体和孢子囊的畸形率显著高于对照。
In order to further understand the potential of bacterial HT-6 in preventing and controlling potato late blight,methods of prolonging the dual culture time and pre-cultivating Phytophthora infestans before adding HT-6 strain were used to evaluate anti-oomycetes activity of HT-6 living body and bacterial fluid against P. infestans. The results showed that there was a significant difference in colony radius of P. infestans between dual culture(11.0 mm) and control(40.0 mm) after culture 15 d. The mycelia growth inhibition effect of HT-6 in rye medium with sucrose was similar to that with soluble starch and glucose. Anti-oomycetes effects of HT-6 bacterial fluid on pathogen were closely related with its concentrations. The optimal concentrations were those with OD1.088 and OD1.928, and the inhibitory rates were 95.33% and 91.78%, respectively. After 10 d of being added into rye medium in pre-culture P. infestans, HT-6 bacterial fluid still strongly inhibited the mycelia growth of P. infestans. Microscopic examination showed that the sporangia number of suppressed P. infestans was significantly lower than that of the control, and deformity rates of mycelia and sporangia were sig-nificantly higher than that of the control.
In order to further understand the potential of bacterial HT-6 in preventing and controlling potato late blight,methods of prolonging the dual culture time and pre-cultivating Phytophthora infestans before adding HT-6 strain were used to evaluate anti-oomycetes activity of HT-6 living body and bacterial fluid against P. infestans. The results showed that there was a significant difference in colony radius of P. infestans between dual culture(11.0 mm) and control(40.0 mm) after culture 15 d. The mycelia growth inhibition effect of HT-6 in rye medium with sucrose was similar to that with soluble starch and glucose. Anti-oomycetes effects of HT-6 bacterial fluid on pathogen were closely related with its concentrations. The optimal concentrations were those with OD1.088 and OD1.928, and the inhibitory rates were 95.33% and 91.78%, respectively. After 10 d of being added into rye medium in pre-culture P. infestans, HT-6 bacterial fluid still strongly inhibited the mycelia growth of P. infestans. Microscopic examination showed that the sporangia number of suppressed P. infestans was significantly lower than that of the control, and deformity rates of mycelia and sporangia were sig-nificantly higher than that of the control.
引文
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[15]王超,郭坚华,席运官,等.拮抗细菌在植物病害生物防治中应用的研究进展[J].江苏农业科学,2017,45(18):1-6.
[16]Hunziker L,Bonisch D,Groenhagen U,et al.Pseudomonas strains naturally associated with potato plants produce volatiles with high potential for inhibition of Phytophthora infestans[J].Applied and Environmental Microbiology,2015,81(3):821-830.
[17]万安琪,蒋继志,沙海天,等.拮抗细菌W-7的鉴定及其对致病疫霉的抑制作用[J].河南农业科学,2017,46(2):55-59,82.
[18]崔月贞,杨小利,杨成德,等.拮抗马铃薯晚疫病菌的高寒草地牧草内生细菌的鉴定及其生物功能测定[J].植物保护学报,2016,43(05):789-795.
[19]李双东,曹克强.枯草芽孢杆菌EB-28对马铃薯晚疫病菌的抑制作用及防病效果[J].安徽农业科学,2009,37(24):11376-11378.
[2]Axel C,Zannini E,Coffey A,et al.Ecofriendly control of potato late blight causative agent and the potential role of lactic acid bacteria:a review[J].Appllied Microbiological Biotechnology,2012,96(1):37-48.
[3]Giovanna D,Kevin M,Mideros M F,et al.An ephemeral sexual population of Phytophthora infestans in the Northeastern United States and Canada[J].Plos One,2014,9(12):e116354-e116354.
[4]蒋继志,郭俊亭,李丽艳.致病疫霉接抗真菌研究进展[J].河北大学学报,2012(1):105-112.
[5]张蕾,蒋继志,刘璇,等.致病疫霉拮抗内生细菌的筛选及离体防病作用[J].河北农业大学学报,2013,36(2):75-79.
[6]李颖,蒋继志,金鑫,等.3株放线菌对致病疫霉抑制作用的比较研究[J].中国植保导刊,2016,36(9):11-15.
[7]杨立宾,宋瑞清,邓勋,等.致病疫霉生防木霉菌株筛选鉴定及生物学特性[J].东北林业大学学报,2013(7):118-122.
[8]黄英菊,蒋继志,冯丽娜,等.几种拮抗菌对致病疫霉抑制作用的比较研究[J].河北农业大学学报,2014,37(4):81-85.
[9]喻江.玉米和大豆根内生细菌多样性及促生细菌鉴定评价[D].哈尔滨:东北农业大学,2016.
[10]吴彩兰.加工番茄内生细菌种群动态及内生芽孢杆菌抑菌促生作用研究[D].新疆维吾尔自治区:石河子大学,2016.
[11]郭尧,李丽,蒋继志,等.致病疫霉拮抗放线菌Sy11发酵条件的优化[J].河北农业大学学报,2014,37(2):71-75.
[12]惠娇,蒋继志,鲍雪,等.致病疫霉拮抗放线菌NB8发酵条件的优化[J].河北农业大学学报,2011,31(2):86-89.
[13]杨胜先,张绍荣,龙国,等.贵州省马铃薯晚疫病菌生理小种的组成与分布[J].南方农业学报,2015,46(4):597-601.
[14]王鹏,朱荣菊.食品中持久性有机污染物现状及对策研究[J].食品安全质量检测学报,2016,7(11):4557-4561.
[15]王超,郭坚华,席运官,等.拮抗细菌在植物病害生物防治中应用的研究进展[J].江苏农业科学,2017,45(18):1-6.
[16]Hunziker L,Bonisch D,Groenhagen U,et al.Pseudomonas strains naturally associated with potato plants produce volatiles with high potential for inhibition of Phytophthora infestans[J].Applied and Environmental Microbiology,2015,81(3):821-830.
[17]万安琪,蒋继志,沙海天,等.拮抗细菌W-7的鉴定及其对致病疫霉的抑制作用[J].河南农业科学,2017,46(2):55-59,82.
[18]崔月贞,杨小利,杨成德,等.拮抗马铃薯晚疫病菌的高寒草地牧草内生细菌的鉴定及其生物功能测定[J].植物保护学报,2016,43(05):789-795.
[19]李双东,曹克强.枯草芽孢杆菌EB-28对马铃薯晚疫病菌的抑制作用及防病效果[J].安徽农业科学,2009,37(24):11376-11378.