寡雄腐霉发酵参数优化及发酵液的生防效应

doi:10.3864/j.issn.0578-1752.2013.02.008

摘要/Abstract

摘要： 【目的】优化寡雄腐霉（Pythium oligandrum）发酵条件，为烟草黑胫病的生物防治奠定基础。【方法】采用单因素和正交试验研究寡雄腐霉的发酵参数及优化组合，并利用烟草黑胫病菌（Phytophthora nicotiana）、离体烟叶和盆栽烟苗验证发酵液的生防效应。【结果】随培养时间的延长，寡雄腐霉菌丝生物量持续增加，但发酵液对烟草黑胫病菌的抑制作用增加至峰值后降低。在不同培养基、温度、pH值和光照等条件下，寡雄腐霉的生物量越大，发酵液对烟草黑胫病菌的抑制作用愈强，二者的相关系数r =0.77**（n=16）。以黄豆饼粉为培养原料，初始pH值6.0、培养温度30℃、持续光照培养6 d所得的寡雄腐霉菌丝生物量最高，其发酵液对烟草黑胫病菌抑制率分别达到78.8%（菌丝生长）和52.9%（孢子萌发）。此外，寡雄腐霉发酵液显著降低离体烟叶和盆栽烟苗黑胫病的发病率和病情指数，对离体烟叶黑胫病的防效为69.3%—75.3%；对盆栽烟苗黑胫病的防效为55.0%—61.3%。【结论】在寡雄腐霉发酵过程中，选用适宜的培养基、pH值、温度、光照和发酵时间组合，可使发酵液抑制烟草黑胫病的效果达到最佳。

关键词: 寡雄腐霉 , 发酵参数 , 烟草黑胫病菌 , 生防效应

Abstract: 【Objective】 The objective of this study is to optimize the conditions of producing Pythium oligandrum fermentation broth and establish a theoretical foundation for biological control of tobacco black shank. 【Method】 The fermentation parameters were optimized through a single factor and orthogonal experiments, and the biocontrol effects of the fermentation broth were researched by tests on Phytophthora nicotiana, leaves of tobacco in vitro and potted plants in greenhouse. 【Result】 The biomass increased continuously with the culture time, but the inhibition rate increased to the peak then had downward trend. Under the condition of different culture media, temperature, initial pH, illumination time, the more the biomass of mycelia had, the stronger inhibition activity would express, and the correlation coefficients was 0.77**(n=16). Under the condition of culture medium composed of soybean cake powder, initial pH 6.0, 30℃, incubating 6 d and 24 h•d-1 illumination, the biomass of mycelia was the highest and the fermentation broth reached 78.8% for the growth of mycelia and 52.9% for the germination of spore on P. nicotiana. Moreover, the disease incidence and disease index were significantly decreased, the control effect was 69.3%-75.3% in the test on leaves of tobacco in vitro and 55.0%-61.3% in potted plants in greenhouse. 【Conclusion】 In the process of P. oligandrum fermentation, selecting the suitable conditions of the culture medium, pH value, temperature, illumination time and fermentation time, the best inhibition of tobacco black shank can be obtained.

Key words: Pythium oligandrum , fermentation parameter , Phytophthora nicotiana , biocontrol effect

赵建, 袁玲, 黄建国. 寡雄腐霉发酵参数优化及发酵液的生防效应[J]. 中国农业科学, 2013, 46(2): 292-299.

ZHAO Jian, YUAN Ling, HUANG Jian-Guo. Fermentation Parameter Optimization of Pythium oligandrum and Biocontrol Effect of the Fermentation Broth[J]. Scientia Agricultura Sinica, 2013, 46(2): 292-299.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2013.02.008

https://www.chinaagrisci.com/CN/Y2013/V46/I2/292

参考文献

[1]Antonopoulos D F, Melton T, Mila A L. Effects of chemical control, cultivar resistance, and structure of cultivar root system on black shank incidence of tobacco. Plant Disease, 2010, 94(5): 613-620.

[2]楼兵干, 张炳欣. 无致病性腐霉的生防作用和诱导防卫反应. 植物保护学报, 2005, 32(1): 93-96.

Lou B G, Zhang B X. Biocontrol and induction of defense responses by the non-pathogenic Pythium spp. Acta Phytophylacica Sinica, 2005, 32(1): 93-96. (in Chinese)

[3]Brozova J. Exploitation of the mycoparasitic fungus Pythium oligandrum in plant protection. Plant Protection Science, 2002, 38(1): 29-35.

[4]Wulff E G, Pham A T H, Cherif M, Rey P, Tirilly Y, Hockenhull J. Inoculation of cucumber roots with zoospores of mycoparasitic and plant pathogenic Pythium species: differential zoospore accumulation, colonization ability and plant growth response. European Journal of Plant Pathology, 1998, 104(1): 69-76.

[5]Al-Rawahi A K, Hancock J G. Parasitism and biological control of Verticillium dahliae by Pythium oligandrum. Plant Disease, 1998, 82(10): 1100-1106.

[6]Picard K, Ponchet M, Blein J P, Rey P, Tirilly Y, Benhamou N. Oligandrin. A proteinaceous molecule produced by the mycoparasite Pythium oligandrum induced resistance to Phytophthora parasitica infection in tomato plants. Plant Physiology, 2000, 124(1): 379-396.

[7]Takenaka S, Nishia Z, Nakamura Y. Induction of defense reactions in sugar beet and wheat by treatment with cell wall protein fractions from the mycoparasite Pythium oligandrum. Phytopathology, 2003, 93(10): 1228-1232.

[8]Takenaka S, Tamagake H. Foliar spray of a cell wall protein fraction from the biocontrol agent Pythium oligandrum induces defence- related genes and increases resistance against Cercospora leaf spot in sugar beet. Journal of General Plant Pathology, 2009, 75(5): 340-348.

[9]Kurzawinska H, Mazur S. The effect of chitosan and Pythium oligandrum used in protection of potato tubers against late blight and soft rot. Progress on Chemistry and Application of Chitin and Its Derivatives, 2008(8): 117-123.

[10]U. S. Environmental Protection Agency. Biopesticide Registration Action Document for Pythium oligandrum DV 74. 2007, 5, 7: 1-27.

[11]Punja Z K, Utkhede R S. Using fungi and yeast to manage vegetable crop diseases. Trend in Biotechnology, 2003, 21(9): 400-407.

[12]林健文, 罗志兵, 骆星丹, 张素红, 裴炎, 张永军. 莱氏野村菌固体发酵培养基筛选与发酵条件优化. 植物保护学报, 2009, 36(5): 437-442.

Lin J W, Luo Z B, Luo X D, Zhang S H, Pei Y, Zhang Y J. Screen of solid media and optimization of fermentation condition of Nomuraea rileyi. Acta Phytophylacica Sinica, 2009, 36(5): 437-442. (in Chinese)

[13]李芳, 葛慈斌, 刘波, 陈家骅. 淡紫拟青霉次生代谢物质的抑菌效应. 植物保护学报, 2006, 33(1): 94-98.

Li F, Ge C B, Liu B, Chen J H. The inhibition effect of secondary metabolite of Paecilomyces lilacinus against Fusarium oxysporum. Acta Phytophylacica Sinica, 2006, 33(1): 94-98. (in Chinese)

[14]冯志珍, 陈太春, 段军娜, 陈德鑫, 成巨龙, 安德荣. 烟草黑胫病拮抗根际芽孢杆菌FB-16的筛选鉴定及其抑菌活性. 植物保护学报, 2012, 39(3): 224-230.

Feng Z Z, Chen T C, Duan J N, Chen D X, Cheng J L, An D R. Screening, identification and antifungal activity of antagonistic rhizospheric Bacillus FB-16 against tobacco black shank. Acta Phytophylacica Sinica, 2012, 39(3): 224-230. (in Chinese)

[15]胡军华, 周泽扬, 蓝希钳, 毕朝位, 谢洁. 假单胞杆菌20#-5菌株对致病疫霉的作用. 西南农业学报, 2007, 20(5): 1002-1006.

Hu J H, Zhou Z Y, Lan X Q, Bi C W, Xie J. Effect of Pseudomonas spp. Strain 20#-5 on Phytophthora infestants. Southwest China Journal of Agricultural Sciences, 2007, 20(5): 1002-1006. (in Chinese)

[16]杨春林, 席亚东, 谢华蓉, 刘波微, 张敏, 彭化贤. 哈茨木霉Th-30几丁质酶的生产条件及对灰霉病菌的拮抗作用. 植物保护学报, 2009, 36(4): 295-300.

Yang C L, Xi Y D, Xie H Y, Liu B W, Zhang M, Peng H X. Chitinase production conditions of Trichoderma harzianum Th-30 and its antagon istic activ ity against Botrytis cinerea. Acta Phytophylacica Sinica, 2009, 36(4): 295-300. (in Chinese)

[17]Lee S J, Song H, Lee S Y. Genome-based metabolic engineering of Mannheimia succiniciproducens for succinic acidproduction. Applied and Environmental Microbiology, 2006, 72(3): 1939-1948.

[18]张博, 吴晓丽, 吴慧玲. P. syringae pv. glycinea M-18生产冠菌素补料发酵工艺优化研究. 中国农业科学, 2010, 43(12): 2533-2540.

Zhang B, Wu X L, Wu H L. Study on fed batch fermentation of producing coronatine with P. syringae pv. glycinea M-18. Scientia Agricultura Sinica, 2010, 43(12): 2533-2540. (in Chinese)

[19]李春艳, 李艳春, 成小松, 肖晶, 熊明华, 潘俊波. 一株丁草胺降解菌的分离鉴定及培养条件优化. 环境科学学报, 2010, 30(2): 347-353.

Li C Y, Li Y C, Cheng X S, Xiao J, Xiong M H, Pan J B. Isolation and identification of a butachlor degrading bacterium and optimization of its growth conditions. Acta Scientiae Circumstantiae, 2010, 30(2): 347-353. (in Chinese)

[20]王爱英, 楼兵干, 徐同. 寡雄腐霉分泌物对植物病原真菌的抑制作用及其对番茄灰霉病的防治效果. 植物保护学报, 2007, 34(1): 57-60.

Wang A Y, Lou B G, Xu T. Inhibitory effect of the secretion of Pythium oligandrum on plant pathogenic fungi and the control effect against tomato grey mould. Acta Phytophylacica Sinica, 2007, 34(1): 57-60. (in Chinese)

[21]Picard K, Tirilly Y, Benhamou N. Cytological effects of cellulases in the parasitism of Phytophthora parasitica by Pythium oligandrum. Applied and Environmental Microbiology, 2000, 66(10): 4305- 4314.

[22]Takahashi H, Hase S, Kanayama Y, Takenaka S. Identification of a protein that interacts with LeATL6 ubiguitin-protein ligase E3 upregulated in tomato treated with elicitin-like cell wall protein of Pythium oligandrum. Journal of Phytopathology, 2010, 158: 132-136.

[23]Hase S, Shimizu A, Nakaho K, Takenaka S, Takahashi H. Induction of transient ethylene and reduction in severity of tomato bacterial wilt by Pythium oligandrum. Plant Pathology, 2006, 55(4): 537-543.