Optimization of Culture Conditions of Penicillium bilaiae 47M-1 and Its Control Efficacy on Fusarium Wilt of Sesame

doi:10.16409/j.cnki.2095-039x.2023.02.005

Abstract

Abstract: Penicillium bilaiae strain 47M-1 is a highly effective biocontrol strain with broad-spectrum inhibition activity and growth promotion ability, but its biological characteristics, optimum culture conditions and control efficacy on sesame diseases have not been systematically studied. The purpose of this study was to determine the biological characteristics, the optimum culture conditions and control efficacy of Penicillium bilaiae 47M-1. On the basis of identifying the biological characteristics of strain 47M-1, the optimum culture conditions of strain 47M-1 were obtained by single factor test with Fusarium oxysporum as target and the control efficacy was determined by pot test. The results showed that mycelium growth was fast under the conditions of 25 ℃～28 ℃, pH 5.0～7.0 and 0～16 h light in photoperiod, and sporulation was the high under the conditions of 25 ℃～28 ℃, pH 3.0～10.0 and 8 h light in photoperiod. Lethal temperature of conidia was 60 ℃ for minutes. The optimal culture fluid for producing inhibition substances was as follows: sucrose 30 g, yeast powder/beef extract/peptone 5 g, KCl 0.5 g, MgSO₄·7H₂O 0.5 g, FeSO₄·7H₂O 0.01 g, distilled water 1000 mL. When 2% or 3% spore suspension were inoculated with 75～125 mL optimized culture fluid at pH 7.0, and cultured at 25℃～27℃ for 5 days on a constant temperature shaker, the inhibition activity of culture filtrate was the strongest. The results of pot experiment showed that the control efficacy of both spore suspension and 10-fold diluent of fermentation filtrate were more than 70% against sesame Fusarium wilt. Penicillium bilaiae 47M-1 showed excellent potential for biocontrol. The results will lay a foundation for the study of inhibition substances, production of biocontrol agent and application in field.

Key words: Penicillium bilaiae, sesame Fusarium wilt, biological characteristics, optimization of culture conditions, control efficacy

CLC Number:

S476

ZHAO Xinbei, ZHAO Hui, NI Yunxia, LIU Xintao, HE Bipo, Jia Min, ZHANG Chunyan, Li Yongdong, ZHAO Shikui, LIU Hongyan. Optimization of Culture Conditions of Penicillium bilaiae 47M-1 and Its Control Efficacy on Fusarium Wilt of Sesame[J]. Chinese Journal of Biological Control, 2023, 39(1): 167-175.

References

[1] 曾金凤. 青霉Z88菌株对水稻纹枯病菌的抗生作用[J]. 福建农业大学学报, 1995, 24(2):180-183.
[2] Hamdi N B, Salem I B, Hamdi M. Evaluation of the efficiency of Trichoderma, Penicillium, and Aspergillus species as biological control agents against four soil-borne fungi of melon and watermelon[J]. Egyptian Journal of Biological Pest Control, 2018, 28(1):25.
[3] 张先富, 相立, 王艳芳, 等. 草酸青霉A1菌株的鉴定及对苹果4种镰孢病菌的拮抗作用[J]. 园艺学报, 2016, 43(5):841-852.
[4] 申光辉, 薛泉宏, 张晶, 等. 草莓根腐病拮抗真菌筛选鉴定及其防病促生作用[J]. 中国农业科学, 2012, 45(22):4612-4626.
[5] 王芳, 李静, 张欢. 青霉菌、放线菌株和石灰水对尖孢镰刀菌抑制作用的研究[J]. 中国农学通报, 2013, 29(12):185-189.
[6] Nicoletti R, De Stefano M, De Stefano S, et al. Antagonism against Rhizoctonia solani and fungitoxic metabolite production by some Penicillium isolates[J]. Mycopathologia, 2004, 158(4):465-474.
[7] 李晓栋. 深海沉积物来源的青霉属和曲霉属真菌次生代谢产物及其生物活性研究[D]. 北京:中国科学院大学, 2015.
[8] 徐蕊. 温特曲霉中抗肿瘤四降二萜培养条件优化及海洋来源真菌简青霉次级代谢产物研究[D]. 北京:中国科学院大学, 2016.
[9] Chambers J W, Yeomans J C. The influence of PB-50 (Penicillium bilaii inoculant) on yield and phosphorus uptake by wheat[C]. In Proceedings of the 33rd Annual Meeting, Manitoba Society of Soil Science. Winnipeg, Manitoba, Canada:University of Manitoba, 1990, 283-293.
[10] Gleddie S C, Hnatowich G L, Polonenko D R. A summary of spring wheat response to Penicillium bilaji, a phosphate inoculant[C]. Proceedings for the Western Phosphate/Sulfur Workshop Conference, Colorado State University, Fort Collins, Colorado, March, 1991, 21-22.
[11] Gleddie S C. Response of pea and lentil to inoculation with the phosphate-solubilizing fungus Penicillium bilaii (PROVIDE^TM)[C]. Proceedings of the Soils and Crops Workshop, Saskatoon, Saskatchewan, 1993, 47-52.
[12] Leggett M E, Gleddie S C, Prusinkiewicz E, et al. The development of a commercial phosphate inoculant for wheat and canola[J]. Developments in Plant and Soil Sciences, 1993, 54:375-379.
[13] Kucey R M N. Increased phosphorus uptake by wheat and field bean inoculated with phosphorus-solubilizing Penicillium bilaji strain and with vesicular arbuscular mycorrhizal fungi[J]. Applied and Environmental Microbiology, 1987, 53:2699-2703.
[14] Wakelin S A, Gupta V V S R, Harvey P R, et al. The effect of Penicillium fungi on plant growth and phosphorus mobilization in neutral to alkaline soils from southern Australia[J]. Canadian Journal of Microbiology, 2007, 53:106-115.
[15] 罗文芳, 魏松红, 王海宁, 等. 水稻纹枯病生防菌的筛选与鉴定[J]. 沈阳农业大学学报, 2017, 48(6):660-665.
[16] 张楠楠, 白净, 张礼文, 等. 拜赖青霉活性产物的抗真菌作用研究[J]. 生物技术进展, 2017, 7(4):315-319.
[17] Zhao X B, Liu X T, Zhao H, et al. Biological control of Fusarium wilt of sesame by Penicillium bilaiae 47M-1[J]. Biological Control, 2021, 158:104601.
[18] 孔华忠. 中国真菌志:第35卷, 青霉属及其相关有性型属[M]. 北京:科学出版社, 2007, 22.
[19] 仇存璞, 张海洋, 常淑娴, 等. 芝麻枯萎病病原菌致病力室内鉴定方法[J]. 植物病理学报, 2014, 44(1):26-35.
[20] Guo X H, Gao W Y, Li K F. Tissue culture of Salvia miltiorrhiza adventitious roots (II) Effects of carbon nitrogen and phosphate sources on culture of Salvia miltiorrhiza adventitious roots[J]. Chinese Traditional and Herbal Drug, 2007, 35(6):907-11.
[21] 张学秋, 余晓斌. 蝉拟青霉液体发酵条件优化[J]. 微生物学杂志, 2019, 39(2):18-24.
[22] 赵龙妹, 张兰, 曹慧, 等. 土壤中产木聚糖酶菌株的筛选及发酵条件优化[J]. 微生物学通报, 2021, 48(10):3506-3519.
[23] Liang X, Liu Y, Zhang R, et al. Optimization of fermentation medium and conditions of Bacillus subtilis PTS-394[J]. Journal of Yangzhou University (Agricultural and Life Science Edition), 2013, 34(3):75-82.
[24] Yang C D, Chang T, Xue L, et al. Optimizing the culture conditions and determining the stability of antibiotic secretion by Polygonum viviparum of the endophytic bacteria Bacillus mojavensis[J]. Acta Prataculturae Sinica, 2015, 24(9):104-112.
[25] 南楠, 戚向阳, 袁勇军. 枯草芽孢杆菌WL17产抗菌物质的分离、分析及其抗菌特性研究[J]. 中国食品学报, 2012, 12(8):17-18.
[26] 王琢. 抑黄曲霉毒素的Hitwh-B05菌株有效成分的分离纯化与鉴定[D]. 哈尔滨:哈尔滨工业大学, 2012.