Isolation, identification and characterization of antagonistic actinomycetes A10 and A17 against Botryodiplodia theobromae

doi:10.3969/j.issn.1004-1524.2020.03.11

›› 2020, Vol. 32 ›› Issue (3): 460-468.DOI: 10.3969/j.issn.1004-1524.2020.03.11

• Plant Protection • Previous Articles Next Articles

Isolation, identification and characterization of antagonistic actinomycetes A10 and A17 against Botryodiplodia theobromae

GUO Xuesong¹, TIAN Libo¹, SHANG Sang^2,*, ZOU Kaixi¹, CHEN Hongrong¹, LI Wanyu¹, YUE Xiaoqi¹

1.College of Horticulture, Hainan University/The Key Laboratory of Tropical Horticultural Crop Quality Regulation of Hainan Province, Haikou 570228, China;
2.College of Life Science and Pharmaceutical Sciences, Hainan University, Haikou 570228, China

Received:2019-11-23 Online:2020-03-25 Published:2020-04-03

Abstract

Abstract: In order to enrich the antagonistic and antibacterial resources of mango stem-end rot and find a non-toxic and non-polluting post-harvest control method instead of chemical fungicide, plate dilution method, scribing purification method and paper diffusion method were used to separate and screen the antagonistic actinomycetes against Botryodiplodia theobromae from mangrove rhizosphere soil. Their taxonomic status was identified, the fermentation conditions were optimized, and the biological control effect was tested. A total of two antagonistic actinomycetes A10 and A17 with strong antibacterial activity against postharvest mango rot fungus (Botryodiplodia theobromae Pat.) were screened. According to the morphological characteristics, physiological and biochemical characteristics and molecular identification, the two actinomycetes were all Streptomyces. The fermentation medium and fermentation conditions of A10 strain were optimized as follows: 2.5% lactose, 2.0% peptone, initial pH 8.0, shaking speed 200 r·min^-1, culture time 120 h. The fermentation medium and fermentation conditions of A17 strain were optimized as follows: 2.0% soluble starch, 2.0% peptone, initial pH 7.0, shaking speed 200 r·min^-1, culture time 120 h. In vivo experiment, on the 4th day, the incidence and disease index of the control group treated with sterile water were 81.7% and 52.5, respectively, 43.3% and 21.7 in the A10 group, 36.7% and 17.5 in the A17 group, and 16.7% and 12.2 in the 450 mg·kg^-1 prochloraz group. Both strains had a certain inhibition effect on the occurrence and expansion of the disease spot, and A17 had a certain inhibition effect on the expansion of the disease spot. There was no significant difference between the inhibitory effect of A17 and that of 450 mg·kg^-1 prochloraz. The results showed that the two strains had some antagonistic effect on the pathogen, which could be further studied.

Key words: mango stem-end rot, antagonistic actinomycetes, screening, identification, control effect

CLC Number:

S436.67

GUO Xuesong, TIAN Libo, SHANG Sang, ZOU Kaixi, CHEN Hongrong, LI Wanyu, YUE Xiaoqi. Isolation, identification and characterization of antagonistic actinomycetes A10 and A17 against Botryodiplodia theobromae[J]. , 2020, 32(3): 460-468.

References

[1] 贺瑞, 赵磊, 符瑞, 等. 海南芒果蒂腐病菌对吡唑醚菌酯的抗药性测定[J]. 植物保护, 2018, 44(4): 188-193.
HE R, ZHAO L, FU R, et al.Resistance of Botryodiplodia theobromae caused mango stem end rot to pyraclostrobin in Hainan[J]. Plant Protection, 2018, 44(4): 188-193.(in Chinese with English abstract)
[2] 胡美姣, 高兆银, 李敏, 等. 杧果果实潜伏侵染真菌种类研究[J]. 果树学报, 2012, 29(1): 105-110.
HU M J, GAO Z Y, LI M, et al.Investigation of latent infection of pathogenic fungi in mango fruits[J]. Journal of Fruit Science, 2012, 29(1): 105-110.(in Chinese with English abstract)
[3] HONG S K, LEE S Y, CHOI H W, et al.Occurrence of stem-end rot on mango fruits caused by Lasiodiplodia theobromae in Korea[J]. The Plant Pathology Journal, 2012, 28(4): 455.
[4] DROBY S, WISNIEWSKI M, MACARISIN D, et al.Twenty years of postharvest biocontrol research: is it time for a new paradigm?[J]. Postharvest Biology and Technology, 2009, 52(2): 137-145.
[5] SPADARO D, GULLINO M L.State of the art and future prospects of the biological control of postharvest fruit diseases[J]. International Journal of Food Microbiology, 2004, 91(2): 185-194.
[6] 王兰英, 王琴, 冉杜, 等. 三桠苦拮抗内生真菌SCK-Y₉筛选及其抑菌活性研究[J]. 中国植保导刊, 2014, 34(8): 10-13.
WANG L Y, WANG Q, RAN D, et al.Screening of antagonistic endophytic fungi SCK-Y9 in Evodia lepta and its fungicidal activities bioassay[J]. China Plant Protection, 2014, 34(8): 10-13.(in Chinese with English abstract)
[7] 赵超, 胡美姣, 李敏, 等. 芒果采后病害拮抗菌的筛选、鉴定及其生防效果研究[J]. 西南农业学报, 2016, 29(2): 385-389.
ZHAO C, HU M J, LI M, et al.Antagonists selection and control effect on postharvest diseases of mango fruit[J]. Southwest China Journal of Agricultural Sciences, 2016, 29(2): 385-389.(in Chinese with English abstract)
[8] LUGTENBERG B, KAMILOVA F.Plant-growth-promoting rhizobacteria[J]. Annual Review of Microbiology, 2009, 63(1): 541-556.
[9] LI R, SHEN Z Z, SUN L, et al.Novel soil fumigation method for suppressing cucumber Fusarium wilt disease associated with soil microflora alterations[J]. Applied Soil Ecology, 2016, 101: 28-36.
[10] 马喆. 根结线虫拮抗放线菌NZ-5菌株的鉴定及其发酵条件优化[J]. 北方园艺, 2019(13): 59-67.
MA Z.Identification of nematicidal actinomycete NZ-5 and optimization of ferment conditions[J]. Northern Horticulture, 2019(13): 59-67.(in Chinese with English abstract)
[11] 张凯, 吴曼莉, 辜柳霜, 等. 胶孢炭疽菌生防放线菌gz-8的鉴定及生物活性初步评价[J]. 农药学学报, 2017, 19(6): 708-715.
ZHANG K, WU M L, GU L S, et al.Identification and preliminary evaluation of a biocontrol actinomycete strain gz-8 against Colletotrichum gloeosporioides[J]. Chinese Journal of Pesticide Science, 2017, 19(6): 708-715.(in Chinese with English abstract)
[12] 曹利, 车程川, 刘金锋, 等. 拮抗黄曲霉海洋放线菌的筛选及发酵条件优化[J]. 中国酿造, 2019, 38(3): 104-109.
CAO L, CHE C C, LIU J F, et al.Screening and fermentation conditions optimization of marine actinomycetes of antagonistic Aspergillus flavus[J]. China Brewing, 2019, 38(3): 104-109.(in Chinese with English abstract)
[13] 赵淑莉, 任飞娥, 刘金亮, 等. 玉米大斑病生防放线菌的筛选鉴定及发酵条件优化[J]. 微生物学报, 2012, 52(10): 1228-1236.
ZHAO S L, REN E R, LIU J L, et al.Screening, identification and optimization of fermentation conditions of an antagonistic actinomycetes strain to Setosphaeria Turcica[J]. Acta Microbiologica Sinica, 2012, 52(10): 1228-1236.(in Chinese with English abstract)
[14] 刘雪梅. 微生物分析仪法与常规纸片扩散法在细菌耐药表型检测中的调查分析[J].中国卫生产业,2017,14(17):51-52.
LIU X M.Investigation and analysis of microbial analyzer method and conventional disc diffusion method in detection of bacterial resistance phenotypes[J].China Health Industry,2017,14(17):51-52. (in Chinese)
[15] 刘建玲, 陈宝宝, 刘永红, 等. 半夏内生菌的分离与初步鉴定[J]. 中国中药杂志, 2009, 34(18): 2305-2307.
LIU J L, CHEN B B, LIU Y H, et al.Isolation and identification of endophytes from Pinellia[J]. China Journal of Chinese Materia Medica, 2009, 34(18): 2305-2307.(in Chinese with English abstract)
[16] XU C N, ZHANG H J, ZHOU Z S, et al.Identification and distribution of Botryosphaeriaceae species associated with blueberry stem blight in China[J]. European Journal of Plant Pathology, 2015, 143(4): 737-752.
[17] ZHOU Y, GONG G S, CUI Y L, et al.Identification of Botryosphaeriaceae species causing kiwifruit rot in Sichuan Province, China[J]. Plant Disease, 2015, 99(5): 699-708.
[18] 徐志周, 王明元, 杜锦鹏, 等. 一株香蕉枯萎病拮抗菌HQB-1的分离鉴定及其发酵条件优化[J]. 微生物学通报, 2019, 46(7): 1611-1618.
XU Z Z, WANG M Y, DU J P, et al.Isolation, identification and fermentation optimization of an antagonistic bacterial strain HQB-1 against banana wilt disease[J]. Microbiology China, 2019, 46(7): 1611-1618.(in Chinese with English abstract)
[19] SADEGHIAN M, SHAHIDI BONJAR G H, SHARIFI SIRCHI G R. Post harvest biological control of apple bitter rot by soil-borne actinomycetes and molecular identification of the active antagonist[J]. Postharvest Biology and Technology, 2016, 112: 46-54.
[20] 孙平平, 贾晓辉, 崔建潮, 等. 梨灰霉病拮抗放线菌L-30的筛选、鉴定及作用机制研究[J]. 园艺学报, 2016, 43(12): 2335-2346.
SUN P P, JIA X H, CUI J C, et al.Selection, identification and characterization of Actinomyces L-30 for the biocontrol of pear gray mold[J]. Acta Horticulturae Sinica, 2016, 43(12): 2335-2346.(in Chinese with English abstract)
[21] VELA GUROVIC M S, OLIVERA N L. Antibacterial producing actinomycetes from Extra-Andean Patagonia[J]. Journal of Arid Environments, 2017, 144: 216-219.
[22] 武峥. 防治苹果采后病害拮抗酵母菌的分离、筛选及其抑菌效果的研究[D]. 武汉: 华中农业大学, 2005.
WU Z.Study on screening yeasts in biocontrol of postharvest diseases of apples and the inhibition effects of the selected antagonists[D]. Wuhan: Huazhong Agricultural University, 2005.(in Chinese with English abstract)
[23] 马龙传. SA、MeJA及O₃增强拮抗酵母对水果采后病害的控制效应[D].镇江:江苏大学,2009.
MA L C.SA, MEJA and O₃ enhanced the biocontrol efficacy of antagonistic yeast to the postharvest disease of fruits[D].Zhenjiang: Jiangsu University,2009. (in Chinese with English abstract)
[24] 石晶盈, 刘爱媛, 李雪萍, 等. 番木瓜果实内生细菌MGP3菌株的鉴定及拮抗作用[J]. 微生物学报, 2011, 51(9): 1240-1247.
SHI J Y, LIU A Y, LI X P, et al.Identification and antagonistic activities of an endophytic bacterium MGP3 isolated from Papaya fruit[J]. Acta Microbiologica Sinica, 2011, 51(9): 1240-1247.(in Chinese with English abstract)
[25] 李敏, 杨谦, 王疏, 等. 哈茨木霉与多菌灵复合使用对水稻苗期立枯病的防治[J]. 浙江大学学报(农业与生命科学版), 2009, 35(1): 65-70.
LI M, YANG Q, WANG S, et al.Trichoderma harzianum in combination with carbendazim for integrated control of rice seedling blight[J]. Journal of Zhejiang University(Agriculture and Life Sciences), 2009, 35(1): 65-70.(in Chinese with English abstract)

Isolation, identification and characterization of antagonistic actinomycetes A10 and A17 against Botryodiplodia theobromae

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	PEI Huimin, WU Mingming, ZHAI Rongrong, YE Jing, JIN Yue, ZHU Yi, HOU Jianjun, ZHU Guofu, YE Shenghai. Research progress on gene function and breeding of low-cadmium rice cultivars [J]. Acta Agriculturae Zhejiangensis, 2025, 37(9): 2012-2020.
[2]	LIU Pengfei, ZHANG Shuhan, HONG Kai, SHAO Yue, LOU Binggan. Isolation and identification of the pathogen causing tomato canker in Zhejiang Province of China [J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1293-1300.
[3]	MA Xian, YOU Yuwei, KANG Juan, WANG Guoqin, ZHENG Rui, SU Jianyu, YUE Sijun. Pathogen identification of post-harvest rotten Lycium barbarum and screening of natural fungicides [J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1327-1335.
[4]	JI Mengting, CHEN Changjiang, ZHU Ling, ZHAN Menglin, XIAO Shun, CAI Xueqing. Identification of etiological bacterium causing leaf spot disease on Ficus carica [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 1097-1106.
[5]	HE Guoxin, LI Sujuan, WANG Jian, TAO Xiaoyuan, YE Zihong, CHEN Guang, XU Shengchun. Screening and identification of soybean germplasm for low nitrogen tolerance during seedling stage [J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 965-976.
[6]	GAO Qiang, WANG Lili, ZHANG Jianlong, YANG Bo, LI Feng, ZHU Xianzhi, LIU Aixin, HAN Chao, TIAN Lei. Screening and identification of Bacillus altitudinis strain CY1 and its control effects against tobacco black shank [J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 405-416.
[7]	LU Na, WANG Weike, LIN Jiayao, ZHOU Zufa. Identification of yellow rot disease of Pleurotus pulmonarius and analysis of its pathogenicity-related transcriptome [J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 426-437.
[8]	GONG Xinxin, LIU Ruiling, HAN Yanchao, MENG Xianghong, GAO Haiyan, CHEN Hangjun. Isolation and identification of main postharvest pathogens of four edible fungi [J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 456-465.
[9]	LI Qiang, LIU Sitong, HUANG Xianbin, JIANG Junlong, DENG Jianyu, WANG Jiaoyu, LI Ling. Identification of pathogenic bacteria of kiwifruit canker in mountainous area and screening of different types of efficient control fungicides [J]. Acta Agriculturae Zhejiangensis, 2025, 37(10): 2116-2128.
[10]	LI Yani, CHEN Weiliang, MAO Bizeng. Pathogen identification of root rot of Curcuma wenyujin [J]. Acta Agriculturae Zhejiangensis, 2024, 36(5): 1086-1093.
[11]	LU Chunxia, LIU Kaili, LIANG Guiqiu, XIAO Xiao, WEI Wei, XU Wenwen, CHEN Jing, MO Bingqiao, LI Xiaoqun, LI Er’na, HUANG Xuhua, LI Anhua, WEI Shini. Morphological observation and molecular identification of 11 sanghuang strains [J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 800-810.
[12]	HE Jiawei, Huang Leqin, LU Zhenyu, FANG Jin, LEI Ziyang, ZHANG Huijuan, JIANG Ming. Isolation and identification of the pathogen causing leaf spot disease on a rare and endangered plant species Rhododendron huadingense [J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 837-845.
[13]	CHAI Rongyao, YOU Yuxin, QIU Haiping, GUO Junning, ZHANG Zhen, LI Bin, SHEN Shengfa, WANG Yanli. Establishment of sweet potato stem rot resistance identification technology and analysis of germplasm resource resistance [J]. Acta Agriculturae Zhejiangensis, 2024, 36(3): 569-578.
[14]	LUO Zhihan, LIU Pengfei, YU Jun, QI He, CHEN Xiaoguang, LOU Binggan. Identification and biological characteristics of pathogen causing branch dieback on Styphnolobium japonicum (L.) Schott [J]. Acta Agriculturae Zhejiangensis, 2024, 36(3): 579-588.
[15]	GUO Aikui, LIANG Zhihao, LI Yuxing, WANG Qiang, CHENG Yifan, XUE Song, YU Wenqing, XU Xiao, ZHANG Yinghu, QIAO Hailong, YANG Hongyan, SHEN Huiquan. Change characteristics of comprehensive traits of barley varieties (strains) in the comparative identification tests in coastal areas of Jiangsu Province of China from 2007 to 2021 [J]. Acta Agriculturae Zhejiangensis, 2024, 36(12): 2666-2675.