Screening, identification and biocontrol effect of antagonistic actinomycetes against the pathogen of Cytospora sp. for apple tree

doi:10.13287/j.1001-9332.201610.010

Abstract

Abstract: The pathogen of Cytospora sp. was considered as the target fungus to evaluate the bio-control potential of antagonistic actinomycetes against Cytospora sp. in the present study, which was isolated from the rhizosphere soil of apple tree. Moreover, the antagonistic effect of actinomycetes against Cytospora sp. growth was determined by the method of confrontation and growth rate, and the screened antagonistic strain was identified by the method of morphology and molecular biology. Also, the inhibitory activity of strain ZZ-9 against the conidium germination and mycelia growth of Cytospora sp. was determined in vivo, and its bio-control effect on Cytospora sp. growth was determined in detached apple twigs. The results showed that fifteen strains of actinomycetes had the inhibitory effect on Cytospora sp. growth, among all the isolated strains, which accounted for 18.8% and especially the inhibitory rate of eight strains was more than 50%. In addition, among all the screened strains, the strain ZZ-9 presented the highest inhibitory rate of 96.4%, which was significantly higher than those of the other isolated strains. The strain ZZ-9 was initially identified as Streptomyces rochei according to cultural characteristics, physiological and biochemical properties and 16S rDNA analysis. The nucleotide sequences in GenBank accession number was registered as KT986228. Different dilution of ZZ-9 fermentation had significant inhibitory effect on Cytospora sp. conidium germination and mycelia growth. The inhibitory rates of the 50 times fermentation on both conidium germination and mycelia growth were more than 80%. The inhibited mycelia’s color was deepened, the mycelia branches were increased, and the ends of hyphae were swelled and deformed, even the protoplasm was concentrated and released. The bio-control effect of the ZZ-9 stock solution on Cytospora sp. growth was more than 75% in detached apple twigs. Thus, our results indicated that the strain ZZ-9 could be used for controlling apple tree Valsa canker in vivo and vitro.

XUE Ying-yu, FAN Wan-ze, ZHANG Shu-wu, XU Bing-liang. Screening, identification and biocontrol effect of antagonistic actinomycetes against the pathogen of Cytospora sp. for apple tree[J]. Chinese Journal of Applied Ecology, 2016, 27(10): 3379-3386.

References

[1] Wang S-T (王树桐), Zhang F-Q (张凤巧), Cao K-Q (曹克强). Bibliometric analysis of apple Valsa canker research in China. Journal of Anhui Agricultural Sciences (安徽农业科学), 2010, 38(25): 13802-13804 (in Chinese)
[2] Chen Z (陈臻), Xu B-L (徐秉良), Pu C-J (蒲崇建), et al. Study on occurrence and biological characteristics of Valsa ceratosperma sexuality in different habitats. Journal of Fruit Science (果树学报), 2013, 30(4): 644-649 (in Chinese)
[3] Wang X-L (王旭丽). Pathogen of Apple Tree Valsa Canker in China: A Combined Analysis of Phenotypic Characteristics and rDNA-ITS Sequences. PhD Thesis. Yangling, Shaanxi: Northwest A＆F University, 2007 (in Chinese)
[4] Kang T-L (康天兰), Wang X-H (王新海). The pre-sent situation and the development countermeasures of fruit industry in Gansu Province. Gansu Agriculture (甘肃农业), 2014(21): 10-11 (in Chinese)
[5] Zhou Q (周启), Wang D-B (王道本). Agricultural Antibiotics and Microbial Pesticides. Beijing: China Agriculture Press, 1995 (in Chinese)
[6] Ruan J-S (阮继生), Liu Z-H (刘志恒), Liang L-R (梁立儒), et al. Research and Application of Actinomycete. Beijing: Sciences Press, 1990 (in Chinese)
[7] Shen H (沈慧). Biocontrol methods of apple tree Valsa canker. Hebei Fruits (河北果树), 2006(4): 42 (in Chinese)
[8] Cui Y-L (崔云龙). Effect of S-921 strain (Streptomyces aureus) and its fermented broth against apple tree canker, Valsa mali. Chinese Journal of Biological Control (生物防治通报), 1992, 8(1): 6-28 (in Chinese)
[9] Gao Z-P (郜佐鹏), Ke X-W (柯希望), Wei J-L (韦洁玲), et al. Biocontrol efficacy of apple tree Valsa canker by endophytic actinomycete. Acta Phytophylacica Sinica (植物保护学报), 2009, 36(5): 410-416 (in Chinese)
[10] Zhan L-R (展丽然), Zhang K-C (张克诚), Ran L-X (冉隆贤), et al. Isolation and identification of the antagonistic actinomycetes against Valsa mali. Hebei Journal of Forestry and Orchard Research (河北林果研究), 2008, 23(2): 182-186 (in Chinese)
[11] Zhang Q-M (张清明), Wang C-X (王彩霞), Wang H-Y (王海艳), et al. Identification of antagonistic endophytic actinomycetes A-2 and evaluation of its activity against Valsa mali. Chinese Journal of Pesticide Science (农药学学报), 2013, 15(3): 286-292 (in Chinese)
[12] Zhang B (张波), Wu W-J (吴文君), Zong Z-F (宗兆峰). Studies on isolation, identification and fungicidal activity of antagonistic actinomycete Z139. Journal of Northwest Sci-Tech University of Agricultural and Forest (Nation Science) (西北农林科技大学学报: 自然科学版), 2005, 33(8): 69-72 (in Chinese)
[13] Zhou D-Q (周德庆). Microbiology Experiment Manual. Shanghai: Shanghai Science and Technology Press, 1986 (in Chinese)
[14] Fang Z-D (方中达). Research Methods of Plant Patho-logy. Beijing: China Agriculture Press, 1998 (in Chinese)
[15] Yan X-C (阎逊初). Classification and Identification of Actinomycetes. Beijing: Science Press, 1992 (in Chinese)
[16] Actinomycetes Classification Group, Chinese Academy of Sciences Institute of Microbiology (中国科学院微生物研究所放线菌分类组). Streptomyces Identification Handbook. Beijing: Science Press, 1975 (in Chinese)
[17] Ruan J-S (阮继生). Classification Basis of Actinomycetes. Beijing: Science Press, 1977 (in Chinese)
[18] Saitou N, Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 1987, 4: 406-425
[19] Wang C-X (王彩霞), Zhang Q-M (张清明), Li G-F (李桂舫), et al. Identification of the antagonistic bacteria BJ1 and its antifungal activity against Valsa ceratosperma. Acta Phytophylacica Sinica (植物保护学报), 2012, 39(5): 431-437 (in Chinese)
[20] Wang W-X (王卫雄), Xu B-L (徐秉良), Xue Y-Y (薛应钰), et al. Identification and antifungal activity of the antagonistic bacteria of Cytospora spp. Chinese Journal of Eco-Agriculture (中国生态农业学报), 2014, 22(10): 1214-1221 (in Chinese)
[21] Liu Q (刘琴), Xu J (徐健), Liu H-A (刘怀阿), et al. Isolation and antagonistic activity of endophytic actinomycete SR-1102 to Fusarium oxysporum f. sp. cucumebrium of cucumber. Journal of Yangzhou University (Agricultural and Life Science) (扬州大学学报: 农业与生命科学版), 2015, 36(2): 83-88 (in Chinese)
[22] Liu C-B (柳成宾), Wan C-X (万传星), Jia X-Y (贾晓宇), et al. Screening of antagonistic actionomycetes against Verticillium dahliae from saline environments in Xinjiang as well as the antagonistic activities and identification of strain TRM42561. Chinese Journal of Biological Control (中国生物防治学报), 2014, 30(3): 408-413 (in Chinese)
[23] Zhang L (张丽), Ji M-S (纪明山), Yu Z-G (于志国). Antifungal activities and stability of metabolites from Streptomyces rochei against Pyricularia oryzae. Journal of Shenyang Agricultural University (沈阳农业大学学报), 2014, 45(2): 143-146 (in Chinese)
[24] Xiong Y-Y (熊瑶瑶), Ao W (敖武), Li Y (李怡), et al. The identification of three antagonistic endophytic actinomycete and their antimicrobial activities. Journal of Jiangxi Normal University (Natural Science) (江西师范大学学报: 自然科学版), 2015, 39(2): 171-176 (in Chinese)
[25] Ruan J-S (阮继生), Huang Y (黄英). Rapid Identification of Actinobacteria. Beijing: Science Press, 2011 (in Chinese)
[26] Gao P (高鹏), Xue Q-H (薛泉宏), Chang X-B (常显波), et al. Study on the media and methods of selecting antimicrobial actinomycetes. Journal of Northwest A＆F University (西北农林科技大学学报), 2005, 33(1): 60-62 (in Chinese)
[27] Yang B (杨斌), Xue Q-H (薛泉宏), Chen Z-Q (陈占全), et al. Effects of microwave irradiation on isolation of soil actinomycetes. Chinese Journal of Applied Ecology (应用生态学报), 2008, 19(5): 1091-1098 (in Chinese)
[28] Wei S-J (魏赛金), Du Y-N (杜亚楠), Ni G-R (倪国荣), et al. Inhibition effect of Ag-antibiotic 702 on plant pathogenic fungi and related mechanisms. Chinese Journal of Applied Ecology (应用生态学报), 2012, 23(12): 3435-3440 (in Chinese)