猕猴桃广谱拮抗内生菌分离鉴定及其对樱桃圣女果采后的保鲜作用
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摘要
筛选具有广谱抑菌效果的内生菌并研究其发酵液对樱桃圣女果采后保鲜效果的影响。采用研磨法从健康猕猴桃的根、茎、叶和果实中分离纯化了18株内生菌,通过平板对峙法选出强活性菌株MR-1,并对其进行菌体、菌落形态的观察、生理生化测定及16S rRNA测序分析,进行菌种鉴定。进一步研究该活性菌株对8种常见致病菌的抑制效果以及樱桃圣女果主要病害菌的抑制作用。结果表明,菌株MR-1对采后致病菌拮抗效果较强,初步鉴定其为解淀粉芽孢杆菌,其对8种植物病原菌均有较强的拮抗作用,对链格孢、灰葡萄孢、辣椒炭疽病菌3种圣女果主要病害菌表现出较强的抑制作用,经内生菌MR-1发酵滤液处理的樱桃圣女果保鲜效果最好,其次为复合果蔬保鲜剂。储藏第8 d,与空白对照组相比,其腐烂直径分别下降了50.6%、50.6%及44.9%。解淀粉芽孢杆菌MR-1次级代谢产物丰富,其在果蔬保鲜中有较好的应用前景。
This article aimed to screen endophytic bacteria with broad-spectrum antibacterial activity and investigate the postharvest preservation effect of their fermentation products on cherry cherries. Eighteen endophytic bacteria were isolated and purified from the roots,stems,leaves or fruits of healthy kiwifruits by grinding method. The strong active strain MR-1 was selected by plate sputum method,and the bacteria,colony morphology,physiological and biochemical determination 16 S rRNA sequencing analysis and strain identification.The inhibitory effect of the active strain on eight common pathogenic bacteria and the inhibitory effect of the main pathogen of cherry sage were further studied. The results showed that the strain MR-1 had strong antagonistic effect on postharvest pathogenic bacteria,and it was initially identified as Bacillus amyloliquefaciens,which had strong antagonistic effect on 8 plant pathogens. It showed strong inhibitory effects on Alternaria nees,Ascomycota and Colletotrichum capsici three main disease bacterium.Cherry cherries treated with endophytic MR-1 fermentation filtrate had the best preservation effect,and the compound fruit and vegetable preservatives followed.On the 8th day of storage,comparing with the blank control group,the decay diameter of MR-1 group decreased by 50.6%,50.6% and 44.9%.The secondary metabolites of Bacillus amyloliquefaciens MR-1 were abundant,which had a good application prospect in the preservation of fruits and vegetables.
This article aimed to screen endophytic bacteria with broad-spectrum antibacterial activity and investigate the postharvest preservation effect of their fermentation products on cherry cherries. Eighteen endophytic bacteria were isolated and purified from the roots,stems,leaves or fruits of healthy kiwifruits by grinding method. The strong active strain MR-1 was selected by plate sputum method,and the bacteria,colony morphology,physiological and biochemical determination 16 S rRNA sequencing analysis and strain identification.The inhibitory effect of the active strain on eight common pathogenic bacteria and the inhibitory effect of the main pathogen of cherry sage were further studied. The results showed that the strain MR-1 had strong antagonistic effect on postharvest pathogenic bacteria,and it was initially identified as Bacillus amyloliquefaciens,which had strong antagonistic effect on 8 plant pathogens. It showed strong inhibitory effects on Alternaria nees,Ascomycota and Colletotrichum capsici three main disease bacterium.Cherry cherries treated with endophytic MR-1 fermentation filtrate had the best preservation effect,and the compound fruit and vegetable preservatives followed.On the 8th day of storage,comparing with the blank control group,the decay diameter of MR-1 group decreased by 50.6%,50.6% and 44.9%.The secondary metabolites of Bacillus amyloliquefaciens MR-1 were abundant,which had a good application prospect in the preservation of fruits and vegetables.
引文
[1]周金伟,周红丽,柏连阳.植物内生菌抗菌活性物质及其在果蔬采后保鲜中的应用进展[J].植物保护,2015,41(6):22-32,37.
[2]Shamala Sundram,Sariah Meon,Idris Abu Seman,et al.Symbiotic interaction of endophytic bacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Ganoderma boninense[J].Journal of Microbiology,2011,49(4):551-557.
[3]陈帮丹,魏洪,黄亚励,等.猕猴桃果实内生菌株的鉴定及其发酵液抑菌活性初探[J].中国病原生物学杂志,2018,13(4):372-378.
[4]田华,韩慢慢,陈乐玲.圣女果保鲜技术研究进展[J].食品工业科技,2016,37(7):396-400.
[5]罗丹,吴昊,陈存坤,等.姜油对番茄致病菌的抑制效果及抑菌机制初探[J].食品安全质量检测学报,2017,8(4):1404-1409.
[6]蔡文韬,夏菠,夏延斌,等.解淀粉芽孢杆菌发酵液处理提高辣椒采后品质[J].农业工程学报,2013,29(23):253-261.
[7]陈超,庞立,张敏,等.猕猴桃葡葡座腔菌拮抗内生菌的筛选及其发酵液稳定性研究[J].食品科技,2018,43(3):8-13,19.
[8]黄敬瑜,张楚军,姚瑜龙,等.植物内生菌生物抗菌活性物质研究进展[J].生物工程学报,2017,33(2):178-186.
[9]Long Xinxian,Chen Xuemei,Chen Yagang,et al.Isolation and characterization endophytic bacteria from hyperaccumulator Sedum alfredii Hance and their potential to promote phytoextraction of zinc polluted soil[J].World Journal of Microbiology and Biotechnology,2011,27(5):1197-1207.
[10]江明明.几株优良内生菌的分离及鉴定[D].南京:南京林业大学,2016.
[11]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[12]RE布坎南,NE吉布斯.伯杰细菌鉴定手册(第8版)[M].北京:科学出版社,1984.
[13]Zhao L F,Xu Y J,Sun R,et al.Identification and characterization of the endophytic plant growth prompter Bacillus cereus strain MQ23 isolated from Sophora alopecuroides root nodules[J].Brazilian Journal of Microbiology,2011,42:567-575.
[14]Tamurak,Petersond.MEGA5:Molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods[J].Molecular Biology and Evolution,2011,28(10):2731-2739.
[15]王世强.打孔法测定抗生素的效价[J].生物学通报,2005(3):2.
[16]唐书谦,叶庆佾,钟白玉,等.姜黄挥发油、水浸出液体外抗真菌试验[J].第三军医大学学报,1997(6):89-91.
[17]赵淑艳,李喜宏,陈丽,等.桃采后病原菌种类及侵染规律研究[J].食品科学,2005(10):253-255.
[18]周金伟.植物内生菌011粗提液的抑菌机理、保鲜作用及抗菌物质的分离鉴定[D].长沙:湖南农业大学,2015.
[19]郭睿文.猕猴桃内生菌多样性及溃疡病拮抗菌筛选[D].成都:成都理工大学,2016.
[20]陈哲,黄静,赵佳,等.解淀粉芽孢杆菌抑菌机制的研究进展[J].生物技术通报,2015,31(6):37-41.
[21]出晓铭,林毅雄,张珅,等.生防菌解淀粉芽孢杆菌抗菌蛋白的研究进展[J].包装与食品机械,2014,32(6):49-54.
[22]洪鹏,安国栋,胡美英,等.解淀粉芽孢杆菌防治果蔬采后病害研究进展[J].中国农学通报,2013,29(12):168-173.
[23]李红晓,张殿朋,卢彩鸽,等.生防解淀粉芽胞杆菌最新研究进展[J].微生物学杂志,2016,36(2):87-92.
[2]Shamala Sundram,Sariah Meon,Idris Abu Seman,et al.Symbiotic interaction of endophytic bacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Ganoderma boninense[J].Journal of Microbiology,2011,49(4):551-557.
[3]陈帮丹,魏洪,黄亚励,等.猕猴桃果实内生菌株的鉴定及其发酵液抑菌活性初探[J].中国病原生物学杂志,2018,13(4):372-378.
[4]田华,韩慢慢,陈乐玲.圣女果保鲜技术研究进展[J].食品工业科技,2016,37(7):396-400.
[5]罗丹,吴昊,陈存坤,等.姜油对番茄致病菌的抑制效果及抑菌机制初探[J].食品安全质量检测学报,2017,8(4):1404-1409.
[6]蔡文韬,夏菠,夏延斌,等.解淀粉芽孢杆菌发酵液处理提高辣椒采后品质[J].农业工程学报,2013,29(23):253-261.
[7]陈超,庞立,张敏,等.猕猴桃葡葡座腔菌拮抗内生菌的筛选及其发酵液稳定性研究[J].食品科技,2018,43(3):8-13,19.
[8]黄敬瑜,张楚军,姚瑜龙,等.植物内生菌生物抗菌活性物质研究进展[J].生物工程学报,2017,33(2):178-186.
[9]Long Xinxian,Chen Xuemei,Chen Yagang,et al.Isolation and characterization endophytic bacteria from hyperaccumulator Sedum alfredii Hance and their potential to promote phytoextraction of zinc polluted soil[J].World Journal of Microbiology and Biotechnology,2011,27(5):1197-1207.
[10]江明明.几株优良内生菌的分离及鉴定[D].南京:南京林业大学,2016.
[11]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[12]RE布坎南,NE吉布斯.伯杰细菌鉴定手册(第8版)[M].北京:科学出版社,1984.
[13]Zhao L F,Xu Y J,Sun R,et al.Identification and characterization of the endophytic plant growth prompter Bacillus cereus strain MQ23 isolated from Sophora alopecuroides root nodules[J].Brazilian Journal of Microbiology,2011,42:567-575.
[14]Tamurak,Petersond.MEGA5:Molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods[J].Molecular Biology and Evolution,2011,28(10):2731-2739.
[15]王世强.打孔法测定抗生素的效价[J].生物学通报,2005(3):2.
[16]唐书谦,叶庆佾,钟白玉,等.姜黄挥发油、水浸出液体外抗真菌试验[J].第三军医大学学报,1997(6):89-91.
[17]赵淑艳,李喜宏,陈丽,等.桃采后病原菌种类及侵染规律研究[J].食品科学,2005(10):253-255.
[18]周金伟.植物内生菌011粗提液的抑菌机理、保鲜作用及抗菌物质的分离鉴定[D].长沙:湖南农业大学,2015.
[19]郭睿文.猕猴桃内生菌多样性及溃疡病拮抗菌筛选[D].成都:成都理工大学,2016.
[20]陈哲,黄静,赵佳,等.解淀粉芽孢杆菌抑菌机制的研究进展[J].生物技术通报,2015,31(6):37-41.
[21]出晓铭,林毅雄,张珅,等.生防菌解淀粉芽孢杆菌抗菌蛋白的研究进展[J].包装与食品机械,2014,32(6):49-54.
[22]洪鹏,安国栋,胡美英,等.解淀粉芽孢杆菌防治果蔬采后病害研究进展[J].中国农学通报,2013,29(12):168-173.
[23]李红晓,张殿朋,卢彩鸽,等.生防解淀粉芽胞杆菌最新研究进展[J].微生物学杂志,2016,36(2):87-92.