不同芽孢杆菌对极早熟桃采后链格孢菌的防治效果及其发酵液挥发性物质成分分析
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摘要
为了明确芽孢杆菌及其发酵液挥发性物质对极早熟桃采后主要致病菌链格孢菌的防治效果,以"津柳早红"桃为试材,通过有伤接种法、采后浸泡法和平板对峙法对三种芽孢杆菌(Bacillus strains) HB-2、B1和X的抑菌活性以及对极早熟桃采后品质保鲜效果的影响进行探究,同时利用平板倒扣法研究三种芽孢杆菌发酵液挥发性物质的抑菌活性,并采用顶空固相微萃取结合气相色谱-质谱联用法(SPME-GC-MS)对发酵液挥发性物质进行成分分析。结果表明:与对照组相比,三种芽孢杆菌发酵液均能显著抑制链格孢菌菌落的生长(p <0.05),同时可有效降低极早熟桃采后腐烂率,延缓果实VC含量、可滴定酸含量和硬度的降低(p <0.05),较好地维持果实品质,且效果HB-2> X> B1;此外,三种芽孢杆菌发酵液产生的挥发性气体对抑制链格孢菌菌落的扩展有明显的抑制作用(p <0.05),抑菌率均在50%以上,并通过对其主要挥发性物质成分进行鉴定分析,从芽孢杆菌HB-2、B1和X发酵液中分别得到28种、31种和30种化合物,主要包括醇类、酯类、酮类、酸类、酚类、烃类和其他类化合物。芽孢杆菌HB-2、B1和X发酵液对极早熟桃采后病害的生物防治具有良好应用潜力。
In order to investigate the control effects of fermented broth of Bacillus strains and volatile organic compounds against postharvest Alternaria alternata of the"JinLiuZaoHong "super-early maturing peach,the injury inoculation method,post-harvest soaking method and plate-to-plate method were studied. The antibacterial activity and the effect of three Bacillus strains HB-2,B1 and X on postharvest quality of super-early maturing peach,and the antibacterial activity of volatile organic compounds of fermented broth was analyzed by the two-sealed-base-plates method and headspace solidphase microextraction combined with gas chromatography-mass spectrometry( SPME-GC-MS) method. The results showed that fermented broth of three Bacillus strains could significantly inhibit the colonization growth of Alternaria alternata( p < 0.05) compared with thecontrol group,and they also significantly delayed the decreasing of vitamin C contents,titratable acid contents and firmness and maintained quality,with the order of HB-2 > X > B1.In addition,the volatile organic compound sproduced by three Bacillus strains fermented broth had significant inhibitory effect on the expansion of Alternaria alternata colonies with the inhibitory rate reached above 50%,and 28,31 and 30 kinds of compounds were identified in HB-2,B1 and X,respectively,such as alcohols,esters,ketones,acids,phenols,alkanes and so on.Bacillus HB-2,B1 and X would have potential application for the biological control of postharvest diseases of super-early maturing peach.
In order to investigate the control effects of fermented broth of Bacillus strains and volatile organic compounds against postharvest Alternaria alternata of the"JinLiuZaoHong "super-early maturing peach,the injury inoculation method,post-harvest soaking method and plate-to-plate method were studied. The antibacterial activity and the effect of three Bacillus strains HB-2,B1 and X on postharvest quality of super-early maturing peach,and the antibacterial activity of volatile organic compounds of fermented broth was analyzed by the two-sealed-base-plates method and headspace solidphase microextraction combined with gas chromatography-mass spectrometry( SPME-GC-MS) method. The results showed that fermented broth of three Bacillus strains could significantly inhibit the colonization growth of Alternaria alternata( p < 0.05) compared with thecontrol group,and they also significantly delayed the decreasing of vitamin C contents,titratable acid contents and firmness and maintained quality,with the order of HB-2 > X > B1.In addition,the volatile organic compound sproduced by three Bacillus strains fermented broth had significant inhibitory effect on the expansion of Alternaria alternata colonies with the inhibitory rate reached above 50%,and 28,31 and 30 kinds of compounds were identified in HB-2,B1 and X,respectively,such as alcohols,esters,ketones,acids,phenols,alkanes and so on.Bacillus HB-2,B1 and X would have potential application for the biological control of postharvest diseases of super-early maturing peach.
引文
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[6]姚良辉,尹京苑,何惠霞,等.生物保鲜剂C06对水蜜桃采后的保鲜效果[J].农业工程学报,2010,26(4):357-361.
[7]毕文慧,姚健,刘学俊,等.微生物在果蔬贮藏保鲜中的应用研究进展[J].食品工业科技,2017,38(20):347-351.
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[9]赵新林,赵思峰.枯草芽孢杆菌对植物病害生物防治的作用机理[J].湖北农业科学,2011,50(15):3025-3028.
[10]魏倩,张娜,张平,等.四种芽孢杆菌对蒜薹灰葡萄孢霉抑制作用的时效性研究[J].生物技术通报,2017,33(6):112-120.
[11]黄运红,王俊明,龙中儿.蜡样芽孢杆菌发酵液对苹果保鲜作用的初步研究[J].食品科学,2006(11):516-519.
[12]吕捷,陈云,朱从一,等.拮抗菌Bs43的鉴定、抑菌机理及其对采后柑橘绿霉病的生防效果[J].果树学报,2014,31(5):885-892,894.
[13]曲慧.解淀粉芽孢杆菌BP30对梨采后灰霉病病害的生物防治研究[D].泰安:山东农业大学,2016:21-22.
[14]张丽,郁志芳,姜丽,等.淀粉液化芽孢杆菌ES-2发酵产物对贮藏期间“湖锦蜜露”水蜜桃品质和生理特性的影响[J].食品科学,2009,30(20):421-425.
[15]谷医林,王远宏,常若葵,等.解淀粉芽孢杆菌LJ1诱导黄瓜抗白粉病的研究[J].农药学学报,2013,15(3):293-298.
[16]郝象2),卢志军,王婧,等.解淀粉芽孢杆菌LJ02抑菌气体分析[J].天津农学院学报,2016,23(3):23-26,30.
[17]Chaurasia B,Pandey A,Palni L M S,et al.Diffusible and volatile compounds produced by an antagonistic Bacillus subtilis strain cause structural deformations in pathogenic fungi in vitro[J].Microbiological Research,2005,160(1):75-81.
[18]Kai M,Haustein M,Molina F,et al.Bacterial volatiles and their action potential[J].Applied Microbiology and Biotechnology,2009,81(6):1001-1012.
[19]Fernando W G D,Ramarathnam R,Krishnamoorthy A S,et al.Identification and use of potential bacterial organic antifungal volatiles in biocontrol[J].Soil Biology and Biochemistry,2005,37(5):955-964.
[20]陈奕鹏,杨扬,桑建伟,等.拮抗内生芽孢杆菌BEB17分离鉴定及其挥发性物质抑菌活性分析[J].植物病理学报,2018,48(4):537-546.
[21]库尔班江,赛丽曼.碘量法测水果蔬菜中维生素C的含量[J].伊犁师范学院学报:自然科学版,2007(3):28-32.
[22]黄华毅,王佳琳,马荣,等.枯草芽孢杆菌STO-12抑菌活性及其抑菌物质分析[J].浙江农业学报,2017,29(1):81-88.
[23]Leelasuphakul W,Hemmanee P,Chuenchitt S.Growth inhibitory properties of Bacillus subtilisstrains and their metabolites against the green mold pathogen(Penicillium digitatum Sacc.)of citrus fruit[J].Postharvest Biology and Technology,2008,48(1):113-121.
[24]周翠.枯草芽孢杆菌(Bacillus subtilis)活性物质的分离和功能测定[D].泰安:山东农业大学,2011:21-22.
[25]Gao Hai-yan,Li Pei-zhong,Xu Xin-xing,et al.Research on volatile organic compounds from Bacillus subtilis CF-3:Biocontrol effects on fruit fungal pathogens and dynamic changes during fermentation[J].Frontiers in Microbiology,2018,9:456.
[26]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.
[27]曹珍珍,周林燕,李淑荣,等.壳聚糖和魔芋精粉复合涂膜对大白杏保鲜效果的影响[J].核农学报,2014,28(11):2051-2057.
[28]李宝庆,鹿秀云,郭庆港,等.枯草芽孢杆菌BAB-1产脂肽类及挥发性物质的分离和鉴定[J].中国农业科学,2010,43(17):3547-3554.
[29]Yuan Jun,Raza W,Shen Qi-rong,et al.Antifungal activity of Bacillus amyloliquefaciens NJN-6 volatile compounds against Fusarium oxysporum f.sp.cubense[J].Applied and Environmental Microbiology,2012,78(16):5942-5944.
[30]Ryu C M,Farag M A,Hu C H,et al.Bacterial volatiles promote growth in Arabidopsis[J].Proceedings of the National Academy of Sciences,2003,100(8):4927-4932.
[31]Almenar E,Del V V,Catala R,et al.Active package for wild strawberry fruit(Fragaria vesca L.)[J].Journal of Agricultural and Food Chemistry,2007,55(6):2240-2245.
[32]Arrebola E,JacobsR,Korsten L.Iturin A is the principal inhibitor in the biocontrol activity of Bacillus amyloliquefaciens PPCB004 against postharvest fungal pathogens[J].Journal of Applied Microbiology,2010,108(2):386-395.
[33]Osman M S,Sivakumar D,Korsten L.Effect of biocontrol agent Bacillus amyloliquefaciens and 1-methyl cyclopropane on the control of postharvest disease and maintenance of fruit quality[J].Crop Protection,2011,30(2):173-178.
[2]冯涛,王庆龙,卢兴霞,等.桃极早熟新品种‘津柳早红’的选育[J].中国果树,2016(1):72-74,101.
[3]赵剑波,郭继英,任飞,等.晚熟蟠桃新品种‘瑞蟠24号’[J].园艺学报,2014,41(11):2359-2360.
[4]张娜,郑香香,于晋泽,等.晚熟桃采后褐腐病致病菌的分离鉴定及拮抗菌对其防治效果的研究[J].食品工业科技,2018,39(2):102-107.
[5]郑琪.核果类果实采后病害拮抗菌筛选及其抑菌作用研究[D].兰州:甘肃农业大学,2013:1-2.
[6]姚良辉,尹京苑,何惠霞,等.生物保鲜剂C06对水蜜桃采后的保鲜效果[J].农业工程学报,2010,26(4):357-361.
[7]毕文慧,姚健,刘学俊,等.微生物在果蔬贮藏保鲜中的应用研究进展[J].食品工业科技,2017,38(20):347-351.
[8]Turner J T,Backman P A.Factors relating to peanut yield increases after seed treatment with Bacillus subtilis[J].Plant Disease,1991,75(4):347-353.
[9]赵新林,赵思峰.枯草芽孢杆菌对植物病害生物防治的作用机理[J].湖北农业科学,2011,50(15):3025-3028.
[10]魏倩,张娜,张平,等.四种芽孢杆菌对蒜薹灰葡萄孢霉抑制作用的时效性研究[J].生物技术通报,2017,33(6):112-120.
[11]黄运红,王俊明,龙中儿.蜡样芽孢杆菌发酵液对苹果保鲜作用的初步研究[J].食品科学,2006(11):516-519.
[12]吕捷,陈云,朱从一,等.拮抗菌Bs43的鉴定、抑菌机理及其对采后柑橘绿霉病的生防效果[J].果树学报,2014,31(5):885-892,894.
[13]曲慧.解淀粉芽孢杆菌BP30对梨采后灰霉病病害的生物防治研究[D].泰安:山东农业大学,2016:21-22.
[14]张丽,郁志芳,姜丽,等.淀粉液化芽孢杆菌ES-2发酵产物对贮藏期间“湖锦蜜露”水蜜桃品质和生理特性的影响[J].食品科学,2009,30(20):421-425.
[15]谷医林,王远宏,常若葵,等.解淀粉芽孢杆菌LJ1诱导黄瓜抗白粉病的研究[J].农药学学报,2013,15(3):293-298.
[16]郝象2),卢志军,王婧,等.解淀粉芽孢杆菌LJ02抑菌气体分析[J].天津农学院学报,2016,23(3):23-26,30.
[17]Chaurasia B,Pandey A,Palni L M S,et al.Diffusible and volatile compounds produced by an antagonistic Bacillus subtilis strain cause structural deformations in pathogenic fungi in vitro[J].Microbiological Research,2005,160(1):75-81.
[18]Kai M,Haustein M,Molina F,et al.Bacterial volatiles and their action potential[J].Applied Microbiology and Biotechnology,2009,81(6):1001-1012.
[19]Fernando W G D,Ramarathnam R,Krishnamoorthy A S,et al.Identification and use of potential bacterial organic antifungal volatiles in biocontrol[J].Soil Biology and Biochemistry,2005,37(5):955-964.
[20]陈奕鹏,杨扬,桑建伟,等.拮抗内生芽孢杆菌BEB17分离鉴定及其挥发性物质抑菌活性分析[J].植物病理学报,2018,48(4):537-546.
[21]库尔班江,赛丽曼.碘量法测水果蔬菜中维生素C的含量[J].伊犁师范学院学报:自然科学版,2007(3):28-32.
[22]黄华毅,王佳琳,马荣,等.枯草芽孢杆菌STO-12抑菌活性及其抑菌物质分析[J].浙江农业学报,2017,29(1):81-88.
[23]Leelasuphakul W,Hemmanee P,Chuenchitt S.Growth inhibitory properties of Bacillus subtilisstrains and their metabolites against the green mold pathogen(Penicillium digitatum Sacc.)of citrus fruit[J].Postharvest Biology and Technology,2008,48(1):113-121.
[24]周翠.枯草芽孢杆菌(Bacillus subtilis)活性物质的分离和功能测定[D].泰安:山东农业大学,2011:21-22.
[25]Gao Hai-yan,Li Pei-zhong,Xu Xin-xing,et al.Research on volatile organic compounds from Bacillus subtilis CF-3:Biocontrol effects on fruit fungal pathogens and dynamic changes during fermentation[J].Frontiers in Microbiology,2018,9:456.
[26]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.
[27]曹珍珍,周林燕,李淑荣,等.壳聚糖和魔芋精粉复合涂膜对大白杏保鲜效果的影响[J].核农学报,2014,28(11):2051-2057.
[28]李宝庆,鹿秀云,郭庆港,等.枯草芽孢杆菌BAB-1产脂肽类及挥发性物质的分离和鉴定[J].中国农业科学,2010,43(17):3547-3554.
[29]Yuan Jun,Raza W,Shen Qi-rong,et al.Antifungal activity of Bacillus amyloliquefaciens NJN-6 volatile compounds against Fusarium oxysporum f.sp.cubense[J].Applied and Environmental Microbiology,2012,78(16):5942-5944.
[30]Ryu C M,Farag M A,Hu C H,et al.Bacterial volatiles promote growth in Arabidopsis[J].Proceedings of the National Academy of Sciences,2003,100(8):4927-4932.
[31]Almenar E,Del V V,Catala R,et al.Active package for wild strawberry fruit(Fragaria vesca L.)[J].Journal of Agricultural and Food Chemistry,2007,55(6):2240-2245.
[32]Arrebola E,JacobsR,Korsten L.Iturin A is the principal inhibitor in the biocontrol activity of Bacillus amyloliquefaciens PPCB004 against postharvest fungal pathogens[J].Journal of Applied Microbiology,2010,108(2):386-395.
[33]Osman M S,Sivakumar D,Korsten L.Effect of biocontrol agent Bacillus amyloliquefaciens and 1-methyl cyclopropane on the control of postharvest disease and maintenance of fruit quality[J].Crop Protection,2011,30(2):173-178.
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