植物提取液对辣椒采后病原菌抑制及对根霉病抗性诱导作用
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摘要
以‘湘研15号’辣椒为试材,分离、鉴定出4株辣椒采后病原菌,采用生长速率法测定丁香、肉桂、花椒等9种植物提取液对尖孢镰刀菌、黑色炭疽菌、红色炭疽菌、根霉菌的抑制效果,通过平板梯度稀释法测定植物提取液对上述4种供试病菌的最小抑菌浓度(minimum inhibitory concentration,MIC)和最小杀菌浓度(minimum bactericidal concentration,MBC);通过测定辣椒的病斑直径、发病率、总酚及类黄酮含量、苯丙氨酸解氨酶(phenylalanine ammonia lyase,PAL)、过氧化物酶(peroxldase,POD)、多酚氧化酶(polyphenol oxidase,PPO)活力,研究复合涂膜剂诱导辣椒对根霉病的抗性作用。结果显示,丁香、肉桂提取液对4种供试病原菌的抑菌率均为100%,对尖孢镰刀菌、黑色炭疽菌、红色炭疽菌和根霉菌的MIC均分别为1.25、2.50、2.50 mg/mL和5.00 mg/mL,MBC均分别为2.50、5.00、5.00 mg/mL和10.00 mg/mL。丁香、肉桂提取液复合涂膜剂能有效抑制辣椒的发病率和减小病斑直径,接种第8天时,与刺伤后接种根霉菌孢子组(CK2)相比,复合涂膜剂组发病率降低了6.67%(P>0.05)、病斑直径减小了30.35%(P<0.05);与刺伤后加无菌水组(CK1)和CK2组相比,复合涂膜剂处理辣椒的PAL活力分别提高了45.91%、24.54%(P<0.05);POD活力分别提高了22.15%(P<0.05)、12.54%(P>0.05);PPO活力分别提高了80.00%、28.57%(P<0.05);总酚含量分别提高了60.00%、43.74%(P<0.05);类黄酮含量分别提高了82.05%、61.36%(P<0.05)。结论:丁香、肉桂提取液复合涂膜剂能诱导辣椒对根霉病产生抗性,本研究可为开发高效、稳定、无毒无害辣椒防腐保鲜剂提供理论和技术支持。
Four species of postharvest pathogens from ‘Xiangyan 15’ hot peppers were isolated and identified as Fusarium oxysporum, Colletotrichum nigrum, Colletotrichum gloeosporioides and Rhizopus spp.. The inhibitory effect of alcoholic extracts from nine plants including clove(Syzygium aromaticum), cinnamon(Cinnamomum cassia) and prickly ash(Zanthoxylunm bungeanum) on these pathogens was evaluated by growth rate method. The minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) were measured by plate gradient dilution method.Furthermore, composite guar gum-pullulan coating incorporated with clove and cinnamon extracts was assessed for its ability to induce resistance to Rhizopus spp. in hot peppers by determination of lesion diameter, disease incidence, total phenolic and total flavonoid contents, and the activities of phenylalanine ammonia lyase(PAL), peroxldase(POD) and polyphenol oxidase(PPO). The results showed that all four pathogens were 100% inhibited by clove and cinnamon extracts. Both plant extracts had an equal MIC of 1.25, 2.50, 2.50 and 5.00 mg/mL; and an equal MBC of 2.50, 5.00, 5.00 and 10.00 mg/mL against Fusarium oxysporum, Colletotrichum nigrum, Colletotrichum gloeosporioides, Rhizopus spp.,respectively. The disease incidence and lesion diameter in hot peppers caused by Rhizopus spp. were effectively inhibited by the composite coating, which were decreases by 6.67%(P > 0.05) and 30.35%(P < 0.05), respectively after 8 d storage,compared with CK2(wounded and inoculated with conidial suspension of Rhizopus spp.). Compared with CK1(wounded and inoculated with sterile water) and CK2, the composite coating enhanced PAL activity by 45.91% and 24.54%(P <0.05), POD activity by 22.15%(P < 0.05) and 12.54%(P > 0.05), and PPO activity by 80.00% and 28.57%(P < 0.05); and increased total phenolic content by 60.00% and 43.74%(P < 0.05), and total flavonoid content by 82.05% and 61.36%(P <0.05), respectively. These results suggested that the composite coating containing clove and cinnamon extracts can induce resistance to Rhizopus spp. in hot peppers. Our data provide experimental support for the development of efficient, stable and safe preservatives for hot peppers.
Four species of postharvest pathogens from ‘Xiangyan 15’ hot peppers were isolated and identified as Fusarium oxysporum, Colletotrichum nigrum, Colletotrichum gloeosporioides and Rhizopus spp.. The inhibitory effect of alcoholic extracts from nine plants including clove(Syzygium aromaticum), cinnamon(Cinnamomum cassia) and prickly ash(Zanthoxylunm bungeanum) on these pathogens was evaluated by growth rate method. The minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) were measured by plate gradient dilution method.Furthermore, composite guar gum-pullulan coating incorporated with clove and cinnamon extracts was assessed for its ability to induce resistance to Rhizopus spp. in hot peppers by determination of lesion diameter, disease incidence, total phenolic and total flavonoid contents, and the activities of phenylalanine ammonia lyase(PAL), peroxldase(POD) and polyphenol oxidase(PPO). The results showed that all four pathogens were 100% inhibited by clove and cinnamon extracts. Both plant extracts had an equal MIC of 1.25, 2.50, 2.50 and 5.00 mg/mL; and an equal MBC of 2.50, 5.00, 5.00 and 10.00 mg/mL against Fusarium oxysporum, Colletotrichum nigrum, Colletotrichum gloeosporioides, Rhizopus spp.,respectively. The disease incidence and lesion diameter in hot peppers caused by Rhizopus spp. were effectively inhibited by the composite coating, which were decreases by 6.67%(P > 0.05) and 30.35%(P < 0.05), respectively after 8 d storage,compared with CK2(wounded and inoculated with conidial suspension of Rhizopus spp.). Compared with CK1(wounded and inoculated with sterile water) and CK2, the composite coating enhanced PAL activity by 45.91% and 24.54%(P <0.05), POD activity by 22.15%(P < 0.05) and 12.54%(P > 0.05), and PPO activity by 80.00% and 28.57%(P < 0.05); and increased total phenolic content by 60.00% and 43.74%(P < 0.05), and total flavonoid content by 82.05% and 61.36%(P <0.05), respectively. These results suggested that the composite coating containing clove and cinnamon extracts can induce resistance to Rhizopus spp. in hot peppers. Our data provide experimental support for the development of efficient, stable and safe preservatives for hot peppers.
引文
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[15]吕明珠,于爽,朱恩俊.肉桂精油对红提葡萄保鲜效果的影响[J].食品科学,2016,37(6):272-277.DOI:10.7506/spkx1002-6630-201606049.
[16]蓬桂华,杨万荣,苏丹,等.中草药处理对辣椒贮藏特性的影响[J].安徽农业科学,2014,42(13):4036-4040.DOI:10.3969/j.issn.0517-6611.2014.13.098.
[17]曾荣,张阿珊,陈金印.植物源防腐剂在果蔬保鲜中应用研究进展[J].中国食品学报,2011,11(4):161-167.DOI:10.3969/j.issn.1009-7848.2011.04.024.
[18]赵恭文,吴建挺,李长松,等.中草药对植物病原真菌的抑菌活性筛选研究[C]//中国植物病理学会2011年学术年会论文集.宜昌:中国农业科学技术出版社,2011:10-15.
[19]罗永兰,张志元,张翼.辣椒果柄表面附生菌与贮藏期病害的关系[J].湖北农业科学,2004(5):93-95.DOI:10.3969/j.issn.0439-8114.2004.05.034.
[20]肖晶,陈维信,刘爱媛,等.辣椒采后病害发生情况[J].中国蔬菜,2008(6):13-16.DOI:10.3249/j.issn.0439-8114.2008.06.018.
[21]张志元,官春云.不同栽培方式对辣椒采后病害的影响[J].植物病理学报,2003,33(2):183-186.DOI:10.3321/j.issn:0412-0914.2003.02.018
[22]余东坡.中草药醇提物抑菌活性筛选及其在青椒保鲜上的应用[D].郑州:河南农业大学,2008:41-48.
[23]王博宇.青椒采后黑斑病的诱导抗病性研究[D].天津:天津科技大学,2009:24-29.
[24]余辰.γ-氨基丁酸对梨果实青霉病抗性的诱导作用及相关机理研究[D].杭州:浙江大学,2014:32-38.
[25]CHEN S,NUSSINOVITCH A.Galactomannans in disturbances of structured wax-hydrocolloid-based coatings of citrus fruit(easypeelers)[J].Food Hydrocolloids,2000,14(6):561-568.DOI:10.1016/s0268-005x(00)00036-9.
[26]张美丽,何玲,苑希蕊,等.银杏叶提取液诱导采后猕猴桃对青霉病的抗性机制[J].食品科学,2016,37(6):242-247.DOI:10.7506/spkx1002-6630-201606044.
[27]CHLEBOWSKA-SMIGIEL A,GNIEWOSZ M,SWICZAK E.An attempt to apply a pullulan and pullulan-protein coatings to prolong apples shelf-life stability[J].Acta Scientiarum Polonorum Technologia Alimentaria,2007,6(1):49-56.DOI:10.1016/b978-0-08-100596-5.21129-3.
[28]NANTAWANIT N,CHANCHAICHAOVIVAT A,PANIPAN B,et al.Induction of defense response against Colletotrichum capsici in chili fruit by the yeast Pichia guilliermondii strain R13[J].Biological Control,2010,52(2):145-152.DOI:10.1016/j.biocontrol.2009.10.011.
[2]ALI A,NOH N M,MUSTAFA M A.Antimicrobial activity of chitosan enriched with lemongrass oil against anthracnose of bell pepper[J].Food Packaging and Shelf Life,2015,3:56-61.DOI:10.1016/j.fpsl.2014.10.003.
[3]DIAZ-PEREZ J C,MUY-RANGEL M D,MASCORRO A G.Fruit size and stage of ripeness affect postharvest water loss in bell pepper fruit(Capsicum annuum L.)[J].Journal of the Science of Food and Agriculture,2007,87(1):68-73.DOI:10.21273/hortsci.41.3.504a.
[4]FALLIK E,GRINBERG S.Hinokitiol:a natural substance that controls postharvest diseases in eggplant and pepper fruits[J].Postharvest Biology and Technology,1992,2(2):137-144.DOI:10.1016/0925-5214(92)90016-i.
[5]闫师杰,梁丽雅,郑伟.中草药提取液贮藏青椒实验[J].食品科学,2002,23(3):138-140.DOI:10.3321/j.issn:1002-6630.2002.03.038.
[6]余东坡,王兰菊,司芳,等.21种中草药醇提物抑菌活性研究[J].安徽农业科学,2008,36(3):1086-1087.DOI:10.3969/j.issn.0517-6611.2008.03.103.
[7]EDIRISINGHE M,ALI A,MAQBOOL M,et al.Chitosan controls postharvest anthracnose in bell pepper by activating defense-related enzymes[J].Journal of Food Science and Technology,2014,51(12):4078-4083.DOI:10.1007/s13197-012-0907-5.
[8]POVERENOV E,ZAISEV Y,AMON H,et al.Effects of a composite chitosan-gelatin edible coating on postharvest quality and storability of red bell peppers[J].Postharvest Biology and Technology,2014,96:106-109.DOI:10.1016/j.postharvbio.2014.05.015.
[9]ILIC Z S,TRAJKOVIC R,PERZELAN Y,et al.Influence of1-methylcyclopropene(1-MCP)on postharvest storage quality in green bell pepper fruit[J].Food and Bioprocess Technology,2012,5(7):2758-2767.DOI:10.1007/s11947-011-0614-7.
[10]魏景超.真菌鉴定手册[M].上海:上海科学技术文献出版社,1979:2-15.
[11]侯田莹,王福东,寇文丽,等.温度变化和1-MCP处理对青椒贮藏品质的影响[J].保鲜与加工,2012,12(5):8-13.DOI:10.3969/j.issn.1009-6221.2012.05.002.
[12]哈斯格根,艾启俊,魏月琴,等.中草药提取物对蔬菜两种采后病原真菌的抑菌活性研究[J].北京农学院学报,2009,24(1):20-23.DOI:10.3969/j.issn.1002-3186.2009.01.005.
[13]罗程印,李高阳,柏连阳,等.植物源活性成分诱导果蔬的抗病性研究[J].食品工业,2016,37(10):237-242.DOI:10.3969/j.issn.1025-45371.2016.10.331.
[14]曹建康,姜微波,赵玉梅.果蔬采后生理生化实验指导[M].北京:中国轻工业出版社,2007:21-22.
[15]吕明珠,于爽,朱恩俊.肉桂精油对红提葡萄保鲜效果的影响[J].食品科学,2016,37(6):272-277.DOI:10.7506/spkx1002-6630-201606049.
[16]蓬桂华,杨万荣,苏丹,等.中草药处理对辣椒贮藏特性的影响[J].安徽农业科学,2014,42(13):4036-4040.DOI:10.3969/j.issn.0517-6611.2014.13.098.
[17]曾荣,张阿珊,陈金印.植物源防腐剂在果蔬保鲜中应用研究进展[J].中国食品学报,2011,11(4):161-167.DOI:10.3969/j.issn.1009-7848.2011.04.024.
[18]赵恭文,吴建挺,李长松,等.中草药对植物病原真菌的抑菌活性筛选研究[C]//中国植物病理学会2011年学术年会论文集.宜昌:中国农业科学技术出版社,2011:10-15.
[19]罗永兰,张志元,张翼.辣椒果柄表面附生菌与贮藏期病害的关系[J].湖北农业科学,2004(5):93-95.DOI:10.3969/j.issn.0439-8114.2004.05.034.
[20]肖晶,陈维信,刘爱媛,等.辣椒采后病害发生情况[J].中国蔬菜,2008(6):13-16.DOI:10.3249/j.issn.0439-8114.2008.06.018.
[21]张志元,官春云.不同栽培方式对辣椒采后病害的影响[J].植物病理学报,2003,33(2):183-186.DOI:10.3321/j.issn:0412-0914.2003.02.018
[22]余东坡.中草药醇提物抑菌活性筛选及其在青椒保鲜上的应用[D].郑州:河南农业大学,2008:41-48.
[23]王博宇.青椒采后黑斑病的诱导抗病性研究[D].天津:天津科技大学,2009:24-29.
[24]余辰.γ-氨基丁酸对梨果实青霉病抗性的诱导作用及相关机理研究[D].杭州:浙江大学,2014:32-38.
[25]CHEN S,NUSSINOVITCH A.Galactomannans in disturbances of structured wax-hydrocolloid-based coatings of citrus fruit(easypeelers)[J].Food Hydrocolloids,2000,14(6):561-568.DOI:10.1016/s0268-005x(00)00036-9.
[26]张美丽,何玲,苑希蕊,等.银杏叶提取液诱导采后猕猴桃对青霉病的抗性机制[J].食品科学,2016,37(6):242-247.DOI:10.7506/spkx1002-6630-201606044.
[27]CHLEBOWSKA-SMIGIEL A,GNIEWOSZ M,SWICZAK E.An attempt to apply a pullulan and pullulan-protein coatings to prolong apples shelf-life stability[J].Acta Scientiarum Polonorum Technologia Alimentaria,2007,6(1):49-56.DOI:10.1016/b978-0-08-100596-5.21129-3.
[28]NANTAWANIT N,CHANCHAICHAOVIVAT A,PANIPAN B,et al.Induction of defense response against Colletotrichum capsici in chili fruit by the yeast Pichia guilliermondii strain R13[J].Biological Control,2010,52(2):145-152.DOI:10.1016/j.biocontrol.2009.10.011.