槐糖脂对蜜橘果实采后青霉病的控制及其机理
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摘要
从自然发病的蜜橘果实上分离出一株致病真菌,通过形态学特征、rDNA-内转录间隔区(rDNA-internal transcribed spacer,rDNA-ITS)分析以及致病性检测,发现该真菌为柑橘意大利青霉(Penicillium italicum)。离体实验发现,当槐糖脂质量浓度为0.1 g/L时,青霉的生长被完全抑制;在果实实验中,当槐糖脂质量浓度为35 g/L时,蜜桔果实的病斑直径最小,且发病率最低。采用质量浓度为35 g/L的槐糖脂对蜜桔果实进行接种和浸泡处理,均能有效提高蜜桔果实的抗病性,且蜜桔果皮的过氧化物酶、多酚氧化酶和苯丙氨酸解氨酶的活力得到了提高。该实验为蜜桔果实采后青霉病的防治提供了一定的理论依据。
A pathogenic fungal strain was isolated from naturally infected satsuma mandarin fruit. On the basis of morphological characteristics, rDNA-internal transcribed spacer(rDNA-ITS) sequence analysis and pathogenicity test,the pathogen was identi?ed as Penicillium italicum. The in vitro test showed that the growth of Penicillium italicum was completely inhibited at a sophorolipid concentration of 0.1 g/L; in the fruit test, the smallest lesion diameter and the lowest incidence of blue mold were observed at a sophorolipid concentration of 35 g/L. Treatment with 35 g/L sophorolipid effectively enhanced disease resistance in inoculated satsuma mandarin fruit by increasing the activities of peroxidase(POD),polyphenol oxidase(PPO) and phenylalanine ammonialyase(PAL). Our experimental results offer a theoretical basis for the prevention and control of postharvest blue mold in satsuma mandarin fruit.
A pathogenic fungal strain was isolated from naturally infected satsuma mandarin fruit. On the basis of morphological characteristics, rDNA-internal transcribed spacer(rDNA-ITS) sequence analysis and pathogenicity test,the pathogen was identi?ed as Penicillium italicum. The in vitro test showed that the growth of Penicillium italicum was completely inhibited at a sophorolipid concentration of 0.1 g/L; in the fruit test, the smallest lesion diameter and the lowest incidence of blue mold were observed at a sophorolipid concentration of 35 g/L. Treatment with 35 g/L sophorolipid effectively enhanced disease resistance in inoculated satsuma mandarin fruit by increasing the activities of peroxidase(POD),polyphenol oxidase(PPO) and phenylalanine ammonialyase(PAL). Our experimental results offer a theoretical basis for the prevention and control of postharvest blue mold in satsuma mandarin fruit.
引文
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[21]KIM K,YOO D,KIM Y,et al.Characteristics of sophorolipid as an antimicrobial agent[J].Journal of Microbiology&Biotechnology,2002,12(2):235-241.
[22]路来风.海洋拮抗酵母Rhodosporidium paludigenum对柑橘果实抗性的增强效应及其生物学机理研究[D].杭州:浙江大学,2015:31.
[23]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,2007:142-147.
[24]SRIVASTAVA M K,DWIVEDI U N.Delayed ripening of banana fruit by salicylic acid[J].Plant Science,2000,158(1/2):87-96.DOI:10.1016/S0168-9452(00)00304-6.
[25]ASSIS J S,MALDONADO R,MU?OZ T,et al.Effect of high carbon dioxide concentration on PAL activity and phenolic contents in ripening cherimoya fruit[J].Postharvest Biology and Technology,2001,23(1):33-39.DOI:10.1016/S0925-5214(01)00100-4.
[26]YOO D S,LEE B S,KIM E K.Characteristics of microbial biosurfactant as an antifungal agent against plant pathogenic fungus[J].Journal of Microbiology&Biotechnology,2005,15(6):1164-1169.
[27]胡静,赵小慧,朱春玉,等.槐糖脂对金黄色葡萄球菌的抑菌机理[J].食品科学,2012,33(5):33-36.
[28]毛淑波.罗伦隐球酵母结合热空气处理对草莓采后病害的防治及其机理研究[D].南京:南京农业大学,2013:8-9.
[29]SCHNEIDER S,ULLRICH W R.Differential induction of resistance and enhanced enzyme activities in cucumber and tobacco caused by treatment with various abiotic and biotic inducers[J].Physiological and Molecular Plant Pathology,1994,45(4):291-304.DOI:10.1016/S0885-5765(05)80060-8.
[30]ABELES F B,BILES C L.Characterization of peroxidases in lignifying peach fruit endocarp[J].Plant Physiology,1991,95(1):269-273.DOI:10.1104/pp.95.1.269.
[31]PELLEGRINI L,ROHFRITSCH O,FRITIG B,et al.Phenylalanine ammonia-lyase in tobacco.molecular cloning and gene expression during the hypersensitive reaction to tobacco mosaic virus and the response to a fungal elicitor[J].Plant Physiology,1994,106(3):877-886.DOI:10.1104/pp.106.3.877.
[32]苏晶.采后苹果果实机械伤诱导的抗病性机理研究[D].南京:南京农业大学,2011:2.
[2]孙萍.粘红酵母对柑橘采后病害的生物防治及其防治机理的研究[D].杭州:浙江大学,2003:1.
[3]李红叶,曹若彬.果蔬产后病害生物防治研究进展[J].中国生物防治通报,1993,9(4):176-180.
[4]BOUBAKER H,SAADI B,BOUDYACH E H,et al.Sensitivity of Penicillium digitatum and P.italicum to imazalil and thiabendazole in Morocco[J].Plant Pathology Journal,2009,8(4):152-158.DOI:10.3923/ppj.2009.152.158.
[5]HOLMES G J,ECKERT J W.Sensitivity of Penicillium digitatum and P.italicum to postharvest citrus fungicides in California[J].Phytopathology,1999,89(9):716-721.DOI:10.1094/PHYTO.1999.89.9.716.
[6]WISNIEWSKI M E,WILSON C L.Biological control of postharvest diseases of fruits and vegetables:recent advances[J].Hortscience,1992,27(2):94-98.DOI:10.1007/BF00041276.
[7]ITO S,INOUE S.Sophorolipids from Torulopsis bombicola:possible relation to alkane uptake[J].Applied&Environmental Microbiology,1982,43(6):1278-1283.
[8]HOMMEL R,STIIWER O,STUBER W,et al.Production of watersoluble surface-active exolipids by Torulopsis apicola[J].Applied Microbiology and Biotechnology,1987,26(3):199-205.DOI:10.1007/BF00286308.
[9]DAVEREY A,PAKSHIRAJAN K.Kinetics of growth and enhanced sophorolipids production by Candida bombicola using a low-cost fermentative medium[J].Applied Biochemistry&Biotechnology,2010,160(7):2090-2101.DOI:10.1007/s12010-009-8797-3.
[10]PEKIN G,VARDAR-SUKAN F,KOSARIC N.Production of sophorolipids from Candida bombicola ATCC 22214 using Turkish corn oil and honey[J].Engineering in Life Sciences,2005,5(4):357-362.DOI:10.1002/elsc.200520086.
[11]ZERKOWSKI J A,SOLAIMAN D K Y.Polyhydroxy fatty acids derived from sophorolipids[J].Journal of the American Oil Chemists’Society,2007,84(5):463-471.DOI:10.1007/s11746-007-1054-4.
[12]BOGAERT I N A V,ROELANTS S,DEVELTER D,et al.Sophorolipid production by Candida bombicola on oils with a special fatty acid composition and their consequences on cell viability[J].Biotechnology Letters,2010,32(10):1509-1514.DOI:10.1109/20.133698.
[13]BOGAERT I N A V,DEVELTER D,SOETAERT W,et al.Cerulenin inhibits de novo sophorolipid synthesis of Candida bombicola[J].Biotechnology Letters,2008,30(10):1829-1832.DOI:10.1007/s10529-008-9764-8.
[14]SMYTH T J,PERFUMO A,MARCHANT R,et al.Directed microbial biosynthesis of deuterated biosurfactants and potential future application to other bioactive molecules[J].Applied Microbiology and Biotechnology,2010,87(4):1347-1354.DOI:10.1007/s00253-010-2592-5.
[15]BANAT I M,MAKKAR R S,CAMEOTRA S S.Potential commercial applications of microbial surfactants[J].Applied Microbiology and Biotechnology,2000,53(5):495-508.DOI:10.1007/s002530051648.
[16]CAMEOTRA S S,MAKKAR R S.Synthesis of biosurfactants in extreme conditions[J].Applied Microbiology and Biotechnology,1998,50(5):520-529.DOI:10.1007/s002530051329.
[17]NAPOLITANO L M.Sophorolipids in sepsis:antiinflammatory or antibacterial?[J].Critical Care Medicine,2006,34(1):258-259.DOI:10.1097/01.CCM.0000196082.26483.F0.
[18]ISODA H,KITAMOTO D,SHINMOTO H,et al.Microbial extracellular glycolipid induction of differentiation and inhibition of the protein kinase C activity of human promyelocytic leukemia cell line HL60[J].Bioscience Biotechnology&Biochemistry,1997,61(4):609-614.DOI:10.1271/bbb.61.609.
[19]SHAH V,DONCEL G F,SEYOUM T,et al.Sophorolipids,microbial glycolipids with anti-human immunodeficiency virus and spermimmobilizing activities[J].Antimicrobial Agents and Chemotherapy,2005,49(10):4093-4100.DOI:10.1128/AAC.49.10.4093-4100.2005.
[20]宋欣,曲音波,李慧.槐糖脂在制备抗宫颈癌药物中的应用:CN101703514A[P].2010-05-12[2017-11-15].https://www.ixueshu.com/document/5a40c305487775dd318947a18e7f9386.html.
[21]KIM K,YOO D,KIM Y,et al.Characteristics of sophorolipid as an antimicrobial agent[J].Journal of Microbiology&Biotechnology,2002,12(2):235-241.
[22]路来风.海洋拮抗酵母Rhodosporidium paludigenum对柑橘果实抗性的增强效应及其生物学机理研究[D].杭州:浙江大学,2015:31.
[23]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,2007:142-147.
[24]SRIVASTAVA M K,DWIVEDI U N.Delayed ripening of banana fruit by salicylic acid[J].Plant Science,2000,158(1/2):87-96.DOI:10.1016/S0168-9452(00)00304-6.
[25]ASSIS J S,MALDONADO R,MU?OZ T,et al.Effect of high carbon dioxide concentration on PAL activity and phenolic contents in ripening cherimoya fruit[J].Postharvest Biology and Technology,2001,23(1):33-39.DOI:10.1016/S0925-5214(01)00100-4.
[26]YOO D S,LEE B S,KIM E K.Characteristics of microbial biosurfactant as an antifungal agent against plant pathogenic fungus[J].Journal of Microbiology&Biotechnology,2005,15(6):1164-1169.
[27]胡静,赵小慧,朱春玉,等.槐糖脂对金黄色葡萄球菌的抑菌机理[J].食品科学,2012,33(5):33-36.
[28]毛淑波.罗伦隐球酵母结合热空气处理对草莓采后病害的防治及其机理研究[D].南京:南京农业大学,2013:8-9.
[29]SCHNEIDER S,ULLRICH W R.Differential induction of resistance and enhanced enzyme activities in cucumber and tobacco caused by treatment with various abiotic and biotic inducers[J].Physiological and Molecular Plant Pathology,1994,45(4):291-304.DOI:10.1016/S0885-5765(05)80060-8.
[30]ABELES F B,BILES C L.Characterization of peroxidases in lignifying peach fruit endocarp[J].Plant Physiology,1991,95(1):269-273.DOI:10.1104/pp.95.1.269.
[31]PELLEGRINI L,ROHFRITSCH O,FRITIG B,et al.Phenylalanine ammonia-lyase in tobacco.molecular cloning and gene expression during the hypersensitive reaction to tobacco mosaic virus and the response to a fungal elicitor[J].Plant Physiology,1994,106(3):877-886.DOI:10.1104/pp.106.3.877.
[32]苏晶.采后苹果果实机械伤诱导的抗病性机理研究[D].南京:南京农业大学,2011:2.