宁夏枸杞鲜果采后病原菌的分离、鉴定及水杨酸的抑病效果研究
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摘要
目的分离、鉴定宁夏中宁地区引起枸杞鲜果采后病害的主要病原菌,并研究水杨酸(salicylic acid,SA)的抑病效果。方法采用常规方法从自然发病果实中分离病原真菌,通过形态学和ITS(internal transcribed spacer)序列信息进行鉴定。使用不同浓度的水杨酸处理病原菌和果实,观察SA对病原菌体外和体内的抑制效果。结果共分离到4株病原真菌菌株,它们均能引起枸杞鲜果的采后腐烂。其中, WB-1鉴定为梨黑斑链格孢(Alternariagaisen),WB-2鉴定为互隔链格孢(Alternariaalternata);WB-3和WB-4分别鉴定为镰刀霉属(Fusarium)和枝孢属(Cladosporium)真菌。SA对3种供试病原真菌的生长均有明显抑制作用,且SA浓度越高抑制效果越明显。SA浸泡处理对常温和低温贮藏的枸杞鲜果病害有显著的控制效果,并且SA处理对枸杞果实固形物含量(soluble solids content, SSC)无显著影响。结论枸杞鲜果采后腐烂由多种病原真菌引起, SA处理对枸杞采后病害具有较好的控制效果,具有潜在的应用前景。
Objective To isolate and identify the main pathogens causing postharvest decay of fresh wolfberry(Lycium barbarum L) fruits, and study the inhibitory effect of salicylic acid(SA). Methods Fungal pathogens were isolated from decayed wolfberry fruits by routine methods and identified according to morphology and ITS(internal transcribed spacer) sequence information. Different concentrations of salicylic acid were used to treat pathogens and fruits, and the inhibitory effects of SA on pathogens in vitro and in vivo were determined. Results A total of 4 fungal pathogen strains were isolated, all of which could cause postharvest decay in wolfberry fruits. Among them, WB-1 was identified as Alternariagaisen, WB-2 as Alternariaalternata, WB-3 and WB-4 as Fusarium spp. and Cladosporium spp., respectively. SA could inhibit the growth of 3 tested pathogen strains, and the inhibitory effects increased along with the concentration of SA. SA treatment showed significant control effect on postharvest decay in fruits stored at both room temperature and low temperature. Moreover, SA treatment had no significant effect on the soluble solids content(SSC). Conclusion Postharvest decay of fresh wolfberry fruits is caused by a variety of fungal pathogens. SA treatment has a good control effect on postharvest diseases of wolfberry and shows a potential application prospect.
Objective To isolate and identify the main pathogens causing postharvest decay of fresh wolfberry(Lycium barbarum L) fruits, and study the inhibitory effect of salicylic acid(SA). Methods Fungal pathogens were isolated from decayed wolfberry fruits by routine methods and identified according to morphology and ITS(internal transcribed spacer) sequence information. Different concentrations of salicylic acid were used to treat pathogens and fruits, and the inhibitory effects of SA on pathogens in vitro and in vivo were determined. Results A total of 4 fungal pathogen strains were isolated, all of which could cause postharvest decay in wolfberry fruits. Among them, WB-1 was identified as Alternariagaisen, WB-2 as Alternariaalternata, WB-3 and WB-4 as Fusarium spp. and Cladosporium spp., respectively. SA could inhibit the growth of 3 tested pathogen strains, and the inhibitory effects increased along with the concentration of SA. SA treatment showed significant control effect on postharvest decay in fruits stored at both room temperature and low temperature. Moreover, SA treatment had no significant effect on the soluble solids content(SSC). Conclusion Postharvest decay of fresh wolfberry fruits is caused by a variety of fungal pathogens. SA treatment has a good control effect on postharvest diseases of wolfberry and shows a potential application prospect.
引文
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[17]Hickert S,Bergmann M,Ersen S,et al.Survey of Alternaria toxin contaminationin food from the German market,using a rapid HPLC-MS/MS approach[J].Mycotoxin Res,2016,(32):7-18.
[18]López P,Venema D,De-Rijk T,et al.Occurrence of Alternaria toxins in food products in the Netherlands[J].Food Contr,2016,(60):196-204.
[19]Zhao K,Shao B,Yang DJ,et al.Natural occurrence of four Alternaria mycotoxins in tomato-and citrus-based foods in China[J].J Agric Food Chem,2015,(63):343-348.
[20]Prelle A,Spadaro D,Garibaldi A.A new method for detection of five Alternaria toxins in food matrices based on LC-APCI-MS[J].Food Chem,2013,(140):161-167.
[21]黄晓静,王少敏,毛丹,等.镰刀菌属真菌毒素的毒性研究进展[J].食品安全质量检测学报,2017,8(8):3117-3128.Huang XJ,Wang SM,Mao D,et al.Research progress on toxicity of Fusarium mycotoxins[J].J Food Saf Qual,2017,8(8):3117-3128.
[2]Amagase H,Farnsworth NR.A review of botanical characteristics,phytochemistry,clinical relevance in efficacy and safety of Lyciumbarbarum fruit(Goji)[J].Food Res Int,2011,44(7):1702-1717.
[3]张曦燕.枸杞鲜果采后贮藏保鲜技术研究进展[J].宁夏农林科技,2010,(6):81-82.Zhang XY.Advances on postharvest storage technology of fresh fruit of Lyciumbarbarum L[J].Ningxia J Agric For Sci Technol,2010,(6):81-82.
[4]魏天军,窦云萍.宁夏枸杞采后呼吸强度和水分变化研究初报[J].北方园艺,2008,(9):210-211.Wei TJ,Dou YP.Preliminary study on respiration and water content of harvested Ningxia wolfberry fruits[J].Northern Hortic,2008,(9):210-211.
[5]葛玉萍,曹有龙,许关,等.构杞鲜果采后品质变化初探[J].北方园艺,2008,(5):227-229.Ge YP,Cao YL,Xu X,et al.Preliminary study on changes in quality of of fresh fruit of Lyciumbarbarum L[J].Northern Hortic,2008,(5):227-229.
[6]兰璞,高芙蓉,陈存坤,等.枸杞采后病害及主要病原真菌的分离与鉴定[J].中国果菜,2014,34(12):9-12.Lan P,Gao FR,Chen CK,et al.Separation and identification of pathogenic fungi fromthe postharvest Lycium Barbarmu[J].Chin Fruit Veg,2014,34(12):9-12.
[7]Wei D,Wang Y,Jiang D,et al.Survey of Alternariatoxins and other mycotoxins in dried fruits in China[J].Toxins,2017,(9):200-211.
[8]田世平,罗云波,王贵禧.园艺产品采后生物学基础[M].北京:中国科学出版社,2011.Tian SP,Luo YB,Wang GX.Postharvest biological basis of horticultural products[M].Beijing:China Science Press,2011.
[9]Asghari M,Aghdam MS.Impact of salicylic acid on post-harvest physiology of horticultural crops[J].Trend Food Sci Technol,2010,21(10):502-509.
[10]Chan Z,Wang Q,Xu X,et al.Functions of defense-related proteins and dehydrogenases in resistance response induced by salicylic acid in sweet cherry fruits at different maturity stages[J].Proteomics,2008,8(22):4791-4807.
[11]Mo Y,Gong D,Liang G,et al.Enhanced preservation effects of sugar apple fruits by salicylic acid treatment during post-harvest storage[J].JSci Food Agric,2008,88(15):2693-2699.
[12]马凌云,赵亮.采后水杨酸处理对油桃果实抗病性的影响[J].河南工业大学学报:自然科学版,2014,35(6):93-96.Ma LY,Zhao L.Effect of postharvest salicylic acid treatment on disease resistance of nectarine fruits[J].J Henan Univ Technol(Nat Sci Ed),2008,88(15):2693-2699.
[13]郭科燕.水杨酸对杏果实采后抗病性和后熟软化的影响[D].乌鲁木齐:新疆农业大学,2012.Guo KY.Effect of salicylic acid treatment on disease resistance and softening of postharvest apricot fruits[D].Urumqi:Xinjiang Agriculture University,2012.
[14]徐祥彬,赖童飞,景云飞,等.山西壶瓶枣缩果病病原菌分离和鉴定[J].植物病理学报,2009,39(3):225-230.Xu XB,Lai TF,Jing YF,et al.Isolation and identification of the pathogens of jujube(Zizyphusjujuba cv.Huping)fruit shrink disease in Shanxi[J].Acta Phytopathol Sinica,2009,39(3):225-230.
[15]Sambrook J,Fritsch E,Maniati Y.Molecular cloning:a laboratory manual[M].New York:Cold Spring Harbor Laboratory Press,1989.
[16]赵游丽,冯美,康建宏.不同温度处理对采后枸杞果实呼吸强度和品质的影响[J].农业科学研究,2010,31(4):34-36.Zhao YL,Feng M,Kang JH.Effect of temperature on respiratory intensity and fruit qualityin Lyciumbarbarum L.post-harvest fruit[J].J Agric Sci,2010,31(4):34-36.
[17]Hickert S,Bergmann M,Ersen S,et al.Survey of Alternaria toxin contaminationin food from the German market,using a rapid HPLC-MS/MS approach[J].Mycotoxin Res,2016,(32):7-18.
[18]López P,Venema D,De-Rijk T,et al.Occurrence of Alternaria toxins in food products in the Netherlands[J].Food Contr,2016,(60):196-204.
[19]Zhao K,Shao B,Yang DJ,et al.Natural occurrence of four Alternaria mycotoxins in tomato-and citrus-based foods in China[J].J Agric Food Chem,2015,(63):343-348.
[20]Prelle A,Spadaro D,Garibaldi A.A new method for detection of five Alternaria toxins in food matrices based on LC-APCI-MS[J].Food Chem,2013,(140):161-167.
[21]黄晓静,王少敏,毛丹,等.镰刀菌属真菌毒素的毒性研究进展[J].食品安全质量检测学报,2017,8(8):3117-3128.Huang XJ,Wang SM,Mao D,et al.Research progress on toxicity of Fusarium mycotoxins[J].J Food Saf Qual,2017,8(8):3117-3128.