反式-2-己烯醛对猕猴桃贮藏过程扩展青霉生长的抑制作用
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摘要
为研究反式-2-己烯醛对扩展青霉生长及展青霉素产生的抑制作用。选取反式-2-己烯醛和研究报道的8种具有抑菌效果的物质对5株扩展青霉进行体外抑菌试验,研究反式-2-己烯醛的抑菌效果并测定其最小抑菌浓度,然后进行猕猴桃活体试验,通过测定硬度、可溶性固形物、可滴定酸、维生素C、pH值等指标研究反式-2-己烯醛对猕猴桃品质的影响,并采用高效液相检测抑菌试验前后猕猴桃活体中展青霉素含量,最终通过扫面电子显微镜观察反式-2-己烯醛对5株扩展青霉形态的影响。结果反式-2-己烯醛与8种抑菌物质相比,反式-2-己烯醛抑菌效果最好,能够完全抑制5株扩展青霉生长且得到对5株试验菌的最小抑菌浓度(MIC,minimuminhibitoryconcentration);反式-2-己烯醛能够极显著降低扩展青霉对猕猴桃果实硬度、可滴定酸、维生素C的的影响(P<0.01),并且对猕猴桃硬度、可溶性固形物、pH值和还原糖及维生素C含量等没有极显著影响(P>0.01),极显著降低了染菌猕猴桃果实的腐烂率(P<0.01),较好的保持了猕猴桃品质,经反式-2-己烯醛处理的染菌猕猴桃均没有展青霉素检出;扫面电子显微镜显示5株试验菌出现菌丝皱缩、折叠、干瘪,孢子生成减少现象。研究反式-2-己烯醛对扩展青霉生长及展青霉素产生均具有很好的抑制作用,可为开发具有抑菌作用可替代农药的天然产物提供理论基础。
Patulin is a neurotoxic secondary metabolite that causes a series of acute and chronic conditions such as enteritis, dyspnea and cancer when the body consumes excessive amounts. China is the birthplace of kiwifruit and the largest producer of kiwifruit in the world. A large number of studies had shown that kiwifruit is susceptible to infection with penicillium expansum(P.expansum) and patulin accumulation during harvesting, storage and processing which not only causes waste, but also harms human health. Therefore, it is necessary to study a safe and effective method to control P.expansum and patulin accumulation of kiwifruit and its products. Trans-2-hexenal is a characteristic aroma component naturally present in kiwi, apple and other fruits. Studies had shown that trans-2-hexenal has a certain inhibitory effect on microorganisms. This article studied the inhibitory effect of hexenal on the growth of P.expansum and the production of patulin. Firstly, the five strains of P.expansum(CICC40658, CGMCC3.3703, LPH6, LPH5, WY) were inoculated into PDA medium plates containing bacteriostatic substance concentration of 5 mg/mL which kept at 28 ℃and the diameter of the colony was measured by the cross method every day. Study the antibacterial effect of trans-2-hexenal and eight antibacterial substances reported in the literature(citric acid, limonene, malic acid, γ-aminobutyric acid, quercetin, catechin, sodium diacetate, oxalic acid) on 5 strains of P.expansum. Determined the minimum inhibitory concentration of trans-2-hexenal by antibacterial test of 5 strains of P.expansum when trans-2-hexenal at a concentration of 40, 60, 80, 100 μL/L. Then 3 mm of sterile iron nails were used to perforate the equator of kiwifruit and 10 μL 1.0×106 cfu/mL spore suspension of 5 test strains were inoculated which simulating injury of kiwifruit secondary infection P.expansum. The kiwifruit in vivo test was carried out by adding trans-hexenal aldehyde at a concentration of 100 μL/L and determined the kiwifruit hardness, soluble solids, titratable acid, reducing sugar, vitamin C, pH value and fruit decay rate to study the effect of trans-2-hexenal on the quality of kiwifruit and the patulin content in kiwifruit was measured by HPLC. Lastly, the effect of trans-2-hexenal on the morphology of 5 strains of P.expansum was observed by scanning electron microscope. The results shown: Compared with the eight antibacterial substances, trans-2-hexenal had the best antibacterial effect and it could completely inhibit the growth of 5 strains of Penicillium expansum. The minimum inhibitory concentration(MIC) of the five strains was 100, 80, 80, 80 and 80 μL/L, respectively; trans-2-hexenal significantly reduced the effect of P. expansum on kiwi fruit firmness, titratable acid and vitamin C(P<0.01)and had no significant effect(P>0.01) on the fruit firm, soluble solids, pH value and reducing sugar content of kiwifruit. The rot rate of kiwi fruit with P. expansum was reduced and the quality of kiwifruit was better maintained by Trans-2-hexenal. All the aldehyde-treated kiwifruits were not detected by patulin; the scanning electron microscope showed that the five strains showed hyphal shrinkage, folding, drying, and sporulation reduction. Trans-2-hexenal has a good inhibitory effect on the growth of P.expansum and the production of patulin and provides a theoretical basis for the development of natural products with antibacterial effects.
Patulin is a neurotoxic secondary metabolite that causes a series of acute and chronic conditions such as enteritis, dyspnea and cancer when the body consumes excessive amounts. China is the birthplace of kiwifruit and the largest producer of kiwifruit in the world. A large number of studies had shown that kiwifruit is susceptible to infection with penicillium expansum(P.expansum) and patulin accumulation during harvesting, storage and processing which not only causes waste, but also harms human health. Therefore, it is necessary to study a safe and effective method to control P.expansum and patulin accumulation of kiwifruit and its products. Trans-2-hexenal is a characteristic aroma component naturally present in kiwi, apple and other fruits. Studies had shown that trans-2-hexenal has a certain inhibitory effect on microorganisms. This article studied the inhibitory effect of hexenal on the growth of P.expansum and the production of patulin. Firstly, the five strains of P.expansum(CICC40658, CGMCC3.3703, LPH6, LPH5, WY) were inoculated into PDA medium plates containing bacteriostatic substance concentration of 5 mg/mL which kept at 28 ℃and the diameter of the colony was measured by the cross method every day. Study the antibacterial effect of trans-2-hexenal and eight antibacterial substances reported in the literature(citric acid, limonene, malic acid, γ-aminobutyric acid, quercetin, catechin, sodium diacetate, oxalic acid) on 5 strains of P.expansum. Determined the minimum inhibitory concentration of trans-2-hexenal by antibacterial test of 5 strains of P.expansum when trans-2-hexenal at a concentration of 40, 60, 80, 100 μL/L. Then 3 mm of sterile iron nails were used to perforate the equator of kiwifruit and 10 μL 1.0×106 cfu/mL spore suspension of 5 test strains were inoculated which simulating injury of kiwifruit secondary infection P.expansum. The kiwifruit in vivo test was carried out by adding trans-hexenal aldehyde at a concentration of 100 μL/L and determined the kiwifruit hardness, soluble solids, titratable acid, reducing sugar, vitamin C, pH value and fruit decay rate to study the effect of trans-2-hexenal on the quality of kiwifruit and the patulin content in kiwifruit was measured by HPLC. Lastly, the effect of trans-2-hexenal on the morphology of 5 strains of P.expansum was observed by scanning electron microscope. The results shown: Compared with the eight antibacterial substances, trans-2-hexenal had the best antibacterial effect and it could completely inhibit the growth of 5 strains of Penicillium expansum. The minimum inhibitory concentration(MIC) of the five strains was 100, 80, 80, 80 and 80 μL/L, respectively; trans-2-hexenal significantly reduced the effect of P. expansum on kiwi fruit firmness, titratable acid and vitamin C(P<0.01)and had no significant effect(P>0.01) on the fruit firm, soluble solids, pH value and reducing sugar content of kiwifruit. The rot rate of kiwi fruit with P. expansum was reduced and the quality of kiwifruit was better maintained by Trans-2-hexenal. All the aldehyde-treated kiwifruits were not detected by patulin; the scanning electron microscope showed that the five strains showed hyphal shrinkage, folding, drying, and sporulation reduction. Trans-2-hexenal has a good inhibitory effect on the growth of P.expansum and the production of patulin and provides a theoretical basis for the development of natural products with antibacterial effects.
引文
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[2]张承,王秋萍,冉飞,等猕猴桃适宜采摘期确定及其贮藏性能[J].农业工程学报,2018,34(17):266-275.Zhang Cheng,Wang Qiuping,Ran Fei,et al.Determination of suitable harvest period of kiwifruit and its storage performance[J].Transactions of the Chinese Society of Agricultural Engineering(Transactions of the CSAE),2018,34(17):266-275.
[3]薛华丽,毕阳,宗元元,等.果蔬及其制品中真菌毒素的污染与检测研究进展[J].食品科学,2016,37(23):285-290.Xue Huali,Bi Yang,Zong Yuanyuan,et al.Progress in mycotoxins contamination and detection in fruits and vegetables and their processed products[J].Food Science,2016,37(23):285-290.(in Chinese with English abstract)
[4]Zong Yuanyuan,Li Boqiang,Qin Guozheng,et al.Toxicity of patulin on fruit quality and its research progress[J].Journal of Agricultural Science and Technology,2013,15(4):36-41.
[5]张欣怡,王威浩,邓丽莉,等.水果及其制品中展青霉素的研究进展[J].食品工业科技,2017,38(11):379-384.Zhang Xinyi,Wang Weihao,Deng Lili,et al.Research progress in patulin of fruits and its products[J].Food Industry Technology,2017,38(11):379-384.(in Chinese with English abstract)
[6]Cassano A,Donato L,Conidi C,et al.Recovery of bioactive compounds in kiwifruit juice by ultrafiltration[J].Innovative Food Science&Emerging Technologies,2008,9(4):556-562.
[7]张业芳,唐诗,周艳,等.猕猴桃乳酸菌饮料的研制[J].中国酿造,2017(12):186-190.Zhang Yefang,Tang Shi,Zhou Yan,et al.Preparation of kiwifruit lactic aid bacteria begerave[J].China Brewing,2017(12):186-190.(in Chinese with English abstract)
[8]Zhang J B,Gao Z P,Liu X H,et al.The Effect of RFtreatment combined with Nisin against alicyclobacillus,spores in kiwi fruit juice[J].Food&Bioprocess Technology,2016,10(2):1-9.
[9]李可,李华佳,袁怀瑜,等.“金艳”猕猴桃贮期腐烂病病原菌的分离鉴定[J].安徽农业科学,2018,46(1):160-164.Li Ke,Li Huajia,Yuan Huaiyu,et al.Isolation and identification of rot disease pathogens during storage of“Jinyan”kiwifruit[J].Anhui Agric.Sci,2018,46(1):160-164.(in Chinese with English abstract)
[10]Appell M,Jackson M A,Dombrink-Kurtzman M A.Removal of patulin from aqueous solutions by propylthiol functionalized SBA-15[J].Journal of Hazardous Materials,2011,187(1/2/3):150-156.
[11]Dong Q F,Manns D C,Feng G P,et al.Reduction of patulin in apple cider by UV radiation[J].Journal of Food Protection,2010,73(1):69-74.
[12]Zhu Y,Koutchma T,Warriner K,et al.Kinetics of patulin degradation in model solution,apple cider and apple juice by ultraviolet radiation[J].Food science and technology international,2013,19(4):291-303
[13]高振鹏,刘瑞,张德举,等.超声波降解苹果汁中展青霉素动力学研究[J].农业机械学报,2015,46(11):230-235.Gao Zhenpeng,Liu Rui,Zhang Deju,et al.Kinetics study of ultrasonic degradation of patulin in apple juice[J].Transactions of the Chinese Society for Agricultural Machinery,2015,46(11):230-235.(in Chinese with English abstract)
[14]Khorrami A R,Taherkhani M.Synthesis and evaluation of a molecularly imprinted polymer for pre-concentration of patulin from apple juice[J].Chromatographia,2011,73(1):151-156.
[15]Zhang W,Yong H,Chen X,et al.Surface molecularly imprinted polymer capped Mn-doped ZnS quantum dots as a phosphorescent nanosensor for detecting patulin in apple juice[J].Food Chemistry,2017,232:145-154.
[16]潘春青,岳田利,王铁成,等.气体沿面放电低温等离子体对扩展青霉孢子杀灭效果[J].食品科学,2017,38(21):1-7.Pang Chunqing,Yue Tianli,Wang Tiecheng,et al.Killing effect of low-temperature plasma generated by gas phase surface discharge on Penicillium expansum spores[J].Food Science,2017,38(21):1-7.(in Chinese with English abstract)
[17]Calvo J,Calvente V,de Orellano M E,et al.Biological control of postharvest spoilage caused by Penicillium expansum and Botrytis cinerea in apple by using the bacterium Rahnella aquatilis[J].International Journal of Food Microbiology,2007,113(3):251-257.
[18]姜楠,王蒙,韦迪哲,等.果蔬中真菌毒素污染及臭氧防治研究进展[J].食品安全质量检测学报,2016,7(11):4415-4420.Jiang Nan,Wang Meng,Wei Dizhe,et al.Progress on mycotoxins contamination and ozone degradation in fruits and vegetables[J].Journal of Food Safety and Quality,2016,7(11):4415-4420.(in Chinese with English abstract)
[19]师俊玲,张小平,李元瑞,等.果汁护色剂JPX对苹果汁中棒曲霉素的降解作用及护色效果[J].农业工程学报,2006,22(10):237-239.Shi Junling,Zhang Xiaoping,Li Yuanrui,et al.Effects of color protection agent JPX on patulin degradation and color protection of apple juice[J].Transactions of the Chinese Society of Agricultural Engineering(Transactions of CSAE),2006,22(10):237-239.(in Chinese with English abstract)
[20]Cao J,Zhang H,Yang Q,et al.Efficacy of Pichia caribbica in controlling blue mold rot and patulin degradation in apples[J].International Journal of Food Microbiology,2013,162(2):167-173.
[21]Yang Q,Zhang H,Zhang X,et al.Phytic acid enhances biocontrol activity of rhodotorula mucilaginosa against penicillium expansum contamination and patulin production in apples[J].Frontiers in Microbiology,2015,6(8):1-9.
[22]Fuchs S,Sontag G,Stidl R,et al.Detoxification of patulin and ochratoxin A,two abundant mycotoxins,by lactic acid bacteria[J].Food&Chemical Toxicology,2008,46(4):1398-1407.
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