茶树精油对扩展青霉线粒体功能的影响研究

摘要/Abstract

摘要： 为了揭示茶树精油（TTO）对果蔬采后病原真菌扩展青霉（Penicillium expansum）的抑菌作用机制，以TTO处理的P. expansum孢子、菌丝和线粒体为研究对象，测定真菌活性氧（ROS）积累及6种线粒体功能相关酶类的活性变化，并采用扫描电镜、透射电镜观察精油处理对线粒体形态和超微结构的影响。结果表明：TTO处理导致P. expansum线粒体发生严重的皱缩，干瘪，胞内有空泡状结构，线粒体基质流失，严重破坏了P. expansum的线粒体结构。同时TTO处理诱导了P. expansum孢子内ROS的大量积累，导致菌丝细胞内三磷酸腺苷（ATP）含量下降和三磷酸腺苷酶（ATPase）、柠檬酸合酶（CS）、异柠檬酸脱氢酶（ICDH）、α-酮戊二酸脱氢酶（α-KGDH）、苹果酸脱氢酶（MDH）和琥珀酸脱氢酶（SDH）活性下降，破坏了P. expansum的三羧酸循环（TCA）。结果表明TTO处理引起P. expansum孢子内ROS的积累，破坏菌丝体内的线粒体形态和超微结构，TCA循环和能量代谢异常。可见，线粒体功能的严重受损是TTO抑制P. expansum的重要机制。

关键词: 茶树精油, 扩展青霉, 活性氧, 线粒体

Abstract: In order to study the antifungal effect of tea tree oil (TTO) on the Penicillium expansum in postharvest fruits and vegetables, the mycelia and mitochondria of P. expansum were treated with TTO, and the accumulation level of reactive oxygen species (ROS) and the changes in activities of enzymes associated with mitochondrial function were then measured. Scanning electron microscope (SEM) and transmission electron microscope (TEM) were also used to observe the morphology and ultrastructure of mitochondria for evaluating TTO effect on mitochondria. The results showed that TTO treatment caused severe shrinking, drying, and vesicular mitochondria, along with loss of mitochondrial matrix. Meanwhile, TTO treatment induced a large amount of ROS accumulated in P. expansum spores, resulting in decreases in intracellular adenosine triphosphate (ATP) content and activities of several important enzymes, including citrate synthetase (CS), isocitrate dehydrogenase (ICDH), α-ketoglutarate dehydrogenase (α-KGDH), malic dehydrogenase (MDH), and succinodehydrogenase (SDH). Thus, the tricarboxylic acid cycle (TCA) of P. expansum was destroyed. This study showed that TTO treatment affected the structure and function of mitochondria in P. expansum, caused ROS highly accumulated, disrupted the TCA cycle of P. expansum, and eventually hinder the normal metabolism of mitochondria.

Key words: Tea Tree Oil (TTO), Penicillium expansum, reactive oxygen species, Mitochondria

中图分类号:

TS225.1

陈雪昱邹秀容韦莹莹许凤邵兴锋. 茶树精油对扩展青霉线粒体功能的影响研究[J]. 食品科学, 0, (): 0-0.

Xing-feng SHAO. Effect of Tea Tree Oil on the Mitochondrial Function of Penicillium expansum[J]. FOOD SCIENCE, 0, (): 0-0.

参考文献

[1] Bhaskara Reddy M V, Belkacemi K, Corcuff R, et al. Effect of pre-harvest chitosan sprays on post-harvest infection by Botrytis cinerea and quality of strawberry fruit[J]. Postharvest Biology and Technology, 2000, 20(1): 39-51. DOI: 10. 1016/ S0925-5214(00)00108-3
[2] Yu T, Yu C, Lu H, et al. Effect of Cryptococcus laurentii and calcium chloride on control of Penicillium expansum and Botrytis cinerea infections in pear fruit[J]. Biological Control, 2012, 61(2): 169-175. DOI: 10. 1016/ j. biocontrol. 2012. 01. 012
[3] 陈倩茹. 植物精油对樱桃番茄主要病原菌抑制效果的研究[D]. 浙江大学, 2014
[4] 李殿鑫, 江琳琳, 朱娜, 等. 植物精油在食品保鲜中的研究进展[J]. 食品工业科技, 2012, 33(12): 396-400. DOI: CNKI: 11-1759/ TS. 20111230. 1118. 011
[5] Feng W, Zheng X. Essential oils to control Alternaria alternata in vitro and in vivo[J]. Food Control, 2007, 18(9): 1126-1130. DOI: 10. 1016/ j. foodcont. 2006. 05. 017
[6] 曾晓房, 高苏娟, 林衍宗, 等. 肉桂精油对紫金春甜桔贮藏保鲜的影响[J]. 现代食品科技, 2012, 28(10): 1281-1284. DOI: CNKI: SUN: GZSP. 0. 2012-10-004
[7] 王丹, 张静, 贾晓曼, 等. 蓝莓采后主要病原菌的分离鉴定及肉桂精油抑菌效果[J]. 食品科学, 2019, 40(24): 167-172. DOI: 10. 7506/ spkx1002-6630-20181029-338
[8] Swords G, Hunter G L K. Composition of Australian tea tree oil (Melaleuca alternifolia) [J]. Journal of Agricultural and Food Chemistry, 1978, 26(3): 734-737. 8DOI: 10. 1021/ jf60217a031
[9] Carson C F, Hammer K A, Riley T V. Melaleuca alternifolia (Tea Tree) Oil: a review of Antimicrobial and Other Medicinal Properties[J]. Clinical Microbiology Reviews, 2006, 19(1): 50-62. DOI: 10. 1128/ CMR. 19. 1. 50-62. 2006
[10] Shi C, Zhao X, Yan H, et al. Effect of tea tree oil on Staphylococcus aureus growth and enterotoxin production[J]. Food Control, 2016, 62: 257-263. DOI: 10. 1016/ j. foodcont. 2015. 10. 049
[11] 静玮, 苏子鹏, 朱德明, 等. 茶树油熏蒸处理对香蕉采后炭疽病害的影响[J]. 农业工程学报, 2011, 27(05): 378-384. DOI: CNKI: SUN: NYGU. 0. 2011-05-067
[12] 钟业俊, 刘伟, 刘成梅, 等. 自然条件下乳化茶树油在香蕉保鲜中的应用[J]. 农业工程学报, 2009, 25(06): 280-284. DOI: 10. 3969/ j. issn. 1002-6819. 2009. 06. 052
[13] 程赛, 邵兴锋, 郭安南, 等. 茶树油熏蒸对草莓采后病害和品质的影响[J]. 农业工程学报, 2011, 27(04): 383-388. DOI: CNKI: SUN: NYGU. 0. 2011-04-069
[14] Shao X, Cheng S, Wang H, et al. The possible mechanism of antifungal action of tea tree oil on Botrytis cinerea[J]. Journal of Applied Microbiology, 2013, 114(6): 1642-1649. DOI: 10. 1111/ jam. 12193
[15] Li Y, Shao X, Xu J, et al. Tea tree oil exhibits antifungal activity against Botrytis cinerea by affecting mitochondria[J]. Food Chemistry, 2017, 234: 62-67. DOI: 10. 1016/ j. foodchem. 2017. 04. 172
[16] Tian J, Ban X, Zeng H, et al. The mechanism of antifungal ction of essential oil from dill (Anethum graveolens L. ) on Aspergillus flavus[J]. PLoS ONE, 2012, 7(1): e30147. DOI: 10. 1371/ journal. pone. 0030147
[17] Li Y, Shao X, Xu J, et al. Effects and possible mechanism of tea tree oil against Botrytis cinerea and Penicillium expansum in vitro and in vivo test[J]. Canadian Journal of Microbiology, 2017, 63(3): 219-227. DOI: 10. 1139/ cjm-2016-0553
[18] 罗曼, 蒋立科. 柠檬醛损伤黄曲霉线粒体生化机理的研究[J]. 微生物学报, 2002(02): 226-231. DOI: CNKI: SUN: WSXB. 0. 2002-02-014
[19] Yu D, Wang J, Shao X, et al. Antifungal modes of action of tea tree oil and its two characteristic components against Botrytis cinerea[J]. Journal of Applied Microbiology, 2015, 119(5): 1253-1262. DOI: 10. 1111/ jam. 12939
[20] Jiang W, Luo Y, Liu H, et al. A Simple and Rapid Determination of ATP, ADP and AMP Concentrations in Pericarp Tissue of Litchi Fruit by High Performance Liquid Chromatography[J]. Food Technology and Biotechnology, 2006, 44(4): 531-534. DOI: doi: 10. 1016/ j. fm. 2005. 11. 001
[21] OuYang Q, Tao N, Jing G. Transcriptional profiling analysis of Penicillium digitatum, the causal agent of citrus green mold, unravels an inhibited ergosterol biosynthesis pathway in response to citral[J]. BMC Genomics, 2016, 17(1). DOI: 10. 1186/ s12864-016-2943-4
[22] Tao N, Jia L, Zhou H. Anti-fungal activity of Citrus reticulata Blanco essential oil against Penicillium italicum and Penicillium digitatum[J]. Food Chemistry, 2014, 153: 265-271. DOI: 10. 1016/ j. foodchem. 2013. 12. 070
[23] Zheng S, Jing G, Wang X, et al. Citral exerts its antifungal activity against Penicillium digitatum by affecting the mitochondrial morphology and function[J]. Food Chemistry, 2015, 178: 76-81. DOI: 10. 1016/ j. foodchem. 2015. 01. 077
[24] McBride H M, Neuspiel M, Wasiak S. Mitochondria: More Than Just a Powerhouse[J]. Current Biology, 2006, 16(14): R551-R560. DOI: 10. 1016/ j. cub. 2006. 06. 054
[25] Fernie A R, Carrari F, Sweetlove L J. Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport[J]. Current Opinion in Plant Biology, 2004, 7(3): 254-261. DOI: 10. 1016/ j. pbi. 2004. 03. 007
[26] Zou L, Hu Y, Chen W. Antibacterial mechanism and activities of black pepper chloroform extract[J]. Journal of Food Science and Technology, 2015, 52(12): 8196-8203. DOI: 10. 1007/ s13197-015-1914-0
[27] Liao C H, Yao L L, Ye B C. Three genes encoding citrate synthases in Saccharopolyspora erythraea are regulated by the global nutrient‐sensing regulators GlnR, DasR, andCRP[J]. Molecular Microbiology, 2014, 94(5): 1065-1084. DOI: 10. 1111/ mmi. 12818
[28] Garnak M, Reeves H C. Phosphorylation of Isocitrate dehydrogenase of Escherichia coli[J]. Science, 1979, 203(4385): 1111-1112. DOI: 10. 1126/ science. 34215
[29] Huang H, Zhang H, Xu H, et al. Inhibition of the α-ketoglutarate dehydrogenase complex alters mitochondrial function and cellular calcium regulation[J]. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2003, 1637(1): 119-126. DOI: 10. 1016/ S0925-4439(02)00222-3
[30] Yao Y, Li M, Zhai H, et al. Isolation and characterization of an apple cytosolic malate dehydrogenase gene reveal its function in malate synthesis[J]. Journal of Plant Physiology, 2011, 168(5): 474-480. DOI: 10. 1016/ j. jplph. 2010. 08. 008
[31] Kobayashi D, Kondo K, Uehara N, et al. Endogenous Reactive Oxygen Species Is an Important Mediator of Miconazole Antifungal Effect[J]. Antimicrobial Agents and Chemotherapy, 2002, 46(10): 3113-3117. DOI: 10. 1128/ AAC. 46. 10. 3113-3117. 2002
[32] Wu X, Cheng A, Sun L, et al. Plagiochin E, an antifungal bis(bibenzyl), exerts its antifungal activity through mitochondrial dysfunction-induced reactive oxygen species accumulation in Candida albicans[J]. Biochimica et Biophysica Acta (BBA) - General Subjects, 2009, 1790(8): 770-777. DOI: 10. 1016/ j. bbagen. 2009. 05. 002
[33] Yaru Li Y N L Z. The possible mechanism of antifungal activity of cinnamon oil against Rhizopus nigricans [J]. Journal of Chemical and Pharmaceutical Resrarch, 2014, 6(5): 12-20
[34] Yang S, Liu L, Li D, et al. Use of active extracts of poplar buds against Penicillium italicum and possible modes of action[J]. Food Chemistry, 2016, 196: 610-618. DOI: 10. 1016/ j. foodchem. 2015. 09. 101
[35] King A, Selak M A, Gottlieb E. Succinate dehydrogenase and fumarate hydratase: linking mitochondrial dysfunction and cancer[J]. Oncogene, 2006, 25(34): 4675-4682. DOI: 10. 1038/ sj. onc. 1209594
[36] Pozniakovsky A I, Knorre D A, Markova O V, et al. Role of mitochondria in the pheromone- and amiodarone-induced programmed death of yeast[J]. The Journal of Cell Biology, 2005, 168(2): 257-269. DOI: 10. 1083/ jcb. 200408145
[37] Shi X, Li B, Qin G, et al. Mechanism of antifungal action of borate against Colletotrichum gloeosporioides related to mitochondrial degradation in spores[J]. Postharvest Biology and Technology, 2012, 67: 138-143. DOI: 10. 1016/ j. postharvbio. 2012. 01. 003