管婉;储婷;鲍大鹏;张建;李福后;唐利华;

• 1:江苏海洋大学食品科学与工程学院
• 2:上海市农业科学院食用菌研究所
• 3:上海海洋大学食品学院

摘要(Abstract)：

为了解活性氧(reactive oxygen species,ROS)在香菇菌丝后熟转色形成中的作用及其自噬细胞学特征,以香菇工厂化菌株KS11为研究材料,分析其在菌丝后熟转色过程中4个时间点(30、45、60、75 d)的活性氧含量(ROS)、丙二醛(MDA)含量、NADPH氧化酶浓度、抗氧化酶活性以及外源活性氧和DPI对其影响的表型试验,利用透射电镜观察该过程菌丝细胞自噬特征变化,并运用实时荧光定量PCR对自噬基因Atg8的表达水平进行比较分析。结果表明:(1) H_2O_2作为主要的活性氧因子在菌丝后熟转色形成中呈现显著动态变化,后熟转色过程中不断升高,并在转色中第60天呈高峰值。(2) NADPH氧化酶浓度与H_2O_2含量变化呈紧密正相关。(3)外源施加一定浓度H_2O_2显著促进香菇菌丝后熟转色,且DPI作为NADPH氧化酶抑制剂显著抑制了香菇菌丝后熟转色的发生。(4)香菇菌丝后熟转色过程中,细胞自噬特征逐渐增强,并在转色中后期最显著。上述结果表明以H_2O_2为主的活性氧在香菇菌丝后熟转色过程中起重要作用,并且可能参与了香菇菌丝后熟转色过程中的自噬进程。

关键词(KeyWords)： 香菇;后熟转色;活性氧;自噬;实时荧光定量PCR

Abstract:

Keywords:

基金项目(Foundation): 上海自然科学基金(19ZR1446500)~~

作者(Authors): 管婉;储婷;鲍大鹏;张建;李福后;唐利华;

DOI: 10.13346/j.mycosystema.210174

参考文献(References)：

• Baba E,Uluk?y G,?nta?C,2015.Effects of feed supplemented with Lentinula edodes mushroom extract on the immune response of rainbow trout,Oncorhynchus mykiss,and disease resistance against Lactococcus garvieae.Aquaculture,448:476-482
• Deng BL,2020.Research progress on the mechanism of reactive oxygen species regulating crop seed germination.Anhui Agricultural Sciences,48(13):21-22(in Chinese)
• Fang LL,Yang L,Li CX,Liu QH,2017.Changes of Atg8expression in Lentinus edodes before and after color change.Chinese Edible Fungi,36(3):44-49 (in Chinese)
• Hao HB,Huang JC,Wang Q,Jun JX,Xiao TT,Song XX,Chen H,Zhang JJ,2021.Effects of heat stress on differential expression of antioxidant enzymes and heat shock protein genes in Agaricus bisporus.Acta Mycologica Sinica,40(3):616-625 (in Chinese)
• Huang JL,Ma HY,Tao F,Zhang Y,2011.Effects of regulatory treatment on characteristic enzymes of three edible fungi.Journal of Food and Biotechnology,30(2):200-206 (in Chinese)
• Jiang S,Yu ZJ,Lan SC,2013.Changes of reactive oxygen species metabolism and transcriptional expression of disease course related protein 1 during the interaction between bryophyte and Botrytis cinerea.Chinese Journal of Aeronautics,37(4):349-354 (in Chinese)
• Karuppanapandian T,Moon JC,Kim C,Manoharan K,Kim W,2011.Reactive oxygen species in plants:their generation,signal transduction,and scavenging mechanisms.Australian Journal of Crop Science,5(6):709-725
• Kaur H,Nyochembeng LM,Mentreddy SR,Banerjee P,Cebret E,2016.Assessment of the antimicrobial activity of Lentinula edodes against Xanthomonas campestris pv.vesicatoria.Crop Protection,89:284-288
• Li QZ,Jiang N,Li ZP,Song CY,Liu JY,Dong HR,Jiang J,Shang XD,2021.Establishment of evaluation system for nutritional quality of Lentinus edodes.Acta Nucifera Sinica,35(4):881-890 (in Chinese)
• Li XD,Lu JP,Li HJ,Lin FC,2011.Apoptosis of filamentous fungi.Microbiology Bulletin,38(2):242-249 (in Chinese)
• Lisanti MP,Martinez-Outschoorn UE,Chiavarina B,Pavlides S,Whitaker-Menezes D,Tsirigos A,Witkiewicz AK,Lin Z,Balliet R,Howell A,Sotgia F,2010.Understanding the‘lethal’drivers of tumor-stroma co-evolution:emerging role(s) for hypoxia,oxidative stress and autophagy/mitophagy in the tumor micro-environment.Cancer Biology&Therapy,10(6):537-542
• Norbert G,Krisztina K,2014.Interconnections of reactive oxygen species homeostasis and circadian rhythm in Neurospora crassa.Antioxidants&Redox Signaling,20(18):3007-3023
• Pang J,Sun GQ,Yu J,Wang HY,Kang LY,Xie YJ,Bao YY,2016.Research progress on the mechanism and influencing factors of color transformation of Lentinus edodes rod.Guizhou Agricultural Sciences,44(3):99-101 (in Chinese)
• Qi YC,Duan QH,Shen XY,Jue C,Shen JW,2012.Effects of high temperature stress on physiological and biochemical characteristics of Pleurotus ostreatus.Acta Edible Fungi Sinica,19(4):14-16 (in Chinese)
• Ren ZZ,Liu WB,Song XL,Qi YR,Zhang C,Gao Z,Zhang JJ,Jia L,2018.Antioxidant and anti-inflammation of enzymatic-hydrolysis residue polysaccharides by Lentinula edodes.International Journal of Biological Macromolecules,120(8):811-822
• Smirnoff N,Arnaud D,2019.Hydrogen peroxide metabolism and functions in plants.New Phytologist,221(3):1197-1214
• Song Y,Liu N,Zhang JJ,Zhang M,2020.Growth characteristics of Shenxiang series of new Lentinus edodes in Liaoning Province.Edible Fungi of China,39(11):22-25 (in Chinese)
• Tang DL,Kang R,Livesey KM,Cheh CW,Farkas A,Loughran P,Hoppe G,Bianchi ME,Tracey KJ,Zeh HJ,Lotze MT,2010.Endogenous HMGB1 regulates autophagy.The Journal of Cell Biology,190(5):881
• Tang LH,Jian HH,Song CY,Bao DP,Shang XD,Wu DQ,Tan Q,Zhang XH,2013.Transcriptome analysis of candidate genes and signaling pathways associated with light-induced brown film formation in Lentinula edodes.Applied Microbiology and Biotechnology,97(11):4977-4989
• Tang LH,Tan Q,Bao DP,Zhang XD,Jia HH,Li Y,Yang RH,Wang Y,2016.Comparative proteomic analysis of light-induced mycelial brown film formation in Lentinula edodes.Bio Med Research International,2016:1-8
• Wang SH,Shih YL,Kuo TC,Ko WC,Shih CM,2009.Cadmium toxicity toward autophagy through ROS-activated GSK-3βin mesangial cells.Toxicological Sciences,108(1):124-131
• Wang Y,Huang CS,Jia NQ,2020.Organic small molecule fluorescent probes for monitoring cell microenvironment and active molecules.Chemical Progress,32(Z1):204-218 (in Chinese)
• Wu F,Zhou LW,Yang ZL,Bau T,Li TH,Dai YC,2019.Resource diversity of Chinese macrofungi:edible,medicinal and poisonous species.Fungal Diversity,98:1-76
• Yan JJ,Tong ZY,Liu YY,Liu ZY,Long Y,Han X,Xu WN,Huang QH,Tao YX,Xie BG,2020.The NADPH oxidase in Volvariella volvacea and its differential expression in response to mycelial ageing and mechanical injury.Brazilian Journal of Microbiology,51(10):87-94
• Yan SY,Jiang XJ,2015.Overview of autophagy and related research in fungi.Mycosystema,34(5):871-879 (in Chinese)
• Yang CH,Ma SX,Bai QY,2010.Color changing of Lentinus edodes rod is the key to produce good mushroom.Chinese Edible Fungi,29(6):63-64 (in Chinese)
• Yao YC,Wei PP,Li M,Zhang KC,Fan GZ,2020.Effects of hydrogen sulfide on different forms of calcium and superoxide anion in Phellinus igniarius mycelium.Chinese Edible Fungi,39(12):106-111 (in Chinese)
• You SJ,Shi JJ,Zhang YL,Liu CF,2011.ROS mediated autophagy and its role in related diseases.Chinese Journal of Pathophysiology,27(1):187-190 (in Chinese)
• Yu HL,Lv BB,Song CY,Shang XD,Zhang LJ,Zhang MY,Tan Q,2016.Effect of compound nitrogen sources on growth and yield of Lentinus edodes.Shanghai Journal of Agriculture,32(3):63-66 (in Chinese)
• Zhou SC,Zou G,Yang ZS,Bao DP,Yao WW,Yang J,Wang Y,Li XL,2019.Physiological and biochemical characteristics of degenerated strains of Cordyceps militaris.Acta Edulis Fungi,26(4):23-28 (in Chinese)
• Zhu J,Wu F,Yue S,Chen C,Song SQ,Wang H,Zhao MW,2019.Functions of reactive oxygen species in apoptosis and ganoderic acid biosynthesis in Ganoderma lucidum.FEMS Microbiology Letters,366:23
• Zhu W,Gong FP,Duan QH,Shang Guan DL,Zhang ZY,Yin CC,Chen J,Zhang YX,2020.Technical regulation for
• autumn cultivation of substitute Lentinus edodes in
• Henan Province.Edible Fungi of China,39(2):19-21 (in
• Chinese)
• 邓本良，2020.活性氧调控作物种子发芽机理的研究进展.安徽农业科学，48(13):21-22
• 房丽丽，杨丽，李翠新，刘庆洪，2017.香菇转色前后Atg8表达的变化规律研究.中国食用菌，36(3):44-49
• 郝海波，黄建春，王倩，隽加香，肖婷婷，宋晓霞，陈辉，张津京，2021.热胁迫对双孢蘑菇抗氧化酶及热激蛋白基因的差异表达的影响.菌物学报，40(3):616-625
• 黄俊丽，马海燕，陶菲，张慜，2011.调控处理对3种食用菌特性酶的影响.食品与生物技术学报，30(2):200-206
• 姜山，余治锦，兰世超，2013.小立碗藓-灰霉菌互作过程中活性氧代谢及病程相关蛋白1转录表达的变化.江西师范大学学报，37(4):349-354
• 李巧珍，姜宁，李正鹏，宋春艳，刘建雨，董浩然，蒋俊，尚晓冬，2021.香菇营养品质评价体系的构建.核农学报，35(4):881-890
• 厉晓东，卢建平，李海娇，林福呈，2011.丝状真菌的细胞凋亡.微生物学通报，38(2):242-249
• 庞杰，孙国琴，于静，王海燕，康立茹，解亚杰，包妍妍，2016.香菇菌棒转色机理及影响因素的研究进展.贵州农业科学，44(3):99-101
• 戚元成，段庆虎，申晓晔，决超，申进文，2012.高温胁迫对糙皮侧耳菌丝生理生化特性的影响.食用菌学报，19(4):14-16
• 宋莹，刘娜，张季军，张敏，2020.申香系列香菇新品种在辽宁地区的生长特性.中国食用菌，39(11):22-25
• 王阳，黄楚森，贾能勤，2020.监测细胞微环境及活性分子的有机小分子荧光探针.化学进展，32(Z1):204-218
• 闫思源，姜学军，2015.细胞自噬及真菌中自噬研究概述.菌物学报，34(5):871-879
• 杨春华，马树新，白琼岩，2010.香菇菌棒转色是出好菇的关键环节.中国食用菌，29(6):63-64
• 姚雨辰，魏盼盼，李茂，张恪诚，范桂枝，2020.硫化氢对桑黄菌丝中不同形态钙含量和超氧阴离子产生的影响.中国食用菌，39(12):106-111
• 尤寿江，石际俊，张艳林，刘春风，2011.ROS介导的自噬及其在相关疾病中的作用.中国病理生理杂志，27(1):187-190
• 于海龙，吕贝贝，宋春艳，2016.复合氮源对香菇生长及产量的影响分析.上海农业学报，32(3):63-66
• 周思池，邹根，杨占山，鲍大鹏，姚威威，杨杰，汪滢，李晓玲，2019.蛹虫草退化菌株的生理生化特征.食用菌学报，26(4):23-28
• 竹玮，龚凤萍，段庆虎，上官端琳，张宗源，尹川川，陈静，张应香，2020.河南省代料香菇秋栽生产技术规程.中国食用菌，39(2):19-21