低温对蒙古口蘑菌丝体漆酶活性及其基因家族表达的影响
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摘要
蒙古口蘑是内蒙古草原久负盛名的野生食用菌,为实现其人工栽培,本研究以蒙古口蘑次生菌丝体为实验材料,利用实时荧光定量PCR技术,研究了低温对蒙古口蘑漆酶活性及其基因家族转录水平的影响。结果表明:蒙古口蘑次生菌丝体漆酶活性普遍较高;随着培养时间的延长,漆酶活性普遍降低;低温刺激可不同程度地影响蒙古口蘑漆酶基因家族的转录,10℃处理菌丝体10天,可显著提高蒙古口蘑漆酶基因的表达水平,主要贡献是lacc1、lacc4、lacc8、lacc10、lacc11。结果可为日后实现蒙古口蘑人工栽培积累资料。
Mongolian Mushroom is a well-known wild edible fungus in Inner Mongolia grasslands, in order to achieve its artificial cultivation, in this study, we use secondary mycelia of Mongolian mushroom as experimental material, and real-time fluorescence quantitative PCR technique, to analyze the effects of low temperature on laccase activity and its gene family transcription level of Mongolian mushroom. The results showed that the laccase activity in secondary mycelia of Mongolian mushroom was generally high. With the prolongation of culture time, laccase activity was generally decreased. Low temperature stimulation affected the transcription of laccase gene family of Mongolian mushroom in varying degrees. Mycelia were treated for 10 days at 10℃, it could improve the expression levels of laccase genes of Mongolian mushroom significantly, the main contribution was lacc1, lacc4, lacc8, lacc10, lacc11. The results could accumulate data to realize artificial cultivation of Mongolian mushroom in the future.
Mongolian Mushroom is a well-known wild edible fungus in Inner Mongolia grasslands, in order to achieve its artificial cultivation, in this study, we use secondary mycelia of Mongolian mushroom as experimental material, and real-time fluorescence quantitative PCR technique, to analyze the effects of low temperature on laccase activity and its gene family transcription level of Mongolian mushroom. The results showed that the laccase activity in secondary mycelia of Mongolian mushroom was generally high. With the prolongation of culture time, laccase activity was generally decreased. Low temperature stimulation affected the transcription of laccase gene family of Mongolian mushroom in varying degrees. Mycelia were treated for 10 days at 10℃, it could improve the expression levels of laccase genes of Mongolian mushroom significantly, the main contribution was lacc1, lacc4, lacc8, lacc10, lacc11. The results could accumulate data to realize artificial cultivation of Mongolian mushroom in the future.
引文
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[17] Zhuo R, Yuan P, Yang Y, et al. Induction of laccase by metal ions and aromatic compounds in Pleurotus ostreatus HAUCC 162 and decolorization of different synthetic dyes by the extracellular laccase[J]. Biochemical Engineering Journal,2017,117:62-72.
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[22] Saparrat M, Balatti P A, Martínez M J, et al. Differential regulation of laccase gene expression in Coriolopsis rigida LPSC No. 232[J].Fungal biology, 2010, 114(11):999-1006.
[23] Islamovic E, García-Pedrajas M D, Chacko N, et al. Transcriptome analysis of a Ustilago maydis ust1 deletion mutant uncovers involvement of laccase and polyketide synthase genes in sporedevelopment[J]. Molecular Plant-Microbe Interactions, 2015,28(1):42-54.
[24]吴领弟,阿拉腾图娅.锡林郭勒盟气候变化研究综述[J].农业与技术, 2016(22):222-226.
[25]李丹.内蒙古锡林郭勒盟近55年地温变化情况研究[J].江西农业,2016(19):42-43.
[26] Llanos A, Fran?ois J M, Parrou J L. Tracking the best reference genes for RT-qPCR data normalization in filamentous fungi[J].BMC genomics, 2015, 16(1):71.
[27] Fan X Z, Zhou Y, Xiao Y, et al. Cloning, expression and phylogenetic analysis of a divergent laccase multigene family in Auricularia auricula-judae[J]. Microbiological research, 2014,169(5):453-462.
[28]刘晓婷,郭九峰,王淑妍,等.蒙古口蘑担孢子萌发及初生菌丝生物学特性研究[J].北方园艺,2016(16):136-141.
[29]税丕容,郑晓冰,林俊芳,等.简便高质量的食用菌总RNA提取方法[J].食用菌学报, 2008(01):32-36.
[30]黄留玉.PCR最新技术原理、方法及应用(第二版)[M].北京:化学工业出版社,2010.
[31]张国良,白羽嘉,张乐,等.低温处理对阿魏侧耳漆酶、酪氨酸酶及同工酶的影响[J].食品科技,2013(07):15-19.
[32]茅文俊,李燕,周陈力,等.草菇漆酶基因在子实体形成过程中的表达分析[J].食用菌学报, 2016(01):1-6.
[33]吴光美,严俊杰,杨志云,等.与金针菇子实体发育有关的漆酶编码基因Fv-lac4的结构及表达差异分析[J].基因组学与应用生物学,2014(06):1254-1260.
[2]董冬,图力古尔.蒙古口蘑分类地位研究[J].菌物研究, 2013(3):172-175.
[3] Wang J, Zhao Y, Li W, et al. Optimization of polysaccharides extraction from Tricholoma mongolicum Imai and their antioxidant and antiproliferative activities[J]. Carbohydrate polymers, 2015,131:322-330.
[4] EI Enshasy H A, Hatti-Kaul R. Mushroom immunomodulators:unique molecules with unlimited applications[J]. Trends inbiotechnology, 2013,31(12):668-677.
[5]刘家扬,焦国宝,有小娟,等.真菌漆酶的性质、生产及应用研究进展[J].生物技术通报,2016(04):24-33.
[6] Li M, Zhang G, Wang H, et al. Purification and characterization of a laccase from the edible wild mushroom Tricholoma mongolicum.[J]. Journal of Microbiology&Biotechnology, 2010, 20(7):1069-1076.
[7]韦丽敏,史兆国,张力,等. 6种白腐真菌的产酶能力及对稻草的降解效果[J].江苏农业科学, 2013(03):319-322.
[8] Eriksson K E L, Blanchette R, Ander P. Microbial and enzymatic degradation of wood and wood components[M]. Springer Science&Business Media, 2012.
[9]杨秀清,文潇玮.白腐真菌Trametes sp. SQ01漆酶的新功能:转化2-羟基-6氧-6-苯基-2,4-己二烯酸[J].微生物学报,2014(08):913-918.
[10] Kuhar F, Papinutti L. Optimization of laccase production by two strains of Ganoderma lucidum using phenolic and metallic inducers[J]. Revista Argentina de Microbiología, 2014, 46(2):144-149.
[11]北本丰,辻山彰一.从生物化学的角度看担子菌子实体形成[J].食用菌学报,2015(04):1-12.
[12]茅文俊,李燕,周陈力,等.草菇漆酶基因在子实体形成过程中的表达分析[J].食用菌学报, 2016(01):1-6.
[13] Wood D A. Production, purification and properties of extracellular laccase of Agaricus bisporus[J]. Microbiology, 1980,117(2):327-338.
[14]茅文俊,李燕,周陈力,等.草菇漆酶基因在子实体形成过程中的表达分析[J].食用菌学报,2016(01):1-6.
[15]吴光美,严俊杰,杨志云,等.与金针菇子实体发育有关的漆酶编码基因Fv-lac4的结构及表达差异分析[J].基因组学与应用生物学,2014(06):1254-1260.
[16]卓睿.白腐真菌漆酶及同工酶基因家族的克隆表达调控研究[D].武汉:华中科技大学,2015.
[17] Zhuo R, Yuan P, Yang Y, et al. Induction of laccase by metal ions and aromatic compounds in Pleurotus ostreatus HAUCC 162 and decolorization of different synthetic dyes by the extracellular laccase[J]. Biochemical Engineering Journal,2017,117:62-72.
[18] Pezzella C, Lettera V, Piscitelli A, et al. Transcriptional analysis of Pleurotus ostreatus laccase genes[J]. Applied microbiology and biotechnology, 2013, 97(2):705-717.
[19] Yang Y, Wei F, Zhuo R, et al. Enhancing the laccase production and laccase gene expression in the white-rot fungus Trametes velutina 5930 with great potential for biotechnological applications bydifferent metal ions and aromatic compounds[J]. PloS one,2013,8(11):e79307.
[20] Periasamy R, Palvannan T. Optimization of laccase production by Pleurotus ostreatus IMI 395545 using the Taguchi DOE methodology[J]. Journal of basic microbiology, 2010,50:548-556.
[21] Arora D S, Rampal P. Laccase production by some Phlebia species[J]. Journal of basic microbiology, 2002,42(5):295-301.
[22] Saparrat M, Balatti P A, Martínez M J, et al. Differential regulation of laccase gene expression in Coriolopsis rigida LPSC No. 232[J].Fungal biology, 2010, 114(11):999-1006.
[23] Islamovic E, García-Pedrajas M D, Chacko N, et al. Transcriptome analysis of a Ustilago maydis ust1 deletion mutant uncovers involvement of laccase and polyketide synthase genes in sporedevelopment[J]. Molecular Plant-Microbe Interactions, 2015,28(1):42-54.
[24]吴领弟,阿拉腾图娅.锡林郭勒盟气候变化研究综述[J].农业与技术, 2016(22):222-226.
[25]李丹.内蒙古锡林郭勒盟近55年地温变化情况研究[J].江西农业,2016(19):42-43.
[26] Llanos A, Fran?ois J M, Parrou J L. Tracking the best reference genes for RT-qPCR data normalization in filamentous fungi[J].BMC genomics, 2015, 16(1):71.
[27] Fan X Z, Zhou Y, Xiao Y, et al. Cloning, expression and phylogenetic analysis of a divergent laccase multigene family in Auricularia auricula-judae[J]. Microbiological research, 2014,169(5):453-462.
[28]刘晓婷,郭九峰,王淑妍,等.蒙古口蘑担孢子萌发及初生菌丝生物学特性研究[J].北方园艺,2016(16):136-141.
[29]税丕容,郑晓冰,林俊芳,等.简便高质量的食用菌总RNA提取方法[J].食用菌学报, 2008(01):32-36.
[30]黄留玉.PCR最新技术原理、方法及应用(第二版)[M].北京:化学工业出版社,2010.
[31]张国良,白羽嘉,张乐,等.低温处理对阿魏侧耳漆酶、酪氨酸酶及同工酶的影响[J].食品科技,2013(07):15-19.
[32]茅文俊,李燕,周陈力,等.草菇漆酶基因在子实体形成过程中的表达分析[J].食用菌学报, 2016(01):1-6.
[33]吴光美,严俊杰,杨志云,等.与金针菇子实体发育有关的漆酶编码基因Fv-lac4的结构及表达差异分析[J].基因组学与应用生物学,2014(06):1254-1260.