不同氮源对刺芹侧耳谷氨酰胺合成酶的酶活力及基因表达影响
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摘要
谷氨酰胺合成酶(GS)是真菌氮素代谢的关键酶之一。为探究氮源的类型和浓度对其活性的调控,采用添加单一氮源成分(硫酸铵、硝酸钠、尿素)的基本培养基培养了刺芹侧耳国森一号菌株,并对培养过程中菌丝生物量和谷氨酰胺合成酶的酶活力进行了测定。荧光定量PCR结果显示尿素和NH_4~+均可以抑制GS基因表达水平以及GS酶活力; NO_3~-可以提高GS基因表达水平和GS酶活力,但是刺芹侧耳利用NO_3~-氮源生产菌丝生物量的能力较差。
Glutamine synthetase(GS) is a central enzyme of nitrogen metabolism in fungi. A single nitrogen of minimal medium was designed to investigate the type and concentration of nitrogen source and the regulation of GS activity in culture the mycelia of Pleurotus eryngii. The transcription of PE-GS was measured by Real Time RT-PCR and the data of enzymatic activity of GS and the biomass of Pleurotus eryngii were analyzed. Fluorescence quantitative PCR showed that both urea and NH_4~+ could inhibit GS gene expression level and GS enzyme activity. NO_3~- could improve GS gene expression level and GS enzyme activity, but the ability to produce mycelium biomass with NO_3~- nitrogen source was poor in Pleurotus eryngii.
Glutamine synthetase(GS) is a central enzyme of nitrogen metabolism in fungi. A single nitrogen of minimal medium was designed to investigate the type and concentration of nitrogen source and the regulation of GS activity in culture the mycelia of Pleurotus eryngii. The transcription of PE-GS was measured by Real Time RT-PCR and the data of enzymatic activity of GS and the biomass of Pleurotus eryngii were analyzed. Fluorescence quantitative PCR showed that both urea and NH_4~+ could inhibit GS gene expression level and GS enzyme activity. NO_3~- could improve GS gene expression level and GS enzyme activity, but the ability to produce mycelium biomass with NO_3~- nitrogen source was poor in Pleurotus eryngii.
引文
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[15]PODGóRSKA A, BURIAN M, GIECZEWSKA K, et al. Altered cell wall plasticity can restrict plant growth under ammonium nutrition[J]. Frontiers in Plant Science, 2017, 8:1344.
[2]CHEN H B, CHEN C H, CHEN M J, et al. The use of mushroom hydrolysate from waste bag-log as the nitrogen source to mycelium biomass and exopolysaccharide production in Pleurotus eryngii cultivation[J]. Journal of the Taiwan Institute of Chemical Engineer, 2013, 44(2): 163-168.
[3]LóPEZ P, GONZáLEZ G M, VALDERAS A, et al. GintAMTi encodes a functional high-affinity ammonium transporter that is expressed in the extthe arbuscular mycorrhizal fungus Glomus intraradices (DAOM 197198) reveals functional tradeoffs in an obligate symbiont[J]. New Phytologist, 2012, 193: 755-769.
[4]ZHANG F, LIU Y, WANG L Y, et al. Molecular cloning and expression analysis of ammonium transporters in different tea (Camellia sinensis (L.) O. Kuntze) cultivars under different nitrogen treatments[J]. Gene, 2018, 658:136-145.
[5]SUZUKI A, KNAFF D B. Glutamate synthase: structural, mechanistic and regulatory properties, and role in the amino acid metabolism[J]. Photosynthesis Research, 2005, 83(2): 191-217.
[6]GARCíA-GUTIéRREZ A, CáNOVAS F M, áVILA C. Glutamate synthases from conifers: gene structure and phylogenetic studies[J]. BMC Genomics, 2018,19: 65.
[7]程向阳, 王莹, 鲍大鹏,等. 刺芹侧耳谷氨酰胺合成酶编码基因克隆和表达分析[J]. 菌物学报, 2014, 33(5): 1036-1044.
[8]何贵香, 宋文明, 谢景文,等. 三种食用菌营养成分的含量测定[J].食药用菌, 2013, 21(3): 156-157.
[9]NUGENT K G, CHOFFE K, SAVILLE B J. Gene espression during Ustilago maydis diploid filamentous growth: EST library creation and analyses[J]. Fungal Genetics and Biology, 2004, 41(3): 349-360.
[10]KONG W W, HUANG C Y, CHEN Q, et al. Nitric oxide is involved in the regulation of trehalose accumulation under heat stress in Pleurotus eryngii var. tuoliensis[J]. Biotechnology Letters, 2012, 34(10): 1915-1919.
[11]马璐, 杜双田, 金凌云, 等. 杏鲍菇营养生理研究[J]. 西北农林科技大学学报(自然科学版), 2010, 38(9): 129-134.
[12]王振河, 武忠伟, 王斌, 等. 杏鲍菇菌丝营养生理特性研究[J]. 河南师范大学学报(自然科学版), 2007, 35(4): 139-143.
[13]HUDSON R C, DANIEL R M. L-glutamate dehydrogenases: distribution, properties and mechanism[J]. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1993, 106(4): 767-792.
[14]KERSTEN M A, MüLLER Y, CAMP H O, et al. Molecular characterization of the glnA gene encoding glutamine synthetase from the edible mushroom Agaricus bisporus[J]. Molecular and General Genetics, 1997, 256(2): 179-186.
[15]PODGóRSKA A, BURIAN M, GIECZEWSKA K, et al. Altered cell wall plasticity can restrict plant growth under ammonium nutrition[J]. Frontiers in Plant Science, 2017, 8:1344.