干旱胁迫对猪苓菌丝生长及多糖合成相关酶活性的影响
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摘要
为探讨干旱胁迫对猪苓(Polyporus umbellatus)菌丝生长与多糖物质积累的影响,采用不同质量浓度(0、50、100、150、200、250 g/L)PEG-6000处理,分析干旱胁迫下猪苓菌丝生长相关指标及多糖合成相关酶活性的变化。结果表明,100 g/LPEG-6000处理下,猪苓菌丝生物量、多糖合成相关酶活性及多糖质量分数最高,其中猪苓菌丝生物量、己糖激酶(HK)活性、磷酸葡萄糖异构酶(GPI)活性、α-磷酸葡萄糖变位酶(α-PGM)活性及多糖质量分数分别较对照组增加60.40%、27.88%、19.89%、28.73%和106.00%。当PEG-6000质量浓度>150 g/L时,猪苓菌丝生物量、过氧化物酶(POD)活性、超氧化物歧化酶(SOD)活性、丙二醛(MDA)质量摩尔浓度、可溶性蛋白质量分数及多糖质量分数均随胁迫程度增大显著降低。因此,适度干旱胁迫有利于猪苓菌丝生长和多糖合成。
To explore the effect of the drought stress on the growth and the accumulation of polysaccharides of Polyporus umbellatus mycelium,with different mass concentrations of PEG-6000(0 g/L, 50 g/L, 100 g/L, 150 g/L, 200 g/L and 250 g/L),the strain of P.umbellatus was treated by PEG-6000 to simulate drought stress. Through liquid culture, the growth index and enzyme activities related to polysaccharide synthesis of mycelium were evaluated. The results indicated that the biomass, enzyme activities and polysaccharide mass fraction of P.umbellatus were the highest in the concentration of PEG-6000(100 g/L). Meanwhile, the biomass, HK activity, GPI activity, α-PGM activity and polysaccharide mass fraction significantly increased by 60.40%, 27.88%, 19.89%, 28.73%, 106.00%, respectively, compared to the CK group. When the mass concentration of PEG-6000 was exceeded 150 g/L, the biomass, POD activity, SOD activity, MDA molality, soluble proteins mass fraction and polysaccharide mass fraction of mycelium significantly decreased with the increase of drought stress. Therefore, moderate drought stress are conducive to growth and polysaccharide synthesis of P.umbellatus.
To explore the effect of the drought stress on the growth and the accumulation of polysaccharides of Polyporus umbellatus mycelium,with different mass concentrations of PEG-6000(0 g/L, 50 g/L, 100 g/L, 150 g/L, 200 g/L and 250 g/L),the strain of P.umbellatus was treated by PEG-6000 to simulate drought stress. Through liquid culture, the growth index and enzyme activities related to polysaccharide synthesis of mycelium were evaluated. The results indicated that the biomass, enzyme activities and polysaccharide mass fraction of P.umbellatus were the highest in the concentration of PEG-6000(100 g/L). Meanwhile, the biomass, HK activity, GPI activity, α-PGM activity and polysaccharide mass fraction significantly increased by 60.40%, 27.88%, 19.89%, 28.73%, 106.00%, respectively, compared to the CK group. When the mass concentration of PEG-6000 was exceeded 150 g/L, the biomass, POD activity, SOD activity, MDA molality, soluble proteins mass fraction and polysaccharide mass fraction of mycelium significantly decreased with the increase of drought stress. Therefore, moderate drought stress are conducive to growth and polysaccharide synthesis of P.umbellatus.
引文
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[18] 余吴梦晓,张薇薇,陈向东,等.蓝光对灵芝子实体生长和糖代谢相关酶的影响[J].时珍国医国药,2017,28(11):2741-2744.YU-WU M X,ZHANG W W,CHEN X D,et al.Effects of blue light on fruiting body growth and glycometabolism-related enzymes of Ganoderma lucidum[J].Lishizhen Medicine and Materia Medica Research,2017,28(11):2741-2744.
[19] 李洁.3种单糖碳源对灵芝多糖合成影响的研究[D].江苏无锡:江南大学,2015.LI J.The effect of three monosaccharide carbon sources on polysaccharides synthesis of Ganoderma lucidum[D].Wuxi Jiangsu:Jiangnan University,2015.
[20] 刘高强,赵艳,王晓玲,等.灵芝多糖的生物合成和发酵调控[J].菌物学报,2011,30(2):198-205.LIU G Q,ZHAO Y,WANG X L,et al.Biosynthesis and fermentation control of polysaccharides from Ganoderma lucidum[J].Mycosystema,2011,30(2):198-205.
[21] 程建徽,谢鸣,蒋桂华,等.植物己糖激酶的信号转导作用[J].中国细胞生物学学报,2004,26(6):594-598.CHENG J H,XIE M,JIANG G H,et al.The signaling role of hexokinase in plants[J].Chinese Journal of Cell Biology,2004,26(6):594-598.
[22] 谢鸣,陈俊伟,秦巧平,等.转化酶和己糖激酶调控草莓聚合果内糖积累[J].植物生理与分子生物学学报,2007,33(3):213-218.XIE M,CHEN J W,QIN Q P,et al.The control of sugar accumulation within strawberry aggregate fruit by invertase and hexokinase[J].Journal of Plant Physiology and Molecular Biology,2007,33(3):213-218.
[23] 张奇茹,丁重阳,周哲敏,等.葡萄糖-1-磷酸对灵芝多糖合成的影响[J].生物加工过程,2017,15(4):9-14.ZHANG Q R,DING CH Y,ZHOU ZH M,et al.Effects of glucose-1-phosphate on the biosynthesis of Ganoderma lucidum polysaccharide[J].Chinese Journal of Bioprocess Engineering,2017,15(4):9-14.
[24] NEUHAUS H E,EMES M J.Nonphotosynthetic metabolism in plastids[J].Annual Review of Plant Physiology and Plant Molecular Biology,2000,51(1):111-140.
[25] SEIFERT G J.Nucleotide sugar interconversions and cell wall biosynthesis:how to bring the inside to the outside[J].Current Opinion in Plant Biology,2004,7(3):277-284.
[26] WANG Y,ZHANG Y H.A highly active phosphoglucomutase from clostridium thermocellum:cloning,purification,characterization and enhanced thermostability[J].Journal of Applied Microbiology,2010,108(1):39-46.
[27] HANSEN T,SCHLICHTING B,FELGENDREHERM,et al.Cupin-type phosphoglucoseisomerases (cupin-PGIs) constitute a novel metal-dependent PGI family representing a convergent line of PGI evolution[J].Journal of Bacteriology,2005,187(5):1621.
[28] KUNZ H H,ZAMANI-NOUR S,H?USLER R E,et al.Loss of cytosolic phosphoglucoseisomerase affects carbohydrate metabolism in leaves and is essential for fertility of Arabidopsis[J].Plant Physiology,2014,166(2):753-765.
[2] 顾雅君,王瑛,李宾.猪苓的化学成分和药用配方[J].食药用菌,2013,21(5):275-276.GU Y J,WANG Y,LI B.Chemical composition and medicinal formula of Polyporus umbellatus[J].Edible and Medicinal Mushrooms,2013,21(5):275-276.
[3] 江泽波,赵晋,李思明,等.猪苓多糖诱导M2亚型巨噬细胞向M1巨噬细胞转化[J].中国免疫学杂志,2015,31(8):1049-1052.JIANG Z B,ZHAO J,LI S M,et al.Transformation of PPS induce M2 subtype to M1 macrophages[J].Chinese Journal of Immunology,2015,31(8):1049-1052.
[4] 谭庆龙,周昌园,刘春萍,等.猪苓多糖对人巨噬细胞形态及免疫功能的影响[J].中华中医药杂志,2018,33(5):1891-1896.TAN Q L ,ZHOU CH Y,LIU CH P,et al.Investigation on the effects of polyporus polysaccharides on morphological changes and immune function in human macrophages[J].China Journal of Traditional Chinese Medicine and Pharmacy,2018,33(5):1891-1896.
[5] 李心群,许文.猪苓多糖通过Toll样受体4对小鼠骨髓来源树突状细胞作用研究[J].中草药,2011,42(1):118-123.LI X Q,XU W.Effect of Polyporus umbellatus polysaccharides on activation of murine bone marrow dendritic cells via Toll-like receptor 4[J].Chinese Traditional and Herbal Drugs,2011,42(1):118-123.
[6] 杨冬野,焦洋,杜志坚,等.猪苓多糖注射液联合顺铂腹腔给药治疗胃癌腹腔积液患者的疗效观察[J].河北医药,2018,40(5):725-727.YANG D Y,JIAO Y,DU ZH J,et al.Therapeutic effects of polyporus polysaccharide injection combined with cisplatinintraperitoneal injection on seroperitoneum due to gastric cancer[J].Hebei Medical Journal,2018,40(5):725-727.
[7] 任拾璞,黄洁,王霞.猪苓多糖联合白细胞介素15增强人外周血单个核细胞对K562细胞杀伤活性的影响[J].临床荟萃,2016,31(12):1352-1356.REN SH P,HUANG J,WANG X.Study of PPS combined with interleukin-15 enhanced killing activity of peripheral blood mononuclear cells to K562 cells[J].Clinical Focus,2016,31(12):1352-1356.
[8] 沈耿,徐谦,黄羽,等.猪苓多糖通过调节HuR介导 bcl-2表达诱导T24细胞凋亡[J].中药新药与临床药理,2016,27(2):160-164.SHEN G,XU Q,HUANG Y,et al.Effect of polyporus polysaccharide on inducing apoptosis of T24 cells by regulating HuR- mediated bcl-2 expression[J].Traditional Chinese Drug Research and Clinical Pharmacology,2016,27(2):160-164.
[9] 许广波,傅伟杰,赵旭奎.我国猪苓研究的进展[J].菌物研究,2003,1(1):58-61.XU G B,FU W J,ZHAO X K.Advances in studies on Polyporus umbellatus in China[J].Journal of Fungal Research,2003,1(1):58-61.
[10] 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000.LI H SH.Expermental Principle and Technology of Plant Biology and Biochemistry[M].Beijing:Higher Education Press,2000.
[11] SCHAFFER A A,PETREIKOV M.Sucrose-to-starch metabolism in tomato fruit undergoing transient starch accumulation[J].Plant Physiology,1997,113(3):739-746.
[12] LOOIJESTEIJN P J,BOELS I C,KLEEREBEZEMM,et al.Regulation of exopolysaccharide production by Lactococcuslactis subsp.cremoris by the sugar source[J].Appl Environ Microbiol,1999,65(11):5003-5008.
[13] 高巧妮,谢淑雨,张跃进,等.猪苓袋料培养条件筛选[J].北方园艺,2017(5):130-133.GAO Q N,XIE SH Y,ZHANG Y J,et al.Screening for bagging culture conditions of polyporus[J].Northern Horticulture,2017(5):130-133.
[14] 刘艳,陈贵林,蔡贵芳,等.干旱胁迫对甘草幼苗生长和渗透调节物质含量的影响[J].西北植物学报,2011,31(11):2259-2264.LIU Y,CHEN G L,CAI G F,et al.Growth and osmoregulation substances accumulation of Glycyrrhiza uralensis seedling under drought stress[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(11):2259-2264.
[15] 高扬.不同土壤水分对丹参耗水特性及有效成分含量的影响[D].陕西杨凌:西北农林科技大学,2004.GAO Y.Effects of different soil moisture on water consumption characteristics and content of active components of Salvia miltiorrhiza Bunge[D].Yangling Shaanxi:Northwest A&F University,2004.
[16] 黄璐琦,郭兰萍.环境胁迫下次生代谢产物的积累及道地药材的形成[J].中国中药杂志,2007,32(4):277-280.HUANG L Q,GUO L P.Secondary metabolites accumulating and geoherbs formation under enviromental stress[J].China Journal of Chinese Materia Medica,2007,32(4):277-280.
[17] 周雪洁,王改利,梁宗锁,等.干旱胁迫及复水对甘草幼苗生理特性和甘草酸积累的影响[J].西北农业学报,2011,20(7):64-70.ZHOU X J,WANG G L,LIANG Z S,et al.Effects of drought stress and rehydration on physiological characteristics and glycyrrhizin accumulation of Glycyrrhiza uralensis Fisch.seedling[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(7):64-70.
[18] 余吴梦晓,张薇薇,陈向东,等.蓝光对灵芝子实体生长和糖代谢相关酶的影响[J].时珍国医国药,2017,28(11):2741-2744.YU-WU M X,ZHANG W W,CHEN X D,et al.Effects of blue light on fruiting body growth and glycometabolism-related enzymes of Ganoderma lucidum[J].Lishizhen Medicine and Materia Medica Research,2017,28(11):2741-2744.
[19] 李洁.3种单糖碳源对灵芝多糖合成影响的研究[D].江苏无锡:江南大学,2015.LI J.The effect of three monosaccharide carbon sources on polysaccharides synthesis of Ganoderma lucidum[D].Wuxi Jiangsu:Jiangnan University,2015.
[20] 刘高强,赵艳,王晓玲,等.灵芝多糖的生物合成和发酵调控[J].菌物学报,2011,30(2):198-205.LIU G Q,ZHAO Y,WANG X L,et al.Biosynthesis and fermentation control of polysaccharides from Ganoderma lucidum[J].Mycosystema,2011,30(2):198-205.
[21] 程建徽,谢鸣,蒋桂华,等.植物己糖激酶的信号转导作用[J].中国细胞生物学学报,2004,26(6):594-598.CHENG J H,XIE M,JIANG G H,et al.The signaling role of hexokinase in plants[J].Chinese Journal of Cell Biology,2004,26(6):594-598.
[22] 谢鸣,陈俊伟,秦巧平,等.转化酶和己糖激酶调控草莓聚合果内糖积累[J].植物生理与分子生物学学报,2007,33(3):213-218.XIE M,CHEN J W,QIN Q P,et al.The control of sugar accumulation within strawberry aggregate fruit by invertase and hexokinase[J].Journal of Plant Physiology and Molecular Biology,2007,33(3):213-218.
[23] 张奇茹,丁重阳,周哲敏,等.葡萄糖-1-磷酸对灵芝多糖合成的影响[J].生物加工过程,2017,15(4):9-14.ZHANG Q R,DING CH Y,ZHOU ZH M,et al.Effects of glucose-1-phosphate on the biosynthesis of Ganoderma lucidum polysaccharide[J].Chinese Journal of Bioprocess Engineering,2017,15(4):9-14.
[24] NEUHAUS H E,EMES M J.Nonphotosynthetic metabolism in plastids[J].Annual Review of Plant Physiology and Plant Molecular Biology,2000,51(1):111-140.
[25] SEIFERT G J.Nucleotide sugar interconversions and cell wall biosynthesis:how to bring the inside to the outside[J].Current Opinion in Plant Biology,2004,7(3):277-284.
[26] WANG Y,ZHANG Y H.A highly active phosphoglucomutase from clostridium thermocellum:cloning,purification,characterization and enhanced thermostability[J].Journal of Applied Microbiology,2010,108(1):39-46.
[27] HANSEN T,SCHLICHTING B,FELGENDREHERM,et al.Cupin-type phosphoglucoseisomerases (cupin-PGIs) constitute a novel metal-dependent PGI family representing a convergent line of PGI evolution[J].Journal of Bacteriology,2005,187(5):1621.
[28] KUNZ H H,ZAMANI-NOUR S,H?USLER R E,et al.Loss of cytosolic phosphoglucoseisomerase affects carbohydrate metabolism in leaves and is essential for fertility of Arabidopsis[J].Plant Physiology,2014,166(2):753-765.