为了明确一氧化碳(CO)对桑黄菌丝的生长和三萜的累积作用。将CO供体氯高铁血红素添加到桑黄菌丝体系中,采用pH计、电导率仪、比色法、高效液相色谱等分析了pH值、电导率、过氧化氢(H2O2)、超氧化物阴离子、抗氧化物酶活性和三萜含量。研究发现,CO处理对桑黄菌丝培养液pH值的影响差异不显著。电导率、H2O2和超氧阴离子含量除30μmol/LCO处理增加外,其他CO浓度处理1~8d内基本呈降低趋势,与之相反,抗氧化物酶活性除30μmol/LCO处理降低外,其他CO浓度处理基本呈增加趋势。除30 μmol/LCO处理下三萜和白桦脂醇含量降低外,CO增加了三萜和白桦脂醇含量,其中10 μmol/LCO处理4d时三萜含量增幅最大,比同期对照增加了 29.63%；20 μmol/LCO处理1d时白桦脂醇含量增幅达最高值,比同期对照增加了30.96%。由此可见,低浓度CO处理对桑黄菌丝未造成胁迫伤害,其生物累积量未受到抑制,但其这种生物效应还需进一步多材料验证。
Abstract
The study aims to explore the effect of carbon monoxide (CO) on the growth of Phellinus linteus mycelium and the accumulation of triterpene. As the donor of carbon monoxide, hemin was added in the Phellinus mycelium culture, then, different methods such as pH instrument, TDS, colorimetry, HPLC were used to analyze the pH value, conductivity, H2O2, O2- , antioxidant enzyme activity and content of triterpene. The results showed that: the change of pH value was not obvious under the influence of CO; only the treatment with 30 μmol/L hemin for 1-8 days had the best inducing effect on the conductivity and accumulation of H2O2, HPLC, O2-, which declined in other concentrations; conversely, antioxidant enzyme activities were reduced in the 30 μmol/L CO treatment, and were increased in other concentrations; triterpene and betulin were decreased only in 30 μmol/L CO treatment, and were increased in other concentrations; the triterpene had the largest growth rate on the fourth day with 10 μmol/L CO, increased by 29.63% compared with CK in the same period; and the betulin had the largest growth rate on the first day with 20 μmol/L CO, increased by 30.96% compared
with CK in the same period. Obviously, low concentration of CO treatment has no damage to Phellinus mycelium, and improves its bio- accumulation, but the biological effect still needs to be verified by many materials.
关键词
一氧化碳;桑黄;三萜;白桦脂醇
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Key words
Carbon monoxide ; P.linteus ; triterpene ; betulin
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参考文献
[1]张维博, 王家国, 李正阔, 等. 药用真菌桑黄的研究进展[J]. 中国中药杂志, 2014, 39(15): 2838-2845.
[2]张林芳,孙婷婷,邹莉. 鲍姆纤孔菌总三萜的提取及其体外抗乳腺癌细胞MCF-7活性[J]. 药物评价研究,2015,38(05):497-502.
[3]孙佳琦,张甘霖,于明薇,等.桑黄成分及其抗肿瘤作用机制研究进展[J]. 中国医药导报,2016,13(33):39-42+47.
[4]戴玉成. 药用担子菌——鲍氏层孔菌(桑黄)的新认识[J]. 中草药,2003,(01):97-98.
[5]黄丽洋,石卉,王晓婷,等. 野生桑黄菌株的分离、鉴定和次生代谢物分析[J]. 中草药,2013,44(23):3394-3399.
[6]史帧婷,包海鹰. 桑黄类真菌有效成分及功效研究进展[J]. 中国实验方剂学杂志,2016,22(22):197-202.
[7]高雯雯,张娜,俞淑文.桑黄抗肿瘤及其作用机制的研究进展[J]. 中国中药杂志,2014,39(21):4165-4168.
[8]Livak K J, Schmittgen T D. Analysis of relative gene expression date using real-time quantitative PCR and the 2-ΔΔCT method[J].method,2001,25(4):402-408
[9]陈明,沈文飚,阮海华,等. 一氧化氮对盐胁迫下小麦幼苗根生长和氧化损伤的影响[J]. 植物生理与分子生物学学报,2004,(05):569-576.
[10]胡淑明,杜强,李青云,等. 外源腐胺和精胺对NaCl胁迫下草莓叶片膜脂过氧化作用的影响[J]. 安徽农业科学,2012,40(04):1989-1991.
[11]作物生理研究法[M]. 农业出版社，1992：208-212.
[12]]翟俏丽. 真菌诱导子促进白桦悬浮细胞中三萜合成机理的初步研究[D].东北林业大学,2011.
[13]Shrishailappa B,Gapta M K,salavanan R. Detection of betulin in Grewia tiliaefolia by HPLC[J]. J Sep Sei,2004,27:197-198
[14]张泽,孙宏. 高效液相色谱法测定白桦树皮中白桦酯醇的含量[J]. 林产化学与工业,2004,(01):61-63.
[15]黄雅婷,詹亚光,马明媚,范桂枝.SNP与K+、Ca2+互作促进白桦悬浮细胞中白桦酯醇累积的初步研究[J]. 林业科学研究,2016,29(06):896-901.
[16]刘瀚旻,谢亮.一氧化碳在心血管系统中的调控作用[J]. 生理科学进展,2017,48(01):52-57.
[17]崔为体,刘锐涛,林玉婷,等.一氧化碳:植物气体信号分子的新成员[J]. 南京农业大学学报,2012,35(05):87-92.
[18]孙菲菲,李春雪,范桂枝. 外源CO对筋骨草β-蜕皮甾酮积累的影响[J]. 草业科学,2015,
[19]孙菲菲.CO和H2S调控碳、硫代谢促进三萜积累的初步研究[D]. 东北林业大学,2016.32(09):1438-1443.
[20]施江. 外源茉莉酸甲酯诱导对茶树鲜叶次生代谢产物的影响[D]. 中国农业科学院,2014，41-42.
[21]陶金华,汪冬庚,濮雪莲,等. 一氧化氮和水杨酸依次介导内生真菌诱导子促进苍术细胞中苍术素生物合成的信号转导[J]. 中草药,2014,45(05):701-708.
[22]王晓东,李晓灿,翟俏丽,等. 真菌诱导子对白桦悬浮体系中N和P的吸收利用和三萜合成的影响[J]. 中草药,2011,42(10):2119-2124.
[23]丁国昌,林思祖,曹光球,等.环境胁迫下邻羟基苯甲酸对杉木幼苗电导率的影响[J]. 福建林学院学报,2008,(04):343-346.
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