真菌激发子对桑黄胞内代谢产物积累的影响
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摘要
利用前期通过大量预试验筛选获得的3种真菌多糖激发子,在桑黄Sanghuangporus sanghuang发酵液中进行添加,研究其对桑黄胞内黄酮、三萜及多糖等代谢产物积累的影响,并对激发效果好的激发子继续进行添加量和激发时间的优化。结果显示:参试的3种真菌多糖激发子在一定程度上均能促进桑黄胞内黄酮和胞内三萜类成分的积累,但对胞内多糖积累的影响不明显;来源菌株为薄多年卧孔菌Perenniporia tenuis的激发子表现最突出,激发效果显著且稳定性良好。激发条件优化的结果表明:在接种的同时按照50mg/L的终浓度进行激发子的添加,激发6d,桑黄胞内黄酮含量达最高值0.7%,比对照提高79.49%;在接种1d后,按照50mg/L的终浓度进行激发子的添加,激发5d,对桑黄胞内三萜类成分代谢的激发效果最显著,胞内三萜含量最高可达5.62%,是对照组的3.43倍。
The effects of fungal elicitor on the accumulation of Sanghuangporus sanghuang intracellular metabolites,such as flavonoids,triterpenoid and polysaccharide,were investigated.Three fungal polysaccharide elicitors were obtained through previous preliminary experiments.The optimal additive amount and stimulation time were optimized by using the best elicitor.The results indicated that the activated effects of the three fungal elicitors on the accumulation of S.sanghuang intracellular flavonoids and triterpenoid were notable,but the effects on the accumulation of intracellular polysaccharide were indistinctive.The elicitor obtained from Perenniporia tenuis mycelia in submerged fermentation owned good excitation effect and stability.After excitation condition optimization the intracellular flavonoid content was 79.49%higher as compared with the control,reaching 0.7%in the sixth day of cultivation under final concentration of 50mg/L and six-day elicitor stimulation.The highest intracellular triterpenoid content was up to 5.62%and became 3.43 times of the control under the final concentration of50mg/L and five-day elicitor stimulation.
The effects of fungal elicitor on the accumulation of Sanghuangporus sanghuang intracellular metabolites,such as flavonoids,triterpenoid and polysaccharide,were investigated.Three fungal polysaccharide elicitors were obtained through previous preliminary experiments.The optimal additive amount and stimulation time were optimized by using the best elicitor.The results indicated that the activated effects of the three fungal elicitors on the accumulation of S.sanghuang intracellular flavonoids and triterpenoid were notable,but the effects on the accumulation of intracellular polysaccharide were indistinctive.The elicitor obtained from Perenniporia tenuis mycelia in submerged fermentation owned good excitation effect and stability.After excitation condition optimization the intracellular flavonoid content was 79.49%higher as compared with the control,reaching 0.7%in the sixth day of cultivation under final concentration of 50mg/L and six-day elicitor stimulation.The highest intracellular triterpenoid content was up to 5.62%and became 3.43 times of the control under the final concentration of50mg/L and five-day elicitor stimulation.
引文
Bahabadi SE,Sharifi M,Behmanesh M,Safaie N,Murata J,Araki R,Yamagaki T,Satake H,2012.Time-course changes in fungal elicitor-induced lignan synthesis and expression of the relevant genes in cell cultures of Linum album.Journal of Plant Physiology,169(5):487-491
Bu L,Zhu ZY,Liang ZQ,Liu AY,2002.Utilization of fungal elicitor in increasing cordycepin of Cordyceps militaris.Mycosystema,21(2):252-256(in Chinese)
Cao CL,Han ML,Cui BK,Yu HY,2013.Antioxidant activities of different fractional extracts from fruiting bodies of three Phellinus species.Mycosystema,32(5):883-890(in Chinese)
Dai YC,2003.The new knowledge on the medicinal fungus-Phellinus baumii.Chinese Traditional and Herbal Drugs,34(1):94(in Chinese)
Dai YC,2010.Hymenochaetaceae(Basidiomycota)in China.Fungal Diversity,45(1):131-343
Dai YC,Cui BK,2014.Progress on the species of medicinal fungus Inonotus sanghuang.Journal of Beijing Forestry University,36(5):1-6(in Chinese)
Dai YC,Li Y,2011.Notes on the nomenclature of six important medicinal fungi in China.Mycosystema,30(4):515-518(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27(6):801-824(in Chinese)
Dai YC,Yang ZL,Cui BK,Yu CJ,Zhou LW,2009.Species diversity and utilization of medicinal mushrooms and fungi in China(review).International Journal of Medicinal Mushrooms,11(3):287-302
Dai YC,Zhou LW,Cui BK,Chen YQ,Decock C,2010.Current advances in Phellinus sensu lato:medicinal species,functions,metabolites and mechanisms.Applied Microbiology and Biotechnology,87(5):1587-1593
Du W,Liang ZQ,Zou X,Han YF,Liang JD,Yu JP,Chen WH,Wang YR,Sun CL,2013.Effects of microbial elicitor on production of hypocrellin by Shiraia bambusicola.Folia Microbiologica,58(4):283-289
Gao XX,Yao Q,Wang L,Liu LD,Wang WW,Han K,2009.Effect of different fungal elicitors on production of polysaccharide and triterpenoid in liquid fermentation of Ganoderma lucidum.Food Science,30(23):309-313(in Chinese)
Li XY,Zhen RY,Zhang RQ,Sheng LM,2009.Effects of fungal elicitors on synthesis of flavonoids in germinating tartary buckwheat.Journal of Tianjin Agricultural University,16(4):34-37(in Chinese)
Liu CJ,Zhang XY,Dai YC,Wei YL,2005.Artifical culture of Phellinus baumii.Mycosystema,24(2):306-309(in Chinese)
Ma XN,2012.Studies on the screening of fungal polysaccharide elicitor and improving the accumulation of bioactive substance from Cordyceps militaris.Master Thesis,Jiangsu University of Science and Technology,Zhenjiang.32-43(in Chinese)
Prasad A,Mathur A,Kalra A,Gupta MM,Lal RK,Mathur AK,2013.Fungal elicitor-mediated enhancement in growth and asiaticoside content of Centella asiatica L.shoot cultures.Plant Growth Regulation,69(3):265-273
Sun J,Chen QJ,Zhu MJ,Wang HX,Zhang GQ,2014.An extracellular laccase with antiproliferative activity from the sanghuang mushroom Inonotus baumii.Journal of Molecular Catalysis B:Enzymatic,99:20-25
Tian XM,2014.Taxonomy and phylogeny of Inonotus linteus complex and liquid fermentation of I.sanghuang.Ph DDissertation,Beijing Forestry University,Beijing.65-67(in Chinese)
Tian XM,Dai YC,Song AR,Xu K,Ng LT,2015.Optimization of liquid fermentation medium for production of Inonotus sanghuang(Higher Basidiomycetes)mycelia and evaluation of their mycochemical contents and antioxidant activities.International Journal of Medicinal Mushrooms,17(7):681-691
Tian XM,Yu HY,Zhou LW,Decock C,Vlasák J,Dai YC,2013.Phylogeny and taxonomy of the Inonotus linteus complex.Fungal Diversity,58(1):159-169
Wu SH,Dai YC,Hattori T,Yu TW,Wang DM,Parmasto E,Chang HY,Shin SY,2012.Species clarification for the medicinally valuable‘Sanghuang’mushroom.Botanical Studies,53(1):135-149
Yang J,Wang N,Yuan HS,Hu JC,Dai YC,2013.A new sesquiterpene from the medicinal fungus Inonotus vaninii.Chemistry of Natural Compounds,49(2):261-263
Yang SH,Liu XF,Guo DA,Zheng JH,2006.Effects of different additives on accumulation of flavonoids in Glycyrrhiza uralensis callus.Chinese Pharmaceutical Journal,41(2):96-99(in Chinese)
Yu HY,Cao CL,Cui BK,2012.Polyphenol content and antioxidant activities of Phellinus lonicericola in liquid culture.Mycosystema,31(6):933-939(in Chinese)
Yu HY,Chen JJ,Tian XM,Cui BK,Liu HG,Dai YC,2014.Effects of light and p H on the liquid culture of Inonotus lonicericola.Mycosystema,33(2):469-476(in Chinese)
Zhang GL,2008.Study on breeding new Taiwanofungus formosanus strains by the way of physical mutagenesis.Master Thesis,Qingdao Agricultural University,Qingdao.47-48(in Chinese)
Zhang WJ,1999.Technology of glycoconjugate biochemical research.2nd edition.Zhejiang University Press,Hangzhou.1-744(in Chinese)
Zhang ZF,Lv GY,Song TT,Jin QL,Huang JB,Fan LF,Cai WM,2015.Comparison of the preliminary characterizations and antioxidant properties of polysaccharides obtained from Phellinus baumii growth on different culture substrates.Carbohydrate Polymers,132:397-399
Zheng WF,Miao KJ,Zhang YX,Pan SY,Zhang MM,Jiang H,2009.Nitric oxide mediates the fungal-elicitor-enhanced biosynthesis of antioxidant polyphenols in submerged cultures of Inonotus obliquus.Microbiology,155(10):3440-3448
Zhou LW,Vlasák J,Decock C,Assefa A,Stenlid J,Abate D,Wu SH,Dai YC,2016.Global diversity and taxonomy of theInonotus linteus complex(Hymenochaetales,Basidiomycota):Sanghuangporus gen.nov.,Tropicoporusexcentrodendri and T.guanacastensis gen.et spp.nov.,and 17 new combinations.Fungal Diversity,77(1):335-347
步岚,朱振元,梁宗琦,刘爱英,2002.真菌激发子对提高蛹虫草虫草菌素的作用.菌物系统,21(2):252-256
曹春蕾,韩美玲,崔宝凯,余海尤,2013.三种木层孔菌子实体不同溶剂提取物抗氧化活性的研究.菌物学报,32(5):883-890
戴玉成,2003.药用担子菌-鲍氏层孔菌(桑黄)的新认识.中草药,34(1):94
戴玉成,崔宝凯,2014.药用真菌桑黄种类研究.北京林业大学学报,36(5):1-6
戴玉成,李玉,2011.中国六种重要药用真菌名称的说明.菌物学报,30(4):515-518
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27(6):801-824
高兴喜,姚强,王磊,刘林德,王雯雯,韩凯,2009.真菌激发子对灵芝液体发酵生产多糖和三萜类物质的影响.食品科学,30(23):309-313
李晓雁,甄润英,张瑞清,盛丽梅,2009.真菌诱导子对苦荞萌发过程中黄酮合成的影响.天津农学院学报,16(4):34-37
刘春静,张晓阳,戴玉成,魏玉莲,2005.鲍氏层孔菌培养条件的研究.菌物学报,24(2):306-309
马晓宁,2012.真菌多糖激发子的筛选及其对蛹虫草活性物质激发作用的研究.江苏科技大学硕士论文,镇江.32-43
田雪梅,2014.桑黄类群真菌系统学及桑黄纤孔菌液体发酵研究.北京林业大学博士论文,北京.65-67
杨世海,刘晓峰,果德安,郑俊华,2006.不同附加物对甘草愈伤组织培养中黄酮类化合物形成的影响.中国药学杂志,41(2):96-99
余海尤,曹春蕾,崔宝凯,2012.忍冬木层孔菌液体培养过程中多酚含量及抗氧化活性研究.菌物学报,31(6):933-939
余海尤,陈佳佳,田雪梅,崔宝凯,刘鸿高,戴玉成,2014.光照和p H对忍冬纤孔菌液体培养的影响.菌物学报,33(2):469-476
张国利,2008.樟芝物理诱变选育新菌株的研究.青岛农业大学硕士论文,青岛.47-48
张惟杰,1999.糖复合物生化研究技术.第2版.杭州:浙江大学出版社.1-744
Bu L,Zhu ZY,Liang ZQ,Liu AY,2002.Utilization of fungal elicitor in increasing cordycepin of Cordyceps militaris.Mycosystema,21(2):252-256(in Chinese)
Cao CL,Han ML,Cui BK,Yu HY,2013.Antioxidant activities of different fractional extracts from fruiting bodies of three Phellinus species.Mycosystema,32(5):883-890(in Chinese)
Dai YC,2003.The new knowledge on the medicinal fungus-Phellinus baumii.Chinese Traditional and Herbal Drugs,34(1):94(in Chinese)
Dai YC,2010.Hymenochaetaceae(Basidiomycota)in China.Fungal Diversity,45(1):131-343
Dai YC,Cui BK,2014.Progress on the species of medicinal fungus Inonotus sanghuang.Journal of Beijing Forestry University,36(5):1-6(in Chinese)
Dai YC,Li Y,2011.Notes on the nomenclature of six important medicinal fungi in China.Mycosystema,30(4):515-518(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27(6):801-824(in Chinese)
Dai YC,Yang ZL,Cui BK,Yu CJ,Zhou LW,2009.Species diversity and utilization of medicinal mushrooms and fungi in China(review).International Journal of Medicinal Mushrooms,11(3):287-302
Dai YC,Zhou LW,Cui BK,Chen YQ,Decock C,2010.Current advances in Phellinus sensu lato:medicinal species,functions,metabolites and mechanisms.Applied Microbiology and Biotechnology,87(5):1587-1593
Du W,Liang ZQ,Zou X,Han YF,Liang JD,Yu JP,Chen WH,Wang YR,Sun CL,2013.Effects of microbial elicitor on production of hypocrellin by Shiraia bambusicola.Folia Microbiologica,58(4):283-289
Gao XX,Yao Q,Wang L,Liu LD,Wang WW,Han K,2009.Effect of different fungal elicitors on production of polysaccharide and triterpenoid in liquid fermentation of Ganoderma lucidum.Food Science,30(23):309-313(in Chinese)
Li XY,Zhen RY,Zhang RQ,Sheng LM,2009.Effects of fungal elicitors on synthesis of flavonoids in germinating tartary buckwheat.Journal of Tianjin Agricultural University,16(4):34-37(in Chinese)
Liu CJ,Zhang XY,Dai YC,Wei YL,2005.Artifical culture of Phellinus baumii.Mycosystema,24(2):306-309(in Chinese)
Ma XN,2012.Studies on the screening of fungal polysaccharide elicitor and improving the accumulation of bioactive substance from Cordyceps militaris.Master Thesis,Jiangsu University of Science and Technology,Zhenjiang.32-43(in Chinese)
Prasad A,Mathur A,Kalra A,Gupta MM,Lal RK,Mathur AK,2013.Fungal elicitor-mediated enhancement in growth and asiaticoside content of Centella asiatica L.shoot cultures.Plant Growth Regulation,69(3):265-273
Sun J,Chen QJ,Zhu MJ,Wang HX,Zhang GQ,2014.An extracellular laccase with antiproliferative activity from the sanghuang mushroom Inonotus baumii.Journal of Molecular Catalysis B:Enzymatic,99:20-25
Tian XM,2014.Taxonomy and phylogeny of Inonotus linteus complex and liquid fermentation of I.sanghuang.Ph DDissertation,Beijing Forestry University,Beijing.65-67(in Chinese)
Tian XM,Dai YC,Song AR,Xu K,Ng LT,2015.Optimization of liquid fermentation medium for production of Inonotus sanghuang(Higher Basidiomycetes)mycelia and evaluation of their mycochemical contents and antioxidant activities.International Journal of Medicinal Mushrooms,17(7):681-691
Tian XM,Yu HY,Zhou LW,Decock C,Vlasák J,Dai YC,2013.Phylogeny and taxonomy of the Inonotus linteus complex.Fungal Diversity,58(1):159-169
Wu SH,Dai YC,Hattori T,Yu TW,Wang DM,Parmasto E,Chang HY,Shin SY,2012.Species clarification for the medicinally valuable‘Sanghuang’mushroom.Botanical Studies,53(1):135-149
Yang J,Wang N,Yuan HS,Hu JC,Dai YC,2013.A new sesquiterpene from the medicinal fungus Inonotus vaninii.Chemistry of Natural Compounds,49(2):261-263
Yang SH,Liu XF,Guo DA,Zheng JH,2006.Effects of different additives on accumulation of flavonoids in Glycyrrhiza uralensis callus.Chinese Pharmaceutical Journal,41(2):96-99(in Chinese)
Yu HY,Cao CL,Cui BK,2012.Polyphenol content and antioxidant activities of Phellinus lonicericola in liquid culture.Mycosystema,31(6):933-939(in Chinese)
Yu HY,Chen JJ,Tian XM,Cui BK,Liu HG,Dai YC,2014.Effects of light and p H on the liquid culture of Inonotus lonicericola.Mycosystema,33(2):469-476(in Chinese)
Zhang GL,2008.Study on breeding new Taiwanofungus formosanus strains by the way of physical mutagenesis.Master Thesis,Qingdao Agricultural University,Qingdao.47-48(in Chinese)
Zhang WJ,1999.Technology of glycoconjugate biochemical research.2nd edition.Zhejiang University Press,Hangzhou.1-744(in Chinese)
Zhang ZF,Lv GY,Song TT,Jin QL,Huang JB,Fan LF,Cai WM,2015.Comparison of the preliminary characterizations and antioxidant properties of polysaccharides obtained from Phellinus baumii growth on different culture substrates.Carbohydrate Polymers,132:397-399
Zheng WF,Miao KJ,Zhang YX,Pan SY,Zhang MM,Jiang H,2009.Nitric oxide mediates the fungal-elicitor-enhanced biosynthesis of antioxidant polyphenols in submerged cultures of Inonotus obliquus.Microbiology,155(10):3440-3448
Zhou LW,Vlasák J,Decock C,Assefa A,Stenlid J,Abate D,Wu SH,Dai YC,2016.Global diversity and taxonomy of theInonotus linteus complex(Hymenochaetales,Basidiomycota):Sanghuangporus gen.nov.,Tropicoporusexcentrodendri and T.guanacastensis gen.et spp.nov.,and 17 new combinations.Fungal Diversity,77(1):335-347
步岚,朱振元,梁宗琦,刘爱英,2002.真菌激发子对提高蛹虫草虫草菌素的作用.菌物系统,21(2):252-256
曹春蕾,韩美玲,崔宝凯,余海尤,2013.三种木层孔菌子实体不同溶剂提取物抗氧化活性的研究.菌物学报,32(5):883-890
戴玉成,2003.药用担子菌-鲍氏层孔菌(桑黄)的新认识.中草药,34(1):94
戴玉成,崔宝凯,2014.药用真菌桑黄种类研究.北京林业大学学报,36(5):1-6
戴玉成,李玉,2011.中国六种重要药用真菌名称的说明.菌物学报,30(4):515-518
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27(6):801-824
高兴喜,姚强,王磊,刘林德,王雯雯,韩凯,2009.真菌激发子对灵芝液体发酵生产多糖和三萜类物质的影响.食品科学,30(23):309-313
李晓雁,甄润英,张瑞清,盛丽梅,2009.真菌诱导子对苦荞萌发过程中黄酮合成的影响.天津农学院学报,16(4):34-37
刘春静,张晓阳,戴玉成,魏玉莲,2005.鲍氏层孔菌培养条件的研究.菌物学报,24(2):306-309
马晓宁,2012.真菌多糖激发子的筛选及其对蛹虫草活性物质激发作用的研究.江苏科技大学硕士论文,镇江.32-43
田雪梅,2014.桑黄类群真菌系统学及桑黄纤孔菌液体发酵研究.北京林业大学博士论文,北京.65-67
杨世海,刘晓峰,果德安,郑俊华,2006.不同附加物对甘草愈伤组织培养中黄酮类化合物形成的影响.中国药学杂志,41(2):96-99
余海尤,曹春蕾,崔宝凯,2012.忍冬木层孔菌液体培养过程中多酚含量及抗氧化活性研究.菌物学报,31(6):933-939
余海尤,陈佳佳,田雪梅,崔宝凯,刘鸿高,戴玉成,2014.光照和p H对忍冬纤孔菌液体培养的影响.菌物学报,33(2):469-476
张国利,2008.樟芝物理诱变选育新菌株的研究.青岛农业大学硕士论文,青岛.47-48
张惟杰,1999.糖复合物生化研究技术.第2版.杭州:浙江大学出版社.1-744