宿主植物对内生真菌产胞外多糖的影响
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摘要
药用植物内生真菌是生物活性物质的新型微生物资源。通过添加宿主植物及优化发酵培养的碳源和氮源,研究宿主植物及发酵条件对绞股蓝内生真菌JY25产胞外多糖的影响。首先优化发酵培养基的氮源和碳源种类及最佳添加量,其次添加不同粒度、不同灭菌方式的宿主植物,检测其对内生真菌JY25产胞外多糖的影响。结果显示:以3 g/L酵母粉为最佳氮源和40 g/L蔗糖为最佳碳源进行发酵,胞外多糖产量达到1.8g/L,相比优化前提升了98%;添加经高压灭菌、粒径约为1.7 mm(12目)、浓度20 g/L的宿主植物绞股蓝,胞外多糖产量达到2.13 g/L,而添加同样粒径、紫外灭菌处理的绞股蓝植物,胞外多糖产量达到2.56 g/L,比优化前提高了180%。研究结果不仅说明适宜的碳源、氮源对植物内生真菌产胞外多糖有明显的促进作用,且宿主植物体内的微生态对内生真菌JY25产胞外多糖也有互作效应。
Endophytic fungus of medicinal plants are new microbial resources of bioactive substances.In this experiment, the carbon and nitrogen sources of fermentation were optimized. And the effect of host plants on the fermentation of endophytic fungus was studied by adding G. pentaphyllum. First, the nitrogen and carbon sources of fermentation medium were optimized and the optimal amount of carbon and nitrogen source was used. Secondly, the effects of different granularity and different sterilizing methods on the production of extracellular polysaccharides by endophytic fungus were observed. The results showed that 3 g/L yeast powder and 40 g/L sucrose were the best carbon and nitrogen source,the output of extracellular polysaccharides reached 1.8 g/L, which was 98% higher than that before optimization. After high pressure sterilization with a diameter of 1.7 mm(12 mesh) with the concentration of 20 g/L, the extracellular polysaccharides production reached 2.13 g/L, while the same diameter of G. pentaphyllum was treated by ultraviolet sterilization. The yield of extracellular polysaccharides reached 2.56 g/L, which was 180% higher than that before optimization. This study showed that the suitable carbon and nitrogen sources could promote the high yield extracellular polysaccharides of plant endophytic fungus, and the microecology of host plants had interaction effect on high yield extracellular polysaccharides.
Endophytic fungus of medicinal plants are new microbial resources of bioactive substances.In this experiment, the carbon and nitrogen sources of fermentation were optimized. And the effect of host plants on the fermentation of endophytic fungus was studied by adding G. pentaphyllum. First, the nitrogen and carbon sources of fermentation medium were optimized and the optimal amount of carbon and nitrogen source was used. Secondly, the effects of different granularity and different sterilizing methods on the production of extracellular polysaccharides by endophytic fungus were observed. The results showed that 3 g/L yeast powder and 40 g/L sucrose were the best carbon and nitrogen source,the output of extracellular polysaccharides reached 1.8 g/L, which was 98% higher than that before optimization. After high pressure sterilization with a diameter of 1.7 mm(12 mesh) with the concentration of 20 g/L, the extracellular polysaccharides production reached 2.13 g/L, while the same diameter of G. pentaphyllum was treated by ultraviolet sterilization. The yield of extracellular polysaccharides reached 2.56 g/L, which was 180% higher than that before optimization. This study showed that the suitable carbon and nitrogen sources could promote the high yield extracellular polysaccharides of plant endophytic fungus, and the microecology of host plants had interaction effect on high yield extracellular polysaccharides.
引文
[1]LI ShouJie,ZHANG Xuan,WANG XiangHua,et al.Novel natural compounds from endophytic fungi with anticancer activity[J].European Journal of Medicinal Chemistry:316-343.
[2]CHRISTINA A,CHRISTAPHER V,BHORE S J.Endophytic bacteria as a source of novel antibiotics:An overview[J].Pharmacognosy Reviews,2013,13(7):11-16.
[3]DING ChangHong,WANG Qian Bo,GUO ShengLei,et al.The improvement of bioactive secondary metabolites accumulation in Rumex gmelini,Turcz through coculture with endophytic fungi[J].Brazilian journal of microbiology,2017,44(1):95-101.
[4]GOSS M J,CARVALHO M,BRITO I,et al.Impacts on Host Plants of Interactions Between AMF and Other Soil Organisms in the Rhizosphere[J].Functional Diversity of Mycorrhiza&Sustainable Agriculture,2017:81-109.
[5]LIU ShengRong,KE BinRong,ZHANG WeiRui,et al.Breeding of new Ganoderma lucidum strains simultaneously rich in polysaccharides and triterpenes by mating basidiospore-derived monokaryons of two commercial cultivars[J].Scientia Horticulturae,2017,216:58-65.
[6]JI XiaoLong,PENG Qiang,YUAN YuePeng,et al.Isolation,structures and bioactivities of the polysaccharides from jujube fruit:A review[J].Food Chemistry,2017,227:349-357.
[7]李传民.绞股蓝内生真菌高产胞外多糖发酵条件的优化及结构分析[D].郑州:河南工业大学,2016.
[8]LI XiaoLi,WANG ZhengHui,ZHAO YongXi,et al.Purification of a polysaccharide from Gynostemma pentaphyllum Makino and its therapeutic advantages for psoriasis[J].Carbohydrate Polymers,2012,89(4):1232-1237.
[9]CHEN Tong,LI Bo,Li YuanYuan,et al.Catalytic synthesis and antitumor activities of sulfated polysaccharide from Gynostemma pentaphyllum Makino[J].Carbohydrate Polymers,2011,83(2):554-560.
[10]程莹,赵骏.桑叶多糖含量测定与成分分析[J].中国临床药理学杂志,2017,33(18):1803-1805,1809.
[11]李良,金文娟.枸杞多糖含量测定方法的比较分析[J].食品研究与开发,2016,37(11):143-146.
[12]JOHN,ROJAN P,NAMPOOTHIRI M,et al.Solidstate fermentation for L-lactic acid production from agro wastes using Lactobacillus delbrueckii[J].Process Bioche mistry,2006,41(4):759-763.
[13]ROJAN P J,NAMPOOTHIRI K M,NAIR A S,et al.L(+)-lactic acid production using Lactobacillus casei in solid[J].Biotechnology Letters,2005,27(21):1685-1688.
[14]武模戈.碳氮源对裂褶菌胞外多糖产量的影响初探[J].濮阳职业技术学院学报,2008,(04):4-5.
[15]MANGANYI M C,REGNIER T,KUMAR A,et al.Phylogenetic analysis and diversity of novel endophytic fungi isolated from medicinal plant Sceletium tortuosum[J].Phytochemistry Letters,2018,27:36-43.
[16]HIRUMA K,KOBAE Y,TOJU H.Beneficial associations between Brassicaceae plants and fungal endophytes under nutrient-limiting conditions:evolutionary origins and host-symbiont molecular mechanisms[J].Current Opinion in Plant Biology,2018,44:145-154.
[17]KUSARI S,PANDEY S P,SPITELLER M.Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites[J].Phytochemistry,2013,91(4):81-87.
[18]MARTINUZ A,SCHOUTEN A,SIKORA R A.Systemically induced resistance and microbial competitive exclusion:implications on biological control[J].Phytopathology,2012,102(3):260.
[19]KUSARI S,HERTWECK C,SPITELLER M.Chemical ecology of endophytic fungi:origins of secondary metabolites[J].Chem Biol,2012,19:792-798.
[20]YU Chao,CAI MengHao,LI Kang,et al.Significance of seed culture methods on mycelial morphology and production of a novel anti-cancer anthraquinone by marine mangrove endophytic fungus Halorosellinia sp[J].Process Biochemistry,2012,47(3):422-427.
[2]CHRISTINA A,CHRISTAPHER V,BHORE S J.Endophytic bacteria as a source of novel antibiotics:An overview[J].Pharmacognosy Reviews,2013,13(7):11-16.
[3]DING ChangHong,WANG Qian Bo,GUO ShengLei,et al.The improvement of bioactive secondary metabolites accumulation in Rumex gmelini,Turcz through coculture with endophytic fungi[J].Brazilian journal of microbiology,2017,44(1):95-101.
[4]GOSS M J,CARVALHO M,BRITO I,et al.Impacts on Host Plants of Interactions Between AMF and Other Soil Organisms in the Rhizosphere[J].Functional Diversity of Mycorrhiza&Sustainable Agriculture,2017:81-109.
[5]LIU ShengRong,KE BinRong,ZHANG WeiRui,et al.Breeding of new Ganoderma lucidum strains simultaneously rich in polysaccharides and triterpenes by mating basidiospore-derived monokaryons of two commercial cultivars[J].Scientia Horticulturae,2017,216:58-65.
[6]JI XiaoLong,PENG Qiang,YUAN YuePeng,et al.Isolation,structures and bioactivities of the polysaccharides from jujube fruit:A review[J].Food Chemistry,2017,227:349-357.
[7]李传民.绞股蓝内生真菌高产胞外多糖发酵条件的优化及结构分析[D].郑州:河南工业大学,2016.
[8]LI XiaoLi,WANG ZhengHui,ZHAO YongXi,et al.Purification of a polysaccharide from Gynostemma pentaphyllum Makino and its therapeutic advantages for psoriasis[J].Carbohydrate Polymers,2012,89(4):1232-1237.
[9]CHEN Tong,LI Bo,Li YuanYuan,et al.Catalytic synthesis and antitumor activities of sulfated polysaccharide from Gynostemma pentaphyllum Makino[J].Carbohydrate Polymers,2011,83(2):554-560.
[10]程莹,赵骏.桑叶多糖含量测定与成分分析[J].中国临床药理学杂志,2017,33(18):1803-1805,1809.
[11]李良,金文娟.枸杞多糖含量测定方法的比较分析[J].食品研究与开发,2016,37(11):143-146.
[12]JOHN,ROJAN P,NAMPOOTHIRI M,et al.Solidstate fermentation for L-lactic acid production from agro wastes using Lactobacillus delbrueckii[J].Process Bioche mistry,2006,41(4):759-763.
[13]ROJAN P J,NAMPOOTHIRI K M,NAIR A S,et al.L(+)-lactic acid production using Lactobacillus casei in solid[J].Biotechnology Letters,2005,27(21):1685-1688.
[14]武模戈.碳氮源对裂褶菌胞外多糖产量的影响初探[J].濮阳职业技术学院学报,2008,(04):4-5.
[15]MANGANYI M C,REGNIER T,KUMAR A,et al.Phylogenetic analysis and diversity of novel endophytic fungi isolated from medicinal plant Sceletium tortuosum[J].Phytochemistry Letters,2018,27:36-43.
[16]HIRUMA K,KOBAE Y,TOJU H.Beneficial associations between Brassicaceae plants and fungal endophytes under nutrient-limiting conditions:evolutionary origins and host-symbiont molecular mechanisms[J].Current Opinion in Plant Biology,2018,44:145-154.
[17]KUSARI S,PANDEY S P,SPITELLER M.Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites[J].Phytochemistry,2013,91(4):81-87.
[18]MARTINUZ A,SCHOUTEN A,SIKORA R A.Systemically induced resistance and microbial competitive exclusion:implications on biological control[J].Phytopathology,2012,102(3):260.
[19]KUSARI S,HERTWECK C,SPITELLER M.Chemical ecology of endophytic fungi:origins of secondary metabolites[J].Chem Biol,2012,19:792-798.
[20]YU Chao,CAI MengHao,LI Kang,et al.Significance of seed culture methods on mycelial morphology and production of a novel anti-cancer anthraquinone by marine mangrove endophytic fungus Halorosellinia sp[J].Process Biochemistry,2012,47(3):422-427.