红景天抗氧化活性内生真菌对宿主化学成分积累的影响

发布时间：2018-04-18 03:15

本文选题：红景天 + 内生真菌　；参考：《山西大学》2015年硕士论文

【摘要】：红景天(Rhodiola. L)是传统而新兴的药用植物,生长于缺氧、高寒、强紫外等恶劣条件的高海拔(2500 m)山区,其抗氧化和抗缺氧双向调节活性是最具研究价值的资源。本研究一方面对从我国东北长白山和青藏高原采集的四种主要红景天中分离出的168株内生真菌进行了抗氧化活性及化合物筛选；另一方面通过抗氧化活性菌株与宿主植物的共培养,研究内生真菌对红景天主要活性成分如红景天苷等的积累规律。结果表明,红景天内生真菌的抗氧化能力显著。DPPH自由基清除能力大于70%的4种红景天内生真菌菌株共62株,占供试菌株的37%；·OH-、Fe2+螯合、02-·及亚硝酸盐清除能力高于70%的菌株分别占总供试菌株的18%、4%、15%、3%。其中编号为Ra-R-32、ZPRa-R-1、Rb-R-1、Rc-L-2、Rs-R-15、ZPRs-S-13的6株菌,表现出明显的抗氧化能力。它们的IC50值分别为1.54、1.41、3.03、0.24、1.91mg/mL,而阳性对照Vc的IC50值分别为0.32、2.72、9.68、0.59、1.70mg/mL。为了考察抗氧化内生真菌对红景天活性成分的影响,实验首先对采集的野生样品和组培苗化学成分的含量进行了分析。结果表明,野生红景天根茎中,库页红景天(R. sachalinensis)中红景天苷(salidroside)、酪醇(p-tyrosol)、洛塞琳(rosarin)、洛塞维(rosavin)和松香(rosin)含量分别为13.6949、1.6389、0.1705、0.0137、0.9564 mg/g,均高于其他品种；大花红景天(R.crenulata)的叶、花、茎、根茎中,根茎中红景天苷含量最高,含量达7.2648 mg/g,花中酪醇含量最高,为11.1653 mg/g,叶中洛塞琳、洛塞维和松香最高,分别为0.0439、0.0853、0.0769 mg/g；大花红景天组培苗中红景天苷和酪醇含量分别为0.0172、0.3324 mg/g,愈伤组织中红景天苷和酪醇含量分别为0.0223、0.1266 mg/g,均低于野生大花红景天的根茎、茎、叶、花部位的含量；不同继代组培苗和愈伤组织培养中,从第2代起各活性成分含量急剧下降,第4代后各成分含量有所上升,至第6代后,含量保持稳定,与第一世代相比,含量均有所降低。实验还将内生真菌与宿主植物共生培养,研究内生真菌对宿主化学成分积累的影响。活性菌ZPRa-R-1与组培苗共生前3d中,红景天苷和酪醇含量逐渐降低,第4d时有所增加；此后随着培养时间的延长,稳步上升。培养至第6d后,与活性菌ZPRa-R-1共生的组培苗中红景天苷和酪醇的含量高于对照组培苗中的含量,最高可分别达到0.0163、0.3207 mg/g。第10d组培苗死亡,其红景天苷和酪醇急剧降低,仅为0.0110、0.3118 mg/g。
[Abstract]:Rhodiola.L) is a traditional and emerging medicinal plant, which grows in high altitude of 2500 m) mountain area with poor conditions such as hypoxia, cold and strong ultraviolet. Its antioxidant and anti-hypoxia bidirectional regulatory activities are the most valuable resources.On the one hand, 168 strains of endophytic fungi isolated from four major Rhodiola species collected from Changbai Mountain and Qinghai-Tibet Plateau in Northeast China were screened for their antioxidant activities and compounds.On the other hand, the accumulation of main active components such as salidroside by endophytic fungi was studied by co-culture of antioxidant active strains and host plants.The results showed that there were 62 strains of endophytic fungi of Rhodiola, whose antioxidant ability was significant. DPPH free radical scavenging ability was more than 70%.The OH-Fe _ 2 chelate 02- and the nitrite scavenging capacity above 70% accounted for 18.4% of the total tested strains, respectively.There were 6 strains of Ra-R-32ZPRa-R-1 Rb-R-1 Rc-L-2Rs-R-15 ZPRs-S-13, which showed obvious antioxidant activity.Their IC50 values were 1.54 卤1.41 ~ 3.01 ~ 0.24 ~ 1.91 mg 路mL ~ (-1), respectively, while the IC50 values of positive control group VC were 0.32 ~ 2.72 ~ 9.68 ~ 0.5991 mg / mL 路mL ~ (-1), and 0.32 卤2.72 ~ (9.68) ~ (0.5) mg / mL ~ (-1) mg 路mL ~ (-1), respectively.In order to investigate the effect of antioxidation endophytic fungi on the active components of Rhodiola, the contents of chemical constituents in wild samples and tissue culture seedlings were analyzed.The results showed that the contents of salidroside, tyrosine p-tyrosola, Roseline rosarinine, Rosevirosavin) and rosin rosin in the rhizome of Rhodiola sachalinensis were 13.6949 91.63890.17050.01370.0.9564 mg / g, higher than those of other species, respectively, and the contents of leaves, flowers, stems, roots and stems of R. crenulata were higher than those of other species, and the contents of leaves, flowers, stems, roots and stems of R. crenulata were higher than those of other species.The content of salidroside in rhizome is the highest, the content is 7.2648 mg / g, the content of tyrosol in flower is the highest, is 11.1653 mg / g, the content of Loseline, Rosin and Rosin in leaves is the highest.The contents of salidroside and tyrosol in the tissue culture seedlings of Rhodiola davidiola were 0.0172 ~ 0.3324 mg / g, and the contents of salidroside and tyrosine in callus were 0.022 ~ (23) ~ 0.1266 mg / g, respectively, which were lower than those in the rhizome, stem, leaf and flower part of Rhodiola davidiola.In different subculture plantlets and callus culture, the contents of each active component decreased sharply from the second generation, increased after the fourth generation, and remained stable after the sixth generation, and decreased compared with the first generation.The effects of endophytic fungi on the accumulation of host chemical components were also studied by symbiotic culture of endophytic fungi and host plants.The contents of salidroside and tyrosol decreased gradually at the 4th day before the symbiosis of the active bacteria ZPRa-R-1 and the plantlets in tissue culture, and then increased steadily with the prolongation of the culture time.After 6 days of culture, the contents of salidroside and tyrosol in the plantlets co-existing with the active bacteria ZPRa-R-1 were higher than those in the control group, and the highest values were 0.0163 mg / g, 0.3207 mg / g, respectively.On the 10th day, the salidroside and tyrosol decreased sharply, only 0.011010 ~ 0.3118 mg / g 路g ~ (-1) 路g ~ (-1) ~ (-1) 路min ~ (-1) ~ (-1).
【学位授予单位】：山西大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S567.239;S182

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