【作者】 赵玥；

【导师】 郭顺星；

【作者基本信息】 北京协和医学院 ， 生药学， 2009， 博士

【摘要】 对两种金线莲组培苗的最佳生根培养基进行了初步研究,结果表明,台湾金线莲和福建金线莲组培苗最佳生根培养基配方分别为0.125倍MS+1.61mg·L-1NAA+1.50mg·L-1 IBA+2.20g·L-1活性炭和0.36倍MS+3.00mg·L-1NAA+1.50mg·L-1活性炭,优化验证试验组培苗平均生根数分别可达2.33条/株和2.45条/株,分别比对照多1.20条和1.30条。栽培基质筛选试验结果显示,腐殖土：蛭石(3：1)的混合栽培基质最适合两种金线莲组培苗的移栽,在该栽培基质中培养90d,台湾金线莲的成活率达96.67%,平均鲜重、干重及株高分别达1.197g/株、0.136g/株及11.302cm/株,福建金线莲的成活率达96.67%,平均鲜重、干重及株高分别达1.177g/株、0.131g/株及10.856g/株。野生金线莲在其整个生活史中都需要与真菌共生。我们利用这一特点,将分离自兰科植物的43株内生真菌回接到两种金线莲组培苗上,在琼脂培养基上筛选出10株对台湾金线莲和8株对福建金线莲有促生作用的兰科内生真菌。温室盆栽试验结果显示,AR18对两种金线莲的促生效果均达显著水平,栽培90d时,接种AR18的台湾金线莲平均鲜重和平均干重分别比对照提高15.080%和19.403%,接种AR18的福建金线莲平均鲜重和平均干重分别比对照提高17.155%和22.584%,接种AR14的福建金线莲平均鲜重和平均干重分别对照提高15.736%和18.652%。生理生化试验结果表明：在共培养的2个月内,随共培养时间延长,对照与接菌处理的4种酶活性均呈升高趋势,真菌的接种使这种上升趋势更为明显。金线莲体内大量元素中钾的含量最高,镁的含量最低,微量元素中铁的含量最高。不同真菌对两种金线莲的元素含量影响不同。次生代谢产物含量检测结果表明,人工栽培金线莲中Kinsenoside的含量随栽培时间延长而提高,促生真菌的接种会促进两种金线莲中Kinsenoside的积累。当栽培时间为8周时,接种AR18的台湾金线莲和福建金线莲中Kinsenoside的含量分别为同时期对照的1.97倍和1.24倍,接种AR14的福建金线莲中Kinsenoside的含量为同时期对照的1.63倍。在人工栽培的福建金线莲中,只检测到1种黄酮单体异鼠李素-3-O-葡萄糖苷,接种AR14对其含量有促进作用,而接种AR18对其含量则是先抑制后促进,但二者的促进作用均不显著。在人工栽培的台湾金线莲中,检测到4种黄酮单体成分,其中两种可定性为异鼠李素-3-O-葡萄糖苷及槲皮素。接种AR18对异鼠李素-3-O-葡萄糖苷的含量影响为先抑制后促进,而对槲皮素的含量影响一直为促进作用。更多还原

【Abstract】 Anoetochilus roxburghii (Wall) Lindl. and A. formosanus, belonging to Orchidaceae, are rare medicinal plants. The most suitable root growth medium for A. formosanus was 0.125MS+1.61mg·L-1+1.50mg·L-1+2.20g·L-1 active carbon, and that for A. roxburghii was 0.36MS+3.00mg·L-1+1.50mg·L-1+0.80g·L-1 active carbon. After 60ds’culture on the root growth medium, the roots number of A. formosanus and A. roxburghii reached 2.33 and 2.45 respectively.A. formosanus and A. roxburghii were transplanted to flowerpots which filled with 5 kinds of culture substrate. The results showed that the most suitable substrate was humus soil:vermiculite (3:1). After 90ds’culture in the substrate, the average fresh weight, dry weight and height of A. formosanus were 1.197g,0.136g and 11.302cm, and that of A. roxburghii were 1.177g,0.131g and 10.856cm.Wild Anoetochilus plants are known to form a symbiotic relationship with fungi. Based on the characteristic, we cultured 43 strains of fungi from orchids together with seedlings of A. roxburghii and A. formosanus on agar medium for 2 months.10 strains could promote seedlings of A. formosanus growth and 8 strains could promote seedlings of A. roxburghii growth.To make sure the promotion activity of the fungi, we inoculated them to the roots of A. formosanus and A. roxburghii. After 90d, the average fresh weight and dry weight of A. formosanus inoculated with AR18 were increased by 15.08% and 19.403% respectively, compared with the non-inoculated ones, while the average fresh weight and dry weight of A. roxburghii inoculated with AR18 or AR14 were increased by 17.155%,22.584% and 15.736%,18.652% respectively, compared with the non-inoculated ones.The element of A. formosanus and A. roxburghii were measured by ICP. The results showed that K content was higher than other macro-elements, and the Mg content was the lowest. Fe content was the highest in the trace elements. Different fungi have different effect on the element contents of A. formosanus and A. roxburghii.The content of Kinsenoside and flavone of A. formosanus and A. roxburghii were detected by HPLC to study the effect of fungi on the secondary metabolite. The content of Kinsenoside in cultivated A. formosanus and A. roxburghii rising over time, and the inoculation of fungi increased the Kinsenoside content. The promotion effect of AR18 on A. roxburghii was more significant than on A. formosanus. The Kinsenoside content of the A. formosanus and A. roxburghii inoculated with AR18 were 1.97 and 1.24 times respectively as much as control at the 8th week. While, the promotion effect of AR14 on A. roxburghii was inferior to AR18, the Kinsenoside content of A. roxburghii inoculated with AR14 was 1.63 times as much as control at the 8th week.The kinds of flavonoid were different in A. formosanus and A. roxburghii, and it didn’t change with fungi inoculation. Only Isorhamnetin-3-O-β-D-glucopyranoside was detected in cultivated A roxburghii, and the inoculation with AR14 could increase its content while AR18 first suppress then increase its content.4 flavonoid were deteced in cultivated A. formosanus and 2 of them could be characterized as Isorhamnetin-3-O-β-D-glucopyranoside and Quercetin. AR18 first suppress then increase the content of Isorhamnetin-3-O-β-D-glucopyranoside, while it increases the content of Quercetin all the time.更多还原