铁皮石斛悬浮培养条件筛选及生产多糖

Research on cell suspension culture of Dendrobium officinale in producing the dendrobium polysaccharides

利用野生铁皮石斛茎尖、种子作外植体,比较所形成的愈伤组织性状、长势,筛选野生铁皮石斛茎尖为最佳诱导愈伤组织的外植体;选用水提醇沉法提取铁皮石斛多糖,并采用改良的硫酸苯酚法测定多糖含量。设计正交试验,筛选得到铁皮石斛愈伤组织生长快速且多糖含量高的培养条件,实验结果 MS+1.0 mg/mL萘乙酸+1.0 mg/mL 6-苄基氨基嘌呤+0.5 mg/mL 2,4-二氯苯氧乙酸,19 d为既能使铁皮石斛愈伤组织生长快且多糖含量高的激素配比。

Stem tips and weeds of Dendrobium officinale were used as explants to produce pultaceous callus.After compared with the growing properties,sites and the rate of abduction, the stem tip is the best-selected explant for inducing callus.Water extraction and ethylalcohol precipitation was used as the method to extract polysaccharides from the Dendrobium officinale and modifred phenol-sulfuric acid is used as the method to determine the polysaccharide content. Orthogonal tests were planed to optimize combination of phytohormones and auxins for inducing pultaceous callus which grew fast and with high content of Dendrobium polysaccharides. The results indicated that MS ＋ 1.0 mg/L naphthalene acetic acid（NAA）＋ 1.0 mg/mL 6-benzylaminopurine（6-BA）＋ 0.5 mg/mL 2,4-dichlorophenoxyacetic acid（2,4-D）＋ 19 d was the most suitable ratio of hormones to induce pultaceous callus to grow fast.The assey of content of Dendrobium moniliforme showed that MS ＋ 1.0 mg/L NAA ＋ 1.0 mg/mL 6-BA ＋ 0.5 mg/mL 2,4-D ＋ 19 d is advantageous to improve the content of Dendrobium moniliforme.MS ＋ 1.0 mg/L NAA ＋ 1.0 mg/mL 6-BA ＋ 0.5 mg/mL 2,4-D ＋ 19 d is good for accelerating the proliferation of pultaceous callus and producing more moniliforme,and we induce a large number of pultaceous callus using this condition.Phenol-sulferic acid method is used to extract the Dendrobium moniliforme,which is totally amount to 300 gram.