Relaxed calli culture of grape and resveratrol content determining from its calli by using HPLC | 葡萄松散型愈伤组织的培养及其白藜芦醇含量的测定

Chinese. 获得葡萄（Vitis vinifera L．）松散型愈伤组织，并了解其白藜芦醇含量的变化。以葡萄的叶、叶柄和茎分别作为外植体，在不同激素组合（6-卞氨基嘌呤（6-BA）、激动素（KT）、吲哚丁酸（IBA）、2,4-二氯苯氧乙酸（2，4-D）的培养基（以B5培养基为基本培养基，含30 g/L蔗糖和6 g/L琼脂）上，进行愈伤组织的诱导和继代培养，并利用高效液相色谱法（HPLC）对不同来源愈伤组织中的白藜芦醇含量进行测定。以葡萄叶为外植体时，诱导的愈伤组织出愈率很低；以叶柄和茎为外植体时，愈伤组织的出愈率均较高。利用叶柄和茎获得松散愈伤组织时，其诱导培养基组成分别是：B5培养基+6-BA 0.5 mg/L+IBA 0.1 mg/L和B5培养基+6-BA 1.0 mg/L+IBA 0.2 mg/L，其继代培养基均为B5培养基+6-BA 1.0 mg/L+2,4-D 0.1 mg/L 。在诱导培养基产生的愈伤组织中，白藜芦醇含量由高到低的顺序为叶柄叶茎。以叶、叶柄和茎为外植体诱导愈伤组织时，其白藜芦醇含量最高的培养基分别为B5培养基+6-BA 0.5 mg/L+IBA 0.2 mg/L、B5培养基+6-BA 1.0 mg/L+IBA 0.2 mg/L和B5培养基+6-BA 0.5 mg/L+IBA 0.2 mg/L。葡萄叶柄和茎易被诱导产生松散的愈伤组织，且经多次继代可以用于建立悬浮培养体系；葡萄愈伤组织中的白藜芦醇含量随外植体及培养基组成的不同而有所差异。

English. Relaxed grape’s（Vitis vinifera L．）calli was acquired and resveratol content change was studied. The leaves, leafstalk, and stems from grapes were used as explants to induce calli and subculture in medium (100 mL B5 medium containing 3 g sucrose and 0.6 g agar) with different plant hormone combinations (6-BA, KT, IBA, 2,4-D), and then the resveratol content from those calli was determinated by using HPLC. The results indicated that, the calli development rate from leaves inducement was lower than that from leafstalk and young stems. The inducement medium was as follows respectively: for the leafstalk, B5 medium with 6-BA 0.5 mg/L+IBA 0.1 mg/L, and for stems, B5 medium with 6-BA 1.0 mg/L+IBA 0.2 mg/L, and subculture medium both were B5 medium with 6-BA 1.0 mg/L+2,4-D 0.1 mg/L. Under the same culture condition, the calli resveratol contents from high to low from different explants were leafstalk, leaves, and young stems in turn. About calli from different explants, there were significant differences in their resveratol content, and their medium of the highest content expressed were respectively: for leaves, B5 medium+6-BA 0.5 mg/L+IBA 0.2 mg/L; for leafstalk, B5 medium+6-BA 1.0 mg/L+IBA 0.2 mg/L;for stems, B5 medium+6-BA 0.5 mg/L+IBA 0.2 mg/L. The leafstalk and stem are facile induced to relaxed callis, and it could be the original of suspension culture by subculture many times. Its content of resveratol is influenced by the different parts of plant and culture condition.