《植物生理学报》 2014, 50(11): 1683-1691

刺葡萄幼胚愈伤组织诱导及其高产原花青素细胞系筛选

赖呈纯*, 范丽华, 黄贤贵*, 谢鸿根

福建省农业科学院农业工程技术研究所, 福州350003

通信作者：赖呈纯；E-mail: lccisland@163.com, hxg323@163.com；Tel: 0591-83839109; 0591-87884297

摘　要：

以刺葡萄幼胚为材料, 研究不同培养方式、培养基配方和培养条件对其愈伤组织诱导的影响, 采用正交试验设计法筛选刺葡萄愈伤组织继代增殖的培养基配方, 并对继代保持的培养条件和方式进行优化, 同时进行了高产原花青素刺葡萄愈伤组织细胞系的筛选研究。结果表明, 刺葡萄幼胚以平放的方式接种到MS+1.0 mg∙L^-1 2,4-D或MS+1.0 mg∙L^-1 2,4-D+0.5 mg∙L^-1 KT的固体培养基上, 在黑暗的条件下, 能有效的诱导出愈伤组织, 诱导效率为80%; 刺葡萄愈伤组织继代增殖以MS+1.5 mg∙L^-1 2,4-D或MS+1.5 mg∙L^-1 2,4-D+0.5 mg∙L^-1 KT的固体培养基为佳, 并且采用此两种培养基交替继代培养, 在光照条件下能长期保持旺盛且生长一致的刺葡萄愈伤组织; 筛选出了紫红色松脆状的高产原花青素的刺葡萄愈伤组织细胞系, 培养35 d后每克鲜样的原花青素含量可达1 671.16 μg。

关键词：刺葡萄; 幼胚; 愈伤组织; 花青素; 原花青素

收稿：2014-08-22   修定：2014-10-13

资助：福建省自然科学基金项目(2012J01102)。

Callus Induction in Brier Grape (Vitis davidii Foëx) from Immature Embryos and Screening of Cell Lines with High-Production of Oligomeric Proanthocyanidins

LAI Cheng-Chun*, FAN Li-Hua, HUANG Xian-Gui*, XIE Hong-Gen

Institute of Agricultural Engineering and Technology, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China

Corresponding author: LAI Cheng-Chun; E-mail: lccisland@163.com, hxg323@163.com; Tel: 0591-83839109; 0591-87884297

Abstract:

The effects of different culture methods, culture medium formula and culture condition on callus induction were studied using immature embryos of brier grape as plant materials. The culture media for brier grape callus subculture multiplication were screened with orthogonal experiment design. The culture condition and culture method for brier grape callus subculture were further optimized. Meanwhile, screening of cell lines with high yield of oligomeric proanthocyanidins was studied in this experiment. The results showed that the brier grape callus was inducted effectively while immature embryos was horizontal onto the medium MS+1.0 mg∙L^-1 2,4-D or MS+1.0 mg∙L^-1 2,4-D+0.5 mg∙L^-1 KT in the dark, with a 80% callus induction rate. The best medium to callus subculture multiplication were MS+1.5 mg∙L^-1 2,4-D or MS+1.5 mg∙L^-1 2,4-D+0.5 mg∙L^-1 KT, and they grew well and conformably through alternative subculture of the two media. Cell lines with high yield of oligomeric proanthocyanidins were screened from these callus, and these cell lines present some characters with aubergine and friable. The yield of oligomeric proanthocyanidins of these cell lines reached up to 1 671.16 μg in 1 g fresh sample after being cultured for 35 days.

Key words: brier grape (Vitis davidii Foëx); immature embryo; callus; anthocyanosides; oligomeric proanthocyanidins