广西植物2016，Vol.36Issue(9)：1082-1086,5.DOI:10.11931/guihaia.gxzw201501038
文心兰类原球茎的愈伤组织诱导及其植株再生
Callus induction and plant regeneration of Oncidium‘Gower Ramsey’ by means of protocorm-like body culture
摘要
Abstract
Oncidium ‘Gower Ramsey ’ is a species of orchid with high ornamental value and strong market competitiveness. Gene engineering breeding technology provides an effective way for the genetic improvement of ornamental plants. And callus is an ideal transformation receptor. Up to now, callus induction and plant regeneration of Onciudium have been successfully achieved using stem segment, root tip and shoot tip. But there are some shortcomings for these in vitro re-generation systems, such as low frequency and long period of callus induction, difficulty in maintaining embryogenic state of callus, etc. In this research, for the first time protocorm-like bodies of Oncidium‘Gower Ramsey’ were used as explants for callus induction and plant regeneration. The effects of different concentrations and combinations of TDZ and 2,4-D on callus proliferation were investigated. The results showed that 1/2MS medium supplemented with 1 mg.L-1 TDZ and 3 mg.L-1 2,4-D could promote the induction of whitish and friable callus, the induction rate was 100%. The proliferation rate of callus was mainly affected by the concentration of TDZ, while the callus subculture medium consisted of TDZ in combination with 2,4-D at the concentrations of 0.5 to 2.0 mg .L-1 . When the concentration of TDZ decreased from 1.0 to 0.5 mg.L-1 , pro-liferation rate of the callus fresh weigh increased significantly. And the optimal medium for callus proliferation contained 1/2MS basal medium, 0.5 mg.L-1 TDZ and 1.0 mg.L-1 2,4-D. After four weeks of subculture in the dark, the highest prolif-eration rate of the callus fresh weigh, which was 6.04 folds, appeared on the optimal medium, while the lowest proliferation rate of the callus fresh weigh, which was 4.50 folds, appeared on the medium containing 1/2MS basal medium, 1.0 mg.L-1 TDZ and 2.0 mg.L-1 2,4-D. The callus subcultured on the optimal proliferation medium for about one month were trans-ferred to T2 medium (3.5 g.L-1 Hyponex 1+20 g.L-1 sweet potato+25 g.L-1 banana+1 g.L-1 tryptone+20 g.L-1 sucrose + 3.5 g.L-1 phytagel) and cultured in the dark for PLBs induction. Approximately, 1 328.67 protocorm-like bodies could be generated in one month from an initial culture of 1 g callus fresh weight. Subsequently, the cultures transferred on-to the fresh T2 medium. The germination rate of protocorm-like bodies was as high as 90.12%after one month of culture un-der the light. When transplanted on 1/2MS medium supplemented with 1 g.L-1 active carbon, all shoots became healthy plants after two months. In this study, the callus induction and plant regeneration system of Oncidium‘Gower Ramsey’ with high frequency was successfully developed to provide an efficient and stable transformation receptor system for the genetic transformation of Oncidium.关键词
文心兰/组织培养/愈伤组织增殖/TDZ/再分化Key words
Oncidium‘Gower Ramsey’/tissue culture/callus proliferation/TDZ/redifferentiation分类
农业科技引用本文复制引用
黄霞,卢禹..文心兰类原球茎的愈伤组织诱导及其植株再生[J].广西植物,2016,36(9):1082-1086,5.基金项目
国家“十二五”科技支撑计划项目(2013BAD01B0702)[Supported by the National Key Technology R&D Program during the Twelfth Five-year Plan Period (2013BAD01B0702)]。 （2013BAD01B0702）
