高羊茅悬浮细胞系的建立及绿色植株的高频再生

Establishment of suspension cell lines and high frequency regeneration of green plants in tall fescue

将5个高羊茅品种的成熟种子经灭菌后,接种于含有不同浓度2,4-D和ABA的N6培养基上进行愈伤组织诱导。结果表明,不同浓度2,4-D和ABA对5个品种的愈伤组织诱导率有显著影响,以培养基中附加9mg/L2,4-D和2mg/LABA的诱导率最高。将经过数次继代改造后获得的高质量胚性愈伤置于AA液体培养基中进行悬浮培养,建立起来自3个品种的8个悬浮细胞系,其中6个悬浮细胞系经高渗预处理后能高频再生出绿色植株,基本克服了以往研究中再生白化植株的现象。

Sterilized mature seeds of five tall fescue (Festuca rubra)varieties were cultured for callus induction on N6 medium containing different concentrations of 2,4D and ABA. The results showed that there were significant influences of different concentrations of 2,4D and ABA on callus induction frequency of these varieties, with the highest induction frequency for supplement of 9 mg/L 2,4D and 2 mg/L ABA in the medium. In addition , marked differences in induction frequency were also observed among the five varieties . High quality embryogenic callus resulting from several times subcultures were transferred to AA medium for suspension culture. Eight suspension cell lines of three varieties were finally established. After high osmosis treatment, green plants were regenerated with high frequency from six lines, largely overcoming the problem of severe albino of regeneration plants in previous cell suspension cultures.