摘要
本研究以草地早熟禾新歌莱德、午夜、纳苏3个品种的成熟种子为材料,研究了浸泡、研磨和接种方式等因素对愈伤组织诱导率和出愈时间的影响以及不同浓度的柠檬酸对其愈伤组织褐化的抑制效果。结果显示,浸泡48h处理有利于种子愈伤组织诱导,随着浸泡时间延长愈伤组织平均诱导率逐渐下降,且3个品种的出愈时间均推迟;在浸泡48h处理下,无论采取机械接种或随机接种方式接种,研磨处理与未经研磨处理所得到的3个品种种子愈伤组织诱导率均无明显差异;在种子浸泡48h,未经研磨处理条件下,分别采用随机接种与机械接种方式所得到的3个品种种子愈伤组织诱导率差异极显著。此外,研究表明在愈伤组织的继代培养中对愈伤组织褐化抑制效果较好的浓度是2mg·L-1,培养2周未见有褐化现象,而随柠檬酸浓度升高褐化率逐渐增加,且对照的褐化率达到25%~37.5%。经试验观察发现2 mg·L-1柠檬酸浓度对愈伤组织的增殖和分化再生无明显影响,新歌莱德、午夜、纳苏的胚性愈伤组织分化率分别为75%、50%、70%,生根率分别为85%、70%、80%。本研究初步探讨了影响草地早熟禾愈伤组织诱导的因素,解决了草地早熟禾愈伤组织继代培养中的褐化现象,完善了草地早熟禾愈伤组织诱导和再生体系建立,为利用愈伤组织再生体系进行基因工程改造研究奠定了重要的技术基础。
Mature seeds of three varieties of Perennial ryegrass （Lolium perenne L. ） ＇ Nuglade＇, ＇Midnight＇ ＇ Nassau＇ were used to investigate the effect of immersion, grinding and transferring way on the callus induction rate and formation time and the inhibition effect of different citric acid concentration on browning of Kentucky bluegrass callus. The results showed that soaking for 48h was in favor of callus induction, and the average callus induction rate gradually decreased with prolonged immersion, while the callus formation time of three varieties were pushed back. Under soaking for 48h treatment, whether inoculated by mechanical or random transferring way, the average callus induction rate of three varieties seeds was no significant different. Under soaking for 48h treatment without grinding treatment, the average callus induction rate of three varieties seeds under random and mechanical transferring way was much significantly different. In addition, the results showed that the inhibition effect of 2mg＂ L-I citric acid was best and there was no browning phenomenon during subculture for 2 weeks. While the browning rate increased with the citric acid concentration increased, the browning rate of the control was 25% to 37.5%. The experiment observed no significant effect of 2mg＂ L-1 citric acid concentration on the proliferation, differentiation and regeneration of callus. The differentiation rate of embryogenic callus of ＇ Nuglade＇ , ＇Midnight＇ , ＇ Nasu＇ was 75% ,50% ,70% , and the rooting rate was 85% ,70% ,80%. This study explored the impact factors of Kentucky Bluegrass callus induction, solved the browning problem in the subculture, improved callus induction and regeneration system of Kentucky bluegrass and laid an important foundation for genetically engineered technology research used callus regeneration system.
出处
《核农学报》
CAS
CSCD
北大核心
2015年第2期270-277,共8页
Journal of Nuclear Agricultural Sciences
基金
"十二五"国家科技计划课题(2011AA100209-2)
关键词
草地早熟禾
组织培养
柠檬酸
褐化
kentucky bluegrass, tissue culture, citric acid, browning
