多年生黑麦草愈伤组织诱导及柠檬酸对其褐化的抑制效应  

Callus Induction and Inhibition Effect of Citric Acid on Browning of Perennial Ryegrass Callus

作　　者：代小梅[1] 孙振元[1] 韩蕾[1] 巨关升[1] 

机构地区：[1]中国林业科学研究院林业研究所,北京100091

出　　处：《核农学报》2015年第10期1893-1900,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家科技计划课题(2011AA100209-2)

摘　　要：为了优化多年生黑麦草愈伤组织诱导培养条件,以多年生黑麦草成熟种子为材料,研究浸泡、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,愈伤组织诱导率逐渐下降,且出愈时间推迟。在浸泡48h处理下,研磨处理的种子愈伤组织诱导率显著高于未研磨处理的种子,随机接种的种子愈伤组织诱导率显著高于机械接种的种子;经研磨处理的种子以随机接种方式接种于愈伤组织诱导培养基上,培养3d后即出现愈伤组织,14-21d后愈伤组织发生达到高峰期,21-30d后胚性愈伤组织大量形成,愈伤组织诱导率高达90%。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柠檬酸浓度是2mg·L^-1,培养2周的褐化率为15%,而随柠檬酸浓度升高,褐化率逐渐增加,而未添加柠檬酸的愈伤褐化率为50%。经试验观察发现2mg·L^-1柠檬酸对愈伤组织的增殖和分化再生无明显影响,胚性愈伤组织分化率为70%,生根率100%。本研究建立的多年生黑麦草的高效再生体系,不仅有利于其基因工程改造,也为其愈伤组织再生体系研究奠定了科学基础。In order to optimize the condition of callus induction,mature seeds of perennial ryegrass（ Lolium perenne L.）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 perennial ryegrass callus. The results showed that soaking for 48 h was in favor of callus induction,the average callus induction rate gradually decreased with prolonged immersion and the time of callus formation was delayed. Under the treatment of soaking for 48 h,the average callus induction rate of seeds under grinding was much significantly higher those without grinding,meantime the average callus induction rate of seeds under random transferring way was much significantly higher than mechanical inoculation way. Under soaking for 48 h and grinding,callus appeared after seeds were inoculated on callus induction medium for 3days by random transferring way,and reached the peak during culture for 14 to 21 days. Calli were subcultured after 21 days and a lot of embryogenic calli formed during culture for 21 to 30 days,the callus induction rate was 90%. In addition,the results showed that the inhibition effect of 2mg·L^- 1Citric acid was better and the browning rate of Perennial ryegrass callus was 15% after subculture for 2 weeks,however,the browning rate increased with the Citric acid concentration increased and the browning rate of the control was 50%. No significant effect of 2 mg·L^- 1Citric acid concentration on the proliferation,differentiation and regeneration of callus were observed. The differentiation rate of embryogenic callus was 70%,and the rooting rate was 100%. The efficient system established for perennial ryegrass in this study would be useful for its gene engineering breeding program,and also be an important theoretical basis for the differentiation theory study in Perennial ryegrass.

关 键 词：多年生黑麦草 愈伤组织诱导 柠檬酸 褐化 

分 类 号：S543.6[农业科学—作物学]