摘要
本论文首次报道了马尾松（Pinus massoniana Lamb.）幼胚体细胞胚胎发生的完整发育过程,并对影响马尾松胚性愈伤组织诱导的因素如球果采种期、球果冷藏处理时间、外植体处理方式等进行了探讨,统计胚性愈伤组织诱导率,进行增殖评价,探讨ABA浓度梯度对马尾松体细胞胚分化成熟的影响,试验数据用SPSS16统计分析软件进行方差分析、差异显著性检验。结果表明：1）2008~2009连续2年内15个采种期得到的幼胚,胚性愈伤组织诱导和增殖有显著性差异,最适宜的马尾松球果采种期是6月下旬至7月下旬,诱导率在9.66%~22.59%之间;2）球果冷藏处理时间,对胚性愈伤组织诱导有显著性差异,其中4℃冷藏球果15d有利于幼胚胚性愈伤组织诱导;3）雌配子体包含幼胚的接种处理方式是可取的;4）胚性愈伤组织经稳定增殖培养后,转入分化成熟培养基,得到体细胞胚状体＂爆发式＂分化成熟,数量多,质量好。适宜体胚成熟转化的培养基为：成熟LP培养基添加ABA5.0mg·L-1＋60.0g·L-1蔗糖,并附加L-谷氨酰胺和水解酪蛋白;5）成熟体细胞胚在无激素萌发型LP培养基上正常萌发,并转化为结构完整的小植株。本研究首次建立了马尾松幼胚体细胞胚胎发生技术平台,为马尾松遗传改良种质创新、缩短育种周期奠定了研究基础。
Somatic embryogenesis （SE） from immature zygotic embryos of masson pine （Pinus massoniana Lamb.） was achieved by megagametophytes containing zygotic embryos on LP medium. Factors influencing somatic embryogenesis （SE） in P. massoniana were studied, such as the cone collection dates, the time of cold treatment on the cone, the concentration of ABA. The initiation rates and the evaluation of embryonic callus were analyzed with SPSS 16.0. The results showed that： 1） The induction of SE were affected by different stages of immature embryos and there was statistically significant differences among the 15 cone collection dates. Considering the evaluation of embryonic callus, the cone collection date between June and July was appropriated with the initiation rates arranged from 9.66% to 22.59%. 2） There was also significant difference among the cold treatments time in initiation of embryonic callus, and the optimum cold treatment time was about 15 d at 4 ℃. 3） Some proliferation callus was fine and which was subcultured to maturation medium. 4） When the LP maturation medium containing 5.0 mg·L-1 ABA and 60.0 g·L-1 sucrose supplemented with both L-glutamine and casein, more mature somatic embryos with high quality were produced compared with 30.0-55.0 mg·L-1 ABA. 5）The somatic embryos were placed on germination medium without PGRs, after 4 to 6 weeks they were germinated and developed into small seedlings. It is the first report about the somatic embryogenesis with immature embryos of masson pine and these findings provide foundation for breeding some elite clones within short period.
出处
《植物生理学报》
CAS
CSCD
北大核心
2011年第9期904-912,共9页
Plant Physiology Journal
基金
湖南省自然科学基金项目(11JJ3038)
湖南省教育厅优秀青年基金项目(09B112)
教育部高等学校博士学科点专项科研基金项目(20104321110002,20060538004)
国家“863”计划课题
“十一五”国家科技支撑计划项目(2006BAD24B0301)
