长白落叶松体胚发生再生体系优化
收稿日期: 2020-02-26
录用日期: 2020-06-05
网络出版日期: 2020-06-05
基金资助
中央高校基金(2572018CL02)
Optimization of the Regeneration System from Somatic Embryogenesis in Larix olgensis
Received date: 2020-02-26
Accepted date: 2020-06-05
Online published: 2020-06-05
以长白落叶松(Larix olgensis)未成熟合子胚为外植体诱导胚性愈伤组织, 通过调节影响体胚发生的营养物质和植物生长调节剂配比, 进行愈伤组织的胚性恢复与保持以及体胚发生再生体系的优化。结果表明: 不同无性系之间胚性愈伤组织诱导率差异显著, 胚性愈伤组织在S+0.2 mg·L -1NAA+0.5 mg·L -1BA+0.5 mg·L -1KT+0.5 g·L -1谷氨酰胺+0.5 g·L -1水解酪蛋白+30 g·L -1蔗糖及3.0 g·L -1植物凝胶培养条件下, 可以恢复胚性并长久保持。在S+20 mg·L -1ABA+60 g·L -1PEG4000+60 g·L -1蔗糖及3.0 g·L -1植物凝胶条件下分化培养6周, 体胚发生率可达100%。将正常发育的体胚先在WPM+ 6 mg·L -1间苯三酚+1.0 g·L -1活性炭+3.0 mg·L -1VB1+20 g·L -1蔗糖及3.0 g·L -1植物凝胶条件下培养2周, 再转接至B5+ 0.4 mg·L -1NAA+1.0 mg·L -1IBA+0.5 mg·L -1GA3+2.0 mg·L -1VB1+1.0 g·L -1活性炭+20 g·L -1蔗糖及3.0 g·L -1植物凝胶条件下培养2周, 可见子叶舒展、下胚轴伸长且根系正常的体胚苗。该研究建立了长白落叶松胚性愈伤组织胚性恢复与保持方法, 并进一步优化了体胚发生的植株再生体系, 为林木资源快速繁育和遗传改良奠定了基础。
刘建飞, 刘炎, 刘克俭, 池阳, 霍志发, 霍永洪, 由香玲 . 长白落叶松体胚发生再生体系优化[J]. 植物学报, 2020 , 55(5) : 605 -612 . DOI: 10.11983/CBB20030
In this study, immature zygotic embryos of Larix olgensis were used as explants to induce embryogenic callus and optimize the regeneration system from somatic embryogenesis. The rejuvenation and preservation of the embryogenic potential of embryogenic callus, the somatic embryogenesis and plant regeneration were investigated through adjusting the nutrition and plant growth regulator. The results showed that the generation rates of embryogenic callus were significantly different among different lines. Under the conditions of S+0.2 mg·L -1NAA+0.5 mg·L -1BA+0.5 mg·L -1KT+0.5 g·L -1glutamine+0.5 g·L -1hydrolyzed casein+30 g·L -1sucrose and 3.0 g·L -1vegetable gel, the embryogenic potential of embryonic callus could be recovered and maintained for a long time. Somatic embryogenesis were induced from embryonic callus cultured in S+20 mg·L -1ABA+60 g·L -1PEG4000+60 g·L -1sucrose and 3.0 g·L -1vegetable gel for 6 weeks, and the generation rate of somatic embryo reached 100%. The normal somatic embryos were first cultured for 2 weeks under the conditions of WPM+6 mg·L -1phloglucinol+1.0 g·L -1active carbon+3.0 mg·L -1VB1+20 g·L -1sucrose and 3.0 g·L -1vegetable gel, and then transferred to B5+0.4 mg·L -1NAA+1.0 mg·L -1IBA+0.5 mg·L -1GA3+2.0 mg·L -1VB1+1.0 g·L -1active carbon+20 g·L -1sucrose and 3.0 g·L -1vegetable gel. After 2 weeks, somatic embryo plantlets with cotyledon stretch, hypocotyl elongation and normal root system were observed. This study established a method for the recovery and maintenance of embryogenic callus from larch, and further optimized the somatic embryogenic pathway, which will lay a foundation for the rapid breeding and genetic improvement of L. olgensis.
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