《植物生理学报》 2014, 50(6): 785-791

蔗糖与肌醇对马尾松胚性细胞系增殖的影响

史昆^1,2, 杨模华^1,2,* , 李志辉², 张冬林^2,3, 丁贵杰⁴

中南林业科技大学¹林业生物技术湖南省重点实验室, ²林学院, 长沙410004; ³乔治亚大学, 美国乔治亚州30602; ⁴贵州大学林学院, 贵阳550000

通信作者：丁贵杰；E-mail: ymh163@163.com；Tel: 13875885952

摘　要：

针对马尾松胚性细胞系增殖困难的问题, 本研究设定了2因素3水平的处理, 分析了增殖培养基中蔗糖与肌醇对马尾松胚性细胞系增殖的影响。研究结果表明, 胚性培养物的增殖倍数在9个处理间存在极显著性差异(P<0.01), 并初步选出马尾松胚性细胞系增殖倍数较高的3个培养基: 5号(蔗糖20 g•L^-1+肌醇1.0 g•L^-1)、7号(蔗糖30 g•L^-1+肌醇0.1 g•L^-1)和8号(蔗糖30 g•L^-1+肌醇1.0 g•L^-1)。胚性细胞在以上3个培养基具有不同分化发育反应, 其中培养基5号中, 培养基胚性细胞发育较慢; 培养基7号中, 胚性细胞发育较快且能形成具有完整结构的正常早期体细胞胚; 在培养基8号中, 胚性细胞易分化形成结构不完整、形态不正常的早期体细胞胚。综合考虑胚性细胞系增殖倍数与胚性细胞分化发育两方面的因素, 在增殖培养基中添加蔗糖30 g•L^-1和肌醇0.1 g•L^-1的组合更适合马尾松胚性细胞系的增殖。

关键词：马尾松; 胚性细胞系; 蔗糖; 肌醇; 增殖

收稿：2013-12-13   修定：2014-04-01

资助：“十二五”国家“863”重点项目(2011AA100203)、国家自然科学基金青年项目(31200481)、湖南省自然科学基金(11JJ-3038)和教育部博士点基金博导类联合资助项目(2010-4321110002)。

Effects of Sucrose and Myo-Inositol on Proliferation of Embryogenic Cell Lines in Pinus massoniana

SHI Kun^1,2, YANG Mo-Hua^1,2,* , LI Zhi-Hui², ZhANG Dong-Lin^2,3, DING Gui-Jie⁴

¹Hunan Provincial Key Laboratory of Forestry Biotechnology, ²College of Forestry, Central South University of Forestry and Technology, Changsha 410004, China; ³University of Georgia, GA 30602, USA; ⁴College of Forestry, Guizhou University, Guiyang 550000, China

Corresponding author: DING Gui-Jie; E-mail: ymh163@163.com; Tel: 13875885952

Abstract:

To address the reduction of embryogenic cultures activity in masson pine (Pinus massoniana), three concentrations of sucrose and myo-inositol were applied in proliferation media with complete random design. The results indicated that proliferation rate had significantly difference among nine treatments (P<0.01). Proliferation rates of three combinations of sucrose and myo-inositol were significantly higher than those of other combinations. Within the three media, the fifth (sucrose 20 g•L^-1+myo-inositol 1.0 g•L^-1), the seventh (sucrose 30 g•L^-1+myo-inositol 0.1 g•L^-1), and the eighth (sucrose 30 g•L^-1+myo-inositol 1.0 g•L^-1), embryogenic cells development was the slowest in the fifth media. Well developed embryogenic cells and the normal somatic embryos were found in the seventh media. In the eighth media, the structure and morphology of early somatic embryos were abnormal and uncompleted. The media of sucrose 30 g•L^-1+myo-inositol 0.1 g•L^-1 should be the most suitable for proliferation of embryogenic cell lines in masson pine somatic embryogenesis.

Key words: masson pine (Pinus massoniana); embryogenic cell lines; sucrose; myo-inositol; proliferation