《植物生理学报》 2018, 54(4): 686-692

抗性赤松体胚发生细胞结构观察与增殖条件的优化

潘珺, 吴小芹^*, 许建秀, 叶建仁

南方现代林业协同创新中心, 南京林业大学林学院, 南京210037

通信作者：吴小芹；E-mail: xqwu@njfu.edu.cn

摘　要：

为了更有针对性地优化原生抗性赤松体胚发生培养体系, 本研究通过体式镜定性观察种室中的合子胚分布、胚性胚柄细胞结构变化, 通过Image-Pro Plus软件定量分析胚性细胞团在不同生长调节物质配方下胚头胚柄的比例变化, 了解体胚发生的营养需求。结果发现, 抗性赤松裂生多胚更易形成胚性细胞团; 改变原有培养条件细胞团会出现胚头胚柄分离、胚头细胞颜色加深的反应; 使用低浓度的混合生长调节物质配比(0.1 mg·L^-1 2,4-D+1 mg·L^-1 NAA+0.5 mg·L^-1 6-BA)能够提高体胚增殖速度; 表明抗性赤松胚性细胞团增殖速度加快的内在机理为增加胚性细胞团中胚头与胚柄的比例。

关键词：赤松; 诱导; 增殖; 多胚现象; 胚性胚柄细胞团; Image-Pro Plus

收稿：2017-12-22   修定：2018-03-22

资助：江苏省科技支撑计划(BE2014405)和江苏高校优势学科建设工程资助项目(PAPD)。

Early stage observation and conditional optimization for proliferation of resistant Pinus densiflora somatic embryogenesis

PAN Jun, WU Xiao-Qin^*, XU Jian-Xiu, YE Jian-Ren

Collaborative Innovation Center of Southern Modern Forestry, Forestry Department, Nanjing Forestry University, Nanjing 210037, China

Corresponding author: WU Xiao-Qin; E-mail: xqwu@njfu.edu.cn

Abstract:

In order to optimize the conditions of somatic embryogenesis of resistant Pinus densiflora, this study investigated the zygotic embryo distribution in the seed chamber and the changes of embryonic suspensor structure by qualitative observation with stereoscopic mirror; and investigated the change of embryo and suspensor ratio in different hormone condition for quantitative observation by Image-Pro Plus software, aiming at the nutritional needs at this stage. The results showed cleaved polyembryony were more easily differentiate into embryogenic cells. The change of the original culture environment would separate embryo from suspensor and the embryo would accumulate more material. The medium (0.1 mg·L^-1 2,4-D+1 mg·L^-1 NAA+0.5 mg·L^-1 6-BA) would improve the proliferatin speed. It improved the mechanism of improve proliferation in resistant P. dnsiflora was the increase of the embryo and suspensor ratio.

Key words: Pinus densiflora; initiation; proliferation; poly embryos; embryonic suspensor cell mass; Image-Pro Plus