南京林业大学学报(自然科学版) ›› 1991, Vol. 15 ›› Issue (03): 31-36.doi: 10.3969/j.jssn.1000-2006.1991.03.006

• 研究论文 • 上一篇    下一篇

杨树细胞悬浮培养及体细胞胚胎发生的研究

王影;黄敏仁;陈道明;许农   

  1. 南京林业大学林学系;南京林业大学林学系;南京林业大学林学系;南京林业大学林学系
  • 出版日期:1991-09-18 发布日期:1991-06-18

A STUDY ON CELL SUSPENSION CULTURE AND SOMATIC EMBRYOGENESIS OF POPULUS SIMONII

Wang Ying Huang Minren Chen Daoming Xu Nong   

  1. Department of Forestry
  • Online:1991-09-18 Published:1991-06-18

摘要: <正>通过调整培养基中还原态氮的含量和变换使用不同浓度的2,4-D,以小叶杨无菌苗的茎段作为外植体,诱导并建立了愈伤组织细胞无性系,将其放入液体培养基中进行振荡培养,建立起分散程度较好的悬浮细胞系。培养基中加入MES有利于pH的稳定,培养基的pH变化与细胞生长速率之间有密切关系。将悬浮培养产生的胚性愈伤组织转移至固体培养基上,经诱导产生大量胚状体。不同的激素浓度、不同的无性系和不同的渗透势,胚状体的发生能力表现出明显的差异。通过细胞学观察,胚状体的发育具有胚胎发生的全部过程。

Abstract: By regulating NH+4 and 2, 4-D content in medium, Populus simoom callus lines were established from its stems. Those callus produced a large number of friable sub-callus on its surface on the callus maintenance medium, which contains 4mg/L 2,4-D. So far, the callus lines could be subcultured. Five suspension cell lines were established from 3-month-old embrycgenic friable callus in liquid medium. The medium, which contains mineral salts, vitamius and inositol of MS medium and 2mg/L 2,4-D, 500mg/L casein hydrolysat, 1. Og/L 2-(N-morpholino) ethanesulfonic acid, 3% coconut milk, and 3 % sucrose, was optimal for growth of the suspension cells. Addition of CM, CH and MES into the liquid medium was benifical for the cells growth. There was a direct relationship between the growth rate of suspension cells and medium pH changes. The growth rate was slow when culture medium pH decreased within initial 3 days. Then it was fast when medium pH rose. The medium, which contained 2. 0mg/L BA and 0. 01 -0. 1mg/L NAA, could produce a large number of somatic embryogenesis from embryogenetic cell complex. The difference of ability is somatic embryogenesis among different cells lines was very remarkable. And different osmotic pressures could affected the produce of somatic embryogenesis. Plant regeneration on 1/2MS medium.