摘要
采用果用香蕉（Ｍｕｓａ ｓｐｐ．）的薄片外植体作为转化受体；通过对ＧＵＳ基因瞬间表 达的研究，找出了较适合的轰击条件和外植体培养条件．研究表明，高渗处理对转化的影响 较大，轰击前后对外植体进行高渗处理的瞬间表达率（１４．１１％）是对照（３．６８％）的３．８６ 倍。预培养外植体转化的瞬间表达率（１７．５％～４２．１１％）普遍高于没有预培养的外植体 （１２．２０％），并以预培养６ｄ为好．最适压力和射程分别为７．５８４ ＭＰａ，９ｃｍ．ＤＮＡ沉淀剂的组成以ＣａＣｌ_２和亚精胺同时存在较佳、对不同ｐＨ（５．０，１０．０）下的Ｃａ ＮＯ_３）_２或ＣａＣｌ_２对瞬间表 达率的影响进行研究发现正常ｐＨ（５．０）的ＣａＣｌ_２优于其他组合．利用最佳条件组合，瞬间表达率可达４８．７２％，初步建立了果用香蕉薄片培养的转化体系。
Using the transient expression of GUS gene, we optimized the physical and biological condition of bombardment for micro-cross section of fruit banana (Musa spp.). The results suggested that osmotic treatment influence expression obviously. Transient expression rates of control and osmotic treatment from 4 h before to 16 h after the bombardment were 3 .68% and 14. 11 %, respectively. Osmotic treatment caused 3. 86 folds increase of the transient expression. The GUS expression can be improved several folds by the preculture of explants. The best pressure and the best distance from micro- carrier to target cell were 7.584 MPa and 9 cm, respectively. Calcium ions and sperimidine were nec- essary to the transient expression. The effect of CaCl_2 or Ca (NO_3 )_2 at either pH 5. 0 or 10. 0 on GUS expression were also studied. CaCl_2 solution at pH 5. 0 in the mixture was superior to the others. Under the above optimal conditions, 48.72% of transient GUS expression rate could he obtained.
出处
《中山大学学报（自然科学版）》
CAS
CSCD
北大核心
2000年第2期87-91,共5页
Acta Scientiarum Naturalium Universitatis Sunyatseni
基金
广州市科委高科技资助项目!(960448252)
关键词
基因枪法
香蕉
薄片外植体
particle bombardment
Musa spp.
micro-cross section
