利用GUS基因瞬时表达探索佛手叶盘基因枪转化参数  

Exploration of the Transformation Parameters of the Leaf Discs of Bergamot by Particle Bombardment in Virtue of Transient Expression of GUS Gene

作　　者：周春丽[1] 郭卫东[2] 王德解[1] 余初浪[2] 路梅[2] 李玉萍[1] 

机构地区：[1]江西科技师范学院生命科学学院 [2]浙江师范大学生命与环境科学学院,浙江金华321004

出　　处：《西北植物学报》2005年第11期2145-2150,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：浙江省科技计划项目资助课题(2003C32043)

摘　　要：佛手是芳香科枸橼类植物,有很好的药用和观赏价值,但其品种较为单一,对其进行遗传改良和品种选育显得十分必要.为了将基因枪转化方法应用于佛手的遗传改良和品种选育,本实验以GUS基因为报告基因,利用基因枪方法将其转入金华佛手外植体内,通过检测GUS基因在佛手叶盘中的瞬时表达情况,分析了外植体的幼嫩程度、射程、氦气压力、真空度、轰击次数等参数对GUS瞬时表达的影响,并研究了最佳检测时间.结果表明,以幼叶为受体,在射程为6 cm、氦气压力为7.584×106Pa,真空度为88.046 kPa条件下,2 d后可检测出GUS基因的最佳表达活性.轰击次数多少对GUS基因瞬时表达影响不明显.Bergamot is a plant of high medicinal and ornamental values that belongs to Citrus medica L, var. sarcodactylis （Noot.） Swingle,but does not have many varieties. Thus it is necessary to conduct its genetic improvement and variety breeding. In order to apply particle bombardment to the genetic improvement and variety breeding of bergamot,this study ,using GUS gene as the report gene,transformed GUS gene into the explants of Jinhua bergamot by particle bombardment ;the transient expression of GUS gene in the leaf discs of bergamot were examined to analyze the effects of explant tenderness ,gunshot, helium pressure, vacuum degree and bombardment frequency on the expression of GUS gene,and explore the optimal examination time. The results revealed that under the circumstances that the explants were young leaves, the gunshot was 6 cm,the helium pressure was 7. 584×10^6 Pa and the vacuum degree was 88. 046 kPa,the most aetive transient expression of GUS gene eould be deteeted two days after partiele bombarding, The bombarding frequeney did not show signifieant influenee on the transient expression of GUS gene.

关 键 词：佛手 基因枪 GUS基因 瞬时表达 

分 类 号：Q785[生物学—分子生物学] Q919.752.7[天文地球—古生物学与地层学]