四川农业大学学报 ›› 2006, Vol. 24 ›› Issue (01): 1-6,24.doi: 10.16036/j.issn.1000-2650.2006.01.001

• 研究论文 •    下一篇

农杆菌介导转化小麦几个影响因素的再研究

张志清1,2, 郑有良1, 王丕武3, 魏育明1, 颜泽洪1, 刘虹1   

  1. 1. 四川农业大学 小麦研究所, 四川 都江堰 611830;
    2. 四川农业大学 信息与工程技术学院, 四川 雅安 625014;
    3. 四川农业大学 吉林农业大学 生物中心, 长春 130118
  • 收稿日期:2005-10-19 出版日期:2006-03-31 发布日期:2017-03-04
  • 基金资助:
    四川省科技厅应用基础项目(2002A008);四川省教育厅重点项目(03JY029-028-2863)

Some Factors Affecting Agrobacterium-mediated Wheat Transformation

ZHANG Zhi-qing1,2, ZHENG You-liang1, WANG Pi-wu3, WEI Yu-ming1, YAN Ze-hong1, LIU Hong1   

  1. 1. Triticeae Research Institute, Sichuan Agricultural University, Dujiangyan 611830, Sichuan, China;
    2. College of Information and Engineering Technology, Sichuan Agricultural University, Yaan 625014, Sichuan, China;
    3. Biotechnology Center, Jilin Agricultural University, Changchun 130118, China
  • Received:2005-10-19 Online:2006-03-31 Published:2017-03-04

摘要: 选用四川省近年来选育的3个小麦新品种川农16、川麦32和川育16幼胚及愈伤组织,针对农杆菌介导遗传转化中筛选剂种类、剂量,预处理方式,侵染时间等几个重要影响因素进行了研究。结果表明:Kan不能抑制幼胚愈伤的生长,平均愈伤诱导率达87%,愈伤组织生长良好。川农16的幼胚在含不同G418剂量的MS2中生长受抑制,但愈伤诱导率不随浓度变化而变化,说明G418不能很好地抑制住幼胚生长,未能筛选出合适的选择浓度。川农16和川麦32的幼胚在含不同L-PPT剂量的MS2培养基中,生长受到明显的抑制,L-PPT浓度为3mg/L时就能完全抑制幼胚的生长,因此在小麦遗传转化诱导愈伤和继代阶段L-PPT合适的筛选浓度是2~3mg/L。侵染时间对不同基因型幼胚愈伤诱导率表现不一致,川农16在1h时表现较高的诱导率;侵染前对幼胚进行高渗处理和乙酰丁酰酮(AS)处理后,愈伤诱导率与未经处理相比并未表现出明显的增加趋势,但基因型对不同处理的反应也表现较大差异。

关键词: 小麦, 农杆菌转化, 筛选剂, 预处理, 侵染时间

Abstract: Three selectable agents, kanamycin(Kan), geneticin(G418) and phosphinothricin(PPT) were tested for optimal select concentrations for callus induction derived from immature embryos of three genotypes, Chuannong16, Chuanmai32 and Chuanyu16. The results indicate that kanamycin is not available to wheat transformation, because higher survival frequency of the explants(87%) is observed in this study. Although different concentrations of G418 retrain the growth of callus derived from immature embryo of Chuannong16, the regular relationship between the callus survival frequency and G418 different concentrations is not detected. So the suitable select concentration of G418 is not obtained. L-PPT with concentration 3mg/L can restrain completely growth of callus derived from Chuannong16 and Chuanyu16, and this level can be used for the selection of bar gene. Immature embryos from these materials are transformed mediated by Agrobacterium strain EHA101, the highest survival frequency is obtained from the immature embryos of Chuannong16 infected for 1h. The clearly positive correlation between two preculture methods of higher osmotic or acetosyringone(AS) treatment and explant survival frequency is not detected. But the different tissue culture response of genotypes for different treatments is observed.

Key words: wheat, Agrobacterium-mediated transformation, selectable agent, pre-culture, infection time

中图分类号: 

  • S512.1