摘要
以啤酒大麦品种"晋引6号"的幼胚为转化起始材料,用基因枪法将分别携带有目的基因(TrxS)和除草剂基因(筛选基因,Bar)的两个质粒进行了共转化,同时对基因转化的相关技术和植株再生的培养方案进行了优化。结果表明,受体材料宜选用预培养15d的幼胚;在培养前2周添加1mg/LABA可抑制胚芽萌发而且有助于胚性愈伤组织的形成;1.0mg/LZT与0.1mg/LIAA激素配比可有效促进愈伤组织的分化。利用优化的培养条件,经在含3～5mg/L筛选剂PPT的培养基上筛选、再生及生根培养。共在178块抗性愈伤组织上获得11株再生植株,再生率达到6.2%,经对T0、T1、T2代PCR、nestedPCR和Southern杂交检测表明,TrxS基因已经稳定整合到大麦基因组中且遗传稳定、结构完整。
The immature embryo of beer barley cultivar Jinyin 6 was used as biolistics transferring acceptor.The plasmids with thioredoxin s gene (target gene) and bar gene (selective gene) were co-transformed into barley bombed by particle bombardment.Selecting,regeneration and rooting were carried out on media with 3~5 mg/L PPT.The effective method of transformation and plant regeneration were established.The resulted indicated that using immature embryos pre-cultured for 15 days could improve gene transformation frequency.Meanwhile it was approved that immature embryo germination was not only inhibited,but also embryonic callus development could significantly improved by culturing callus on the medium containing ABA (1.0 mg/L) for two weeks.The efficient hormone proportions were ZT 1.0 mg/L+IAA 0.1 mg/L for bud differentiation.Using the optimize condition,11 regenerated plantlets were obtainted from 178 PPT resistant calli.The regeneration frequency was 6.2%.The TrxS gene in T_0,T_1 and T_2 plants were assayed by PCR,nested-PCR and Southern blotting.The result showed that foreign TrxS gene had been integrated into the barley genomes,inherited to the progenies and structure integrity.
出处
《西北植物学报》
CAS
CSCD
2004年第9期1662-1668,共7页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家948项目
