甜荞高效离体再生与GUS基因瞬时表达分析

Regeneration of Buckwheat Plantlets(Jinqiao 1 Cultivars) and Transient GUS Gene Expression in vitro

以晋荞1号叶片为外植体,研究了不同类型(2,4-D,6-BA和NAA)及不同剂量的生长调节剂对愈伤组织诱导、幼苗分化及植株再生的影响;利用Ca MV35S启动子驱动GUS基因PBI121质粒转化EHA105农杆菌侵染愈伤组织,进行GUS染色鉴定。结果表明,愈伤组织诱导培养基中加入3.0 mg/L 2,4-D和1.0 mg/L 6-BA诱导愈伤组织的效果最好,出愈率达到69.5%;再分化培养基中加入1.0 mg/L的6-BA和1.0 mg/L的NAA分化率最高(47.0%);农杆菌侵染愈伤组织经过GUS染色鉴定证明为阳性。晋荞1号高效离体再生体系的建立为下一步转基因选育优良品种提供了理论依据和技术指导。

We developed an in vitro regeneration system from Fagopyrum esculentum （Jinqiao 1 cultivars） leaves and studied the influence of different types and concentrations of growth regulators （2,4-D, 6-BA, NAA ） on the induction of callus and the differentia- tion and regeneration of plantlets. We utilized EHA105 that had been transformed with PBII21, which contained the CaMV35S promoter controlling the GUS gene, to infect callus and studied gene expression by GUS staining. The highest callus induction rate （69.5%） was obtained when leaves were incubated on MS solid medium supplemented with 3.0 mg/L 2,4-D and 1.0 mg/L 6-BA. Supplying 1.0 mg/L 6-BA and 1 mg/L NAA to the culture medium induced adventitious shoots by up to 47.0%. When the callus was co-cultured with E- HA105 （transformed with PBI121 ）, GUS gene was active, as indicated by GUS staining. These results provide an improved regeneration and transformation system for F. esculentum, which will be useful for utilizing transformation technologies and breeding new buckwheat varieties.