普通小麦品种Alondra＇s遗传转化体系的建立

小麦（Triticum aestivum）幼胚愈伤组织的诱导和分化再生有高度依赖基因型特征。为了建立和优化Alondra’s的高效再生及遗传转化体系,为小麦遗传转化提供更多的受体基因型,以Alondra’s的幼胚为外植体,研究了培养基种类、不同激素配比等对其幼胚愈伤组织诱导及再生的影响。结果表明,在使用N6培养基时,添加3mg·L^-1的2,4-D并附加1000mg·L^-1的CH对愈伤组织的诱导效果较好;添加4mg·L^-1的ZT、不附加IAA对愈伤组织的分化效果最好。通过构建植物表达载体pCAMBIA1301-220.6,利用基因枪法将HYG基因导入Alondra’s幼胚愈伤组织中,以建立Alondra’s的高效遗传转化体系。结果在含100mg·L^-1潮霉素的选择培养基上进行筛选、分化,获得了30棵抗性植株。经PCR检测,其中5株为阳性转基因植株,转化率为0.5%。Alondra＇s遗传转化体系的建立丰富了小麦遗传转化的基因型,为小麦品种的转基因改良和在不同背景下研究基因的功能奠定了良好的基础。

Establishment of High-efficiency Genetic Transformation System for the Wheat Variety Alondra's

Callus induction and regeneration of wheat is highly genotype dependent.To establish an optimal system for callus induction,regeneration and highly efficient genetic transformation of the wheat variety Alondra＇s,we studied the effects of culture medium,hormones on the callus induction and plant regeneration of immature embryos.Use of the N6 medium,3 mg·L^-1 2,4-D,together with 1 000 mg·L^-1 CH,achieved the best callus induction,and use of 4 mg·L^-1 ZT without indoleacetic acid had the highest regeneration frequency.The vector pCAMBIA1301-220.6 was constructed and transformed into Alondra＇s by particle bombardment.Thirty hygromycin-resistant plants were selected by use of 100 mg·L-1 hygromycin and regenerated.Among them,5 plants were identified to be positive by PCR,with transformation frequency of 0.5%.This system has further enriched the wheat variety Alondra＇s for transformation and will be helpful for both genetic improvement and gene function analysis in various backgrounds by genetic transformation.