论文部分内容阅读
随着植物抗逆性研究和植物转基因技术的发展,通过异源目的基因转化培育耐盐碱苜蓿品种的研究已引起人们的关注,植物受体高频再生体系的建立是异源转化高效的基础。选取新疆大叶紫花苜蓿种子萌发5～7d无菌苗的子叶、下胚轴及根为外植体,诱导愈伤培养基为MS+2,4-D0.1～3.0mg/L(8种不同水平)或MS+2,4-D2.0mg/L+KT0.01～0.5mg/L(10种不同水平),诱导芽培养基为MS+6-BA0.5mg/L+NAA0.05mg/L,生根培养基为MS。结果表明,外植体在MS+2,4-D2.0mg/L+KT0.2mg/L培养基中能够产生状态较好可再分化的愈伤组织,子叶、下胚轴、根的平均出愈率分别为93.1%、100%、100%。愈伤组织在MS+6-BA0.5mg/L+NAA0.05mg/L培养基中培养40～80d中均可分化芽,子叶、下胚轴、根的芽平均分化率为50%、78%、50%,将2cm以上的芽转入MS培养基中诱导生根,14d后,生根的小植株炼苗移入花土中,成活率达90%以上。子叶、下胚轴、根在该体系中均能获得再生植株,根也是一种较好的植株再生材料,以根为外植体进行植株再生的研究报道还较少。
With the research of plant stress resistance and the development of plant transgenic technology, the research of cultivating salt-tolerant alfalfa varieties by heterologous gene transformation has drawn much attention. The establishment of high-frequency plant regeneration system is the basis of efficient heterologous transformation . Cotyledons, hypocotyls and roots of the sterile seedlings of Xinjiang alfalfa (Medicago sativa) germinated for 5-7 days were selected as explants. The callus induction medium was MS + 2,4-D0.1 ~ 3.0 mg / L Different levels) or MS + 2,4-D2.0mg / L + KT0.01 ~ 0.5mg / L (10 different levels), the induction buds culture medium was MS + 6-BA0.5mg / L + NAA0.05mg / L, rooting medium for MS. The results showed that the explants could produce better redifferentiable calli, cotyledons, hypocotyls and roots in MS + 2,4-D2.0mg / L + KT0.2mg / L medium The cure rates were 93.1%, 100% and 100% respectively. Average callus bud differentiation rate was 50% and callus 78% in buds, cotyledons, hypocotyls and roots at 40 ~ 80d in MS + 6-BA0.5mg / L + NAA0.05mg / , 50%. The shoots of more than 2cm were transferred into MS medium to induce rooting. After 14 days, the rooted plantlets were moved to flower soil with the survival rate of more than 90%. Cotyledons, hypocotyls, and roots can be regenerated in this system. Roots are also a good plant regeneration material. There are few reports about the regeneration of plants with roots as explants.