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苎麻(BoehmerianiveaL.Guad)是荨麻科苎麻属宿根型草本植物,主要分布在热带和亚热带地区,为我国特产,是重要的纺织纤维和饲料作物。由于苎麻为杂合体,其遗传背景十分复杂,有性杂交等常规方法难以取得育种上的突破,因此,探讨利用基因工程技术改良苎麻品种具有重要意义。本研究以苎麻品种圆叶青叶盘为外植体,选用MS培养基为基本培养基,附加激素BA、IAA、GA3和NAA,通过研究不同激素浓度组合对外植体愈伤诱导及不定芽分化的影响以及不同激素配比对生根的作用,建立了较好的遗传转化再生体系。试验结果表明圆叶青叶盘最适诱导愈伤培养基为MS+6-BA2.0mg/L+GA30.4mg/L+IAA0.2mg/L+蔗糖30g/L+琼脂7.5g/L;分化培养基为MS+6-BA2.0mg/L+GA30.4mg/L+蔗糖30g/L+琼脂7.5g/L;小苗的最佳生根培养基为MS+NAA0.2mg/L+IAA0.1mg/L+蔗糖30g/L+琼脂7.5g/L。在抗生素浓度梯度筛选实验中获得50mg/L卡那霉素可以抑制叶盘的分化;40mg/L卡那霉素可以抑制再生植株的生根。筛选出的遗传转化再生体系,为进一步利用基因工程技术对苎麻进行遗传改良打下基础。
Ramie (Boehmerianivea L.Guad) is a Urticaceae ramie is a perennial herb, mainly in the tropical and subtropical regions, for our specialty, is an important textile fiber and forage crops. Because ramie is a heterozygote, its genetic background is very complicated, and conventional methods such as sexual crossbreeding are hard to achieve breeding breakthrough. Therefore, it is of great significance to explore the improvement of ramie varieties by genetic engineering. In this study, the ramie leaf discs were used as explants. MS medium was selected as the basic medium and the hormones BA, IAA, GA3 and NAA were added. The effects of different hormone combinations on callus induction and adventitious bud differentiation The effects of different hormones on rooting and the establishment of a better genetic transformation system. The results showed that the optimal medium for callus induction of leaf discs was MS + 6-BA 2.0 mg / L + GA30.4 mg / L + IAA 0.2 mg / L + sucrose 30 g / L + agar 7.5 g / L and differentiation medium The best medium for rooting was MS + NAA0.2mg / L + IAA0.1mg / L + sucrose 30g / L and sucrose30g / L + GA3.4mg / L + agar 7.5 g / L. In the antibiotic concentration gradient screening experiment obtained kanamycin 50mg / L can inhibit leaf disc differentiation; 40mg / L kanamycin can inhibit the regeneration of plant roots. The genetic transformation and regeneration system screened out laid the foundation for further genetic improvement of ramie using genetic engineering technology.