‘温莎甜荞’再生体系建立的研究

Research on the Regeneration System Establishment of'Windsor Sweet Buckwheat'

为了荞麦的进一步研究和转基因体系的建立以及农杆菌介导转化的研究奠定基础,为基因工程改良荞麦芦丁生物合成提供了理论依据及技术保证。以‘温莎甜荞’为研究材料,建立组织培养再生体系,以子叶和下胚轴为外植体,基本培养基为MS,添加不同浓度的生长素和细胞分裂素,设计不同激素配比组合,在相同的培养条件下,研究‘温莎甜荞’子叶和下胚轴愈伤组织诱导和生长特征,筛选出诱导‘温莎甜荞’愈伤组织的最适培养基,为基因工程提供参考和理论依据。研究表明:‘温莎甜荞’子叶产生愈伤组织的最佳培养基为MS+2,4-D2mg/L+6-BA0.4mg/L。诱导芽的最佳培养基为MS+6-BA1.5mg/L+IBA0.3mg/L。下胚轴诱导愈伤组织的最佳培养基为MS+2,4-D3mg/L+6-BA0.1mg/L。诱导芽的最佳培养基为MS+6-BA3mg/L+IBA0.5mg/L。生根的最佳培养基为1/2MS。

The research as the transgene system establishment,the agricultural bacillus leads transformed the research to lay the foundation which has further provided the theory basis and the technical guarantee for the genetic engineering improvement of the buckwheat biosynthesis.Selecting the buckwheat which is’windsor sweet buckwheat’for establishing regeneration system and antibiotic screening system,We know explants are cotyledon and hypocotyls.The basic culture medium is called MS.We can add into the different auxin and cytokinin under the different condition auxin and cytokinin under the different condition,we studied’Jiujiang bitter buckwheat’of the seed leaf and under the hypocotyls injuries the organization the induction as well as does not decide the bud differentiation and the growth situation.We can establish the best regeneration system through researching the different explants,as the transgene system establishment has further provided the theory basis.The research indicates that the’windsor sweet buckwheat’seed leaf produces injuries the organization the best culture medium is MS ＋ 2,4-D 2 mg/L ＋ 6-BA 0.4 mg/L.The induction bud’s best culture medium is MS ＋ 6-BA 1.5 mg/L ＋ IBA 0.3 mg/L.The next hypocotyls induction injuries the organization the best culture medium is MS ＋ 2,4-D 3 mg/L ＋ 6-BA 0.5 mg/L,the induction bud’s best culture medium is MS ＋ 6-BA 0.05 mg/L ＋ IBA 0.1 mg/L.The rooting induction the best culture medium is 1/2 MS.