亚麻品种范妮再生体系的建立和抗生素阈值筛选

Establishment of Regenerational Flax Species FANY System and Screening of the Antibiotics Threshold

【目的】建立亚麻品种范妮器官发生途径的高频再生芽、再生根体系,确定抗生素筛选阈值,为遗传转化改良亚麻品质奠定基础。【方法】以亚麻品种范妮幼苗下胚轴为外植体,用软件SPSS17.0进行正交试验设计,在不同激素组合的20种培养基上进行愈伤和芽诱导,9种生根培养基上进行再生根诱导。用不同浓度抗生素Kan、Hyg确定适于抑制范妮下胚轴切段出愈的阈值。【结果】20种不同激素组合的培养基均能诱导出愈伤并直接得到再生芽,以1/2MS+0.5 mg/L 6-BA+0.9 mg/L KT+0.2 mg/L IAA诱导的外植体出芽块数最多,出愈率达(99.7±0.65)%,褐化率(0.83±1.44)%,出芽块数占总数的(93.2±5.89)%,诱导再生根最佳培养基组成为:1/2MS+0.3 mg/L NAA+0.05 mg/L IBA。Kan和Hyg适于范妮的筛选阈值为120和40mg/L。【结论】不同基因型的亚麻品种所需的最佳再生体系不同,通过直接器官发生途径确立的范妮高频再生体系,为加快亚麻品质改良的遗传转化和筛选工作奠定了基础。

[ Objective] Through this research, the high frequency in organogenesis regeneration system of adventitious buds, root regeneration and antibiotic selection threshold of flax species FANY was established and the foundation for genetic transformation to improve the quality of flax laid. [ Method ] The experiment was carried out with hypocotylar segments of the FANY seeding as explants, using SPSS statistiss17.0 to design the orthogonal experiment in 20 different hormone combinations callus and shoot induction medium, and 9 kinds of root induction medium, induction callus, buds, and roots. With different concentrations of antibiotics Kan, Hyg were used to determine FANY hypocotyls segments to show a higher threshold. [ Result ] All the 20 kinds of combination of medium could induce callus and buds. The optimum medium for inducing adventitious buds was 1/2MS ＋0.5 mg/L 6 - BA ＋0.9 mg/L KT ＋0.2 mg/L IAA, induction callus rate reached （99.7 -0. 65 ） % , browning rate reached （0.83 -1.44） % , and budding number reached （93.2 - 5.89 ） %. The opti- mum medium for root inducing was 1/2MS ＋ 0.3 mg/L NAA ＋ 0.05 mg/L IBA. Kan and Hyg for FANY screening threshold was 120 mg/L and 40 mg/L. [ Conclusion ] Different genotypes of flax species required different optimal culture condition of regeneration system. In this research we established the FANY direct or- ganogenesis system, laid the foundation for improving the quality of flax by genetic transformation and screen-ing efforts.