苦荞毛状根的诱导及高频再生体系的建立

发布时间：2017-12-28 19:33

本文关键词：苦荞毛状根的诱导及高频再生体系的建立　出处：《四川农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：苦荞是一种古老的一年生双子叶草本植物,它广泛分布于世界各地,具有适应性强、耐干旱、耐贫瘠、抗盐碱等特点。苦荞起源于我国,其具有极高的营养保健功能,同时兼具优秀的食疗作用以及卓越的药用价值,是许多绿色食品和重要功能性食品的生产原料。本研究以园子荞、川荞1号和九江苦荞三种不同苦荞品种为试验材料,建立了一个高效的经由发根农杆菌诱导的苦荞毛状根培养体系。同时,深入探讨了影响毛状根诱导的诸多因素。试验发现,发根农杆菌A4对苦荞毛状根的诱导能力最好,园子荞较其他两种苦荞更易诱导苦荞毛状根的产生。经过两天的预培养,利用发根农杆菌A4菌液侵染园子荞下胚轴,其毛状根诱导效果最好,诱导率可达到83.6%。本研究还以园子荞和九江苦荞两种不同苦养品种为试验材料,利用不同外植体,不同培养基进行诱导培养,成功建立了经由体细胞胚胎途径的苦荞高频再生体系。结果表明：苦荞下胚轴的再生能力最好,园子荞的再生能力较强；最佳愈伤组织诱导培养基为MS培养基中添加2.0 mg/L 2,4-D和1.0 mg/L KT,其诱导率可达98.94%;最佳芽分化培养基为MS培养基中添加1.0 mg/L KT和2.0 mg/L 6-BA,其诱导率可达55.53%；最佳生根培养基为1/2MS培养基中添加1.0 mg/L IBA,其生根率高达86%。同时,我们还以苦荞顶端分生组织为外植体,通过直接诱导丛生芽的产生,首次建立了经由组织器官途径的苦荞高频再生体系。其最佳丛生芽诱导培养基为MS培养基中添加2.0 mg/L 6-BA和0.5 mg/L TDZ,最高芽分化率达69.05%,平均外植体出芽数最高可达18个。
[Abstract]:Tartary buckwheat is an ancient annual dicotyledonous plant. It is widely distributed all over the world, and has strong adaptability, drought resistance, barren resistance and salt and alkali resistance. Tartary buckwheat originated in China. It has very high nutritional and health functions. It also has excellent dietary therapy and excellent medicinal value. It is the raw material for many green foods and important functional foods. In this study, the garden of buckwheat chuanqiao No. 1 and Jiujiang three different varieties of tartary buckwheat as experimental materials, the establishment of an efficient via Agrobacterium rhizogenes induced by buckwheat hairy root culture system. At the same time, many factors that affect the induction of hairy root are discussed in depth. The test found that the inducing ability of Agrobacterium rhizogenes A4 on Buckwheat hairy root of the best garden, compared to the other two kinds of tartary buckwheat are more susceptible to induction of hairy roots of buckwheat. After two days of pre culture, by Agrobacterium rhizogenes A4 infection garden buckwheat hypocotyl, the hairy root inducing effect is the best, the induction rate can reach 83.6%. The study also used the garden of Jiujiang buckwheat buckwheat and two different from Buckwheat Varieties, using different explants, different media were cultured successfully established via somatic embryogenesis pathway of Tartary Buckwheat high frequency regeneration system. The results show that the regeneration of Hypocotyl of Tartary Buckwheat under the best garden buckwheat regeneration ability is strong; the addition of 2 mg/L 2,4-D and 1 mg/L KT medium was MS medium induced by the best callus, the induction rate of up to 98.94%; the best bud differentiation with 1 mg/ L and 2 KT mg/L 6-BA based MS in the culture medium. The induction rate was 55.53%; 1 mg/L IBA added the best rooting medium for 1/2MS culture medium, the rooting rate up to 86%. At the same time, we also used buckwheat top meristem as explant, and directly induced the emergence of clustered buds, and first established a high frequency regeneration system of buckwheat through tissue and organ. The best bud induction medium was MS, adding 2 mg/L 6-BA and 0.5 mg/L TDZ, the highest bud differentiation rate was 69.05%, and the average explant sprouting number reached 18.
【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S517

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