甘蓝小孢子培养技术体系的优化与植株再生技术研究

发布时间：2018-03-29 19:28

本文选题：甘蓝　切入点：小孢子培养　出处：《西北农林科技大学》2017年硕士论文

【摘要】：探究甘蓝小孢子培养效果,非子叶型胚再生不定芽和不定芽成苗的影响因素,提高甘蓝小孢子胚状体诱导和再生植株效果,增强DH植株生长势和翌年抽苔率,加快甘蓝单倍体育种进程。本试验研究了整枝方式,NLN-14培养基中添加外源激素褪黑素(MT)和对氯苯氧异丁酸(PCIB)以及生化物阿拉伯半乳聚糖(AG)的浓度对甘蓝诱导出胚效果的影响。MS培养基中分别添加KT和NAA浓度不同比例、PCIB浓度和AgNO_3浓度对非子叶型胚分化不定芽的影响。MS培养基中添加MT和NAA浓度对甘蓝胚状体不定芽成苗的影响。研究获得以下结果:1.小孢子供体植株抽苔显蕾时,整枝保留20%、50%和80%的一级分枝,花蕾中单核靠边期小孢子所占比率和出胚率均比对照显著性增加。整枝保留50%的一级分枝,与对照相比,花蕾中小孢子发育处于单核靠边期所占比率增加122.38%,出胚率增加3.2胚/蕾,子叶型胚比率增加0.77%。2.NLN-14小孢子培养基中添加适宜的MT、AG和PCIB浓度比对照能显著性地提高胚状体诱导效果,且不同添加剂对不同材料影响效果不同。添加1.0μmol/LMT,早熟材料新绿洲出胚率最高达19.5胚/蕾;添加20mg/LAG,中熟材料C64出胚率最高达9.8胚/蕾;添加40μmol/LPCIB,晚熟材料C69出胚率最高达18.5胚/蕾。3.材料惠2非子叶型胚MS分化培养基中分别添加1.0mg/L KT和0.02mg/L NAA促使不定芽分化率高达26.67%,比对照提高20.0%;添加40μmol/L PCIB促使不定芽分化率高达16.67%;添加2~10mg/L AgNO_3不利于非子叶型胚转绿,但适宜浓度能促进转绿胚分化不定芽。4.不定芽成苗MS培养基中添加5μmol/L MT和0.1mg/L NAA比对照能显著性地促进DH植株生根和快速健壮生长,且对DH-A、DH-B和DH-C材料均有促进作用,但材料间差异性不显著;三种材料DH植株翌年抽苔率分别比对照提高33.4%,20.0%和20.0%。
[Abstract]:To investigate the effect of microspore culture on the regeneration of adventitious buds and adventitious buds of non-cotyledonous embryos, to improve the induction and regeneration of microspore embryoid, to enhance the growth potential of DH plants and the rate of sprouting and coating the next year. The experiment was conducted to study the effects of the concentration of exogenous hormones melatonin (MTM), p-chlorophenoxy isobutyrate (PCIBB) and the biodegradable arabinogalactan (AGG) on the induction of embryos from cabbage. Effects of different concentrations of KT and NAA on the differentiation of adventitious buds from non-cotyledonous embryos. Effects of MT and NAA on shoot formation of Brassica oleracea embryoid adventitious buds in MS medium. The following results were obtained: 1. When the microspore donor plants were moss and showed buds, The percentage of microspore and embryo production in bud in the floral bud was significantly higher than that in the control, and 50% of the first branch was retained in the pruning branch, compared with the control, the percentage of primary branches in the bud was 50% and 80% respectively, and the percentage of microspore in the bud was significantly higher than that in the control. The percentage of microspore development in the uninucleated stage increased 122.38%, the embryogenesis rate increased 3.2 embryo / bud ratio, the cotyledon embryo ratio increased in 0.77%.2.NLN-14 microspore medium, and the suitable concentration of MTAG and PCIB could significantly improve the embryoid induction effect compared with the control. The effects of different additives on different materials were different. When 1.0 渭 mol / L MTT was added, the embryogenic rate of the early maturing material reached 19.5 embryos / buds in the new oasis, and that of the medium maturing material C64 reached 9.8 embryos / buds when 20mg / L tag was added. When 40 渭 mol / L LPCIBS was added, the embryo yield of late maturing material C69 was up to 18.5 / bud. 3. Addition of 1.0mg/L KT and 0.02mg/L NAA to MS differentiation medium of non-cotyledonous embryos of line Hui2 promoted adventitious bud differentiation rate up to 26.67%, which was 20.0% higher than that of control, and 40 渭 mol/L PCIB promoted adventitious bud differentiation. The differentiation rate was up to 16.670.The addition of 2~10mg/L AgNO_3 was not conducive to the green transformation of non-cotyledonous embryos. However, the suitable concentration could promote the differentiation of adventitious buds. 4. Addition of 5 渭 mol/L MT and 0.1mg/L NAA to MS medium could significantly promote the rooting and rapid and robust growth of DH plants, and could also promote the growth of DH-An DH-B and DH-C materials. But there was no significant difference among the three materials, and the bolting rate of DH plants in the following year was 33.4% and 20.0% higher than that of the control, respectively.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S635

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