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裂叶垂枝桦离体培养与植株再生

发布时间:2019-01-11 21:18  文章来源:笔耕文化传播
【摘要】:垂枝桦(BetulapendulaRoth.)是桦木科桦木属落叶乔木。目前,桦木科植物大部分都采用实生繁殖方式,但实生繁殖易导致后代产生大量遗传变异,为获得更多后代遗传性状一致植株,植物组织培养成为最佳的繁殖方式之一。本试验以裂叶垂枝桦木质化茎段、幼嫩茎段和幼嫩叶片为试验材料,经过初代培养、继代培养、增殖培养、生根培养、移栽和驯化,最后建立了裂叶垂枝桦离体快繁再生体系。通过试验得出以下结论:(1)在建立无菌培养体系中,用70%的酒精30s,0.1%HgCl2对裂叶垂枝桦木质化茎段(6、8、10、12、15 min)和幼嫩茎段(2、3、4min)进行消毒处理,筛选出裂叶垂枝桦木质化茎段最佳消毒时间为10 min,幼嫩茎段最佳消毒时间为3 min。(2)初代培养中以MS、WPM、1/2MS三种培养基为基本培养基,分别附加不同浓度的NAA、2,4-D、6-BA和GA3对裂叶垂枝桦不同外植体进行离体培养,筛选出裂叶垂枝桦木质化茎段最适培养基为:MS + 0.2 mg/L 6-BA + 0.2 mg/L NAA + 0.2 mg/L GA3+20 g/L蔗糖+6g/L琼脂,分化率可达83.3%;幼嫩茎段最适培养基为:MS+0.5 mg/L 6-BA + 0.05 mg/L NAA + 0.2 mg/L GA3 + 20 g/L蔗糖 + 6 g/L 琼脂,其分化率可达73.3%;裂叶垂枝桦叶片诱导最适培养基及激素种类浓度为:1/2 MS+ 0.5 mg/L 6-BA+0.2 mg/LNAA+2.0mg/L2,4-D + 20 g/L 蔗糖 + 6g/L 琼脂,出愈率最高为 82%。(3)对裂叶垂枝桦无菌苗进行增殖培养时分别选择MS和WPM培养基,附加不同种类及浓度的激素,筛选出裂叶垂枝桦增殖培养最适培养基和激素组合为:WPM + 1.0 mg/L 6-BA + 0.5 mg/L NAA + 20 g/L蔗糖+ 6 g/L琼脂,增殖系数可达到4。(4)将裂叶垂枝桦无菌苗接种在1/2 MS培养基上,添加NAA和IBA,浓度分别为0、0.05、0.1、0.5 mg/L,筛选出裂叶垂枝桦无菌苗生根最适培养基和激素组合为1/2 MS+ 0.1 mg/LNAA;将生根的无菌苗移栽至蛭石:珍珠岩:草炭土为1:1:2的已灭菌的基质中,15 d后成活率达到80%以上。
[Abstract]:Birch (BetulapendulaRoth.) It is a deciduous tree of the family Betulaceae. At present, most birch plants adopt the method of seed reproduction, but seed propagation is easy to cause a large amount of genetic variation in offspring. In order to obtain more progeny plants with the same genetic traits, plant tissue culture is one of the best breeding methods. In this experiment, the lignified stem segments, young stem segments and young leaves of Betula platyphylla were used as experimental materials. After primary culture, subculture, multiplication culture, rooting culture, transplanting and acclimation, the regeneration system of Betula platyphylla in vitro was established. The following conclusions were obtained from the experiment: (1) in the establishment of aseptic culture system, 70% alcohol 30sCl2 was used to sterilize the lignified stem segments of Betula lobata (6 ~ 810 ~ 10 ~ (12) min) and young stem segments (2 ~ 3 ~ 3 ~ (4 min). The best disinfection time for the lignified stem segment of Betula platyphylla was 10 min,. The best disinfection time was 3 min. (2) the three kinds of MS,WPM,1/2MS medium were used as the basic culture medium and the different concentrations of NAA, were added respectively in the primary culture. In vitro culture of different explants of Betula lobata was carried out with 2-D 6-BA and GA3. The optimum medium for lignetized stem segments of Betula lobata was selected as follows: MS 0.2 mg/L 6-BA 0.2 mg/L NAA 0.2 mg/L GA3 0.2 g / L sucrose 6g/L Agar with a differentiation rate of 83.3%; The optimum medium for young stem segments was: MS 0.5 mg/L 6-BA 0. 05 mg/L NAA 0. 2 mg/L GA3 20 g / L sucrose 6 g / L Agar, and the differentiation rate was 73. 3%; The optimum medium and hormone concentration of 1 / 2 MS 0.5 mg/L 6-BA 0.2 mg/LNAA 2.0 mg / L 2-D 4-D 20 g / L sucrose 6g/L Agar were suitable for leaf induction of Betula lobata. The highest callus rate was 82. (3) MS medium and WPM medium were selected for multiplication and culture of aseptic plantlets of Betula lobata, supplemented with different kinds and concentrations of hormones. The optimum culture medium and hormone combination were obtained as follows: WPM 1.0 mg/L 6-BA 0.5 mg/L NAA 20 g / L sucrose 6 g / L Agar. The multiplication coefficient can reach 4. (4) the sterile plantlets of Betula platyphylla cleft were inoculated on 1 / 2 MS medium, and the concentration of NAA and IBA, were 0 0. 05% 0. 1 0. 1 mg/L, respectively. The optimum rooting medium and hormone combination for aseptic plantlets of Betula platyphylla was 1 / 2 MS 0.1 mg/LNAA;. The rooting sterile seedlings were transplanted into sterilized substrates with vermiculite, perlite and peat soil at 1:1:2. The survival rate was more than 80% after 15 days.
【学位授予单位】:沈阳农业大学
【学位级别】:硕士
【学位授予年份】:2017
【分类号】:S792.15

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