裂葉垂枝樺離體培養(yǎng)與植株再生
發(fā)布時間：2019-01-11 21:18
【摘要】：垂枝樺(BetulapendulaRoth.)是樺木科樺木屬落葉喬木。目前,樺木科植物大部分都采用實生繁殖方式,但實生繁殖易導致后代產生大量遺傳變異,為獲得更多后代遺傳性狀一致植株,植物組織培養(yǎng)成為最佳的繁殖方式之一。本試驗以裂葉垂枝樺木質化莖段、幼嫩莖段和幼嫩葉片為試驗材料,經(jīng)過初代培養(yǎng)、繼代培養(yǎng)、增殖培養(yǎng)、生根培養(yǎng)、移栽和馴化,最后建立了裂葉垂枝樺離體快繁再生體系。通過試驗得出以下結論:(1)在建立無菌培養(yǎng)體系中,用70%的酒精30s,0.1%HgCl2對裂葉垂枝樺木質化莖段(6、8、10、12、15 min)和幼嫩莖段(2、3、4min)進行消毒處理,篩選出裂葉垂枝樺木質化莖段最佳消毒時間為10 min,幼嫩莖段最佳消毒時間為3 min。(2)初代培養(yǎng)中以MS、WPM、1/2MS三種培養(yǎng)基為基本培養(yǎng)基,分別附加不同濃度的NAA、2,4-D、6-BA和GA3對裂葉垂枝樺不同外植體進行離體培養(yǎng),篩選出裂葉垂枝樺木質化莖段最適培養(yǎng)基為:MS + 0.2 mg/L 6-BA + 0.2 mg/L NAA + 0.2 mg/L GA3+20 g/L蔗糖+6g/L瓊脂,分化率可達83.3%;幼嫩莖段最適培養(yǎng)基為: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 瓊脂,其分化率可達73.3%;裂葉垂枝樺葉片誘導最適培養(yǎng)基及激素種類濃度為: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 瓊脂,出愈率最高為 82%。(3)對裂葉垂枝樺無菌苗進行增殖培養(yǎng)時分別選擇MS和WPM培養(yǎng)基,附加不同種類及濃度的激素,篩選出裂葉垂枝樺增殖培養(yǎng)最適培養(yǎng)基和激素組合為:WPM + 1.0 mg/L 6-BA + 0.5 mg/L NAA + 20 g/L蔗糖+ 6 g/L瓊脂,增殖系數(shù)可達到4。(4)將裂葉垂枝樺無菌苗接種在1/2 MS培養(yǎng)基上,添加NAA和IBA,濃度分別為0、0.05、0.1、0.5 mg/L,篩選出裂葉垂枝樺無菌苗生根最適培養(yǎng)基和激素組合為1/2 MS+ 0.1 mg/LNAA;將生根的無菌苗移栽至蛭石:珍珠巖:草炭土為1:1:2的已滅菌的基質中,15 d后成活率達到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.
【學位授予單位】：沈陽農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S792.15
本文編號：2407458
[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.
【學位授予單位】：沈陽農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S792.15
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