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新疆雪莲组培快繁体系优化的研究
傅晓春
学位类型硕士
导师王晓军
2007-06
学位授予单位中国科学院研究生院
学位授予地点北京
学位专业有机化学
关键词新疆雪莲 高频再生 条件优化 瓶外生根
摘要

本文通过对新疆雪莲组培快繁过程中不定芽发生、增殖、复壮和生根等多种影响因子的研究,建立了稳定、重复性好的新疆雪莲植株再生体系,并首次实现了新疆雪莲的瓶外生根,简化了组培快繁的程序,缩短了快繁周期,提高了试管苗移
栽成活率,为新疆雪莲的大规模人工种植提供优良种苗奠定了技术基础。建立了稳定、重复性好的新疆雪莲高频不定芽再生体系。研究了激素种类和配比、外植体类型、培养条件对新疆雪莲再生时间和再生频率的影响。结果表明:以新疆雪莲25d 苗龄无菌苗的不同部位为外植体,在MS+6-BA 1.0mg/L + NAA
0.1mg/L+2,4-D 0.1mg/L 上进行初代培养,30d 后转接到MS+6-BA 1.0mg/L+ 2,4-D0.01mg/L 上继续培养15~20d,培养温度25±1℃,在45d 的培养周期内均可诱导出大量不定芽丛。其中以真叶为外植体芽诱导率最高,达到66%,且每个外植体上平均出芽数达到6.1。研究了不同环境因子对新疆雪莲增殖和复壮的影响,确定了最佳增殖及复壮条件为:在MS+6-BA 0.5mg/L+NAA 0.05mg/L 上进行增殖扩繁,接种密度为12~16 芽/瓶,每继代2~3 次,转移到MS+多效唑0.05mg/L 释放激素复壮一次。控制温度在20±1℃,平均增殖倍数可达到5.2,且试管苗健壮。首次实现了新疆雪莲的瓶外生根。与瓶内生根的试管苗进行了对比发现:新疆雪莲瓶外生根率略低于瓶内生根,但是移栽成活率明显高于后者。瓶苗质量是瓶外生根能否成功的关键,最佳的瓶外生根条件为:将生长健壮的试管苗敞口炼苗3d后取出,洗净底部培养基,在含有1mg/L NAA,1g/L 多菌灵的溶液中浸泡30min,扦插入灭菌过的蛭石中,蛭石与1/2MS 大量元素溶液体积比在2:1 至3:1 之间。控制温度在20±1℃,光强3500lx,经过30d,瓶外生根率达到80%左右。再生植株移栽入花卉营养土中在温室中进行培养,遮阴、保湿,成活率100%。

其他摘要

This study focused on different relevant factors on adventitious shoots formation, multiplication, rejuvenation and rooting of Saussurea involucrata Kar. et Kir. In the experiments we established a stable plantlet regeneration system of this endangered plant. Furthermore, we succeeded in out test-tube rooting in this plant for the first time. Our established regeneration system simplified the process of operation, shortened the period of rapid propagation and increased the survival rate of transplant seedlings. This research laid the technical foundation for large scale artificial propagation of S. involucrata Kar. et Kir. The establishment of high frequency adventitious bud regeneration system of S. involucrata Kar. et Kir was stable and repetitive. The effects of ratio of differnent hormones, explant types, culture conditions on regeneration time and regeneration frequency were analyzed. The results were as follows: different parts of 25-day old seedlings served as explants and were cultured on MS medium with 1.0mg/L 6-benzyladenine (6-BA), 0.1mg/L α-naphthalene acetic acid (NAA) and 0.1mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) for 30 days; then the explants were transferred to MS medium with 1.0mg/L 6-BA and 0.1mg/L 2,4-D for another 15~20 days. Large quantities of adventitious buds were induced in the 45-day terms. The budding ratio was at the highest frequency when using leaf as explant (up to 66%) and each explant could get 6.1 adventitious buds. Through the research on effects of different environmental factors, we determined the best conditions for proliferation and rejuvenation of S. involucrata Kar. et Kir. The buds were proliferated on MS medium supplemented with 0.5mg/L 6-BA and 0.05mg/L NAA. The best inoculation density was 12~16 buds each bottle. The materials should be transferred to MS medium supplemented with 0.05mg/L paclobutrazol (PP333) to release hormone and rejuvenation every 2~3 generations. The proliferation multiple of adventitious buds reached 5.2 at 20±1℃ and the buds grew well. Out Test-tube rooting of Saussurea involucrata Kar. et Kir was realized for the first time. Compared with inner Test-tube rooting rate, out Test-tube rooting rate was a bit lower but the livability after transferring-planting was obviously higher. The quality of Test-tube seedlings was a crucial factor for out Test-tube rooting. Our results indicated that the perfected culture process for out Test-tube rooting was as follows: Firstly, the strong buds was brought out from bottle after trained for 3 days and washed thoroughly clean, then immersed for 30 min in the solution supplemented with 1mg/LNAA and 1g/L chlorothalonil. Secondly, the treated strong buds were cuttaged in the sterilized vermiculite. The volume ratio of vermiculite and solution supplemented with half-strength MS macroelementsand was between 2:1 to 3:1. Out Test-tube rooting ratio reached 80% after 30d at 20±1℃ and 3500lx. The regenerated plants was transplanted into artificial soils in greenhouse, the transplanting survival rate was 100% with shading and moisture retention.

文献类型学位论文
条目标识符http://ir.xjipc.cas.cn/handle/365002/3486
专题资源化学研究室
作者单位中国科学院新疆理化技术研究所
推荐引用方式
GB/T 7714
傅晓春. 新疆雪莲组培快繁体系优化的研究[D]. 北京. 中国科学院研究生院,2007.
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