【作者】 石乐娟；

【导师】 张放；

【作者基本信息】 浙江大学 ， 果树学， 2006， 硕士

【摘要】 线艺兰是指兰科（Orchidaceae）,兰属（Cymbidium）中的春兰（Cym.goer ingii（Rchb.f.））,建兰（Cym.emsifolium（L.）SW.）,寒兰（Cym.Kanran Ma kino）,墨兰(Cym.Sinense（Andr.Wild）,蕙兰（Cym.faberi Rolfe）五种国兰线艺品种的总称。线艺兰花是因受到外界环境因素（如光照、雷击）的刺激,导致其遗传基因产生变异,叶片上出现黄、白色线条或斑纹等艺象的变异品种。线艺兰因叶片色彩亮丽,比一般兰花更具观赏价值,较受市场欢迎。但沿用传统的分株繁殖法,线艺兰繁殖系数极低,且由于其生态环境及养护条件的多变,线艺不能稳定的遗传下去,致使大量兰花珍品丧失。中国兰的组织培养已有一定进展,但有关线艺兰的组培研究尚未见报道。本文以线艺春兰‘绿云’的类原球茎为材料,分析建立了线艺春兰的高效再生体系,研究了其根状茎增殖与分化的起源。此外通过对兰花叶片的细胞超微结构的观察初步探讨了其叶片线艺产生原因。为线艺兰珍稀品种的保存及大规模工厂化生产提供理论依据。主要研究结果如下: （一）线艺春兰再生体系的优化 1.细胞分裂素BA、KT对根状茎的增殖有明显的促进作用;但KT浓度超过1.0mg·L^-1后根状茎易玻璃化;0-3.0mg·L^-1范围内生长素浓度越高越利于根状茎的形成,IBA促进根状茎的生长较NAA效果好。 2.基本培养基B₅上的根状茎增殖较快,但培养50d后根状茎呈深绿色,部分开始褐化。1/2 MS固体培养基上的根状茎培养50d后直径较大,生长粗壮,颜色嫩绿,生长状况明显优于B₅。因此B₅培养基有利于根状茎短期的快速增殖,长期继代培养及保存宜选用1/2 MS固体培养基。研究表明1/2 MS与B₅的转换培养在实际生产中比任何一种培养基单独培养更有利于根状茎的增殖。 此外,在MS的基础上微量元素的改变(硫酸锌8.6→15,硫酸铁27.8→20,EDTA二钠37.3→26.83,硫酸锰 22.3→12;单位:mg·L^-1)更利于根状茎不定芽的分化。 3.BA 1.0mg·L^-1+IBA 1.0mg·L^-1是根状茎分化的最佳激素组合,芽分化率达到36.9%。更多还原

【Abstract】 Cymbidium with verge line pattern leaves is the total appellation of five Oriental Orchids varieties with verge line pattern, which are Cym.goerymgii, Cym.emsifolium, Cym.faberi, Cym.Sinense, and Cym.KanranMakino in Orchidaceae Cymbidium. Cymbidium with verge line pattern leaves is the aberrance variety .The lamina has art phenomena such as yellow, white or stripe verge line pattern. Its genes are brought aberrance because of environment stimulate such as illumination and lightning strike. This cymbidium is more valuable than other orchids, as its leaf has beautiful color.But its propagating coefficient is very low by traditional method of crown division. And art phenomena on the lamina aren’t inherited steadily on account of levity on environments and planting condition. So many rare cymbidium breeds are lost.Tissue culture of Oriental Orchids has some work over such as Cym Sinense.But there isn’t study on the tissue culture of Cymbidium with verge line pattern leaves yet. The ideal rapid multiplication system of Cymbidium goeringii cv. Luyun with verge line pattern leaves by protocorm like-body was established. Then origin of proliferation and differentiation of rhizomes was studied. Effects of plant growth regulators on rhizome ’ s multiplication and differentiation were studied. Furthermore, causation of verge line pattern leaves was discussed ordinarily by observation on cell ultrastructure of verge line pattern leaves.The main results are summarized as follows:1 .Optimization of regenerated system in Cymbidium goeringii cv. Luyun with verge line pattern leaves(1) Effects of cytokinins KT and BA are favorable to the multiplication of rhizomes, but rhizomes were vitrifiable easily when KT concentration over 1.0 mg L~-1;the number of rhizomes increased with the increasing of auxin concentration (0-3 mg L~-1), thereinto IBA was more effective than NAA.(2) Rhizomes on the basic culture medium B5 proliferate rapidly, but they began tobrown after 50 days cultured. Growth of rhizomes on 1/2 MS solid medium was better than on B5.S0 B5 is favor of short-term fast multiplication, and 1/2 MS is propitious to long subculture and preservation. Study showed: It was better for rhizomes’ multiplication and growth in culture converted of 1/2 MS and B5 than in any.(3) 1/2MS supplemented with BA 1.0 mg-L"1 and IBA 1.0 mg-L’1 was successful for rhizome’s differentiation, Number of buds differentiation after 80 days unto 36.9%.(4) Optimal subculture time is 80 days to accelerate differentiation of rhizomes. The reason is dedifferentiation accelerated via restraining vegetative growth of rhizomes.(5) Dasheen paste added to the culture medium was better to the multiplication of rhizomes than coconut milk and banana mud .The rhizomes were well by the way, which had broken off with fingers and thumb, each conglomeration with 3-5 rhizomes.2. Rise and genesis of rhizome’s multiplication and bud’s differentiationApical differentiating tissue divides continuously and prolongates to become to the rhizome;internode cortex cells of rhizome divides continuously to form adnation differentia tissue, then develop directly to bud or branch of rhizome. Clustered rhizomes with multi differentia tissues subdifferentiate buds.3. Plant structure anatomise of tube-plantlet and study of lamina cells’ ultrastructure(1) Verge line pattern on the leaf of Cymbidium goeringii cv.’Luyun’ test-tube plantlets was clear, and younger leaf ’s verge line was so more than old one that it could be stably inherited.(2) Lamina surfaces of Cymbidium goeringii with verge line pattern leaves are covered with thick cutin.Epidermis cells array tightly and there are many vascular bundles clinging epidermises.The mesophyll is not differentiated into spongy and palisade tissues, and clearance is smaller. Vascular bundles of leaf with parallel venation are collateral bundles.(3) Vascular boundless in pseudobulb are scattered in ground tissue. In cross section view, from outer to inner part, root includes velamen, cortex, and vascular cylinder. Pseudobulb and root all have the function of storing and keeping water. It’s typical shade-demanding and drought-enduring plant. So water management is key to the young plant surviving in nursing management of cymbidium.(4) Cells’ultrastructures were viewed of one lamina’s art segment (meaning: yellow segment) and normal green segment. Pilot study indicated the primary causation of etiolating : (T).amount of chloroplast decreases;(2). chloroplast ateliosis, some swell, mostly is former-body of chloroplast -proplastid, few thylakoids, just grana.更多还原