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桂林小花苣苔的快速繁殖及器官发生研究

Studies on Rapid Reproduction and Organ Formation of Chiritopsis Repanda Var. Guilinensis

【作者】 苏以丽

【导师】 王任翔;

【作者基本信息】 广西师范大学 , 生态学, 2012, 硕士

【摘要】 苦苣苔科植物种类繁多,全世界约有150属3700余种。而中国有56属463种,均属于苦苣苔亚科。桂林小花苣苔(Chiritopsis repanda var. guilinensis)是苦苣苔科小花苣苔属(Chiritopsis)小花苣苔(Chiritopsis repanda)的变种之一,是广西特有特有物种。通过本研究初期在桂林对桂林小花苣苔生物学特性进行的粗略调查,发现其主要分布在山体背阴处及山洞口附近的石灰岩基质上,有些生于干旱石岩壁间的缝隙,长势较差,且受病虫害严重。桂林小花苣苔受人类活动影响,种群数量减少,当前迫切需要建立起人工繁育和保护技术体系,以获得大规模的种苗和植物材料。本文以桂林小花苣苔的叶片为外植体,主要研究了桂林小花苣苔的扦插繁殖、快速繁殖及其器官发生。主要研究结果如下:1、建立了桂林小花苣苔快速繁殖体系:外植体消毒选用0.1%HgCl2,野生苗的叶片以消毒时间为6min的效果最佳,而盆栽苗的叶片最佳消毒时间为4min;从野生苗的叶片、盆栽苗的叶片及试管苗的叶柄、叶片、茎段五种材料中筛选出不定芽诱导的最佳外植体为试管苗叶片,其分化系数最高;以试管苗叶片为外植体,筛选出不定芽最佳诱导的的激素组合是6-BA和NAA,其分化系数和长势最佳;以试管苗叶片为外植体,诱导不定芽增值的最佳培养基为MS+6-BA0.1mg/L+NAA0.1mg/L+蔗糖30/L+琼脂7g/L,增值系数为23.79:适宜的生根培养基是1/2MS+30g/L蔗糖+5g/L活性炭+琼脂7g/L,生根率为100%;移栽基质为腐殖土,其成活率最高,为90%。2、对桂林小花苣苔不定芽进行组织学观察,结果发现桂林小花苣苔的不定芽发生有两种发生方式,一是由离体叶片表皮细胞及其下面几层细胞直接产生不定芽,即直接不定芽发生;二是间接不定芽发生,外植体先脱分化形成愈伤组织,瘤状愈伤组织再分化形成不定芽原基,最后发育形成不定芽。3、对桂林小花苣苔不定根进行组织学观察,结果发现桂林小花苣苔的不定根原基起源于形成层细胞,属于诱导型。4、以桂林小花苣苔的全叶(无叶柄)为材料进行扦插繁殖,结果表明以粗河沙为基质的扦插效果最佳。

【Abstract】 Gesneriaceae has many varieties, including150genus and3700species in the world. And there are56genus and463species in China, which belong to Cyrtandroideae. Chiritopsis repanda var. guilinensis is one of the most rare species and endemic in Guangxi, which grew poorly in the shady side of hills and the limestone substrate near the cave, or in the dry gap between the drought stone walls, damaged by diseases and insects, according to a rough survey in Guilin. It is necessary to build a technology system of artificial breeding and protection to get the seeds and plants in quantity for the reducing population of Chiritopsis repanda var. guilinensis affected by human activities significantly.This study mainly researched cutting propagation, rapid reproduction and organ formation anatomy of the Chiritopsis repanda var. Guilinensis, using its leaves as explants. The results were as follows:1. Rapid reproduction system of Chiritopsis repanda var. Guilinensis was set up. HgCl2was selected as the surface disinfectant. It was optimum to disinfect the leaves of wilding with0.1%HgCl2for6min, and the leaves in pot culture with0.1%HgCl2for4min. Compared with the leaves of wild plants and potted plants, and the petiole, stem and leaves of test-tube plantlets, the leaves of test-tube plantlets was the optimal explants for adventitious buds induction, with the highest coefficient of differentiation. The optimum hormone for adventitious buds induction was6-BA and NAA, using tue plantlets as explants. The optimum medium for adventitious buds induction was MS+6-BA0.1mg/L+NAA mg/L+sucrose30g/L+arga7g/L, the enhancement factor was23.79.1/2MS+sucrose30g/L+Ac5g/L+arga7g/L were suitable to promote the growth of root, with the rooting rate reached100%. The humus soil was used as medium when transplanting, and the survival rate was90%for Chiritopsis repanda var. guilinensis, reached highest.2. The histological examination of adventitious buds indicated that there were two ways to produce them. One was directly from epidermal cells and the following several layers of cells in vitro leaves. And the other was indirectly from primordium, which was differentiated from callus. 3. The primordium of adventitious root originated in the cambial cells in the type of the induced radication.4. The study on cutting propagation revealed that the thick river sand for the matrix gave the best result, which was using the whole leaf as materal but without petiole.

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