【作者】 朱红莲；

【导师】 汪李平； 柯卫东；

【作者基本信息】 华中农业大学 ， 蔬菜学， 2006， 硕士

【摘要】 本研究以慈姑(Sagittaria trifolia Linn．var．sinensis Makino)茎尖为外植体，进行了组织培养快速繁殖技术的研究，筛选出诱导茎尖分化的最适启动培养基，并探讨了继代增殖培养基的最佳激素配比；同时，以试管苗为基础，对试管慈姑诱导形成的主要影响因素进行了初步研究。试验主要研究结果如下：1．外植体的消毒处理试验结果表明，慈姑茎尖消毒的最佳方法是：以72％农用硫酸链霉素处理20min，再用70％乙醇表面消毒30s，最后用0.1％升汞处理5min。可使成活率达到53.7％，而污染率降至43.9％。2．建立了有效的慈姑组培扩繁体系。以MS为基本培养基，通过调节6-BA和NAA的不同浓度配比，得出适宜慈姑茎尖分化的启动培养基为MS+6-BA1.0mg／L+NAA0.3mg／L，分化率为88.1％；继代增殖的最适培养基为MS+6-BA3.0mg／L+NAA0.1mg／L，最佳的继代周期为6周，增殖系数达8.87。3．通过正交设计研究蔗糖浓度、激素配比、温度和光照时间4因子对株高、根状茎、试管慈姑等的影响，结果表明，4因子中仅蔗糖浓度对株高有显著影响，随着蔗糖浓度的增加试管苗高度逐渐降低；4因子中仅蔗糖浓度和温度2因子对单株试管苗抽生根状茎的条数有显著影响，以蔗糖浓度30g／L或60g／L和温度22℃时效果较好，可得到数量较多的根状茎；4因子对结慈姑株率和单株结慈姑数都有显著影响，以组合蔗糖浓度=60g／L，激素配比6-BA：NAA=0：0，温度=22℃，光照时间=0h，结慈姑株率和单株结慈姑数均为最高值，为诱导试管慈姑的最佳组合。4．组培苗移栽试验结果显示：组培苗所结慈姑较种球繁殖苗所结慈姑在平均单个球茎鲜重上增加57.63％。更多还原

【Abstract】 The shoot tips of Sagittaria trifolia Linn. var. sinensis Makino were used as explant in the study to establish an effective mass propagation technique of tissue culture. We gained the favorable induction medium for shoot tips, and studied the best plant growth regulator proportion for regeneration medium. Meanwhile, we preliminarily studied the major influence factors to microtubers of Sagittaria trifolia Linn. var. sinensis. The main results were as the following:1. The best surface sterilization method for inoculation shoot tips was immersed in 72% streptomycin sulphate for 20 minutes, then dipped in 70% ethanol for 30s, and finally agitated in 0.1% HgCl₂ for 2 minutes. The effect was excellent and the survival rate could up to 53.7% and the pollution rate reduced to 43.9%.2. Effective rapid propagation system in vitro of Chinese Arrowhead was set up. Through regulating the concentration of 6-BA and NAA in MS medium, We found out that the favorable induction medium for shoot tips was MS + 6-BA 1.0 mg/L + NAA 0.3mg/L, and the differentiation rate was 88.1%; The optimum medium for regeneration was MS + 6-BA 3.0 mg/L + NAA 0.1mg/L, and the best multiplicative period was 6 weeks, the propagation coefficient could up to 8.87.3. Through orthogonal experiment, carried on research on sucrose concentration. 6-BA:NAA, temperature and illumination time 4 factors to the influence of height of plantlet, microrhizome and microtuber. The results showed that only sucrose concentration could influence to height of plantlet significantly, and with the rising of sucrose concentration, height of plantlet presented the downward trend; Both sucrose concentration and temperature 2 factors could influence to number of microrhizome per plantlet significantly, when we used 30g/L or 60g/L sucrose concentration and 22℃temperature, we got much more microrhizome; All of 4 factors could influence to ratio of microtuber and number of microtuber per plantlet significantly, and the combination of sucrose concentration=60g/L, 6-BA:NAA=0:0, temperature=22℃, illumination time =0h was the best combination to induce tubers in vitro of Sagittaria trifolia Linn. var. sinensis Makino, then we could gain the highest value of both ratio of microtuber and number of microtuber per plantlet. 4. Transplantation experiment of planlets showed that the fresh weight on per tuber formed by plantlets increased 57.63% compared with by seedlings.更多还原