【作者】 唐红；

【导师】 孙吉雄；

【作者基本信息】 甘肃农业大学 ， 草业科学， 2002， 硕士

【摘要】 草坪草种的品种真实性鉴定是评定萆种质量的重要指标，是保证草坪安全生产和商业流通顺利进行的重要前提。传统的利用植物形态学特征鉴定的方法已被证明具有局限性，研究新的有效的品种鉴定技术已是草坪种业面临的首要任务。为此，本文采用等电聚焦电泳技术对适于西北地区的几种冷季型草坪草种进行了品种真实性鉴定研究，并使用电泳谱带分析软件ImageMaster 1D对草坪草种的过氧化物同工酶谱带进行了分析。通过多方验证，评定此技术作为草坪草种品种鉴定方法的可靠性。结果如下： 1、等电聚焦电泳技术鉴定品种真实性的关键是针对不同植物种类选用不同pH值范围的两性电解质(Ampholine)。通过大量实验，证明几类草坪草种在pH值为4-6范围的凝胶中过氧化物同工酶谱带更为清晰、易于鉴定。 2、在不同的生长期，植物体内同工酶的组分及含量产生变化。草坪草幼苗不同苗龄对电泳谱带的清晰程度产生影响，进而影响鉴定结果。实验证明，多年生黑麦草、高羊茅、紫羊茅及白三叶的最佳取材是发芽6-8天的幼苗，草地早熟禾的最佳取材是发芽10-12天的幼苗。 3、在实验过程中发现，这几种草坪草整株幼苗(除根)在统一加水25微升提取的条件下，酶提取液浓度有所差异，进而影响鉴定结果。经试验，早熟禾与白三叶在加水20微升提取酶液时，获得理想效果，其余种加水25微升提取效果较佳。 4、通过软件分析，对检测结果进行定性定量评定，充分表明了品种间的相关性及差异性。通过聚类分析测得各品种间遗传关系的远近程度。并 证明试验结果与植物分类学及室内种植结果基本一致。说明等电聚焦电泳 方法是鉴别草坪草品种的有效测定技术。更多还原

【Abstract】 The identification of turfgrass seed cultivar truthfulness is an important index to assess the quality of grass seed, and important premise to assure the lawn produce safely and circulate smoothly. The traditional identify method for using plant morphologic character has been proved is not enough. The research on new and effective technology of breeding identify has been the chiefly task of turfgrasses seeding production. So in this paper, the isoelectric focusing electrophoresis technology has been used to identify several kinds of the cold-season turf grasses breeding truthfulness in west -north region, and the electrophoresis isozyme bands analysis software (ImageMaster ID) has been used to analysis the peroxidase isozyme bands of the turfgrasses. The reliability of this technology to identify turfgrasses cultivar has been proved in different way. The result as following:1 -. The key of using the isoelectric focusing electrophoresis technology to identify truthfulness, is to choose different pH value ampholyte for different plant species. Lots of experiments have proved that, the peroxidase isozyme bands of several kinds of the turfgrasses are more clear and more easily to identify at pH4-6 value gel.2. At the different growth period, the composition and content of isozyme have changed. The age of the turfgrasses cultivar effects the electrophoresis bands clear degree, so effects the identify results. The experiment proves that, The best experiment materials of Lolicem perenne, Festuca rubra, Festuca arundinacea Schreb and Trifolium repens are the seedlings of 6-8 days after germination. The best experiment materials of Poa pratensis is the seedlings of 10-12 days after germinate.3. During the experiment we discover that, under all added 25 ul water extract condition, the enzyme extract liquids concentrations of these surfgrasses cultivars (the roots been cutted) are different, so affect the results. The experiment shows that, under the condition of added 20wl water to extract the enzyme, the results of Poa pratensis and Trifolium repens are better, the others are less.4. Using software analysis, the examine results have been quality and quantity assessed. It fully indicates the relativity and diversity among different cultivars. Through clustering analysis, the far and near degrees of the heredity relations between breeds have been measured, and the experiment result has been proved basically accord with the results of plant taxology and indoor planting. The results show that the isoelectric focusing electrophoresis technology is effective measure technology to identify turfgrasses cultivar.更多还原