【作者】 杜雪玲；

【导师】 符义坤； 杨中艺；

【作者基本信息】 甘肃农业大学 ， 草业科学， 2004， 硕士

【摘要】 多年生黑麦草（Lolium perenne L.）和结缕草(Zoysia japonica L.)是重要的牧草和草坪草，也是冷季型和暖季型的代表性草种。本文以这两种草为材料，进行了组织培养、植株再生与遗传转化的研究。本试验研究结果如下： 1.建立了黑麦草和结缕草的再生体系。从黑麦草的11个品种中选出再生能力较强的三个品种：绿宝石、百尼牡和旋风作为转化受体。在MS基本培养基上附加0.05 mg/l 6-BA、5 mg/l 2，4-D，诱导胚性愈伤组织效果较好；在糖分和固化剂的选择上，通过对结缕草的愈伤组织诱导，得出糖分和固化剂分别选用蔗糖和进口植物凝胶时，诱导愈伤率较高。分化培养基上MS附加1 mg/lNAA、1 mg/l KT，3 mg/l 6-BA，黑麦草和结缕草分化率较高；在愈伤组织的继代时间的选择上，得出继代80天的愈伤组织分化率较高。2.建立了多年生黑麦草农杆菌和结缕草遗传转化体系。在农杆菌为介导的基因转化中，确定并建立了遗传优化体系：性状良好的愈伤组织预培养3天，在OD≈0.3-0.5时浸染时间15分钟，共培养2天，脱菌采用300 mg/l头孢霉素(cef)，附加0.1 mol/l的乙酰丁香酮(AS )。3 运用GUS瞬间检测和对转化30天后的愈伤组织统计，获得了抗草甘瞵抗性愈伤组织。更多还原

【Abstract】 Perennial ryegrass(Lolium perenne L.) and Zoysia grass(Zoysia japonica Steud.) are important forage and turf grasses,and are representatives of cool season and warm season grass as well.The two species were used as materials to study plant regeneration system in diverse cultivars of perennial ryegrassand zoysia grass genetictransferring system of the two plant species by Agrobacterium tumefaciens with Gly gene.The results are as follows:1. The regeneration system of perennial ryegrass and zoysia grass have been estabilished.The three varieties as lvbaoshi ,bainimu,xuanfeng were selected out from eleven ones of perennial ryegrassand were used as acceptors for transformation.MS was used as basic medium aiding with 0.05mg/l 6-BA and 5 mg/l 2,4-D to induce embryogenic callus and subculture as well.Meanwhile,in the selecting of sugar and arga,sucrose and phytagel are the best compared with others by infected zoysia callus,the induced rate is high.At the same time,1 mg/lNAA,1 mg/l KT and 3 mg/l 6-BA are preferable concentration of plants hormone during in differentiation medium.In addition,80 days are appropriate time for getting higher differentiation frequency.2.Genetic transformation has been established.By Agrobacterium tumefaciens mediated,genetic transformation is optimized,using the Agrobacterium strain LBA4404:Embryogenic callus subcultured 80 days,which precultivated 3 days ;Immersed in the bacterial suspension for 15 minute and co-cultivated on medium for 2 days;300 mg/l Cef appending 0.1 mol/l AS is better than Carb in controlling bacterium .3.Transgenic resistant GLY callus are detected by GUS detecting.更多还原