寒兰离体化学诱变研究..doc

编号： 论 文 题 目 寒兰离体化学诱变研究 The Research on Chemical Mutation in Vitro of Cymbidium Kanran Makino 专业名称： 植物学 专业代码： 071001 种类（在相应方框内打√）： 统招博士□ 统招硕士□同等学力人员申请硕士学位□ 工程硕士□ 高校教师在职攻读硕士学位□ 公共管理□ 工商管理□ 摘要 本文以甲基磺酸乙酯(EMS)、硫酸二乙酯(DES)、秋水仙素(colchicines)、咖啡碱(caffein)、5-溴尿嘧啶(5-BU)、马来酰肼(MH)、亚硫酸钠(Na2SO3)为化学诱变剂，以寒兰(Cymbidium kanran Makino)原球茎为材料进行离体化学诱变研究，探讨这7种化学诱变剂对寒兰原球茎生长、增殖和分化的效应，筛选适宜的诱变浓度和时间，以形态学方法鉴定再生植株的表型变异，以细胞学方法观察化学诱变剂对寒兰根尖细胞核的影响，并分析诱变后的再生植株在生理生化指标的变化，以ISSR–PCR 扩增 ABSTRACT Using EMS, DES, colchicines, caffeine, 5-BU, MH, Na2SO3 as mutagenic agents, and using the PLBs of Cymbidium kanran Makino as material, we studied the tech- nique of mutagen in vitro. This article researched the effects on growth, proliferation and differentiation of PLBs treated with mutagenic agents, chosed appropriate concentration and time, used technique of morphologic to check up phenotypic mutation, used cytological technique to observe the influences on karyons of root tip, analyzed the physiological and biochemical change of plants from PLBs treated with mutagenic agents, at last, used ISSR–PCR amplified system to analyze the change on genome of plants from PLBs treated with mutagenic agents. The results were as follows: Each mutagenic agent could resrain the growth of PLBs, with the increase of concentration and the prolonged time, the brown ratio and death ratio rised, the brown ratio were always less than death ratio. Each mutagenic agents could resrain the proliferation of PLBs, with the increase of concentration and the prolonged time, the death ratio rised, the connection between proliferation rate and absolute growth rate were not simple correlativity. Analyzing the results, we calculated the LD50 on PLBs of mutagenic agents, it related to concentration and time, the results could be the referenced dosage for chemical mutation of Cymbidium kanran Makino. Some phenotypic mutation appeared on plants