【作者】 于福科；

【导师】 张广军；

【作者基本信息】 西北农林科技大学 ， 环境科学， 2003， 硕士

【摘要】 开展玫瑰组织培养技术研究，是实现良种快繁和产业化发展的客观需求。 所开展的工作及取得的结论或成果包括以下八个方面： 1 外植体选取 以带芽茎段作为外植体，一年内有两个适宜选取期，一是3月中旬至4月底之间，二是10月间。各时期的利弊是：前一时期的外植体侧芽萌动快，生长也快，但污染及褐化较严重；后一时期，情况恰好相反。 2 外植体灭菌 以外植体成活率与未污染率之和作为指标，通过正交试验优选，结果表明：70％（m/m）酒精浸泡6s后，再用0.1％～0.2％（m/m）Hgcl₂浸泡8min左右，灭菌效果理想。 3 培养基琼脂用量计算的依据 通过试验，提出用胨力强度（g/cm²）表示培养基硬度的观点，并确定了初代培养基胨力强度为160～170g/cm²，继代培养基、生根培养基均为150～160g/cm²。 4 培养基糖源试验 分别对外植体分化、不定芽分化及生根过程进行了蔗糖、麦芽糖和乳糖的培养效果研究，发现卡氏玫瑰初代培养以30g·L^-1蔗糖，继代培养以40g·L^-1蔗糖，生根培养以30g·L^-1蔗糖为好；百氏玫瑰初代培养以40g·L^-1蔗糖，继代培养以30g·L^-1麦芽糖，生根培养以40g·L^-1麦芽糖为宜。走出了培养基专选蔗糖的传统。 5 激素用量组合优化 用均匀法设计试验，进行多项式回归分析，结果表明，卡氏玫瑰的MS培养基最佳激素含量组合为：初代培养1.50mg·L^-16-BA+0.02mg·L^-1NAA，继代培养1.0mg·L^-16-BA+0.1mg·L^-1 NAA+0.1mg·L^-1PP₃₃₃，生根培养0.4g·L^-16-BA+0.1mg·L^-1 NAA+0.2g·L^-1AC；百氏玫瑰MS培养基最佳激素含量组合为：初代培养2.02mg·L^-16-BA+0.11mg·L^-1 NAA，继代培养1.8mg·L^-16-BA+0.5mg·L^-1NAA+0.5mg·L^-1PP₃₃₃，生根培养0.68mg·L^-16-BA+0.27mg·L^-1 NAA+0.40g·L^-1AC。 6 专用培养基pH值的优选 两类供试玫瑰初代培养基pH值均以5.9为宜，继代和生根培养基的pH值均以5.8为宜。 7 影响培养基pH值因素的研究 培养基pH值校正试剂、胨力强度、温度、高温高压灭菌都是影响pH值变动的重要因素；灭菌时间、培养基体积、以及培养基内脱脂棉、NaCl、CaCl₂、蔗糖及活性炭等的含量等，能在高温高压灭菌过程中引起MS培养基pH值下降；各因素对pH值变动的影响趋势和程度明显不同。 8 培养基简化试验的研究 试验结果表明，用自来水代替蒸馏水、普通绵白糖代替分析用蔗糖、普通果酱瓶代替三角瓶制备培养基，对试管苗分化和增殖的影响甚微，但是却能大大降低成本，提高经济效益。更多还原

【Abstract】 It is necessary to study technology of rose tissue culture for good seed cultivation and industrialization.All work and conclusions or results are below:1 Choosing explant. Rose stem is explant. There are two proper choosing periods in a year. One is from the middle of March to the end of April. The other is October. At the former period, axillary buds on explant germinate easier, grow quicker, but contamination and browning is serious; at the later period, circumstance is contrary.2 Sterilization of explant. Sum of live rate and un-contamination rate of explant is statistic index. It revealed that soaking explant by spirit (70%,m/m) for 6 seconds and Hgch (0.1~0.2 %,m/m) for 8 minutes can sterilize explant well.3 Calculative basis of agar dosage. Put forward the viewpoint of denoting the rigidity of media by peptonization intensity(g/cm2).Ascertained that the peptonization intensity of initial media is 160-170 g/cm2,that of subculture and rooting media is 150~160 g/cm2.4 Experiment on sugar source of media. The result of comparison between sucrose, maltose and lactose showed that type K is suitable to sucrose 30g-L-1 in initial culture, 40g-L-1 in subculture and 30g-L-1 in rooting culture; type B is suitable to sucrose 40g-L-1 in initial culture, maltose 30g-L-1 in subculture and 40g-L-1 in rooting culture. Disinherited the tradition of choosing sucrose solely.5 Optimizing combination of hormone content. Experimental results showed that type K is fitting to 1.50mg-L-16-BA + 0.02mg-L-1 NAA in initial culture,1.0mg-L-16-BA + 0.1mg-L-1 NAA + 0.1mg-L-1PP333 in subculture and 0.4g-L-16-BA+0.1mg-L-1 NAA+0.2g-L-1AC in rooting culture; type B is fitting to 2.02mg-L-16-BA+0.11mg-L-1 NAA in initial culture,1.8mg-L-16-BA+0.5mg-L-1 NAA+ 0.5mg-L-1PP333 in subculture and 0.68mg-L-16-BA+0.27mg-L-1 NAA+0.40g-L-1AC in rooting culture.6 Selecting pH for special media. It showed that the proper pH of initial media for two types rose is 5.9,that of multiplying and rooting media is 5.8.7 Study on factors of media’s pH. Dose type used to revision of media’s pH, peptonization intensity of media, temperature, high temperature and pressure pasteurization are factors of media’s pH; Sterilizing time, volume of media, content of pledget, NaCl,CaCl2,sucrose and active carbon can Iead to the lowing of MS media’s pH after high temperature and pressure pasteurization; The influencing trend and extent of every factor is different obviously.8 Study on media simplification experiment. The result showed that the effect of replacing tap water by distilled water, replacing analysable sucrose by common white sugar, replacing cone bottle by general bottle on tissue culture is very little. However, it can reduce the cost of tube culture seeding.更多还原