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新疆薄皮核桃品种组织培养及其影响因素的研究
胡石开
学位类型硕士
导师王晓军
2009-06
学位授予单位中国科学院研究生院
学位授予地点北京
学位专业生物化学
关键词薄皮核桃 温185 新新2 组织培养 影响因素 新疆
摘要论文以温185号和新新2号薄皮核桃品种实生苗和嫁接苗茎段、茎尖和叶片为材料,系统研究了其组织培养快速繁殖的整个过程,并对各个阶段的影响因素进行了详细的探讨,为新疆薄皮核桃品种器官发生植株再生途径奠定了基础,取得如下结果: 1. 适宜的基本培养基为DKW培养基,不定根诱导发生时使用1/4 DKW。2. 外植体表面消毒,使用75%酒精、0.1%升汞和15%双氧水组合,各处理时间依据外植体的种类、状态而定,一般分别在30 s -- 3 min,20 s -- 3 min,15 min -- 25 min。3. 外植体取材时间不同明显的影响了核桃的污染率、分化率,在5月中下旬所取的材料,污染率最低、分化率最高。随着外植体生长时间的推移,外植体的污染率逐渐上升,而分化率逐渐下降。4. 聚乙烯吡咯烷酮(PVP)的抗褐化效果明显优于硫代硫酸钠(Na2S2O3)。培养基中附加250mg/LPVP,以葡萄糖为碳源能有效的抑制褐化发生。5. 茎段和叶柄是诱导愈伤组织的最佳外植体,DKW+0.5 mg/L TDZ +0.1 mg/L IBA激素组合对愈伤诱导和继代都具有明显的优势,愈伤组织诱导率可到100%。愈伤组织再分化较困难。6. 在继代扩繁增殖培养时使用DKW+1.0 mg/L 6-BA +0.01 mg/L IBA+8 g/L琼脂+30 g/L葡萄糖对不定芽增殖的作用效果最好。7. 不定根诱导方法为一步浸蘸生根法,即在80mg/L的无菌IBA溶液中浸蘸处理75分钟,然后转入不含任何激素的1/4 DKW中,黑暗处理12天后置于光照下诱导不定根发生。使用一步浸蘸生根法,不论是生根率、平均生根数还是平均根长方面都能达到较好的生根效果。8. 影响不定根发生的因素较多,基本培养基、外源激素、光周期、黑暗条件,葡萄糖浓度、琼脂浓度等对核桃不定根发生具有重要影响。基本培养基中降低无机盐浓度,即使用1/4 DKW,诱导前期暗培养能促进试管苗不定根发生。黑暗处理条件和使用葡萄糖碳源是不定根发生的必备条件。降低琼脂浓度使用量能促进不定根侧根的发生。不定根发生时,使用培养基1/4 DKW+25 g/L 葡萄糖+4 g/L 琼脂+250 mg/L PVP为宜。9. 生根后的试管苗应及时移栽,试管苗不定根发生后30天移栽时成活率最高,可达36.36%。土壤基质对试管苗移栽成活率具有较大影响,随着土壤基质中土质成分含量的增加,试管苗的成活率下降。土壤基质为土:蛭石:珍珠岩(1:1:1)时试管苗的成活率较高,为33.33%。移栽时,温度控制在18-25℃,湿度保持在85%为宜。
其他摘要This thesis systematically researched the whole process of tissue culture and rapid propagation by taking the stems, shoot tips and leaves of Wen 185 and Xinxin 2 thin-skinned walnut cultivar as explants. And the influence factors of the whole process were researched detailedly, which laid the foundation for the organogenesis of Xinjiang thin-skinned walnut. The foremost results can be briefly illustrated as follows: 1. Appropriate medium is the DKW medium, while using 1/4 DKW at the time of adventitious roots induction. 2. 75% alcohol, 0.1% mercuric chloride and 15% hydrogen peroxide can be used to disinfect the explant surface, in general, the processing time is 30s-3min, 20s-3min, 15min-25min respectively, which based on the type and state of the explants. 3. Explants taken at different times apparently affect the rate of pollution and differentiation, the lowest rate of pollution and the highest rate of differentiation were achieved when explants taken in the mid-to-late of May. With the growth of explants, explant the rate of pollution increase while the the rate of differentiation decrease. 4. The anti-browning effect of polyvinylpyrrolidone (PVP) was superior to sodium thiosulfate (Na2S2O3). Medium supplemented with 250mg/L PVP, glucose as carbon source can effectively inhibit the occurrence of browning. 5. Stems and petioles are the best explants for callus induction. DKW+0.5mg/L TDZ +0.1mg/L IBA hormone combinations has obvious advantages on callus induction and subculture, the callus induction rate was 100%, but callus redifferentiation is difficult. 6. DKW +1.0 mg/L 6-BA +0.01 mg/L IBA +8g/L agar +30g/L glucose can be used to proliferate . 7. The method of adventitious roots induced is a step-induced dip method that was firstly induced in 80 mg/L of IBA solution for 75 minutes, and then transferred to the 1/4 DKW medium which did not contain any hormone, and then took to induce rooting under the light after in dark condition for 12 days. 8. The basic medium, exogenous hormones, photoperiod, dark conditions, glucose concentration and agar concentration have an important influence on the occurrence of adventitious roots. Reducing the concentration of inorganic salts of the basic medium, that is, using 1 / 4 DKW, dark pre-induction training can promote adventitious root occurrence in vitro. Dark conditions and glucose carbon source is a prerequisite for the occurrence of adventitious roots. Reducing the use of agar concentration can promote the occurrence of lateral root. The use of medium 1/4 DKW +25 g/L glucose +4 g/L agar +250 mg/L PVP is appropriate for adventitious roots induction. 9. Rooting seedlings should be transplanted in time. The highest survival rate (36.36%) can be achieved when transplanting after 30 days of the occurrence of adventitious roots. The soil matrix have a important impact on survival, with the increasing of the quality of soil ingredient of the soil matrix, the survival rate decreased. The highest survival rate (33.33%) can be achieved when using the soil matrix of the soil: vermiculite: perlite (1:1:1). Controling the temperature at 18-25 ℃ and humidity at 85% is appropriate for transplanting.
文献类型学位论文
条目标识符http://ir.xjipc.cas.cn/handle/365002/3560
专题资源化学研究室
作者单位中国科学院新疆理化技术研究所
推荐引用方式
GB/T 7714
胡石开. 新疆薄皮核桃品种组织培养及其影响因素的研究[D]. 北京. 中国科学院研究生院,2009.
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