| 油松胚性愈伤组织培养中褐化因素研究及增殖培养基的优化 |
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| 引用本文： | 付双彬,李慧,赵健,符学军,朱松林,张金凤.油松胚性愈伤组织培养中褐化因素研究及增殖培养基的优化[J].华南农业大学学报,2017,38(5):91-96. |
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| 作者姓名： | 付双彬 李慧 赵健 符学军 朱松林 张金凤 |
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| 作者单位： | 1. 林木育种国家工程实验室/林木花卉遗传育种教育部重点实验室/国家林业局树木花卉育种生物工程重点开放实验室/北京林业大学生物科学与技术学院,北京,100083;2. 山西吕梁山国有林管理局国家油松良种基地,山西临汾,041300 |
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| 基金项目： | 国家自然科学基金（31370658）；948项目（2014-4-59）；北京林业大学青年教师科学研究中长期项目（2015ZCQ-SW-02）；北京市园林绿化局计划项目（CEG-2016-01） |
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| 摘 要： | 【目的】调查油松Pinus tabulaeformis胚性愈伤组织增殖培养阶段不同因素对于褐化产生的影响,并结合增殖率对培养基进行调整优化以降低褐化率,提高利用率。【方法】以3个年份的9个细胞系的油松胚性愈伤组织为材料,利用单因素试验设计对放置不同数量愈伤块、pH、植物凝胶含量、糖种类和含量以及培养时间的褐化率和增殖率进行测定计算。采用正交试验选出最佳培养条件。【结果】单因素试验结果表明,褐化率在不同基因型间差异显著,且与增殖继代年龄显著正相关;在培养皿中放置6~7块愈伤组织、pH 5.8~6.0、培养基中添加2.0~3.0 g·L~(-1)植物凝胶、添加10 g·L~(-1)蔗糖能保证较低的褐化率和较高的增殖率,且蔗糖优于葡萄糖和麦芽糖。【结论】结合正交设计,90 mm培养皿中放置7块愈伤组织,培养基中添加2.5 g·L~(-1)植物凝胶、10 g·L~(-1)蔗糖,调节pH为5.9时,褐化率、增殖率和利用率最理想。
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| 关 键 词： | 油松 胚性愈伤 褐化 增殖 正交设计 |
| 收稿时间： | 2016/12/1 0:00:00 |
Investigation of browning factors in embryogenic callus culture of Pinus tabulaeformis and optimization of proliferation medium |
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| FU Shuangbin,LI Hui,ZHAO Jian,FU Xuejun,ZHU Songlin and ZHANG Jinfeng.Investigation of browning factors in embryogenic callus culture of Pinus tabulaeformis and optimization of proliferation medium[J].Journal of South China Agricultural University,2017,38(5):91-96. |
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| Authors: | FU Shuangbin LI Hui ZHAO Jian FU Xuejun ZHU Songlin and ZHANG Jinfeng |
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| Institution: | National Engineering Laboratory for Tree Breeding/Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education/Key Laboratory of Forest Trees and Ornamental Plants Biological Engineering of State Forestry Administration/College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China,National Engineering Laboratory for Tree Breeding/Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education/Key Laboratory of Forest Trees and Ornamental Plants Biological Engineering of State Forestry Administration/College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China,National Engineering Laboratory for Tree Breeding/Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education/Key Laboratory of Forest Trees and Ornamental Plants Biological Engineering of State Forestry Administration/College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China,National Pinus tabulaeformis seed base, State-owned Forest Administration of Lüliang Mountain, Linfen 041300, China,National Pinus tabulaeformis seed base, State-owned Forest Administration of Lüliang Mountain, Linfen 041300, China and National Engineering Laboratory for Tree Breeding/Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education/Key Laboratory of Forest Trees and Ornamental Plants Biological Engineering of State Forestry Administration/College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China |
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| Abstract: | Objective] To study the effects of different factors on browning of embryogenic callus of Pinus tabulaeformis during proliferation, and optimize the culture medium based on proliferation rate for lower browning rate and higher efficiency.Method] Embryogenic callus of P. tabulaeformis from nine cell lines and three different year were used as material.We performed a serious of single factor experiments and calculated the browning rates and proliferation rates of embryogenic callus under the treatments of different callus number, pH, phytagel content, type and content of sugars, and culturing time. Orthogonal experiment was then conducted to screen out the best culture conditions.Result] of the single factor experiments showed that browning rate was significantly different among different genotypes, and it was significantly positively correlated with the age of subculture. The browning rate was relatively low and the proliferation rate was relatively high when the callus number in each petri dish was 6–7, medium pH was 5.8–6.0, phytagel was 2.0–3.0 g·L–1, and sucrose was 10 g·L–1. Sucrose was better than glucose and maltose.Conclusion] Based on the orthogonal design, placing 7 callus in each 90 mm petri dish, adding 2.5 g·L–1 phytagel and 10 g·L–1 sucrose to the medium, and adjusting the pH to 5.9 could achieve the preferred browning rate,proliferation rate and efficiency. |
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| Keywords: | Pinus tabulaeformis embryogenic callus browning proliferation lorthogonal design |
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