华南农业大学学报2017，Vol.38Issue(5)：91-96,6.DOI:10.7671/j.issn.1001-411X.2017.05.016
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
Investigation of browning factors in embryogenic callus culture of Pinus tabulaeformis and optimization of proliferation medium
摘要
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 ofP.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] Results 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.关键词
油松/胚性愈伤/褐化/增殖/正交设计Key words
Pinus tabulaeformis/embryogenic callus/browning/proliferation/lorthogonal design分类
农业科技引用本文复制引用
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):91-96,6.基金项目
国家自然科学基金(31370658) （31370658）
948项目(2014-4-59) （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) （2015ZCQ-SW-02）
北京市园林绿化局计划项目(CEG-2016-01) （CEG-2016-01）
