棉花多胺HPLC的测定方法优化及其在体细胞胚胎发生过程中的变化规律  

Method Optimization of Polyamine Content by High-Performance Liquid Chromatography and Its Changes in the Process of Somatic Embryogenesis in Cotton

作　　者：程文翰[1] 朱华国[1] 李鹏飞[1] 王凡龙[1] 朱守鸿 赵兰杰[1] 郭丽雪[1] 孙杰[1] 

机构地区：[1]石河子大学农学院/新疆兵团绿洲生态农业重点实验室,新疆石河子832003

出　　处：《棉花学报》2014年第2期138-144,共7页Cotton Science

基　　金：兵团种质资源创新与功能基因发掘利用专项(2012BB049);农业部转基因生物新品种培育重大专项(2011ZX08005);兵团博士基金项目(2011BB004)

摘　　要：利用HPLC法测定棉花组织培养物中的多胺含量,以棉花品种新陆早33号愈伤组织为材料进行多胺含量的测定,结果表明：冰浴浸提时间及衍生反应时间优化后,可将内源腐胺（Putrescine,Put）、亚精胺（Spermidine,Spd）、精胺（Spermine,Spm）在15min完全分离并定量测定,线性关系良好（r＞0.99）,回收率高（96.8％～103.1％）;建立了棉花多胺HPLC测定方法.利用该方法对棉花体细胞胚胎发生各时期培养物进行多胺含量测定,结果显示胚性愈伤组织形成后多胺含量显著上升,且腐胺与亚精胺和精胺的比例随着体细胞胚胎发生持续下降,表明非胚性细胞向胚性细胞转化过程中,二胺（腐胺）不断向三胺（亚精胺）和四胺（精胺）转化,特别是在胚状体形成期,亚精胺和精胺的积累显著增加,初步揭示了棉花体细胞胚胎发生过程中多胺的变化规律.The high-performance liquid chromatography method for the determination of the polyamine content of cotton cultures was established through relevant and recovery analysis. Polyamine contents in different state callus of variety Xinluzao 33 were detected. The results showed that endogenous putrescine, spermidine, and spermine can be completely separated and quantified in 15 min by optimizing the ice-bath extraction time and derivative reaction time, and good linear relationships （r values greater than 0.99） and accurate recovery rates （96.8% - 103.1%） were obtained. Polyamine contents increased significantly while those of putrescine, spermidine, and spermine decreased significantly, and the conversion fi＇om putrescine to spermidine and spermine occurred continuously during somatic embryogenesis. In particular, the contents of spermidine and spermine obviously accumu- lated during the formation stage of the somatic embryos. Thus, we preliminarily revealed the change rule of polyamines in the process of somatic embryogenesis in cotton.

关 键 词：棉花 愈伤组织 高效液相色谱 多胺 体细胞胚胎发生 

分 类 号：S562[农业科学—作物学]