利用高效液相色谱法分离和测定棉花组织培养过程中4种内源激素

目的：优化利用高效液相色谱法测定棉花组织培养过程中吲哚-3-乙酸（IAA）、玉米素（ZT）、赤霉素（GA3）和脱落酸（ABA）等4种植物内源激素的条件,了解棉花胚性愈伤组织发生过程中4种内源激素及激素含量比例的规律性变化,以及添加不同外源激素对内源激素和愈伤组织的影响,为棉花组织培养由经验型变为理论型提供基础。方法：采用高效液相色谱法。结果：棉花胚性愈伤组织发生过程中4种内源激素及激素含量比例呈规律性变化：ZT、ABA、ABA/GA3、ABA/IAA呈现先上升后下降的趋势,GA3呈现先上升后下降再上升的趋势,ZT/IAA呈现明显的上升-下降-上升-下降的趋势;不同激素组合诱导下愈伤组织中内源激素含量及愈伤组织状态也有较大差别。结论：研究结果对指导组织培养过程中激素的调整和配比、制定适宜的培养计划有一定的指导意义;4种不同的内源激素都是愈伤组织生长的重要因子,各自适宜的浓度和恰当的比例调节着外植体的脱分化和再分化,各种激素协同作用促进细胞的生长和分化。

Separation and Determination of Four Plant Hormones During Somatic Embryogenesis of Gossypium hirsutum L. with HPLC

Objective： To optimize the chromatographic conditions of HPLC for the separation and determination of plant hormones indoles-3-acetic acid（IAA）, zeatin（ZT）, gibberellic acid（GA3） and abscisic acid（ABA）. To understand the regular Changes of the content of four endogenous hormones as well as the rates of different plant hormones, and to discuss the effect of different extraneous hormones to the endogenous hormones during somatic embryogenesis of Gossypium hirsutum L.. Methods： HPLC was used including a system of Surveyor LC Pump Plus, a Surveyor PDA Plus Detector. Results： The content of four endogenous hormones changed regularly during somatic embryogenesis： ZT, ABA, ABA/GA3 and ABA/IAA （low-high）; GA3（low-high-low）; ZT/IAA（high-low-high-low）. The endogenous hormones content and growth situation changed differently under the effect of different hormone combination in the callus induction. Conclusions： The results have a certain sense of adjusting and allocating the proportion of different plant hormones in the tissue culture. These four kinds of different endogenous hormones are all important factors to the callus induction; suitable content and appropriate proportion control the differentiation and dedifferentiation of the explants; each hormone cooperate with each other to promote the cell growth and differentiation.