应用正交试验法优化玉米幼胚诱导胚性愈伤的研究

Study on Optimization for Embryonic Callus Initiation from Maize Immature Embryo Using the Orthogonal Experiment

[目的]为提高不同玉米自交系幼胚胚性愈伤组织诱导提供参考。[方法]以玉米幼胚为外植体,对其进行诱导和继代,通过正交试验设计L16(45),研究不同品种、蔗糖浓度、pH值、2,4-D浓度4个因素及水平对非胚性愈伤组织转化为胚性愈伤组织的影响。[结果]各因素对诱导胚性愈伤组织的影响次序是:品种>pH值>2,4-D浓度>蔗糖浓度,其中品种和pH值均达到极显著水平。各品种产生胚性愈伤由易到难的顺序依次是:H99>D988>7922>D598。诱导产生胚性愈伤组织的最优组合为A1B4C4D1,即玉米自交系H99、蔗糖浓度35.0 g/L、pH值5.8、2,4-D浓度1.0 mg/L。[结论]基因型是影响胚性愈伤组织产生的最主要因素,培养基酸碱度对胚性愈伤组织的产生影响较大。

[Objective] The aim was to provide the reference for enhance embryonic callus induction of immature embryos from different maize inbred lines.[Method] Taking maize immature embryos as explants,inducing and subculturing them,the effects of different varieties,sucrose concentration,pH value,2,4-D concentration on the transformation from non-embryonic callus to embryonic callus were researched through the orthogonal test design L16（4^5）.[Result] The effects of each factor on the embryonic callus induction were genotype pH value2,4-D concentrationsucrose concentration,and the effects of genotype and pH value reached at extremely significant level.Induction capacities of different maize varieties from easy to difficulty in turn was H99D9887922D598.The optimum combination for embryonic callus induction was A1B4C4D1,namely,maize inbred lines H99,sucrose concentration 35.0 g/L,pH value 5.8,2,4-D concentration 1.0 mg/L.[Conclusion] Genotype is the most primary factor which affects embryonic callus induction,and pH value also affects greatly embryonic callus induction.