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1 材料与方法 1.1 材料 供试材料为玉米不同基因型(386,315,517等)的未成熟胚。MET是江苏建湖农药厂产品,有效成分≥99％。 1.2 方法 将受粉后8～12d的玉米幼胚取出,接种在诱导培养基上,当愈伤组织长到直径约1cm时,转入继代和分化培养基中,并不断挑选胚性愈伤组织。胚性愈伤组织形成后用MET处理,1个月后进行生理测定。在分化培养中,幼苗高约2cm时转入壮苗培养基中。诱导培养基为N_6+2,4-D2mg/L+NAA2mg/L+蔗糖3％+琼脂0.7％;继代、分化和壮苗培养基中均附加不同浓度的MET。培养室温度为25±1℃,光照强度为2000lx,14h/d。移栽试验在自然条件下进行,15d后统计成活率。 叶绿素含量的测定参照Arnon法,IAA氧化酶活性的测定参照Gordon et al.法,过氧化物酶活性的测定参照Kochba et al.法,测定仪器为岛津UV-206紫外分光光度计。乙烯释放率用气相色谱法测定,仪器为103型气相层析仪。
1 Materials and methods 1.1 Materials The test material for the different genotypes of maize (386,315,517, etc.) of immature embryos. MET is Jiangsu Jianhu Pesticide Plant products, the active ingredient ≥ 99%. 1.2 Method After 8 ~ 12d maize immature embryos removed, inoculated on the induction medium, when the callus grow to a diameter of about 1cm, transferred to subculture and differentiation medium, and continue to select embryogenic callus . Embryogenic callus was treated with MET and physiologically measured one month later. In the differentiation culture, seedlings transferred to strong seedling medium about 2cm high. Induction medium was N6 + 2,4-D2mg / L + NAA2mg / L + sucrose 3% + agar 0.7%; different concentrations of MET were added in subculture, differentiation and seedling culture medium. Incubation room temperature 25 ± 1 ℃, light intensity 2000lx, 14h / d. Transplanting test under natural conditions, statistical survival rate after 15d. Determination of chlorophyll content According to the Arnon method, the determination of IAA oxidase activity was performed according to the method of Gordon et al., And the peroxidase activity was measured by the method of Kochba et al., Shimadzu UV-206 ultraviolet spectrophotometer. Ethylene release rate was determined by gas chromatography, the instrument is a type 103 gas chromatograph.