丙酯草醚胁迫下靶标基因在油菜子叶胚状体中的表达特征

EXPRESSION CHARACTERISTICS OF TARGET GENES UNDER HERBICIDE ZJ0273 STRESS IN COTYLEDONARY EMBRYOS OF Brassica napus L.

以甘蓝型油菜品种浙双758为材料,利用小孢子离体培养和实时荧光定量PCR技术,研究了油菜子叶胚状体中ALS酶和KARI酶在不同浓度(0、1、10 mg/L)油菜田新型除草剂丙酯草醚(ZJ0273)处理下的生理活性、基因相对表达量差异及其动态变化特征。结果表明,ZJ0273处理后,ALS酶和KARI酶的活性发生了变化,均显著低于对照。说明其在支链氨基酸合成过程具有相应的作用。荧光定量PCR结果显示,随着胚状体分化时间的延长,ALS和KARI基因在子叶胚状体中的表达量在低浓度(1mg/L)ZJ0273处理下无显著影响,而在高浓度(10mg/L)下的相对表达量则明显降低,无论分化时间延长与否,2个基因均表现为下调表达。研究初步表明,ALS基因和KARI基因参与了除草剂ZJ0273处理下油菜胚状体分化发育过程中的表达调控,可用来作为ZJ0273的靶标基因。

The response of acetolactate synthase（ALS） and ketol-acid reductoisomerase（KARI） enzymes as well as ALS and KARI genes to different concentrations of the herbicide ZJ0273 at cotyledonary embryo stage of Brassica napus were investigated,and effect of this novel herbicide on physiological activity and gene expression differences was clarified.Oilseed rape variety Zheshuang 758,with good response to microspore culture,was subjected to various ZJ0273 treatments（0,1,10 mg/L） and the expressions of two genes（ALS and KARI） encoding ALS and KARI in leaves were detected by using real-time fluorescence quantitative PCR（FQ-PCR）.The results showed that the effects of low ZJ0273 concentration（1 mg/L） on the expression of ALS and KARI genes were not significant,while down regulation was observed for ALS and KARI genes under high ZJ0273 concentration（10mg/L）.These results indicated that ALS and KARI genes were involved in the regulation of differentiation of cotyledonary embryos under herbicide ZJ0273 stress,and ALS and KARI genes are the possible target genes for herbicide ZJ0273.