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以松散型玉米自交系沈137为材料,利用同源克隆法成功克隆与水稻Dwarf4基因同源的玉米Dwarf4基因。序列分析发现,该基因cDNA序列全长1 626 bp,开放阅读框1 521 bp,编码506个氨基酸,与高粱、水稻、拟南芥的氨基酸同源性分别为95%、89%和71%。Real-Time PCR分析表明,ZmDwarf4基因的表达量为3~11叶期表达量逐渐增加,9~11叶期达到最大量,12~19叶期表达量逐渐降低,推测ZmDwarf4基因正向调控玉米叶夹角的大小。ZmDwarf4基因的表达量在叶片中最高,在叶枕中最低。不同激素处理的结果表明,用植物激素(IAA)处理的样品ZmDwarf4基因的表达量始终高于对照;用Brassinolide(BR)处理的样品6叶期ZmDwarf4基因的表达量高于对照,7~10叶期该基因的表达趋势与对照基本一致,但始终低于对照;用IAA+BR处理的样品,该基因的表达趋势与对照基本一致,但高于对照。ZmDwarf4参与玉米BR生物合成途径,进而调控其相关株型建成。
A loose maize inbred line Shen 137 was used as material to clone the Dwarf4 gene homologous to Dwarf4 in rice by homology cloning method. Sequence analysis revealed that the cDNA sequence of this gene was 1 626 bp in length with an open reading frame of 1 521 bp encoding a protein of 506 amino acids with 95%, 89% and 71% amino acid identity with sorghum, rice and Arabidopsis thaliana respectively. Real-time PCR analysis showed that the expression level of ZmDwarf4 gene increased gradually from 3 to 11 leaf stage, reaching the maximum at 9 to 11 leaf stage, and gradually decreased at 12 to 19 leaf stage. ZmDwarf4 gene positively regulated maize leaf The angle between the size. The expression of ZmDwarf4 gene was the highest in leaves and the lowest in leaves. The results of different hormones treatment showed that the expression level of ZmDwarf4 gene was always higher than that of the control with plant hormone (IAA). The expression level of ZmDwarf4 gene in 6-leaf stage of Brassinolide (BR) The expression trend of this gene was basically the same as that of the control, but it was always lower than that of the control. The expression of this gene in the sample treated with IAA + BR was basically the same as that of the control but higher than that of the control. ZmDwarf4 is involved in the BR biosynthesis pathway of maize, and then the related plant type is completed.