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玉米矮秆基因d15的克隆及表达分析 被引量:1

Cloning and Expression Analysis of Dwarf Gene d15 in Maize
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摘要 发掘矮秆基因并解析其调控机制,为玉米矮化育种提供基因资源和理论依据。利用形态观察和石蜡切片方法,研究了玉米矮秆突变体K15d与野生型K15的矮化特征差异;通过等位性鉴定并利用PCR扩增克隆了矮秆基因d15,分析了3个时期茎节间中d15的表达模式。与野生型K15比较,突变体K15d的株高、穗位高和穗下节间数分别降低39.22%、69.75%和38.83%,差异均达显著或极显著水平;茎秆横切面细胞大小差异不明显,纵切面细胞变短,排列不规则。矮秆基因d15与br2等位,第5外显子5485~5685 bp区间缺失200 bp,编码区全长3983 bp。d15编码蛋白的跨膜结构域为10个,比D15编码蛋白的跨膜结构域减少2个,负责底物结合和转运功能的第2个保守功能域缺失。d15启动子较br2仅有2个SNPs差异。在拔节前、拔节期和拔节后3个时期,突变体中d15基因的表达量与野生型间均无显著差异。由此可见,矮秆基因d15的矮化特征及表达模式均与br2相似,是1个新的br2等位基因,丰富了玉米矮秆基因资源。 Exploring dwarf genes and analyzing their regulatory mechanisms may provide germplasm resources and theoretical bases for maize dwarf breeding.The differences of dwarfing characteristics between maize dwarf mutant K15d and its wild type K15 were analyzed by morphological observation and paraffin section.Based on the result of allelism test,the functional gene in K15d mutant(designated d15)was cloned by PCR amplification,and the expression pattern of d15 gene in stem internodes were analyzed at three stages.Compared with wild type K15,the plant height,ear height and the internodes number below ear position of mutant K15d significantly decreased by 39.22%,69.75% and 38.83%,respectively.There was no significant difference in cell size in the transverse section of the stem,and the cells in the longitudinal section became shorter and arranged irregularly.The dwarf gene d15 was allelic to br2,showing a 200 bp deletion(5485-5685 bp)at the fifth exon,with the coding region of 3983 bp.The d15-encoded protein had ten transmembrane domains,two less than that of D15-encoded protein,and a second conserved functional domain responsible for substrate binding and transport functions was missing.Compared to the allele br2,only two SNPs differences were observed in the promoter sequence of d15.There were no significant differences in the expression levels of d15 gene between the mutant and the wild type at the three stages of pre-elongation,elongation and post-elongation.In conclusion,the dwarfing characteristics and expression pattern of dwarf gene d15 are similar to br2,which is a new br2 allele and enriches maize dwarf gene resources.
作者 赵长云 白光庭 何少勇 余学杰 李仁飞 夏伟 许秀兰 柯永培 ZHAO Chang-yun;BAI Guang-ting;HE Shao-yong;YU Xue-jie;LI Ren-fei;XIA Wei;XU Xiu-lan;KE Yong-pei(College of Agronomy,Sichuan Agricultural University,Chengdu 611130;Sichuan Zhenghong Bio Co.,Ltd.,Chengdu 610213;Gansu Yasheng Agricultural Research Institute Co.,Ltd.,Lanzhou 730000)
出处 《植物遗传资源学报》 CAS CSCD 北大核心 2023年第4期1141-1150,共10页 Journal of Plant Genetic Resources
基金 四川省重点研发项目(2021YFYZ0017) 四川省重大科技专项(2022ZDX0013) 成都市科技创新平台资助项目(2019-YF04-00022-JH) 成都市重点研发项目(2021-YF05-00527-SN)。
关键词 玉米 矮秆 br2基因 等位突变 基因表达 maize dwarf br2 gene allelic mutation gene expression
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