Ogawa M, Kusano T, Koizumi N, Katsumi M, Sano H
Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Ikoma, Japan.
Plant Mol Biol. 1999 Jul;40(4):645-57. doi: 10.1023/a:1006291917591.
In order to identify genes that are related to the gibberellin (GA) response in maize (Zea mays L.), mRNA species from wild-type and single-gene dwarf mutants, d5 and D8, were compared by fluorescent differential display. The d5 mutant is unable to produce biologically active GA, but recovers its tall stature on exogenous application of GA. D8 is insensitive to GA, despite the accumulation of a high level of endogenous GA, suggesting it to be a receptor mutant or a mutant in signal transduction pathway(s). After screening 7000 cDNA populations, one clone was isolated, for which transcripts were rare in d5 shoots but accumulated within 1 h after GA3 application. This clone, designated as ZmGR1a, encodes a polypeptide with a relative molecular mass of ca. 13 kDa, which shows significant homology to proline-rich proteins from several plant species. A similar experiment with D8 identified a clone, ZmGR2a, with low transcript levels, but accumulation within 6 h after GA3 treatment of d5 shoots. ZmGR2a encodes a polypeptide with a relative molecular mass of ca. 19 kDa, which shows no significant homology with any known protein. Southern blot analysis indicated that ZmGR1a and ZmGR2a form a small multigene family within the maize genome. In situ hybridization with wild-type seedlings showed transcripts on both to be abundant in leaf sheath meristematic tissue, in which GA enhances cell elongation and cell division.
为了鉴定与玉米(Zea mays L.)中赤霉素(GA)反应相关的基因,通过荧光差异显示比较了野生型和单基因矮化突变体d5和D8的mRNA种类。d5突变体无法产生具有生物活性的GA,但在外源施加GA后可恢复其高株型。尽管积累了高水平的内源GA,但D8对GA不敏感,这表明它是受体突变体或信号转导途径中的突变体。在筛选了7000个cDNA群体后,分离出一个克隆,其转录本在d5芽中很少见,但在施加GA3后1小时内积累。这个克隆被命名为ZmGR1a,编码一种相对分子质量约为13 kDa的多肽,与几种植物物种的富含脯氨酸的蛋白质具有显著同源性。对D8进行的类似实验鉴定出一个克隆ZmGR2a,其转录水平较低,但在对d5芽施加GA3处理后6小时内积累。ZmGR2a编码一种相对分子质量约为19 kDa的多肽,与任何已知蛋白质均无显著同源性。Southern印迹分析表明,ZmGR1a和ZmGR2a在玉米基因组中形成一个小的多基因家族。对野生型幼苗进行原位杂交显示,两者在叶鞘分生组织中均有丰富的转录本,在该组织中GA可促进细胞伸长和细胞分裂。