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Loss-of-Function Mutations of the Rice GAMYB Gene Impair α-Amylase Expression in Aleurone and Flower Development

机译:水稻GAMYB基因的功能丧失突变会损害α-淀粉酶在花粉中的表达和花朵发育

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摘要

GAMYB was first isolated as a positive transcriptional regulator of gibberellin (GA)-dependent α-amylase expression in barley aleurone cells, and its molecular and biochemical properties have been well characterized. However, the role of GAMYB elsewhere in the plant is not well understood. To investigate the molecular function of GAMYB outside of the aleurone cells, we isolated loss-of-function mutants from a panel of rice mutants produced by the insertion of a retrotransposon, Tos17. Through PCR screening using primers for rice GAMYB (OsGAMYB) and Tos17, we isolated three independent mutant alleles that contained Tos17 inserted in the exon region. No α-amylase expression in the endosperm was induced in these mutants in response to GA treatment, indicating that the Tos17 insertion had knocked out OsGAMYB function. We found no significant defects in the growth and development of the mutants at the vegetative stage. After the phase transition to the reproductive stage, however, shortened internodes and defects in floral organ development, especially a defect in pollen development, were observed. On the other hand, no difference was detected in flowering time. High-level OsGAMYB expression was detected in the aleurone cells, inflorescence shoot apical region, stamen primordia, and tapetum cells of the anther, but only low-level expression occurred in organs at the vegetative stage or in the elongating stem. These results demonstrate that, in addition to its role in the induction of α-amylase in aleurone, OsGAMYB also is important for floral organ development and essential for pollen development.
机译:GAMYB首先被分离为大麦糊粉细胞中赤霉素(GA)依赖性α-淀粉酶表达的正转录调节因子,其分子和生化特性已得到很好的表征。但是,GAMYB在植物中其他部位的作用尚不十分清楚。为了研究糊粉细胞外GAMYB的分子功能,我们从插入反转录转座子Tos17的水稻突变体中分离了功能缺失突变体。通过使用水稻GAMYB(OsGAMYB)和Tos17引物的PCR筛选,我们分离了三个独立的突变等位基因,这些等位基因包含插入外显子区域的Tos17。响应GA处理,在这些突变体中未诱导胚乳中的α-淀粉酶表达,表明Tos17插入已敲除OsGAMYB功能。我们发现营养期突变体的生长和发育没有明显的缺陷。然而,在过渡到生殖阶段后,观察到节间缩短和花器官发育缺陷,特别是花粉发育缺陷。另一方面,未发现开花时间的差异。在糊粉细胞,花序顶生区,雄蕊原基和花药的绒毡层细胞中检测到高水平的OsGAMYB表达,但在营养期或伸长的茎中仅发生低水平的表达。这些结果表明,OsGAMYB除了在糊粉中诱导α-淀粉酶方面发挥作用外,对花器官发育也很重要,对花粉发育也很重要。

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