Almagro Anabel, Lin Shan Hua, Tsay Yi Fang
Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
Plant Cell. 2008 Dec;20(12):3289-99. doi: 10.1105/tpc.107.056788. Epub 2008 Dec 2.
This study of the Arabidopsis thaliana nitrate transporter NRT1.6 indicated that nitrate is important for early embryo development. Functional analysis of cDNA-injected Xenopus laevis oocytes showed that NRT1.6 is a low-affinity nitrate transporter and does not transport dipeptides. RT-PCR, in situ hybridization, and beta-glucuronidase reporter gene analysis showed that expression of NRT1.6 is only detectable in reproductive tissue (the vascular tissue of the silique and funiculus) and that expression increases immediately after pollination, suggesting that NRT1.6 is involved in delivering nitrate from maternal tissue to the developing embryo. In nrt1.6 mutants, the amount of nitrate accumulated in mature seeds was reduced and the seed abortion rate increased. In the mutants, abnormalities (i.e., excessive cell division and loss of turgidity), were found mainly in the suspensor cells at the one- or two-cell stages of embryo development. The phenotype of the nrt1.6 mutants revealed a novel role of nitrate in early embryo development. Interestingly, the seed abortion rate of the mutant was reduced when grown under N-deficient conditions, suggesting that nitrate requirements in early embryo development can be modulated in response to external nitrogen changes.
对拟南芥硝酸盐转运蛋白NRT1.6的这项研究表明,硝酸盐对早期胚胎发育很重要。对注射了cDNA的非洲爪蟾卵母细胞进行的功能分析显示,NRT1.6是一种低亲和力硝酸盐转运蛋白,不转运二肽。逆转录聚合酶链反应(RT-PCR)、原位杂交和β-葡萄糖醛酸酶报告基因分析表明,仅在生殖组织(角果和珠柄的维管组织)中可检测到NRT1.6的表达,且授粉后该表达立即增加,这表明NRT1.6参与了将硝酸盐从母体组织输送到发育中的胚胎。在nrt1.6突变体中,成熟种子中积累的硝酸盐量减少,种子败育率增加。在这些突变体中,异常情况(即过度的细胞分裂和细胞膨压丧失)主要出现在胚胎发育一细胞期或二细胞期的胚柄细胞中。nrt1.6突变体的表型揭示了硝酸盐在早期胚胎发育中的新作用。有趣的是,在缺氮条件下生长时,突变体的种子败育率降低,这表明早期胚胎发育中的硝酸盐需求可根据外部氮变化进行调节。