Instituto de Biotecnología, Instituto Nacional de Tecnología Agropecuaria (IB-INTA), and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), PO Box 25, B1712WAA Castelar, Argentina.
Max Planck Institute for Molecular Plant Physiology, Wissenschafts Park Golm, Am Mühlenberg 1, D-14476 Potsdam-Golm, Germany.
J Exp Bot. 2016 Jul;67(14):4091-103. doi: 10.1093/jxb/erw185. Epub 2016 May 18.
Improving carbon fixation in order to enhance crop yield is a major goal in plant sciences. By quantitative trait locus (QTL) mapping, it has been demonstrated that a vacuolar invertase (vac-Inv) plays a key role in determining the radical length in Arabidopsis. In this model, variation in vac-Inv activity was detected in a near isogenic line (NIL) population derived from a cross between two divergent accessions: Landsberg erecta (Ler) and Cape Verde Island (CVI), with the CVI allele conferring both higher Inv activity and longer radicles. The aim of the current work is to understand the mechanism(s) underlying this QTL by analyzing structural and functional differences of vac-Inv from both accessions. Relative transcript abundance analyzed by quantitative real-time PCR (qRT-PCR) showed similar expression patterns in both accessions; however, DNA sequence analyses revealed several polymorphisms that lead to changes in the corresponding protein sequence. Moreover, activity assays revealed higher vac-Inv activity in genotypes carrying the CVI allele than in those carrying the Ler allele. Analyses of purified recombinant proteins showed a similar K m for both alleles and a slightly higher V max for that of Ler. Treatment of plant extracts with foaming to release possible interacting Inv inhibitory protein(s) led to a large increase in activity for the Ler allele, but no changes for genotypes carrying the CVI allele. qRT-PCR analyses of two vac-Inv inhibitors in seedlings from parental and NIL genotypes revealed different expression patterns. Taken together, these results demonstrate that the vac-Inv QTL affects root biomass accumulation and also carbon partitioning through a differential regulation of vac-Inv inhibitors at the mRNA level.
提高固碳能力以提高作物产量是植物科学的主要目标。通过数量性状位点(QTL)作图,已经证明液泡转化酶(vac-Inv)在决定拟南芥根长方面起着关键作用。在这个模型中,通过对来自两个不同品系(Ler 和 Cape Verde Island,CVI)杂交的近等基因系(NIL)群体的 vac-Inv 活性进行检测,发现 CVI 等位基因赋予了更高的 Inv 活性和更长的根。本研究的目的是通过分析来自两个品系的 vac-Inv 的结构和功能差异来了解该 QTL 的机制。通过定量实时 PCR(qRT-PCR)分析相对转录丰度显示,两个品系的表达模式相似;然而,DNA 序列分析揭示了几个导致相应蛋白质序列改变的多态性。此外,活性测定显示,携带 CVI 等位基因的基因型的 vac-Inv 活性高于携带 Ler 等位基因的基因型。对纯化的重组蛋白的分析表明,两个等位基因的 K m 相似,而 Ler 等位基因的 V max 略高。用泡沫处理植物提取物以释放可能相互作用的 Inv 抑制蛋白,导致 Ler 等位基因的活性大大增加,但携带 CVI 等位基因的基因型没有变化。对来自亲本和 NIL 基因型的幼苗中的两种 vac-Inv 抑制剂的 qRT-PCR 分析显示出不同的表达模式。综上所述,这些结果表明,vac-Inv QTL 通过在 mRNA 水平上对 vac-Inv 抑制剂的差异调节,影响根生物量的积累和碳分配。