González-Guzmán Miguel, Apostolova Nadezda, Bellés José M, Barrero José M, Piqueras Pedro, Ponce María R, Micol José L, Serrano Ramón, Rodríguez Pedro L
Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia-Consejo Superior de Investigaciones Científicas, Camino de Vera, E-46022 Valencia, Spain.
Plant Cell. 2002 Aug;14(8):1833-46. doi: 10.1105/tpc.002477.
Mutants able to germinate and perform early growth in medium containing a high NaCl concentration were identified during the course of two independent screenings and named salt resistant (sre) and salobreño (sañ). The sre and sañ mutants also were able to germinate in high-osmoticum medium, indicating that they are osmotolerant in a germination assay. Complementation analyses revealed that sre1-1, sre1-2, sañ3-1, and sañ3-2 were alleles of the abscisic acid (ABA) biosynthesis ABA2 gene. A map-based cloning strategy allowed the identification of the ABA2 gene and molecular characterization of four new aba2 alleles. The ABA2 gene product belongs to the family of short-chain dehydrogenases/reductases, which are known to be NAD- or NADP-dependent oxidoreductases. Recombinant ABA2 protein produced in Escherichia coli exhibits a K(m) value for xanthoxin of 19 micro M and catalyzes in a NAD-dependent manner the conversion of xanthoxin to abscisic aldehyde, as determined by HPLC-mass spectrometry. The ABA2 mRNA is expressed constitutively in all plant organs examined and is not upregulated in response to osmotic stress. The results of this work are discussed in the context of previous genetic and biochemical evidence regarding ABA biosynthesis, confirming the xanthoxin-->abscisic aldehyde-->ABA transition as the last steps of the major ABA biosynthetic pathway.
在两项独立筛选过程中,鉴定出了能够在含有高浓度氯化钠的培养基中发芽并进行早期生长的突变体,分别命名为抗盐(sre)和萨洛布雷尼奥(sañ)。sre和sañ突变体也能够在高渗培养基中发芽,这表明它们在发芽试验中具有渗透耐受性。互补分析表明,sre1-1、sre1-2、sañ3-1和sañ3-2是脱落酸(ABA)生物合成基因ABA2的等位基因。基于图谱的克隆策略使得能够鉴定出ABA2基因,并对四个新的aba2等位基因进行分子特征分析。ABA2基因产物属于短链脱氢酶/还原酶家族,已知该家族为NAD或NADP依赖性氧化还原酶。通过高效液相色谱-质谱联用分析确定,在大肠杆菌中产生的重组ABA2蛋白对黄氧素的Km值为19微摩尔,并且以NAD依赖性方式催化黄氧素转化为脱落醛。ABA2 mRNA在所有检测的植物器官中组成性表达,并且在渗透胁迫下不会上调。本文的研究结果结合之前关于ABA生物合成的遗传和生化证据进行了讨论,证实了黄氧素→脱落醛→ABA的转变是主要ABA生物合成途径的最后步骤。