Delmas Frédéric, Séveno Martial, Northey Julian G B, Hernould Michel, Lerouge Patrice, McCourt Peter, Chevalier Christian
INRA (Institut National de la Recherche Agronomique), Unité Mixte de Recherche 619 sur la Biologie du Fruit, Institut Fédératif de Recherche 103, F-33883 Villenave d'Ornon, France.
J Exp Bot. 2008;59(10):2639-47. doi: 10.1093/jxb/ern118. Epub 2008 May 23.
Despite a very complex structure, the sugar composition of the rhamnogalacturonan II (RG-II) pectic fraction is extremely conserved. Among its constituting monosaccharides is the seldom-observed eight-carbon sugar 3-deoxy-D-manno-octulosonic acid (Kdo), whose phosphorylated precursor is synthesized by Kdo-8-P synthase. As an attempt to alter specifically the RG-II structure in its sugar composition and assess the consequences on the function of RG-II in cell wall and its relationship with growth, Arabidopsis null mutants were sought in the genes encoding Kdo-8-P synthase. Here, the isolation and characterization of one null mutant for the isoform 1 (AtkdsA1-S) and two distinct null mutants for the isoform 2 of Arabidopsis Kdo-8-P synthase (AtkdsA2-V and AtkdsA2-S) are described. Evidence is provided that AtkdsA2 gene expression is preferentially associated with plantlet organs displaying a meristematic activity, and that it accounts for 75% of the mRNAs to be translated into Kdo-8-P synthase. Furthermore, this predominant expression of AtKDSA2 over AtKDSA1 was confirmed by quantification of the cytosolic Kdo content in the mutants, in a variety of ecotypes. The inability to identify a double knockout mutant originated from pollen abortions, due to the inability of haploid pollen of the AtkdsA1- AtkdsA2- genotype to form an elongated pollen tube properly and perform fertilization.
尽管鼠李半乳糖醛酸聚糖II(RG-II)果胶部分的结构非常复杂,但其糖组成却极为保守。其组成单糖中有一种很少见的八碳糖3-脱氧-D-甘露辛酮糖酸(Kdo),其磷酸化前体由Kdo-8-P合酶合成。为了特异性改变RG-II结构的糖组成,并评估其对细胞壁中RG-II功能及其与生长关系的影响,研究人员在编码Kdo-8-P合酶的基因中寻找拟南芥缺失突变体。本文描述了拟南芥Kdo-8-P合酶同工型1的一个缺失突变体(AtkdsA1-S)以及同工型2的两个不同缺失突变体(AtkdsA2-V和AtkdsA2-S)的分离和鉴定。有证据表明,AtkdsA2基因表达优先与具有分生组织活性的幼苗器官相关,并且它占可翻译成Kdo-8-P合酶的mRNA的75%。此外,通过对多种生态型突变体中胞质Kdo含量的定量分析,证实了AtKDSA2相对于AtKDSA1的这种优势表达。由于AtkdsA1-AtkdsA2-基因型的单倍体花粉无法正常形成伸长的花粉管并完成受精,因此无法鉴定出源自花粉败育的双敲除突变体。