Avelange-Macherel Marie-Hélène, Payet Nicole, Lalanne David, Neveu Martine, Tolleter Dimitri, Burstin Judith, Macherel David
Agrocampus Ouest, UMR 1345 Institut de Recherche en Horticulture et Semences, Angers, F-49045, France.
INRA, UMR 1345 Institut de Recherche en Horticulture et Semences, Angers, F-49045, France.
Plant Cell Environ. 2015 Jul;38(7):1299-311. doi: 10.1111/pce.12480. Epub 2014 Dec 26.
LEAM, a late embryogenesis abundant protein, and HSP22, a small heat shock protein, were shown to accumulate in the mitochondria during pea (Pisum sativum L.) seed development, where they are expected to contribute to desiccation tolerance. Here, their expression was examined in seeds of 89 pea genotypes by Western blot analysis. All genotypes expressed LEAM and HSP22 in similar amounts. In contrast with HSP22, LEAM displayed different isoforms according to apparent molecular mass. Each of the 89 genotypes harboured a single LEAM isoform. Genomic and RT-PCR analysis revealed four LEAM genes differing by a small variable indel in the coding region. These variations were consistent with the apparent molecular mass of each isoform. Indels, which occurred in repeated domains, did not alter the main properties of LEAM. Structural modelling indicated that the class A α-helix structure, which allows interactions with the mitochondrial inner membrane in the dry state, was preserved in all isoforms, suggesting functionality is maintained. The overall results point out the essential character of LEAM and HSP22 in pea seeds. LEAM variability is discussed in terms of pea breeding history as well as LEA gene evolution mechanisms.
胚胎后期丰富蛋白LEAM和小分子热激蛋白HSP22在豌豆(Pisum sativum L.)种子发育过程中会在线粒体中积累,预计它们有助于提高种子的耐脱水性。在此,通过蛋白质免疫印迹分析检测了89种豌豆基因型种子中它们的表达情况。所有基因型中LEAM和HSP22的表达量相似。与HSP22不同,LEAM根据表观分子量显示出不同的异构体。89种基因型中的每一种都含有单一的LEAM异构体。基因组和逆转录聚合酶链反应分析揭示了四个LEAM基因,它们在编码区存在一个小的可变插入缺失。这些变异与每种异构体的表观分子量一致。发生在重复结构域中的插入缺失并未改变LEAM的主要特性。结构建模表明,在干燥状态下允许与线粒体内膜相互作用的A类α螺旋结构在所有异构体中均得以保留,这表明其功能得以维持。总体结果表明LEAM和HSP22在豌豆种子中具有重要作用。从豌豆育种历史以及胚胎后期丰富蛋白基因进化机制的角度对LEAM的变异性进行了讨论。