Characterization of a germin-like protein gene expressed in somatic and zygotic embryos of pine (Pinus caribaea Morelet)

被引:48
作者
Neutelings, G
Domon, JM
Membré, N
Bernier, F
Meyer, Y
David, A
David, H
机构
[1] Univ Sci & Tech Lille Flandres Artois, UFR Biol, Lab Physiol Parois Vegetales, F-59655 Villeneuve Dascq, France
[2] Fac Sci, Lab Biotechnol & Physiol Vegetales, UPRES EA 2085, F-80039 Amiens 1, France
[3] Univ Perpignan, Lab Physiol & Biol Mol Plantes, CNRS, UMR 5545, F-66860 Perpignan, France
[4] Inst Bot, CNRS, IBMP, F-67083 Strasbourg, France
关键词
apoplast; cDNA cloning; conifer embryogenesis; gene expression; germin-like proteins; Pinus caribaea;
D O I
10.1023/A:1006033622928
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Germin-like proteins (GLPs) ionically bound to the walls of preglobular somatic embryos of Pinus caribaea Morelet are markers of this early developmental stage. In order to reveal the physiological implications of such markers during early embryo development, we isolated a cDNA clone from somatic embryos predicted to encode a protein with sequence similarity to GLPs. PcGER1 has an open reading frame corresponding to a 220 amino acid polypeptide with a putative N-glycosylation site on Asn-69. The presence of a 24 amino acid putative signal peptide supports the hypothesis of an apoplastic location. The N-terminal 20 amino acid sequence of the predicted mature protein is identical to the amino terminal sequence of GP111, one of the extracellular pine GLPs previously identified. Southern blot hybridizations indicate that PcGER1 is probably unique in the pine genome. Transcripts homologous to PcGER1 are abundant in all embryogenic lines, absent from nonembryogenic lines, and present in quiescent zygotic embryos but not in the female gametophyte, the haploid storage tissue of conifers. Their abundance sharply decreases during germination. Isolation of gf-0.8, a genomic fragment identical to PcGER1 cDNA sequence, confirms that no introns disrupt the coding region as it has been already described for wheat gf-2.8 and gf-3.8 genomic clones. Recombinant PcGER1, produced in Escherichia coli, is recognized by antibodies raised against the GP111 N-terminal nonapeptide and the unglycosylated wheat germin monomer. The implications of GLPs in pine embryogenesis are discussed.
引用
收藏
页码:1179 / 1190
页数:12
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