Identification of new gene expression regulators specifically expressed during plant seed maturation.
A cDNA-AFLP approach on Linum usitatissimum (flax) was used to identify genes specifically expressed during the seed maturation process. Among the 20,000 cDNA-AFLP tags produced, 486 were selected for their seed-specific expression during maturation. When compared with the publicly available databases, half of them presented some significant similarity with known plant sequences. The results obtained confirmed the accuracy of the approach as numerous genes previously described as being expressed exclusively in plant seeds were identified in this screen. The focus was on sequences similar to plant regulators involved in the control of gene expression, either at the transcriptional, post-transcriptional, or post-translational levels. Using a real-time RT-PCR approach, seed-specific expression kinetics were confirmed for 13 of these regulators that were never characterized for being expressed during seed maturation. Among these, a flax gene of the non-LEC1-like HAP3 family and a flax MYB factor were shown to be expressed in specialized tissues of flax embryo using an in situ hybridization approach. By expression kinetic comparison between these flax genes and their Arabidopsis counterparts, it was found that the new HAP3 gene should be related to a ubiquitous seed maturation mechanism, while a new MYB factor appears to be related to a more seed-specific maturation mechanism. These results demonstrate the utility of the flax database in not only identifying new genes expressed during seed maturation but also in being able to highlight the distinction between conserved and non-conserved seed maturation mechanisms.
Gutierrez
Laurent
L
Laboratoire de Génomique Fonctionnelle des Plantes, Université de Picardie Jules Verne, Faculté des Sciences, Amiens, France.
Conejero
Geneviève
G
Castelain
Mathieu
M
Guénin
Stéphanie
S
Verdeil
Jean-Luc
JL
Thomasset
Brigitte
B
Van Wuytswinkel
Olivier
O
D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000378:metabolism; D018076:DNA, Complementary; D018744:DNA, Plant; D020224:Expressed Sequence Tags; D019597:Flax / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D018506:Gene Expression Regulation, Plant; D017403:In Situ Hybridization; D018613:Microscopy, Confocal; D010940:Plant Proteins / Q000378:metabolism / Q000502:physiology; D012150:Polymorphism, Restriction Fragment Length; D046188:Protein Modification, Translational; D012323:RNA Processing, Post-Transcriptional; D018749:RNA, Plant / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000254:growth & development* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology
D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D018744:DNA, Plant; C491930:HAP3a protein, Arabidopsis; C500589:Myb protein, Arabidopsis; D010940:Plant Proteins; D018749:RNA, Plant; D014157:Transcription Factors