A 6374 unigene set corresponding to low abundance transcripts expressed following fertilization in Solanum chacoense Bitt, and characterization of 30 receptor-like kinases.
In order to characterize regulatory genes that are expressed in ovule tissues after fertilization we have undertaken an EST sequencing project in Solanum chacoense, a self-incompatible wild potato species. Two cDNA libraries made from ovule tissues covering embryo development from zygote to late torpedo-stage were constructed and plated at high density on nylon membranes. To decrease EST redundancy and enrich for transcripts corresponding to weakly expressed genes a self-probe subtraction method was used to select the colonies harboring the genes to be sequenced. 7741 good sequences were obtained and, from these, 6374 unigenes were isolated. Thus, the self-probe subtraction resulted in a strong enrichment in singletons, a decrease in the number of clones per contigs, and concomitantly, an enrichment in the total number of unigenes obtained (82%). To gain insights into signal transduction events occurring during embryo development all the receptor-like kinases (or protein receptor kinases) were analyzed by quantitative real-time RT-PCR. Interestingly, 28 out of the 30 RLK isolated were predominantly expressed in ovary tissues or young developing fruits, and 23 were transcriptionaly induced following fertilization. Thus, the self-probe subtraction did not select for genes weakly expressed in the target tissue while being highly expressed elsewhere in the plant. Of the receptor-like kinases (RLK) genes isolated, the leucine-rich repeat (LRR) family of RLK was by far the most represented with 25 members covering 11 LRR classes.
Germain
Hugo
H
Institut de Recherche en Biologie Végétale (IRBV). Département de sciences biologiques, Université de Montréal, 4101 Sherbrooke Est, H1X 2B2, Montréal, QC, Canada.
Rudd
Stephen
S
Zotti
Corine
C
Caron
Sébastien
S
O'Brien
Martin
M
Chantha
Sier-Ching
SC
Lagacé
Marie
M
Major
François
F
Matton
Daniel P
DP
D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D015723:Gene Library; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010940:Plant Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics; D012098:Reproduction / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D017422:Sequence Analysis, DNA; D032322:Solanum / Q000196:embryology / Q000235:genetics*; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics
D018076:DNA, Complementary; D010940:Plant Proteins; D012333:RNA, Messenger; D020794:Receptor Protein-Tyrosine Kinases