ECHENIQUE Carmen Viviana
congresos y reuniones científicas
Analysis and functional clasification of transcripts from diploid wheat with different growth habits.
MARINA DÍAZ; VIVIANA ECHENIQUE; PANIEGO N; HELGUERA M; DUBCOVSKY J
Congreso; V Encuentro Latinoamericano y del Caribe de Biotecnología Agrícola, REDBIO 2004; 2004
Wheat is an allohexaploid species (2n=6x=42, AABBDD). With a genome size of 16,000 Mb is one of the biggest and more complex plant genomes. As an alternative to waiting for whole-genome sequencing, collections of transcript sequences from cDNA libraries (ESTs) can provide a valuable alternative to access most of the Triticeae genes. The aim of ourwork was the characterization of three cDNA libraries from shoot apexes from spring (DV92) and winter (G3116) lines of Triticum monococcum (2n=2x=14, AA). In the first case vegetative (TMO11XXX) and reproductive (TMO43XXX) apexes were used. In the second one, apexes in different vernalization stages (TMO46XXX) were used. Apexes were collected under stereoscopic microscope, total RNA was extracted using TRIZOL and purified with the PolyATract mRNA system. The cDNAs were directionally cloned into the Uni-ZAP XR vector. The primary lambda cDNA libraries were mass excised in vivo using the host strain XL1-Blue-MRF´ and the helper phage ExAssist to produce pBluescript phagemid populations. The titer and amplification fold were: primary library (TMO11XXX): 1,564,000 pfu/ml, amplified library 1.038x 108, amplification fold 141; TM043E1X: 290,000 pfu/ml, 4.02x108, 5,544; TM046E1X: 9.6 106 pfu/ml, 1.5 1010, 14,934. For this communication a total of 5917 sequencies longer than 100 nucleotides were considered, 2024 (TMO11XXX), 530 (TMO43EIX) y 3363 (TMO46EIX). The mean size of the sequences was 450 nucleotides. The redundancy was analyzed using the CAP3 software defining 1764, 509 and 2444 unigenes for the libraries TMO11XXX, TMO43EIX y TMO46EIX respectively. The sequencies were analyzed for similarity using BLASTX and the SwissProt database obtaining identity with sequences from the databases for 1,276 of 4,717 compared sequences. The functional categorization of the transcripts was based on the nomenclature and the classification outline proposed by Gene Ontology (GO; www.geneontology.org): biological processes (BP), cellular components (CC) and molecular function (MF).