Encoding Elongation Factor 1-alpha Research Articles

Candidate Genes for Freezing and Drought Tolerance Selected on the Basis of Proteome Analysis in Doubled Haploid Lines of Barley.

Plant tolerance to environmental stress is determined by a very complicated network composed of many intra- and extracellular factors. The aim of this study was to select candidate genes involved in responses to freezing and drought in barley on the basis of previous proteomic studies and to analyze changes in their expression caused by application of both stress factors. Six candidate genes for freezing tolerance (namely the genes encoding elongation factor 1 alpha (EF1A), ferredoxin-NADP reductase, a 14-3-3a protein, β-fructofuranosidase, CBF2A and CBF4B) and six for drought tolerance (encoding transketolase, periplasmic serine protease, triosephosphate isomerase, a protein with a co-chaperon region (GroEs), pfam14200 and actin) were chosen arbitrarily on the basis of in silico bioinformatic analyses. The expression levels of these genes were measured under control and stress conditions in six DH (doubled haploid) lines with differing freezing and drought tolerance. The results of gene expression analysis confirmed the roles of the candidate genes preselected in this study on the basis of previous proteome analysis in contributing to the differences in freezing and drought tolerance observed in the studied population of DH lines of winter barley.

A new aphid genus and species (Hemiptera: Aphididae: Macrosiphini) living on ferns in Costa Rica and Mexico

AbstractAphid species colonising ferns belong to the subfamily Aphidinae (Hemiptera: Aphididae) and the majority of these to the tribe Macrosiphini. A new genus in this tribe and its type species:Gibbomyzus pteridophytorumnew genus,new species, are established. Apterous and alate viviparous females are described from specimens collected onBlechnum buchtieniiRosenstock (Blechnaceae) in Costa Rica and onPteridium aquilinum(Linnaeus) Kuhn (Dennstaedtiaceae) and an unidentified fern in Mexico. The taxonomic validity of the two new taxa is discussed based on morphological and molecular data. Morphologically, the new genus is compared with genera with swollen siphunculi recorded in the New World, and also with genera living on ferns anywhere in the world. The identification key by Blackman and Eastop to aphids living on ferns is modified. Molecular analyses were carried out on the sequences of a fragment of the mitochondrial gene encoding for cytochromecoxidase subunit 1 and of a fragment of the nuclear gene encoding elongation factor-1 alpha. In both analyses,G. pteridophytorumnewspeciessequences showed considerable divergence (∼4% or more) from those of 23 other species from diverse genera of Macrosiphini, supporting the conclusions of the morphological study and justifying the establishment of the new genus.

Molecular cloning and tissue specific expression of an elongation factor 1A gene in sugarcane stalks

Genomic reconstruction done by using the cDNA insert containing the ORF of GZ-1-alpha as the probe indicated the presence of about 5 copies of eEF1A gene per haploid sugarcane genome. Northern hybridization studies revealed the presence of eEF1A transcripts in Saccharum officinarum stalks at late growth stage. A full-length cDNA clone (GZ-1-alpha) encoding elongation factor 1-alpha has been isolated from stalks of S. officinarum L. (accession number EF581011). The 1736 bp nucleotide sequence and the deduced 447 amino acid sequence shared 65–92 % and 95–99 % identity, respectively, to the other plant elongation factor 1As. The 400 mg/L ethephon treatment significantly induced the expression of GZ-1-alpha and maintained higher level in the top internodes for a long time, but did not induce the expression in the mid and bottom internodes. The results suggest that ethephon mediates the tissue and growth stage specific expressions of eEF1A gene isolated from S. officinarum stalks.

Cloning of aSchizosaccharomyces pombehomologue of elongation factor 1 alpha by two-hybrid selection of calmodulin-binding proteins

This study reports the cloning and characterization of a cDNA encoding elongation factor 1-alpha (EF1alpha) from the yeast Schizosaccharomyces pombe. The cDNA was cloned from an Schizosaccharomyces pombe expression library by a two-hybrid selection for clones encoding calmodulin (CaM)-binding proteins. The predicted protein is highly homologous to mammalian EF1alpha, indicating a strong tendency towards conservation of the primary amino acid sequence. The protein was expressed as a glutathione S-transferase fusion in both bacteria and in Schizosaccharomyces pombe. The bacterial protein was shown by solution assay to compete with CaM kinase II for CaM. The CaM binding domain was localized to the C-terminus of the protein by this method. Expression of full-length EF1alpha in vivo caused an increase in cell cycle length and a decreased rate of growth as evidenced by a lack of elongated cells in slowly dividing cultures. This effect appears to involve CaM binding because a truncation mutant version of EF1alpha lacking the CaM binding domain did not cause cell cycle delay.

Cloning of a <i>Schizosaccharomyces pombe</i> homologue of elongation factor 1 alpha by two-hybrid selection of calmodulin-binding proteins

This study reports the cloning and characterization of a cDNA encoding elongation factor 1-alpha (EF1alpha) from the yeast Schizosaccharomyces pombe. The cDNA was cloned from an Schizosaccharomyces pombe expression library by a two-hybrid selection for clones encoding calmodulin (CaM)-binding proteins. The predicted protein is highly homologous to mammalian EF1alpha, indicating a strong tendency towards conservation of the primary amino acid sequence. The protein was expressed as a glutathione S-transferase fusion in both bacteria and in Schizosaccharomyces pombe. The bacterial protein was shown by solution assay to compete with CaM kinase II for CaM. The CaM binding domain was localized to the C-terminus of the protein by this method. Expression of full-length EF1alpha in vivo caused an increase in cell cycle length and a decreased rate of growth as evidenced by a lack of elongated cells in slowly dividing cultures. This effect appears to involve CaM binding because a truncation mutant version of EF1alpha lacking the CaM binding domain did not cause cell cycle delay.

Molecular phylogeny of the major arthropod groups indicates polyphyly of crustaceans and a new hypothesis for the origin of hexapods.

A phylogeny of the arthropods was inferred from analyses of amino acid sequences derived from the nuclear genes encoding elongation factor-1 alpha and the largest subunit of RNA polymerase II using maximum-parsimony, neighbor-joining, and maximum-likelihood methods. Analyses of elongation factor-1 alpha from 17 arthropods and 4 outgroup taxa recovered many arthropod clades supported by previous morphological studies, including Diplopoda, Myriapoda, Insecta, Hexapoda, Branchiopoda (Crustacea), Araneae, Tetrapulmonata, Arachnida, Chelicerata, and Malacostraca (Crustacea). However, counter to previous studies, elongation factor-1 alpha placed Malacostraca as sister group to the other arthropods. Branchiopod crustaceans were found to be more closely related to hexapods and myriapods than to malacostracan crustaceans. Sequences for RNA polymerase II were obtained from 11 arthropod taxa and were analyzed separately and in combination with elongation factor-1 alpha. Results from these analyses were concordant with those derived from elongation factor-1 alpha alone and provided support for a Hexapoda/Branchiopoda clade, thus arguing against the monophyly of the traditionally defined Atelocerata (Hexapoda + Myriapoda).