Gene expression analysis in the roots of salt stressed wheat and the cytogenetic derivatives of wheat combined with the salt-tolerant wheatgrass, Lophopyrum elongatum

Lophopyrum elongatum is among one of the most salt tolerant members of the Triticeae; important genetic stocks developed from crosses between wheat and L. elongatum provide a unique opportunity to compare gene expression in response to salt stress between these highly related species. The octaploid amphiploid contains the entire genome of T. aestivum and L. elongatum, and the wheat disomic substitution line DS3E(3A) has chromosome 3A of wheat replaced by chromosome 3E of L. elongatum. In this study, microarray analysis was used to characterize gene expression profiles in the roots of three genotypes, Triticum aestivum, the octaploid amphiploid, and the DS3E(3A) substitution line, in response to salt stress. We first examined changes in gene expression in wheat over a time course of three days of salt stress, and then compared changes in gene expression in wheat, the T. aestivum x L. elongatum amphiploid and in the DS3E(3A) substitution line after three days of salt stress. In the time course experiment, 237 genes had a 1.5 fold or greater change at least once out of three time points assayed in the experiment. The comparison between the three genotypes revealed 304 genes with significant differences in changes of expression between the genotypes. Forty two of these genes had at least a two-fold change in expression in response to salt treatment; 18 of these genes have signaling or regulatory function. Genes with significant differences in induction or repression between genotypes included transcription factors, protein kinases, ubiquitin ligases and genes related to phospholipid signaling

Wheat EST resources for functional genomics of abiotic stress

BACKGROUND: Wheat is an excellent species to study freezing tolerance and other abiotic stresses. However, the sequence of the wheat genome has not been completely characterized due to its complexity and large size. To circumvent this obstacle and identify genes involved in cold acclimation and associated stresses, a large scale EST sequencing approach was undertaken by the Functional Genomics of Abiotic Stress (FGAS) project. RESULTS: We generated 73,521 quality-filtered ESTs from eleven cDNA libraries constructed from wheat plants exposed to various abiotic stresses and at different developmental stages. In addition, 196,041 ESTs for which tracefiles were available from the National Science Foundation wheat EST sequencing program and DuPont were also quality-filtered and used in the analysis. Clustering of the combined ESTs with d2_cluster and TGICL yielded a few large clusters containing several thousand ESTs that were refractory to routine clustering techniques. To resolve this problem, the sequence proximity and "bridges" were identified by an e-value distance graph to manually break clusters into smaller groups. Assembly of the resolved ESTs generated a 75,488 unique sequence set (31,580 contigs and 43,908 singletons/singlets). Digital expression analyses indicated that the FGAS dataset is enriched in stress-regulated genes compared to the other public datasets. Over 43% of the unique sequence set was annotated and classified into functional categories according to Gene Ontology. CONCLUSION: We have annotated 29,556 different sequences, an almost 5-fold increase in annotated sequences compared to the available wheat public databases. Digital expression analysis combined with gene annotation helped in the identification of several pathways associated with abiotic stress. The genomic resources and knowledge developed by this project will contribute to a better understanding of the different mechanisms that govern stress tolerance in wheat and other cereals

Unique and conserved MicroRNAs in wheat chromosome 5D revealed by next-generation sequencing

MicroRNAs are a class of short, non-coding, single-stranded RNAs that act as post-transcriptional regulators in gene expression. miRNA analysis of Triticum aestivum chromosome 5D was performed on 454 GS FLX Titanium sequences of flow sorted chromosome 5D with a total of 3,208,630 good quality reads representing 1.34x and 1.61x coverage of the short (5DS) and long (5DL) arms of the chromosome respectively. In silico and structural analyses revealed a total of 55 miRNAs; 48 and 42 miRNAs were found to be present on 5DL and 5DS respectively, of which 35 were common to both chromosome arms, while 13 miRNAs were specific to 5DL and 7 miRNAs were specific to 5DS. In total, 14 of the predicted miRNAs were identified in wheat for the first time. Representation (the copy number of each miRNA) was also found to be higher in 5DL (1,949) compared to 5DS (1,191). Targets were predicted for each miRNA, while expression analysis gave evidence of expression for 6 out of 55 miRNAs. Occurrences of the same miRNAs were also found in Brachypodium distachyon and Oryza sativa genome sequences to identify syntenic miRNA coding sequences. Based on this analysis, two other miRNAs: miR1133 and miR167 were detected in B. distachyon syntenic region of wheat 5DS. Five of the predicted miRNA coding regions (miR6220, miR5070, miR169, miR5085, miR2118) were experimentally verified to be located to the 5D chromosome and three of them : miR2118, miR169 and miR5085, were shown to be 5D specific. Furthermore miR2118 was shown to be expressed in Chinese Spring adult leaves. miRNA genes identified in this study will expand our understanding of gene regulation in bread wheat

A computational-based update on microRNAs and their targets in barley (Hordeum vulgare L.)

<p>Abstract</p> <p>Background</p> <p>Many plant species have been investigated in the last years for the identification and characterization of the corresponding miRNAs, nevertheless extensive studies are not yet available on barley (at the time of this writing). To extend and to update information on miRNAs and their targets in barley and to identify candidate polymorphisms at miRNA target sites, the features of previously known plant miRNAs have been used to systematically search for barley miRNA homologues and targets in the publicly available ESTs database. Matching sequences have then been related to Unigene clusters on which most of this study was based.</p> <p>Results</p> <p>One hundred-fifty-six microRNA mature sequences belonging to 50 miRNA families have been found to significantly match at least one EST sequence in barley. As expected on the basis of phylogenetic relations, miRNAs putatively orthologous to those of <it>Triticum </it>are significantly over-represented inside the set of identified barley microRNA mature sequences. Many previously known and several putatively new miRNA/target pairs have been identified. When the predicted microRNA targets were grouped into functional categories, biological processes previously known to be regulated by miRNAs, such as development and response to biotic and abiotic stress, have been highlighted and most of the target molecular functions were related to transcription regulation. Candidate microRNA coding genes have been reported and genetic variation (SNPs/indels) both in functional regions of putative miRNAs (mature sequence) and at miRNA target sites has been found.</p> <p>Conclusions</p> <p>This study has provided an update of the information on barley miRNAs and their targets representing a foundation for future studies. Many of previously known plant microRNAs have homologues in barley with expected important roles during development, nutrient deprivation, biotic and abiotic stress response and other important physiological processes. Putative polymorphisms at miRNA target sites have been identified and they can represent an interesting source for the identification of functional genetic variability.</p

Boron Stress Responsive MicroRNAs and Their Targets in Barley

Boron stress is an environmental factor affecting plant development and production. Recently, microRNAs (miRNAs) have been found to be involved in several plant processes such as growth regulation and stress responses. In this study, miRNAs associated with boron stress were identified and characterized in barley. miRNA profiles were also comparatively analyzed between root and leave samples. A total of 31 known and 3 new miRNAs were identified in barley; 25 of them were found to respond to boron treatment. Several miRNAs were expressed in a tissue specific manner; for example, miR156d, miR171a, miR397, and miR444a were only detected in leaves. Additionally, a total of 934 barley transcripts were found to be specifically targeted and degraded by miRNAs. In silico analysis of miRNA target genes demonstrated that many miRNA targets are conserved transcription factors such as Squamosa promoter-binding protein, Auxin response factor (ARF), and the MYB transcription factor family. A majority of these targets were responsible for plant growth and response to environmental changes. We also propose that some of the miRNAs in barley such as miRNA408 might play critical roles against boron exposure. In conclusion, barley may use several pathways and cellular processes targeted by miRNAs to cope with boron stress

Discovery of barley miRNAs through deep sequencing of short reads

Background: MicroRNAs are important components of the regulatory network of biological systems and thousands have been discovered in both animals and plants. Systematic investigations performed in species with sequenced genomes such as Arabidopsis, rice, poplar and Brachypodium have provided insights into the evolutionary relationships of this class of small RNAs among plants. However, miRNAs from barley, one of the most important cereal crops, remain unknown. Results: We performed a large scale study of barley miRNAs through deep sequencing of small RNAs extracted from leaves of two barley cultivars. By using the presence of miRNA precursor sequences in related genomes as one of a number of supporting criteria, we identified up to 100 miRNAs in barley. Of these only 56 have orthologs in wheat, rice or Brachypodium that are known to be expressed, while up to 44 appear to be specifically expressed in barley. Conclusions: Our study, the first large scale investigation of small RNAs in barley, has identified up to 100 miRNAs. We demonstrate that reliable identification of miRNAs via deep sequencing in a species whose genome has not been sequenced requires a more careful analysis of sequencing errors than is commonly performed. We devised a read filtering procedure for dealing with errors. In addition, we found that the use of a large dataset of almost 35 million reads permits the use of read abundance distributions along putative precursor sequences as a practical tool for isolating miRNAs in a large background of reads originating from other non-coding and coding RNAs. This study therefore provides a generic approach for discovering novel miRNAs where no genome sequence is available.Andreas W Schreiber, Bu-Jun Shi, Chun-Yuan Huang, Peter Langridge, Ute Bauman

Comparative analysis of local governments of Turkey and Bulgaria in the context of local government reform

Yerel yönetimler, ülkelerin hem yönetim yapısı içinde önemli bir role sahip olup hem de demokratikleşmenin ve halkın yönetime katılımının en iyi şekilde temsil edilebileceği birimlerdir. Günümüzde yerel yönetimler, ülkelerin çoğunda, anayasa ile düzenlenmiş, demokratik ve özerk kuruluşlar olma özelliği ile ülkelerin yönetim sistemleri içerisinde her geçen gün ağırlıklarını arttırmaktadırlar. Çalışmamızın amacı Türkiye ve Bulgaristan yerel yönetim sistemlerini karşılaştırmaktır. Bu bağlamda her iki ülkenin yerel yönetim sistemlerinin tarihi süreci, yerel yönetimlerin yasal mevzuatı ve yapısı, yerel yönetim alanında yapılan reform çalışmaları incelenmiştir. Çalışmamızın en son kısmı ise, bu iki ülkenin yerel yönetim sistemlerinin karşılaştırılıp değerlendirilmesini içermektedir. Çalışma sonucunda iki ülkenin yerel yönetim yapıları arasında bazı benzerlik ve farklılıklar ortaya konulmuştur. Bununla birlikte her iki ülkede de yapılan yerel yönetim reformları sonucunda elde edilen olumlu sonuçlara rağmen, hem Türkiye'de hem Bulgaristan'da merkeziyetçi yapının hala devam ettiği sonucuna ulaşılmıştır.Local government are the units which both have an important role in countries administration structure and also can be represented in the best way of democratization and participation of people in administration. Today, local government most of which are organized by the constitution of countries as democratic and autonomous institutions has been gaining more importance in their administrative systems day by day. The aim of our study is to compare the local government systems of Turkey and Bulgaria. İn this respect, the historical process of local government systems of both countries, legal regulations and structure of local government, local government reform studies in this area have been explained. The last part of this study includes comparing and evaluating of local government systems of these two countries. As a result of this study, it was proved that are some similarities and differences between local governments of these countries. However, despite positive results obtained from the local government reform, in both countries, it is concluded that a centralized structure still exists in both Turkey and Bulgaria