Computational Discovery of a miRNA and its Putative Target Genes in Ziziphus Jujuba using Genome-wide Expressed Sequence Tags

Background Ziziphus jujuba is an important fruit crop which is increasingly becoming popular among consumers due to its medicinal properties. Increasing worldwide demand for the fruit poses new challenges to the industry which includes the need for accelerated cultivar development of jujubes. To embark on cultivar development with improved traits such as high yield and disease resistance, molecular and conventional breeding, and genetic engineering become imperative. But inadequate trait-enhancing alleles or gene pleiotropism limit the direct use of several identified genes. To overcome these issues, microRNAs (miRNAs) can be utilized in breeding of jujubes as genetic modulators to fine-tune the regulation of gene expression, thus the discovery of miRNAs becomes important. Methods and results In this study using a computational approach, we identified one potential miRNA (zju-miR-215-3p) from 2904 expressed sequence tags. The miRNA showed down regulation of five target proteins (AP-2 complex subunit alpha, C2H2-type domain-containing protein, sentrin-specific protease 1, hydrolase_4 domain-containing protein and putative alpha-ketoglutarate-dependent dioxygenase) and their suppression appears to be helpful to the plant to overcome stress conditions. Conclusion The miRNA identified in this study is associated with five potential target proteins, most of which are implicated in metabolic and developmental processes associated with plant growth and reproduction. Future studies are necessary to validate the miRNA by RNA sequencing and to confirm the molecular functions of the down regulations of target proteins.

Data from expressed sequence tags from the organs and embryos of parthenogenetic Haemaphysalis longicornis

Objectives Haemaphysalis longicornis is the most important tick species in Japan and has a wide range of vector capacity. Due to its veterinary and medical importance, this tick species has been used as a model for tick/vector biological studies. To identify the key molecules associated with physiological processes during blood feeding and embryogenesis, full-length cDNA libraries were constructed using the fat body, hemocytes-containing hemolymph, midgut, ovary and salivary glands of fed females and embryos of the laboratory colony of parthenogenetic H. longicornis. The sequences of cDNA from the salivary glands had been already released. However, the related information is still poor, and the other expressed sequence tags have not yet been deposited. Data description A total of 39,113 expressed sequence tags were obtained and deposited at the DNA DataBank of Japan. There were 7745 sequences from embryos, 7385 from the fat body, 8303 from the hemolymph including hemocytes, 7385 from the midgut, and 8295 from the ovary. The data, including expressed sequence tags from the salivary glands was summarized into Microsoft Excel files. Sharing this data resource with the tick research community will be valuable for the identification of novel genes and advance the progress of tick research.

Molecular diversity and evolutionary trends of cysteine-rich peptides from the venom glands of Chinese spider Heteropoda venatoria

Heteropoda venatoria in the family Sparassidae is highly valued in pantropical countries because the species feed on domestic insect pests. Unlike most other species of Araneomorphae, H. venatoria uses the great speed and strong chelicerae (mouthparts) with toxin glands to capture the insects instead of its web. Therefore, H. venatoria provides unique opportunities for venom evolution research. The venom of H. venatoria was explored by matrix-assisted laser desorption/ionization tandem time-of-flight and analyzing expressed sequence tags. The 154 sequences coding cysteine-rich peptides (CRPs) revealed 24 families based on the phylogenetic analyses of precursors and cysteine frameworks in the putative mature regions. Intriguingly, four kinds of motifs are first described in spider venom. Furthermore, combining the diverse CRPs of H. venatoria with previous spider venom peptidomics data, the structures of precursors and the patterns of cysteine frameworks were analyzed. This work revealed the dynamic evolutionary trends of venom CRPs in H. venatoria: the precursor has evolved an extended mature peptide with more cysteines, and a diminished or even vanished propeptides between the signal and mature peptides; and the CRPs evolved by multiple duplications of an ancestral ICK gene as well as recruitments of non-toxin genes.

In Silico Investigation of Conserved miRNAs and Their Targets From the Expressed Sequence Tags in Neospora Caninum Genome

Neospora caninum is a protozoan parasite, the etiologic agent of Neosporosis—a common cause of abortion in cattle worldwide. Herd level prevalence of Neosporosis could be as high as 90%. However, there is no approved treatment and vaccines available for Neosporosis. MicroRNA (miRNA) based prophylaxis and therapeutics could be options for Neosporosis in cattle and other animals. The current study aimed to investigate the genome of Neospora caninum to identify and characterize the conserved miRNAs through Expressed Sequence Tags (ESTs) dependent homology search. A total of 1,041 mature miRNAs of reference organisms were employed against 336 non-redundant ESTs available in the genome of Neospora caninum. The study predicted one putative miRNA “nca-miR-9388-5p” of 19 nucleotides with MFEI value -1.51 kcal/mol and (A + U) content% 72.94% corresponding with its pre-miRNA. A comprehensive search for specific gene targets was performed and discovered 16 potential genes associated with different protozoal physiological functions. Significantly, the gene “Protein phosphatase” was found responsible for the virulence of Neospora caninum. The other genes were accounted for gene expression, vesicular transport, cell signaling, cell proliferation, DNA repair mechanism, and different developmental stages of the protozoon. Therefore, this study finding will provide pivotal information to future aspirants upon Bovine Neosporosis. It will also serve as the baseline information for further studies of the bioinformatics approach to identify other protozoal miRNAs.

VfODB: a comprehensive database of ESTs, EST-SSRs, mtSSRs, microRNA-target markers and genetic maps in Vicia faba

Faba bean (Vicia faba) is an essential food and fodder legume crop worldwide due to its high content of proteins and fibres. Molecular markers tools represent an invaluable tool for faba bean breeders towards rapid crop improvement. Although there have historically been few V. faba genome resources available, several transcriptomes and mitochondrial genome sequence data have been released. These data in addition to previously developed genetic linkage maps represent a great resource for developing functional markers and maps that can accelerate the faba bean breeding programmes. Here, we present the Vicia faba Omics database (VfODB) as a comprehensive database integrating germplasm information, expressed sequence tags (ESTs), expressed sequence tags-simple sequence repeats (EST-SSRs), and mitochondrial-simple sequence repeats (mtSSRs), microRNA-target markers and genetic maps in faba bean. In addition, KEGG pathway-based markers and functional maps are integrated as a novel class of annotation-based markers/maps. Collectively, we developed 31 536 EST markers, 9071 EST-SSR markers and 3023 microRNA-target markers based on V. faba RefTrans V2 mining. By mapping 7940 EST and 2282 EST-SSR markers against the KEGG pathways database we successfully developed 107 functional maps. Also, 40 mtSSR markers were developed based on mitochondrial genome mining. On the data curation level, we retrieved 3461 markers representing 12 types of markers (CAPS, EST, EST-SSR, Gene marker, INDEL, Isozyme, ISSR, RAPD, SCAR, RGA, SNP and SSR), which mapped across 18 V. faba genetic linkage maps. VfODB provides two user-friendly tools to identify, classify SSR motifs and in silico amplify their targets. VfODB can serve as a powerful database and helpful platform for faba bean research community as well as breeders interested in Genomics-Assisted Breeding.

Identification of genes involved in flowering in Stevia rebaudiana using expressed sequence tags (ESTs)

Stevia rebaudiana, or stevia, is a perennial herb native to the northern region of South America. It is commercially important due to the high level of non-caloric sweetening compound, which has been consumed worldwide as food and medicine. Stevioside is considered as the most important compound of steviol glycosides that functions as the main sweetener. Exposure of the stevia plant to long-day conditions (increased exposure to light) will delay the flowering process, which led to an increase in glycoside accumulation. The bioinformatics analysis of 5548 expressed sequence tags (ESTs) obtained from dbEST (NCBI) using the Blast2GO software and manual curation identified 7 ESTs that contain domains involved in the flowering process. The domains were the heat shock protein (Hsp) domain, squamosa promoter-binding protein (SBP) domain, CONSTANS, CO-like, and TOC1 (CCT) domains; K homology (KH) domain; and ubiquitin-conjugating enzyme domain (UBC).