Isaria farinosa (Hypocreales, Ascomycota) is a cosmopolitan entomopathogenic fungus affecting a wide range of arthropod hosts. It has mainly been studied as a insecticidal agent to control the agricultural pests. To investigate the useful secondary metabolite(SM) genes in Isaria farinosa C1012 strain, de novo assembly and genome mining were carried out. A whole genome sequencing with PacBio RSII system generated NGS reads greater than 4Gb, which were assembled into 16 contigs using FALCON program. The total size of genome was 33.36Mb. The N50 and N90 were 6,686,213 and 1,912,865bp, respectively. The assembled genome data was analyzed with antiSMASH3 program with a default setting to localize the gene region responsible for synthesizing SMs, such as non-ribosomal peptide synthetases (NRPS) and polyketide synthase (PKS). In this study, we predicted 16 NRPS, 13 PKS, and 9 PKS-NRPS hybrid gene clusters in I. farinosa genome.
The small brown planthopper (SBPH), Laodelphax striatellus Fallén (Hemiptera: Delphacidae) is one of the major insect pest against rice, Oryza sativa L. in Korea. High density of SBPH could cause severe damage on rice plant by directly sucking and indirectly transmitting viral pathogens, Rice stripe virus and Rice streaked dwarf virus. As a preliminary study for de novo whole-genome sequencing of SBPH, we investigated 6 transcriptomes isolated from different developmental stages, sex, and tissue (egg, 1st ~ 3rd nymphs, 4th ~ 5th nymphs, female and male adults, salivary gland). Clean-sequence data of 19.3 Gb were obtained from total 47.8 Gb raw data after adaptor and quality trimming (Q30) and overlapped reads joining. As a suitable assembler, Bridger was selected based on the results of reference mapping (93.45%) and CEGMA completeness (95.97%). Finally, we obtained 158,207 reads (size range: 201 ~ 22,162 bp; Mean size: 1,048.04 bp; N50: 2,417 bp) after clustering the assembly results by CD-HIT-EST (similarity threshold: 99%). Based on these results, we are conducting further studies such as transcript expression pattern among different developmental stages and gene annotation.
Rice stripe virus disease (RSVD), one of the most serious disease of rice is mediated through the sucking by small brown planthopper, Laodalphax striatellus. So far, the studies have been mainly focused on the interaction between the host plant and the virus. In this study, for better comprehension of the interactions among the host plant, vector insect and plant-pathogenic virus, we investigated transcriptome of the vector insect and the differences between viruliferous and naïve L.striatellus. For this, naïve L. striatellus were collected from non-infected rice field and 50 L.striatellus of them were fed RSV-infected rice for 5 days. With the RSV-viruliferous and the naïve insects, we conducted Illumina RNA sequencing (Hiseq 2000) and obtained 175,243,488 and 146,031,348 reads from viruliferous and naïve L.striatellus, respectively. These reads were assembled into contigs and two transcriptome databases were generated. The transcriptome of naïve and RSV-viruliferous L. striatellus were campared to figure out up-regulated or down-regulated genes. These RSV-dependently regulated genes may have important function in the behavior of planthoppers or the transmission of RSV.
The high quality of gene set is necessary to study the functional research of genes. Although perilla is cultivated as an oil crop and as a vegetable crop in Asian countries such as Korea, Japan, northeast China and Nepal, the reference genome is absent. To assembly perilla gene set, we sequenced the various tissues of perilla (Perilla citriodora) RNA-seq with Illumina HiSeq platform, generating 548,549,314 short reads. When de novo transcriptome assembly was performed with five samples, 86,396 and 38,413 transcripts were assembled as total and representative transcripts, respectively. Using 1,917,424 proteins at Phytozome ver. 9.1, we annotated the perilla assembled transcripts, and 66,139(76.55%) and 24,030(62.55%) transcripts showed the similarity with known plant proteins (E-value < 1e-10) as total and representative transcripts, respectively. Among the diverse molecular functions, we were interested in the regulatory components, such as transcription factor and transcription regulator. Using this data, we identified 499 transcripts annotated the putative transcription factor differentially expressed transcripts. 165 putative transcription factors were significantly expressed in perilla flower and 121 putative transcription factors in both leaf and flower. This study provides the perilla reference gene set and the understanding of the molecular regulation of transcription factor dependent on the tissue.
Bulb onion (Allium cepa) is one of the second most widely cultivated and consumed vegetable crops in the world. During winter where the temperature can be as low, plant could get cold injury and limit the production of bulb onion. However, the genomic resources available for bulb onion are still very limited. To date, no studies about heritably durable cold and freezing tolerance were carried out in bulb onion genotypes using high-throughput sequencing technology was applied. We sequenced cold (2°C) freezing (-5 and -15°C) treated and control (25°C) samples of contrasting genotypes of A. cepa lines and obtained 4,52,194,370 total high quality reads. After de novo assembly reads were assembled into 54,047 genes finally generated with an average length of 1,331 bp. Based on the similarity search aligning all genes with known public non-redundant (NR) database, including Swiss-prot, KEGG and COG. Differentially expressed genes (DEGs) were investigated using FPKM method. Overall, 92,862 genes were differentially regulated in all libraries were identified. Additionally, increase our understanding of the DEGs, we performed GO and KEGG pathway enrichment analyses. Based on FDR<=0.01 value in cold freezing tolerant line candidate genes were selected and discussed. Finally 25 candidate genes were examined using qRT-PCR were differentially regulated and known to be associated with cold and freezing stresses. Moreover, in silico prediction of putative molecular marker 4,437 SSRs and 6,076 SNPs. Our study is the first to provide the transcriptome sequence resource of Allium spp., for cold and freezing stress. We identified large set of genes to determine its DEGs profile under cold and freezing condition using two different genotypes. These data provides a valuable resource of genetic and genomic studies of Allium spp.
Wheat is the third largest crop in the world behind corn and rice. Wheat is grown over a wide range of environments, and an essential source of carbohydrates. However, the genomics of wheat, a non-model species, is still challenging despite of corn and rice was done. The recent advent of RNA-Sequencing, a massively parallel sequencing method for genome and transcriptome analysis, provides opportunity to identify gene discovery and molecular mechanisms of cellular processes. We performed a RNA-Seq experiment to find differentially expressed genes under high temperature condition. More than 344 million shot reads were generated using Illumina HiSeq technology. A comprehensive and integrated 285,324 transcripts were assembled via Trinity by combining tentative consensus sequences. Transcripts annotated by BLAST2Go and differently expressed transcripts were analyzed. A total of 208 up-regulated and 182 down-regulated transcripts were found that involve in plastid, starch and sucrose metabolism, glycerolipid metabolism and glycerolipid and dicarboxylate metabolism. Our results demonstrate that RNA-Seq can be successfully used for gene identification, transcript profiling in wheat. Furthermore these sequences will provide valuable resources for wheat researchers.