Project description:We used PacBio data to identify more reliable transcripts from hESC, based on which we can estimate gene/transcript abundance better from Illumina data. PacBio long reads and Illumina short reads were generated from the same hESC cell line H1. PacBio reads were error-corrected by Illumina reads to identify transcripts. rSeq is used to estimate gene/transcript abundance of the identified transcriptome.
Project description:Citrus species are among the most important fruit crops. However, gene regulation and signaling pathways related to etiolation in this crop remain unknown. Using Illumina sequencing technology, modification of global gene expression in two hybrid citrus cultivars—Huangguogan and Shiranuhi, respectively—were investigated. More than 834.16 million clean reads and 125.12 Gb of RNA-seq data were obtained, more than 91.37% reads had a quality score of Q30 (sequencing error rate, 0.1%). 124,952 unigenes were finally generated with a mean length of 1,189 bp. Of these unigenes, 98,904 (79.15%), 105,408 (84.35%), 42,016 (33.62%), 78,872 (63.12%), 72,068 (57.67%), 72,464 (57.99%), 72464 (57.99%) and 46,308 (37.06%) had been annotated in NR, NT, KO, SwissProt, PFAM, GO and KOG databases, respectively. Further, we identified 604 differentially expressed genes (DEGs) in multicoloured and etiolated seedlings of Shiranuhi, including 180 up-regulated genes and 424 down-regulated genes. While in Huangguogan, we found 1,035 DEGs, 271 of which were increasing and the others were decreasing. 7 DEGs were commonly up-regulated, and 59 DEGs down-regulated in multicoloured and etiolated seedlings of these two cultivars, suggesting that some genes play fundamental roles in two hybrid citrus seedlings during etiolation. Functional classification of the DEGs in two cultivars using GO term indicated that biological process, cellular component and molecular function were three major groups. Our study is the first to provide the transcriptome sequence resource for seedlings etiolation of Shiranuhi and Huangguogan, and advance our knowledge of the genes involved in the complex regulatory networks of seedling etiolation.
Project description:Purpose: We obtained RNA-seq-based differential expression profile of Valencia sweet orange plants challenged against healthy and CLas-infected psyllid infection at 1 dpi and 5 dpi. The goals of this study are to reveal the interaction between citrus and psyllid/CaLas during the early phase of infection and understand the molecular mechanisms underlying the host-pathogen interactions and the susceptibility of most citrus varieties. Methods: leaf mRNA profiles of in vitro cultured Valencia sweet orange (VAL) budwood (WT) and of VAL fed by healthy and CLas-infected psyllid were generated by RNA-seq, in triplicate (one sample is duplicate), using Illumina HiSeq platform. The sequence reads that passed quality filters were used for gene expression and DEG detection analysis by EBseq algorithms. qRT–PCR validation was performed using SYBR Green assays Results: Using the RNA-seq data analysis workflow, we mapped about 136.80M sequence reads per sample to the reference Citrus clementina v1.0 genome and a total of 32,677 genes were detected. The average total mapping of each library was 71.98%. RNA-seq data were validated with qRT–PCR. Conclusions: Our study obtained the transcriptional profiles of citrus host by feeding of psyllid transmitting Candidatus Liberibacter asiaticus at early stages of infection, with biologic replicates, generated by RNA-seq technology. The RNA-seq data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.