RT Journal Article SR Electronic T1 Chromosomal length reference assembly for Diaphorina citri using single-molecule sequencing and Hi-C proximity ligation with manually curated genes in developmental, structural and immune pathways JF bioRxiv FD Cold Spring Harbor Laboratory SP 869685 DO 10.1101/869685 A1 Prashant S. Hosmani A1 Mirella Flores-Gonzalez A1 Teresa Shippy A1 Chad Vosburg A1 Crissy Massimino A1 Will Tank A1 Max Reynolds A1 Blessy Tamayo A1 Sherry Miller A1 Jordan Norus A1 Kyle Kercher A1 Bec Grace A1 Margaryta Jernigan A1 Doug Harper A1 Sam Adkins A1 Yesmarie DeLaFlor A1 Thomson Paris A1 Sara Vandervoort A1 Rebekah Adams A1 Seantel Norman A1 Jessica Ventura A1 Michael Perry A1 Matthew Weirauch A1 Josh Benoit A1 Wayne B. Hunter A1 Helen Wiersma-Koch A1 Tom D’elia A1 Susan Brown A1 Lukas A. Mueller A1 Surya Saha YR 2019 UL http://biorxiv.org/content/early/2019/12/10/869685.abstract AB Hemipterans include some of the most important insect pests of agricultural systems and vectors of plant pathogens. The vector, Diaphorina citri (Asian citrus psyllid) belonging to the Psylloidae superfamily, is the primary target of approaches to stop the spread of the pathogen Ca. Liberibacter asiaticus that causes Huanglongbing or citrus greening disease. High quality genomic resources enable rapid functional discovery that can target disease transmission and control. The previous psyllid genome (Diaci v1.1) available in NCBI is missing 25% of the single copy markers conserved in other Hemipterans. Manual genome curation helped to identify a significant number of genome anomalies including misassemblies and missing genes. We present an improved and highly contiguous de novo assembly based on PacBio long reads followed by Dovetail Chicago and Hi-C based scaffolding. The current assembly (Diaci v3) has 13 chromosomal length scaffolds with a genome size of 475 Mb. This is the first report of a chromosomal length assembly in the Hemiptera order according to our knowledge. Full-length cDNA transcripts were sequenced with PacBio Iso-Seq technology from diseased and healthy tissue at multiple life stages. Iso-Seq along with diverse Illumina RNA-Seq expression data were used to predict 19,049 protein-coding genes in psyllid using MAKER annotation pipeline. We also generated a genome independent transcriptome with a comprehensive catalog of all genes in the psyllid.Gene-targeting technologies like RNAi, antisense oligos and CRISPR require accurate annotation of genes. Lack of closely related and well characterized model organisms coupled with the diversity of insect genomes impacts the quality of predicted gene models. We used the improved genomic resources to create a high-quality manually curated gene set for developmental, structural and immune pathways. All resources are available on https://citrusgreening.org/, a portal for all omics resources for the citrus greening disease research community. The high quality ACP genome assembly, annotation based on transcriptomics evidence, manual curation of critical pathways and a genome independent de novo transcriptome will provide a foundation for comparative analysis among genomes of agricultural pests and vectors in the Hemiptera.