Abstract
BackgroundLeafhoppers (Hemiptera: Cicadellidae) are plant-phloem feeders that are known for their ability to vector plant pathogens. The black-faced leafhopper (Graminella nigrifrons) has been identified as the only known vector for the Maize fine streak virus (MFSV), an emerging plant pathogen in the Rhabdoviridae. Within G. nigrifrons populations, individuals can be experimentally separated into three classes based on their capacity for viral transmission: transmitters, acquirers and non-acquirers. Understanding the molecular interactions between vector and virus can reveal important insights in virus immune defense and vector transmission.ResultsRNA sequencing (RNA-Seq) was performed to characterize the transcriptome of G. nigrifrons. A total of 38,240 ESTs of a minimum 100 bp were generated from two separate cDNA libraries consisting of virus transmitters and acquirers. More than 60% of known D. melanogaster, A. gambiae, T. castaneum immune response genes mapped to our G. nigrifrons EST database. Real time quantitative PCR (RT-qPCR) showed significant down-regulation of three genes for peptidoglycan recognition proteins (PGRP – SB1, SD, and LC) in G. nigrifrons transmitters versus control leafhoppers.ConclusionsOur study is the first to characterize the transcriptome of a leafhopper vector species. Significant sequence similarity in immune defense genes existed between G. nigrifrons and other well characterized insects. The down-regulation of PGRPs in MFSV transmitters suggested a possible role in rhabdovirus transmission. The results provide a framework for future studies aimed at elucidating the molecular mechanisms of plant virus vector competence.
Highlights
Hemipteran insects such as aphids, whiteflies, planthoppers and leafhoppers are arguably the most important vectors of plantinfecting viruses
Expression of seven genes with known function in insect innate immunity was evaluated in transmitters and leafhoppers raised on healthy plants using Real time quantitative PCR (RT-qPCR): acetylcholine receptor subunit alpha-L1 (AChR) (GnigEST002867), autophagy protein 5 (ATG 5) (GnigEST012478), defensin (GnigEST011432), peptidoglycan recognition protein SB1 (PGRPSB1) (GnigEST015027), peptidoglycan recognition proteins (PGRPs)-SD (GnigEST009213), PGRP-LC (GnigEST006324), tripeptidyl peptidase II (TPP II) (GnigEST021246)
Sequence assembly and annotation A total of 38,240 good quality ESTs of a minimum 100 bp were generated for the G. nigrifrons transcriptome
Summary
RNA sequencing (RNA-Seq) was performed to characterize the transcriptome of G. nigrifrons. A total of 38,240 ESTs of a minimum 100 bp were generated from two separate cDNA libraries consisting of virus transmitters and acquirers. More than 60% of known D. melanogaster, A. gambiae, T. castaneum immune response genes mapped to our G. nigrifrons EST database. Real time quantitative PCR (RT-qPCR) showed significant down-regulation of three genes for peptidoglycan recognition proteins (PGRP – SB1, SD, and LC) in G. nigrifrons transmitters versus control leafhoppers
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