Pyrosequencing of the adult tarnished plant bug, Lygus lineolaris, and characterization of messages important in metabolism and development

Abstract

The adoption of Bt transgenic cotton has practically eliminated lepidopteran pests from this crop and has produced a secondary pest problem, with pierce‐sucking insects such as the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae). The future of cotton genetic pest management is threatened by these insects and their development of resistance to chemical insecticides. Lygus lineolaris is also a pest of more than 100 other crops. The development of transcriptome data for this insect should be transformative in essentially all aspects of research on plant bug biology and the development of control strategies. The first 454 tarnished plant bug whole body (WB) and gut (G) transcriptomes were constructed (half plate for each). A total of 116 163 527 bases were obtained, representing 262 555 WB and 229 919 G reads (SRA048217) of which 232 058 (SRS280903) and 168 069 (SRS280894) reads, respectively, were available for assembly. The average read length was 233.1 and 208.5 bp for WB and G, respectively. The whole body and gut reads were assembled together (WB‐G) to produce the most complete transcriptome possible from our sequencing effort and resulted in 6 970 contigs with an average length of 393 bp. The gut transcriptome alone was assembled into 3 549 contigs with an average length of 349 bp. The smallest contig was 55 bp and the largest was 3 466 bp, and there were 62 484 sequences that could not be assembled (singletons) among both transcriptomes. Overall transcriptome analysis was organized according to the Gene Ontology consortium, enzyme commission, and InerPro using the Blast2GO® program. We further characterized metabolic systems and messages associated with development.

De novo construction of an expanded transcriptome assembly for the western tarnished plant bug, Lygus hesperus

“…To assess the relative quality and completeness of our assembly, we compared core statistics for published Lygus transcriptomes [ 2 , 3 , 11 ] with those of the L. hesperus transcriptome described in this study (Table 2 ). The total number of sequence reads used in the current assembly represent 1660 and 300-fold increases over those used in the L. lineolaris transcriptome [ 11 ] and the initial Roche 454-based L. hesperus transcriptome [ 2 ] respectively. The expansion of read inputs resulted in average transcript lengths increasing from 725 to 2075 bp, and a larger percentage of transcripts with BLAST hits and assigned GO terms.…”

“…transcripts with a BLAST hit 3126 (44.9 %) 19,393 (54 %) - 16,942 (76.9 %) No. transcripts with GO term 2196 (31.5 %) 7898 (21 %) - 12,114 (54.9 %) PFAM - 3705 (22.2 %) - 14,575 (66.1 %) Data from: a Magalhaes et al 2013 [ 11 ]; b Hull et al 2013 [ 2 ]; c Hull et al 2014 [ 3 ] …”

Expression profiles of astakine-like transcripts in the tarnished plant bug,Lygus lineolaris, exposed to fungal spores ofBeauveria bassiana

“…The recent availability of transcriptomes for L. lineolaris (Magalhaes et al ., ) and for Lygus hesperus (Hull et al ., ) has allowed the identification of many possible gene silencing targets suitable for control measures against these pests. Malgalhaes (2013) in particular described several insect chemokines and receptors in the L. lineolaris transcriptome which might be suitable targets for further study in this regard.…”

Molecular Cloning and Characterization of G Alpha Proteins from the Western Tarnished Plant Bug, Lygus hesperus

“…Despite the pest status of the Lygus spp. complex, transcriptional resources have only recently been developed [ 43 , 44 , 45 , 46 ], and our knowledge of chemosensory signal transduction is limited to odorant binding proteins [ 45 , 47 ] and the olfactory receptor co-receptor (Orco) [ 12 ] in L. lineolaris and L. hesperus . Furthermore, while G proteins have been studied in a number of insects with Gα subunits cloned from Drosophila melanogaster [ 17 , 21 , 48 , 49 , 50 ], Anopheles gambiae [ 20 ], Bombyx mori [ 19 , 51 , 52 ], Manduca sexta [ 53 ], Locusta migratoria [ 54 ], Lissorhoptrus oryzophilus [ 55 ], Helicoverpa assaulta [ 56 ], Mamestra brassicae [ 18 ], Bemisia tabaci [ 57 ], and Oncopeltus fasciatus [ 58 ], little progress has been made on the role of these genes in mediating chemosensory behaviors in plant bugs such as Lygus .…”