A reproducible and sensitive method for generating high‐quality transcriptomes from single whitefly salivary glands and other low‐input tissues. (15th February 2022)

• Record Type: Journal Article
• Authors: Gebiola, Marco; Le, Brandon H.; Mauck, Kerry E.
• Is Part Of: Insect science. Volume 29:Number 5(2022)
• Page Start: 1318
• Page End: 1328
• Publication Date: 2022-02-15
• Subjects: Aleyrodidae -- Bemisia tabaci -- bulk RNA‐Seq -- insect vectors -- low‐input RNA‐Seq -- plant pathogens; Insects -- Periodicals; Entomology -- Periodicals; 595.705
• DOI: 10.1111/1744-7917.13008
• Languages: English
• ISSNs: 1672-9609

Abstract

Abstract: Transcriptomic studies are an important tool for understanding the molecular pathways underlying host plant use by agricultural pests, including vectors of damaging plant pathogens. Thus far, bulk RNA‐Seq has been the main approach for non‐model insects. This method relies on pooling large numbers of whole organisms or hundreds of individually dissected organs. The latter approach is logistically challenging, may introduce artifacts of handling and storage, and is not compatible with biological replication. Here, we tested an approach to generate transcriptomes of individual salivary glands and other low‐input body tissues from whiteflies ( Bemisia tabaci MEAM1), which are major vectors of plant viruses. By comparing our outputs to published bulk RNA‐Seq datasets for whole whitefly bodies and pools of salivary glands, we demonstrate that this approach recovers similar numbers of transcripts relative to bulk RNA‐Seq in a tissue‐specific manner, and for some metrics, exceeds performance of bulk tissue RNA‐Seq. Libraries generated from individual salivary glands also yielded additional novel transcripts not identified in pooled salivary gland datasets, and had hundreds of enriched transcripts when compared with whole head tissues. Overall, our study demonstrates that it is feasible to produce high quality, replicated transcriptomes of whitefly salivary glands and other low‐input tissues. We anticipate that our approach will expand hypothesis‐driven research on salivary glands of whiteflies and other Hemiptera, thus enabling novel control strategies to disrupt feeding and virus transmission. Abstract : We used a low‐input RNA workflow to generate transcriptomes from single salivary glands, single heads, and 10‐head pools of whiteflies ( Bemisia tabaci MEAM1) We compared transcriptomes to those derived from pools of several hundred dissected glands or whole insects, and with Smart‐Seq2 data from a single salivary gland We showed that our single‐cell‐like workflow is reproducible, sensitive, and leads to discovery of novel transcripts expressed only in salivary glands