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Double-stranded RNA in the biological control of grain aphid (Sitobion avenae F.)

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Abstract

Grain aphid (Sitobion avenae F.) is the most dominant and destructive pest of wheat, which causes significant yield loss of cereal plants each year by inflicting damage both through the direct effects of feeding and by vectoring debilitating plant viruses. In this study, we performed de novo transcriptome sequencing of grain aphid via Roche 454 GS-FLX pyrosequencing. A total of 1,106,696 reads were obtained and assembled into 32,277 unigenes, of which 25,389, 21,635, and 16,211 unigenes matched the Nt, Nr, and Swiss-Prot databases, respectively. Functional annotation of these unigenes revealed not only the presence of genes that encode the key components of RNAi machinery such as Dicer and Argonaute but also the genes encoding the TAR RNA binding protein (TRBP) and the SID-1 protein, which function in assisting the RNA-induced silencing complex (RISC) formation in microRNA (miRNA) pathway and mediating a systemic RNA interference (RNAi) effect though a cellular uptake mechanism. Furthermore, among a set of 66 unigenes selected for a double-stranded RNA (dsRNA) artificial diet assay, four novel effective RNAi targets, which led to high mortality of aphids due to the down-regulation of the expression of the respective target gene, were identified. Moreover, the expansion of systemic RNAi effect in grain aphid was observed by adding the fluorescently labeled dsRNA in an artificial diet assay.

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Abbreviations

BLAST:

Basic Local Alignment Search Tool

KOG:

Clusters of orthologous groups

dsRNA:

Double-stranded RNA

ESTs:

Expressed sequence tags

GO:

Gene ontology

KEGG:

Kyoto Encyclopedia of Genes and Genomes

Nr:

Non-redundant database

Nt:

Nucleic acid database

qPCR:

Quantitative polymerase chain reaction

RNAi:

RNA interference

RPKM:

Read per kilobase per million

RT-PCR:

Reverse-transcript polymerase chain reaction

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Acknowledgments

The authors thank Prof. Huw Jones at Rothamsted Research, UK, for his critical review of this manuscript. This work was supported by grants from the Research Initiative on Transgenic Plants from the Ministry of Agriculture of China (2014ZX0800201B), Natural Science Foundation of China (grant nos. 31171618 and 31371702), and the Chinese State Key Laboratory for Biology of Plant Diseases and Insects (grant no. SKLOF201307). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Institute of Crop Sciences/The National Key Facility for Crop Gene Resource and Genetic Improvement, Chinese Academy of Agricultural Sciences (CAAS), 12 Zhongguanchun South Street, Beijing, 10081, China

    Dahai Wang, Yongwei Sun, Hui Wang & Lanqin Xia

  2. Beijing Autolab Biotechnology Co. Ltd., Eastern Campus, Beijing Jiaotong University, Beijing, 100081, China

    Dahai Wang

  3. The Medical College of Wenzhou, Wenzhou, 325032, China

    Qi Liu

  4. Institute of Zoology, Chinese Academy of Sciences (CAS), 1 Beichen West Road, Chaoyang District, Beijing, 100101, China

    Xia Li

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  1. Dahai Wang
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  2. Qi Liu
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  3. Xia Li
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Correspondence to Lanqin Xia.

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Table S1

The primer sets of 66 unigenes selected for dsRNA amplification. (XLSX 19 kb)

Table S2

The primer sets of 14 unigenes selected for qPCR amplifying to detecting the expression correlation between RPKM and qPCR of unigenes. (XLSX 22 kb)

Table S3

The annotations of 66 unigenes selected for dsRNA feeding experiments. (XLSX 31 kb)

Figure S1

The length distribution of contigs, singletons, and unigenes, and reads number distribution of the unigenes. (GIF 27 kb)

High Resolution (TIFF 1832 kb)

Figure S2

A Venn diagram illustrating shared and unique unigenes annotated in Nr, Swiss-Prot, KOG, GO, and KEGG public databases. Among 32,277 unigenes assembled, 27,093 unigenes were annotated in at least one of the public databases, including 21,635, 16,211, 15,957, 13,876, and 11,731 in Nr, Swiss-Prot, KOG, KEGG, and GO databases, respectively. (GIF 6 kb)

High Resolution (TIFF 853 kb)

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Wang, D., Liu, Q., Li, X. et al. Double-stranded RNA in the biological control of grain aphid (Sitobion avenae F.). Funct Integr Genomics 15, 211–223 (2015). https://doi.org/10.1007/s10142-014-0424-x

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