Dataset Information

Expression and RNA Interference of Ribosomal Protein L5 Gene in Nilaparvata lugens (Hemiptera: Delphacidae).

ABSTRACT: The ribosomal proteins play important roles in the growth and development of organisms. This study aimed to explore the function of NlRPL5 (GenBank KX379234), a ribosomal protein L5 gene, in the brown planthopper Nilaparvata lugens. The open reading frame of NlRPL5 was cloned from N. lugens based on a previous transcriptome analysis. The results revealed that the open reading frame of NlRPL5 is of 900?bp, encoding 299 amino acid residues. The reverse transcription quantitative PCR results suggested that the expression of NlRPL5 gene was stronger in gravid females, but was relatively low in nymphs, males, and newly emerged females. The expression level of NlRPL5 in the ovary was about twofolds of that in the head, thorax, or fat body. RNAi of dsNlRPL5 resulted in a significant reduction of mRNA levels, ?50% decrease in comparison with the dsGFP control at day 6. Treatment of dsNlRPL5 significantly restricted the ovarian development, and decreased the number of eggs laid on the rice (Oryza sativa) plants. This study provided a new clue for further study on the function and regulation mechanism of NlRPL5 in N. lugens.

SUBMITTER: Zhu J

PROVIDER: S-EPMC5538327 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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Expression and RNA Interference of Ribosomal Protein L5 Gene in Nilaparvata lugens (Hemiptera: Delphacidae).

Zhu Jiajun J Hao Peiying P Lu Chaofeng C Ma Yan Y Feng Yalin Y Yu Xiaoping X

Journal of insect science (Online) 20170501 3

The ribosomal proteins play important roles in the growth and development of organisms. This study aimed to explore the function of NlRPL5 (GenBank KX379234), a ribosomal protein L5 gene, in the brown planthopper Nilaparvata lugens. The open reading frame of NlRPL5 was cloned from N. lugens based on a previous transcriptome analysis. The results revealed that the open reading frame of NlRPL5 is of 900 bp, encoding 299 amino acid residues. The reverse transcription quantitative PCR results sugges ...[more]

PMID: 28973571

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Project description:The brown planthopper (BPH), Nilaparvata lugens (Hemiptera, Delphacidae), is one of the most important rice pests. Abundant genetic studies on BPH have been conducted using reverse-transcription quantitative real-time PCR (qRT-PCR). Using qRT-PCR, the expression levels of target genes are calculated on the basis of endogenous controls. These genes need to be appropriately selected by experimentally assessing whether they are stably expressed under different conditions. However, such studies on potential reference genes in N. lugens are lacking. In this paper, we presented a systematic exploration of eight candidate reference genes in N. lugens, namely, actin 1 (ACT), muscle actin (MACT), ribosomal protein S11 (RPS11), ribosomal protein S15e (RPS15), alpha 2-tubulin (TUB), elongation factor 1 delta (EF), 18S ribosomal RNA (18S), and arginine kinase (AK) and used four alternative methods (BestKeeper, geNorm, NormFinder, and the delta Ct method) to evaluate the suitability of these genes as endogenous controls. We examined their expression levels among different experimental factors (developmental stage, body part, geographic population, temperature variation, pesticide exposure, diet change, and starvation) following the MIQE (Minimum Information for publication of Quantitative real time PCR Experiments) guidelines. Based on the results of RefFinder, which integrates four currently available major software programs to compare and rank the tested candidate reference genes, RPS15, RPS11, and TUB were found to be the most suitable reference genes in different developmental stages, body parts, and geographic populations, respectively. RPS15 was the most suitable gene under different temperature and diet conditions, while RPS11 was the most suitable gene under different pesticide exposure and starvation conditions. This work sheds light on establishing a standardized qRT-PCR procedure in N. lugens, and serves as a starting point for screening for reference genes for expression studies of related insects.

Project description:INTRODUCTION:The brown planthopper (BPH, Nilaparvata lugens Stål, Hemiptera: Delphacidae) is one of the most devastating insect pests of the crucially important cereal crop, rice (Oryza sativa L.). Currently, multiple BPH-resistant rice varieties have been cultivated and generalized to control BPH. However, the defence metabolic responses and their modes of action against BPH in different rice cultivars remain uncharacterized. OBJECTIVE:We used a non-biased metabolomics approach to explore the differences in metabolite profiles in response to BPH infestation in the susceptible TN1 rice cultivar and two resistant cultivars (IR36 and IR56). METHODS:The metabolomic detection based on gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) was performed to investigate the content changes of identified metabolites in TN1, IR36 and IR56 rice varieties at various time points (0 h, 24 h, 48 h and 96 h) post BPH feeding. The differentially expressed metabolites were screened and the corresponding metabolic pathways were further enriched. RESULTS:The results showed that compared to that in TN1, the content changes of most primary metabolites were more stable, but the concentration alterations of some defence-related metabolites were more acute and persistent in IR36 and IR56. Furthermore, the differentially expressed pathways analysis revealed that cyanoamino acids and lipids metabolism was persistently induced in IR36, but changes in thiamine, taurine and hypotaurine metabolism were more significant in IR56 during BPH infestation. Besides, the contents of quercetin and spermidine which were harmful to BPH fitness, were significantly elevated by BPH in TN1 and IR36, and the quercetin level was significantly decreased during BPH feeding in IR56. CONCLUSION:The results of the differences in metabolite profiles in response to BPH infestation in different rice cultivars were useful to clarify the metabolic mechanism of rice plants during BPH infestation and to provide new resources to control this insect pest.

Dataset Information

Expression and RNA Interference of Ribosomal Protein L5 Gene in Nilaparvata lugens (Hemiptera: Delphacidae).

Publications

Expression and RNA Interference of Ribosomal Protein L5 Gene in Nilaparvata lugens (Hemiptera: Delphacidae).

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