Dataset Information

A Sensitive Immunocapture-RT-PCR Assay for Detection of Maize Iranian Mosaic Virus in Individual Small Brown Planthopper Vector, Laodelphax striatellus.

ABSTRACT:

Background

Sensitive detection of Maize Iranian mosaic virus (MIMV) in its insect vector, Laodelphax striatellus is essential for effective forecast and control of this viral disease.

Objectives

Three methods of ELISA, RT-PCR and IC-RT-PCR were compared regarding their sensitivity for detection of MIMV in the single planthopper with a series of various dilutions.

Materials and methods

To detect MIMV from a single insect vector, the sensitivity of three methods including ELISA, RT-PCR and IC-RT-PCR was evaluated.

Results

Compared to the other methods, the IC-RT-PCR showed more sensitivity and detected virus at least at the 160 dilution. While, both ELISA and RT-PCR methods could detect up to the 120.

Conclusion

The results reported herein showed that IC-RT-PCR is a sensitive and simple method to detect MIMV from a single insect vector with high efficiency and reliability. These findings might be useful in the prediction of viral disease incidence in host plants and this method can also be effective to detect other viruses in their insect vectors.

SUBMITTER: Moeini P

PROVIDER: S-EPMC8035424 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND: The small brown planthopper (Laodelphax striatellus) is an important agricultural pest that not only damages rice plants by sap-sucking, but also acts as a vector that transmits rice stripe virus (RSV), which can cause even more serious yield loss. Despite being a model organism for studying entomology, population biology, plant protection, molecular interactions among plants, viruses and insects, only a few genomic sequences are available for this species. To investigate its transcriptome and determine the differences between viruliferous and naïve L. striatellus, we employed 454-FLX high-throughput pyrosequencing to generate EST databases of this insect. RESULTS: We obtained 201,281 and 218,681 high-quality reads from viruliferous and naïve L. striatellus, respectively, with an average read length as 230 bp. These reads were assembled into contigs and two EST databases were generated. When all reads were combined, 16,885 contigs and 24,607 singletons (a total of 41,492 unigenes) were obtained, which represents a transcriptome of the insect. BlastX search against the NCBI-NR database revealed that only 6,873 (16.6%) of these unigenes have significant matches. Comparison of the distribution of GO classification among viruliferous, naïve, and combined EST databases indicated that these libraries are broadly representative of the L. striatellus transcriptomes. Functionally diverse transcripts from RSV, endosymbiotic bacteria Wolbachia and yeast-like symbiotes were identified, which reflects the possible lifestyles of these microbial symbionts that live in the cells of the host insect. Comparative genomic analysis revealed that L. striatellus encodes similar innate immunity regulatory systems as other insects, such as RNA interference, JAK/STAT and partial Imd cascades, which might be involved in defense against viral infection. In addition, we determined the differences in gene expression between vector and naïve samples, which generated a list of candidate genes that are potentially involved in the symbiosis of L. striatellus and RSV. CONCLUSIONS: To our knowledge, the present study is the first description of a genomic project for L. striatellus. The identification of transcripts from RSV, Wolbachia, yeast-like symbiotes and genes abundantly expressed in viruliferous insect, provided a starting-point for investigating the molecular basis of symbiosis among these organisms.