We've updated our Privacy Policy to make it clearer how we use your personal data.

We use cookies to provide you with a better experience. You can read our Cookie Policy here.

Advertisement
Reference Gene Selection for Gene Expression Studies Using RT-qPCR in Virus-Infected Planthoppers
News

Reference Gene Selection for Gene Expression Studies Using RT-qPCR in Virus-Infected Planthoppers

Reference Gene Selection for Gene Expression Studies Using RT-qPCR in Virus-Infected Planthoppers
News

Reference Gene Selection for Gene Expression Studies Using RT-qPCR in Virus-Infected Planthoppers

Read time:
 

Want a FREE PDF version of This News Story?

Complete the form below and we will email you a PDF version of "Reference Gene Selection for Gene Expression Studies Using RT-qPCR in Virus-Infected Planthoppers"

First Name*
Last Name*
Email Address*
Country*
Company Type*
Job Function*
Would you like to receive further email communication from Technology Networks?

Technology Networks Ltd. needs the contact information you provide to us to contact you about our products and services. You may unsubscribe from these communications at any time. For information on how to unsubscribe, as well as our privacy practices and commitment to protecting your privacy, check out our Privacy Policy

Background:
Planthoppers not only severely affect crops by causing mechanical damage when feeding but are also vectors of several plant virus species. The analysis of gene expression in persistently infected planthoppers might unveil the molecular basis of viral transmission. Quantitative real-time RT-PCR (RT-qPCR) is currently the most accurate and sensitive method used for quantitative gene expression analysis. In order to normalize the resulting quantitative data, reference genes with constant expression during the experimental procedures are needed.

Results:
Partial sequences of the commonly used reference genes actin (ACT), alpha1-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1A), ribosomal protein S18 (RPS18) and polyubiquitin C (UBI) from Delphacodes kuscheli, a planthopper capable of persistently transmitting the plant fijivirus Mal de Rio Cuarto virus (MRCV), were isolated for the first time. Specific RT-qPCR primers were designed and the expression stability of these genes was assayed in MRCV-infective and naive planthoppers using geNorm, Normfinder and BestKeeper tools. The overall analysis showed that UBI, followed by 18S and ACT, are the most suitable genes as internal controls for quantitative gene expression studies in MRCV-infective planthoppers, while TUB and EF1A are the most variable ones. Moreover, EF1A was upregulated by MRCV infection.

Conclusions:
A RT-qPCR platform for gene expression analysis in the MRCV-infected planthopper vector Delphacodes kuscheli was developed.

The article is published online in Virology Journal and is free to access.

Advertisement