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<p>The rugose spiraling whitefly (<i>Aleurodicus rugioperculatus</i> Martin) is an invasive pest of several economically important crops, including coconut. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a fundamental and widely used technique in genomic research, particularly for analyzing gene expression levels with high sensitivity and specificity. One of the most critical factors influencing the accuracy and reliability of qRT-PCR results is the choice of internal reference genes, also known as housekeeping genes. These genes are used to normalize the expression levels of target genes. However, the lack of studies validating reference genes in <i>A. rugioperculatus</i> significantly restricts the accurate application of qRT-PCR for gene expression analysis. This study aims to identify suitable reference gene(s) across different biotic and abiotic conditions in <i>A. rugioperculatus</i>. The expression stability of 14 reference genes (<i>18S rRNA</i>, <i>GST</i>, <i>HSP90</i>, <i>GAPDH</i>, <i>RPS17</i>, <i>actin</i>, <i>RPL13</i>, <i>NADH</i>, <i>ETF-QO</i>, <i>EF1α</i>, <i>UBQ</i>, β<i>-TUB</i>, <i>peptidylprolyl isomerase A</i>, and <i>Myosin L</i>) was analyzed using five different computational programs, such as geNorm, NormFinder, BestKeeper, Comparative ΔCT, and RefFinder. The findings suggested that the optimal combinations of reference genes in <i>A. rugioperculatus</i> were <i>18S rRNA</i> and <i>NADH</i> for different developmental stages, <i>18S rRNA</i> and <i>EF1α</i> for different sexes and temperatures, and <i>RPL13</i> and <i>18S rRNA</i> for starvation conditions. This study presents the first report of stable reference genes for normalizing qRT-PCR analysis in <i>A. rugioperculatus,</i> laying the foundation for future expression studies and RNAi-based management of the rugose spiraling whitefly.</p>

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Validation of suitable reference genes for gene expression studies in rugose spiraling whitefly (Aleurodicus rugioperculatus Martin) under various experimental conditions

  • 期刊论文

  • 发表日期：

    2026年3月11日

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  • A. A. Sabana,
  • M. K. Rajesh,
  • Jasmin Habeeb,
  • M. Sujithra,
  • A. Josephrajkumar

摘要

The rugose spiraling whitefly (Aleurodicus rugioperculatus Martin) is an invasive pest of several economically important crops, including coconut. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a fundamental and widely used technique in genomic research, particularly for analyzing gene expression levels with high sensitivity and specificity. One of the most critical factors influencing the accuracy and reliability of qRT-PCR results is the choice of internal reference genes, also known as housekeeping genes. These genes are used to normalize the expression levels of target genes. However, the lack of studies validating reference genes in A. rugioperculatus significantly restricts the accurate application of qRT-PCR for gene expression analysis. This study aims to identify suitable reference gene(s) across different biotic and abiotic conditions in A. rugioperculatus. The expression stability of 14 reference genes (18S rRNA, GST, HSP90, GAPDH, RPS17, actin, RPL13, NADH, ETF-QO, EF1α, UBQ, β-TUB, peptidylprolyl isomerase A, and Myosin L) was analyzed using five different computational programs, such as geNorm, NormFinder, BestKeeper, Comparative ΔCT, and RefFinder. The findings suggested that the optimal combinations of reference genes in A. rugioperculatus were 18S rRNA and NADH for different developmental stages, 18S rRNA and EF1α for different sexes and temperatures, and RPL13 and 18S rRNA for starvation conditions. This study presents the first report of stable reference genes for normalizing qRT-PCR analysis in A. rugioperculatus, laying the foundation for future expression studies and RNAi-based management of the rugose spiraling whitefly.