Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
Metopolophium dirhodum (Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified before th...
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Veröffentlicht in: | Scientific reports 2020-12, Vol.10 (1), p.21951-21951, Article 21951 |
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Hauptverfasser: | , , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Metopolophium dirhodum
(Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified before they can be used to normalize target gene expression levels. In this study, 10 candidate reference genes in
M
.
dirhodum
were analyzed by qRT-PCR under various experimental conditions. Their expression stability was evaluated with delta Ct, BestKeeper, geNorm, and NormFinder methods, and the final stability ranking was determined with RefFinder. The results indicate that the most appropriate sets of internal controls were
SDHB
and
RPL8
across geographic population;
RPL8
,
Actin
, and
GAPDH
across developmental stage;
SDHB
and
NADH
across body part;
RPL8
and
Actin
across wing dimorphism and temperature;
RPL4
and
EF1A
across starvation stress;
AK
and
RPL4
across insecticide treatments;
RPL8
and
NADH
across antibiotic treatments;
RPL8
,
RPL4
,
Actin
, and
NADH
across all samples. The results of this study provide useful insights for establishing a standardized qRT-PCR procedure for
M. dirhodum
and may be relevant for identifying appropriate reference genes for molecular analyses of related insects. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/s41598-020-78974-z |