Selection and validation of reference genes for quantitative real-time PCR in Artemisia sphaerocephala based on...

Area of Science:

• Plant Science
• Molecular Biology
• Ecology

Background:

• Artemisia sphaerocephala is a vital perennial semi-shrub in northwestern China, crucial for rangeland protection and desert ecosystem stability.
• This species holds potential for forage, plant oil extraction, and biofuel production.
• Accurate gene expression analysis using Real-Time quantitative PCR (qPCR) relies on stable internal reference genes.

Purpose of the Study:

• To identify and validate the most stable internal reference genes for qPCR in Artemisia sphaerocephala.
• To ensure reliable gene expression studies in this ecologically and economically important plant species.

Main Methods:

• Transcriptome data of A. sphaerocephala was utilized to identify 21 candidate internal genes.
• Gene expression stability was evaluated across 16 diverse samples.
• The Delta Ct method and three statistical programs (GeNorm, NormFinder, BestKeeper) were employed for analysis.

Main Results:

• Analysis identified UBC9 and TIP41-like as the most stable reference genes.
• These genes demonstrated superior reliability for normalization in qPCR experiments involving A. sphaerocephala.

Conclusions:

• UBC9 and TIP41-like are recommended as optimal internal reference genes for quantitative PCR in Artemisia sphaerocephala.
• This finding will facilitate more accurate molecular research on this significant plant species.