Reference gene selection for real-time quantitative PCR normalization in Hemarthria compressa and Hemarthria...

Area of Science:

• Agricultural Science
• Molecular Biology
• Plant Science

Background:

• Hemarthria compressa and Hemarthria altissima are vital forage crops in tropical and subtropical grasslands.
• These grasses are essential for livestock, ecological balance, and animal husbandry development.

Purpose of the Study:

• To evaluate the expression stability of 12 reference genes in Hemarthria species under abiotic stresses.
• To identify reliable reference genes for accurate gene expression studies in Hemarthria.

Main Methods:

• RT-qPCR was used to measure mRNA expression levels of 12 candidate reference genes.
• Expression stability was analyzed using Delta-Ct, BestKeeper, Genorm, Normfinder, and RefFinder algorithms.
• Stress conditions included drought, salt, acid-aluminum, and cold treatments on leaf tissues.

Main Results:

• Reference gene expression varied significantly across different abiotic stress conditions.
• CL18892, eEF-1α, and CL16384 showed high stability under specific stresses (salt, cold/acid-aluminum, drought, respectively).
• CYP5, BMK.74327, and CL21527 were identified as the most suitable overall reference genes for Hemarthria under abiotic stress.

Conclusions:

• The selection of appropriate reference genes is critical for reliable gene expression studies in Hemarthria.
• This research provides essential technical support for future molecular research on abiotic stress responses in these important forage grasses.