Selection and validation of suitable reference genes for quantitative real time PCR analysis of gene expression studies in patchouli under Meloidogyne incognita attack and PGPR treatment

Abstract

Patchouli (Pogostemon cablin) is an economically important aromatic plant and is often vulnerable to nematode attack (Meloidogyne incognita). The application of PGPR has tremendously changing disease management approach in plants and a systematic understanding the molecular mechanism behind it is needed to control over the disease more precisely. Since, qRT-PCR is the most frequently used molecular tools in studying molecular mechanism but is greatly dependent upon suitable reference or housekeeping genes. This study is aim to select suitable reference genes for qRT-PCR experiments in patchouli under the condition of nematode attack and PGPR treatments. Expression pattern of six reference genes (Actin, EF1, ACP, COX3, GAPDH and Pat18S) were statistically analyzed and validated using different algorithms viz., ΔCt, BestKeeper, GeNorm, NormFinder and RefFinder. The result showed that the Actin and ACP are the most suitable reference genes and the combination of ACP &GAPDH gene pairs can also be used whenever the experiment needed.