Evaluation of Sorghum [Sorghumbicolor(L.)] Reference Genes in Various Tissues and under AbioticStress Conditions for Quantitative Real-Time PCRD at a Normalization

P. Sudhakar Reddy, Dumbala Srinivas Reddy, Sivasakthi Kaliamoorthy, Pooja Bhatnagar - Mathur, Vincent Vadez, Kiran Sharma. (25/4/2016). Evaluation of Sorghum [Sorghumbicolor(L. )] Reference Genes in Various Tissues and under AbioticStress Conditions for Quantitative Real-Time PCRD at a Normalization. Frontiers in Plant Science, 07 (529, pp. 1-14.

Abstract

AccurateandreliablegeneexpressiondatafromqPCRdependsonstablereferencegeneexpressionforpotentialgenefunctionalanalyses.Inthisstudy,15referencegeneswereselectedandanalyzedinvarioussamplesetsincludingabioticstresstreatments(salt,cold,waterstress,heat,andabscisicacid)andtissues(leaves,roots,seedlings,panicle,andmatureseeds).Statisticaltools,includinggeNorm,NormFinderandRefFinder,wereutilizedtoassessthesuitabilityofreferencegenesbasedontheirstabilityrankingsforvarioussamplegroups.Forabioticstress,PP2AandCYPwereidentifiedasthemoststablegenes.Incontrast,EIF4αwasthemoststableinthetissuesampleset,followedbyPP2A;PP2Awasthemoststableinallthesampleset,followedbyEIF4α.GAPDH,andUBC1weretheleaststablyexpressedinthetissueandallthesamplesets.Theseresultsalsoindicatedthattheuseoftwocandidatereferencegeneswouldbesufficientfortheoptimizationofnormalizationstudies.Tofurtherverifythesuitabilityofthesegenesforuseasreferencegenes,SbHSF5andSbHSF13geneexpressionlevelswerenormalizedusingthemostandleaststablesorghumreferencegenesinrootandwaterstressed-leaftissuesoffivesorghumvarieties.ThisisthefirstsystematicstudyoftheselectionofthemoststablereferencegenesforqPCR-relatedassaysinSorghumbicolorthatwillpotentiallybenefitfuturegeneexpressionstudiesinsorghumandothercloselyrelatedspecies.