Excavation and Verification on Internal Reference Genes in Abelmoschus esculentus Seeds Based on Transcriptome Sequencing
To identify stably expressed reference genes in Abelmoschus esculentus seeds at different developmental stages,transcriptomic sequencing was conducted on okra seeds at 3 days,6 days,15 days,25 days and 40 days post-pollination.Based on the whole genome annotation of Abelmoschus es-culentus and transcriptome sequencing data of Abelmoschus esculentus seeds,six new candidate internal reference genes ZCCHC11L,SEU,RPL11,FKBP62,MBR2L and BOB1 were compared for stability with six commonly used housekeeping genes GAPDH,EF1-α,TUA5,YLS8,ACT2 and eIF4A reported in literature.qRT-PCR was employed to assess gene expression in Abelmoschus esculentus seeds at various growth stages.The expression stability of selected internal reference factors was comprehensively evaluated using geNorm,NormFinder,BestKeeper and RefFinder.Subsequently,the chosen internal reference genes were validated against the starch synthetase gene AeAGPL2.Results indicated that ZCCHC11L and SEU exhibited optimal stability while TUA5 displayed the least stable expression.The newly identified internal reference genes ZCCHC11L and SEU as well as their combination ZCCHC11L+SEU could be used as internal reference genes at dif-ferent developmental stages of Abelmoschus esculentus seeds.
万方数据
维普
