Transcriptome-wide effect of DE-ETIOLATED1 (DET1) suppression in embryogenic callus of Carica papaya

J Plant Res. 2019 Mar;132(2):181-195. doi: 10.1007/s10265-019-01086-x. Epub 2019 Jan 16.

Abstract

Papaya is one of the most nutritional fruits, rich in vitamins, carotenoids, flavonoids and other antioxidants. Previous studies showed phytonutrient improvement without affecting quality in tomato fruit and rapeseed through the suppression of DE-ETIOLATED-1 (DET1), a negative regulator in photomorphogenesis. This study is conducted to study the effects of DET1 gene suppression in papaya embryogenic callus. Immature zygotic embryos were transformed with constitutive expression of a hairpin DET1 construct (hpDET1). PCR screening of transformed calli and reverse transcription quantitative PCR (RT-qPCR) verified that DET1 gene downregulation in two of the positive transformants. High-throughput cDNA 3' ends sequencing on DET1-suppressed and control calli for transcriptomic analysis of global gene expression identified a total of 452 significant (FDR < 0.05) differentially expressed genes (DEGs) upon DET1 suppression. The 123 upregulated DEGs were mainly involved in phenylpropanoid biosynthesis and stress responses, compared to 329 downregulated DEGs involved in developmental processes, lipid metabolism, and response to various stimuli. This is the first study to demonstrate transcriptome-wide relationship between light-regulated pathway and secondary metabolite biosynthetic pathways in papaya. This further supports that the manipulation of regulatory gene involved in light-regulated pathway is possible for phytonutrient improvement of tropical fruit crops.

Keywords: DE-ETIOLATED1; Embryogenic callus; HpRNAi; Photomorphogenesis; RNA-seq.

MeSH terms

  • Carica / embryology
  • Carica / growth & development
  • Carica / metabolism*
  • DNA, Plant
  • Down-Regulation
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant
  • Gene Silencing
  • Genes, Plant / physiology
  • Plant Proteins / physiology
  • RNA, Small Interfering
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Seeds / metabolism
  • Transcriptome

Substances

  • DNA, Plant
  • Plant Proteins
  • RNA, Small Interfering