Characterization of three active transposable elements recently inserted in three independent DFR-A alleles and one high-copy DNA transposon isolated from the Pink allele of the ANS gene in onion (Allium cepa L.)

Mol Genet Genomics. 2015 Jun;290(3):1027-37. doi: 10.1007/s00438-014-0973-7. Epub 2014 Dec 17.

Abstract

Intact retrotransposon and DNA transposons inserted in a single gene were characterized in onions (Allium cepa) and their transcription and copy numbers were estimated in this study. While analyzing diverse onion germplasm, large insertions in the DFR-A gene encoding dihydroflavonol 4-reductase (DFR) involved in the anthocyanin biosynthesis pathway were found in two accessions. A 5,070-bp long terminal repeat (LTR) retrotransposon inserted in the active DFR-A (R4) allele was identified from one of the large insertions and designated AcCOPIA1. An intact ORF encoded typical domains of copia-like LTR retrotransposons. However, AcCOPIA1 contained atypical 'TG' and 'TA' dinucleotides at the ends of the LTRs. A 4,615-bp DNA transposon was identified in the other large insertion. This DNA transposon, designated AcCACTA1, contained an ORF coding for a transposase showing homology with the CACTA superfamily transposable elements (TEs). Another 5,073-bp DNA transposon was identified from the DFR-A (TRN) allele. This DNA transposon, designated AchAT1, belonged to the hAT superfamily with short 4-bp terminal inverted repeats (TIRs). Finally, a 6,258-bp non-autonomous DNA transposon, designated AcPINK, was identified in the ANS-p allele encoding anthocyanidin synthase, the next downstream enzyme to DFR in the anthocyanin biosynthesis pathway. AcPINK also possessed very short 3-bp TIRs. Active transcription of AcCOPIA1, AcCACTA1, and AchAT1 was observed through RNA-Seq analysis and RT-PCR. The copy numbers of AcPINK estimated by mapping the genomic DNA reads produced by NextSeq 500 were predominantly high compared with the other TEs. A series of evidence indicated that these TEs might have transposed in these onion genes very recently, providing a stepping stone for elucidation of enormously large-sized onion genome structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / genetics*
  • Alleles
  • Amino Acid Sequence
  • Base Sequence
  • DNA Transposable Elements / genetics*
  • DNA, Complementary / genetics
  • Gene Expression Profiling
  • High-Throughput Nucleotide Sequencing
  • Molecular Sequence Data
  • Onions / enzymology
  • Onions / genetics*
  • Oxygenases / genetics*
  • Phylogeny
  • Pigments, Biological / genetics
  • Plant Proteins / genetics
  • Retroelements / genetics*
  • Sequence Alignment
  • Sequence Analysis, RNA
  • Terminal Repeat Sequences / genetics
  • Transposases / genetics

Substances

  • DNA Transposable Elements
  • DNA, Complementary
  • Pigments, Biological
  • Plant Proteins
  • Retroelements
  • Alcohol Oxidoreductases
  • dihydroflavanol 4-reductase
  • Oxygenases
  • anthocyanidin synthase
  • Transposases

Associated data

  • GENBANK/KM434202
  • GENBANK/KM434203
  • SRA/SRR1686960