Characterization of OsMIK in a rice mutant with reduced phytate content reveals an insertion of a rearranged retrotransposon

Theor Appl Genet. 2013 Dec;126(12):3009-20. doi: 10.1007/s00122-013-2189-3. Epub 2013 Sep 17.

Abstract

The rice low phytic acid (lpa) mutant Os-lpa-XS110-1(XS-lpa) has ~45 % reduction in seed phytic acid (PA) compared with the wild-type cultivar Xiushui 110. Previously, a single recessive gene mutation was shown to be responsible for the lpa phenotype and was mapped to a region of chromosome 3 near OsMIK (LOC_Os03g52760) and OsIPK1 (LOC_Os03g51610), two genes involved in PA biosynthesis. Here, we report the identification of a large insert in the intron of OsMIK in the XS-lpa mutant. Sequencing of fragments amplified through TAIL-PCRs revealed that the insert was a derivative of the LINE retrotransposon gene LOC_Os03g56910. Further analyses revealed the following characteristics of the insert and its impacts: (1) the inserted sequence of LOC_Os03g56910 was split at its third exon and rejoined inversely, with its 5' and 3' flanking sequences inward and the split third exon segments outward; (2) the LOC_Os03g56910 remained in its original locus in XS-lpa, and the insertion probably resulted from homologous recombination repair of a DNA double strand break; (3) while the OsMIK transcripts of XS-lpa and Xiushui 110 were identical, substantial reductions of the transcript abundance (~87 %) and the protein level (~60 %) were observed in XS-lpa, probably due to increased methylation in its promoter region. The above findings are discussed in the context of plant mutagenesis, epigenetics and lpa breeding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • Blotting, Western
  • Chromosome Mapping
  • Chromosomes, Plant / genetics
  • DNA Methylation
  • DNA, Plant / genetics
  • Gene Expression Regulation, Plant
  • Gene Rearrangement*
  • Mutagenesis, Insertional
  • Mutation / genetics*
  • Oryza / genetics*
  • Oryza / metabolism
  • Phenotype
  • Phytic Acid / metabolism*
  • Plant Proteins / genetics*
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Retroelements / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA, Plant
  • Plant Proteins
  • RNA, Messenger
  • Retroelements
  • Phytic Acid