title: Characterization of OsMIK in a rice mutant with reduced phytate content reveals an insertion of a rearranged retrotransposon.
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.
authors.affiliation: State Key Laboratory of Rice Biology and Key Laboratory of Nuclear-Agricultural Sciences of the Ministry of Agriculture and Zhejiang Province, Institute of Nuclear Agricultural Sciences, Zhejiang University, Hangzhou, 310029, China.
authors.forename: Hai-Jun
authors.initials: HJ
authors.lastname: Zhao
authors.forename: Hai-Rui
authors.initials: HR
authors.lastname: Cui
authors.forename: Xiu-Hong
authors.initials: XH
authors.lastname: Xu
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Tan
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Fu
authors.forename: Guo-Zhen
authors.initials: GZ
authors.lastname: Liu
authors.forename: Yves
authors.initials: Y
authors.lastname: Poirier
authors.forename: Qing-Yao
authors.initials: QY
authors.lastname: Shu
mesh_terms: D015139:Blotting, Southern; D015153:Blotting, Western; D002874:Chromosome Mapping; D032461:Chromosomes, Plant / Q000235:genetics; D019175:DNA Methylation; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D015321:Gene Rearrangement; D016254:Mutagenesis, Insertional; D009154:Mutation / Q000235:genetics*; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D010833:Phytic Acid / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D018626:Retroelements / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018744:DNA, Plant; D010940:Plant Proteins; D012333:RNA, Messenger; D018626:Retroelements; D010833:Phytic Acid
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