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JXB Advance Access published online on July 3, 2006
Journal of Experimental Botany, doi:10.1093/jxb/erj203
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© 2006 The Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Received November 3, 2005
Accepted March 19, 2006

RESEARCH PAPER

In planta mobilization of mPing and its putative autonomous element Pong in rice by hydrostatic pressurization

Xiuyun Lin 1 *, Likun Long 2 *, Xiaohui Shan 2, Sanyuan Zhang 3, Sile Shen 4, and Bao Liu 5 *

1 Laboratory of Plant Molecular Epigenetics, Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China; Jilin Academy of Agricultural Sciences, Gongzuling 136100, China
2 Laboratory of Plant Molecular Epigenetics, Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
3 Jilin Academy of Agricultural Sciences, Gongzuling 136100, China
4 The National Key Laboratory of Super-hard Material, Jilin University, Changchun 130020, China
5 Laboratory of Plant Molecular Epigenetics, Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China; Key Laboratory for Applied Statistics of MOE, Northeast Normal University, Changchun 130024, China

* To whom correspondence should be addressed.
Bao Liu, E-mail: baoliu6677{at}yahoo.com.cn


  Abstract

The miniature Ping (mPing) is a recently discovered endogenous miniature inverted repeat transposable element (MITE) in rice, which can be mobilized by tissue culture or irradiation. It is reported here that mPing, together with one of its putative transposase-encoding partners, Pong, was efficiently mobilized in somatic cells of intact rice plants of two distinct cultivars derived from germinating seeds subjected to high hydrostatic pressure, whereas the other autonomous element of mPing, Ping, remained static in the plants studied. mPing excision was detected in several plants of both cultivars in the treated generation (P0), which were selected based on their novel phenotypes. Southern blot analysis and transposon-display assay on selfed progenies (P1 generation) of two selected P0 plants, one from each of the cultivars, revealed polymorphic banding patterns consistent with mobilization of mPing and Pong. Various mPing excisions and de novo insertions, as detected by element-bracketing, locus-specific PCR assays, occurred in the different P1 plants of both cultivars. Pong excision at one locus for each cultivar was also detected by using a Pong internal primer together with locus-specific flanking primers in the P1 plants. In contrast to the pressurized plants, immobility of both mPing and Pong in control plants, and the absence of within-cultivar heterozygosity at the analysed loci were verified by Southern blotting and/or locus-assay. Sequencing at 18 mPing empty donor sites isolated from the pressurized plants indicated properties characteristic of the element excision. Sequence-based mapping of 10 identified mPing de novo insertions from P1 progenies of pressurized plants indicated that all were in unique or low-copy regions, conforming with the targeting propensity of mPing. No evidence for further mPing activity was detected in the P2 plants tested. In spite of the high activity of mPing and Pong in the pressurized plants, amplified fragment length polymorphism (AFLP) analysis denoted their general genomic stability, and several potentially active retrotransposons also remained largely immobile. Further investigation showed that the same hydrostatic pressure treatments also caused mobilization of mPing in the standard laboratory cultivar for japonica rice, Nipponbare. Thus, a simple and robust approach for in planta MITE-mobilization in rice has been established by using high hydrostatic pressure treatment, which may be useful as an alternative for gene-tagging in this important crop plant.

Keywords: Hydrostatic pressure; MITEs; rice; tagging; transposon mobilization.

*These authors contributed equally to this work.


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Y. Ohmori, M. Abiko, A. Horibata, and H.-Y. Hirano
A Transposon, Ping, is Integrated into Intron 4 of the DROOPING LEAF Gene of Rice, Weakly Reducing its Expression and Causing a Mild Drooping Leaf Phenotype
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