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

文献类型： 外文期刊

第一作者： Lin, Xiuyun

作者： Lin, Xiuyun;Long, Likun;Shan, Xiaohui;Zhang, Sanyuan;Shen, Sile;Liu, Bao

作者机构：

关键词： hydrostatic pressure;MITEs;rice;tagging;transposon mobilization

期刊名称：JOURNAL OF EXPERIMENTAL BOTANY （ 影响因子：6.992； 五年影响因子：7.86 ）

ISSN： 0022-0957

年卷期： 2006 年 57 卷 10 期

页码：

收录情况： SCI

摘要： 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 (P-0), which were selected based on their novel phenotypes. Southern blot analysis and transposon-display assay on selfed progenies (P-1 generation) of two selected P-0 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 P-1 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 P, 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 P, 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 P-2 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.

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