Journal Article

Large-scale Systematic Study on Stability of the <i>Ds</i> Element and Timing of Transposition in Rice

Ildiko Szeverenyi, Rengasamy Ramamoorthy, Zhi Wei Teo, Hong Fen Luan, Zhi Gang Ma and Srinivasan Ramachandran

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 47, issue 1, pages 84-95
Published in print January 2006 | ISSN: 0032-0781
Published online January 2006 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci226
Large-scale Systematic Study on Stability of the Ds Element and Timing of Transposition in Rice

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  • Biochemistry
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Activator/Dissociation (Ac/Ds) transposon mutagenesis is a widely used tool for gene identification; however, several reports on silencing of the Ac/Ds element in starter lines and in stable transposants question the applicability of such an approach in later generations. We have performed a systematic analysis on various aspects of the silencing phenomenon in rice (Oryza sativa ssp. japonica cv. Nipponbare). High somatic and germinal transposition frequencies observed in earlier generations were maintained as late as T4 and T5 generations; thus the propagation of parental lines did not induce transposon silencing. Moreover, the stably transposed Ds element was active even at the F5 generation, since Ac could remobilize the Ds element as indicated by the footprint analysis of several revertants. Expression of the bar gene was monitored from F3 to F6 generations in >1,000 lines. Strikingly, substantial transgene silencing was not observed in any of the generations tested. We analyzed the timing of transposition during rice development and provide evidence that Ds is transposed late after tiller formation. The possibility, that the independent events could be the result of secondary transposition, was ruled out by analyzing potential footprints by reciprocal PCR. Our study validates the Ac/Ds system as a tool for large-scale mutagenesis in rice, since the Ds elements were active in the starter and insertion lines even in the later generations. We propose that harvesting rice seeds using their panicles is an alternative way to increase the number of independent transposants due to post-tillering transposition.

Keywords: Ac/Ds transposon; Independent transposition; Rice; Silencing; Timing; Ac/Ds, Activator/Dissociation; CaMV, cauliflower mosaic virus; DIG, digoxigenin; FP, flanking primer; FST, flanking sequence tag; GFP, green fluorescent protein; GUS, β-glucuronidase; Rv, revertant; TAIL-PCR, thermal asymmetric interlaced PCR; TIR, terminal inverted repeat

Journal Article.  8240 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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