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Plant and Cell Physiology Advance Access published online on November 7, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci226
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received August 30, 2005
Accepted October 28, 2005

Regular Paper

Large Scale Systematic Study on Stability of Ds Element and Timing of Transposition in Rice

Ildiko Szeverenyi 1, Rengasamy Ramamoorthy 1, Zhi Wei Teo 1, Hong Fen Luan 1, Zhi Gang Ma 1, and Srinivasan Ramachandran 1*

1 Rice Functional Genomics Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604

* To whom correspondence should be addressed.
Srinivasan Ramachandran, E-mail: sri{at}tll.org.sg


  Abstract

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 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; thereby the propagation of parental lines did not induce transposon silencing. Moreover, the stably transposed Ds element was active even at F5 generation, since Ac could remobilize Ds element as indicated by the footprint analysis of several revertants. The bar gene expression was monitored from F3 to F6 generations in more than thousand lines. Strikingly, substantial transgene silencing was not observed in all the generations tested. We analyzed the timing of transposition during rice development and provide evidence that Ds transpose late after tiller formation. The independent events could be the results of secondary transposition events were ruled out by analyzing possible footprints with reciprocal PCRs. 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 by panicle are an alternative way to increase the number of independent transposants due to post-tillering transposition.

Keywords: Ac/Ds transposon; independent transposition; rice; silencing; timing.
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