Stimulation of the cell cycle and maize transformation by disruption of the plant retinoblastoma pathway - PubMed (original) (raw)

. 2002 Sep 3;99(18):11975-80.

doi: 10.1073/pnas.142409899. Epub 2002 Aug 16.

Brian P Dilkes, Keith Lowe, George Hoerster, Xifan Sun, Margit Ross, Laura Church, Chris Bunde, Jeff Farrell, Patrea Hill, Sheila Maddock, Jane Snyder, Louisa Sykes, Zhongsen Li, Young-min Woo, Dennis Bidney, Brian A Larkins

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Stimulation of the cell cycle and maize transformation by disruption of the plant retinoblastoma pathway

William Gordon-Kamm et al. Proc Natl Acad Sci U S A. 2002.

Abstract

The genome of the Mastreviruses encodes a replication-associated protein (RepA) that interacts with members of the plant retinoblastoma-related protein family, which are putative cell cycle regulators. Expression of ZmRb1, a maize retinoblastoma-related gene, and RepA inhibited and stimulated, respectively, cell division in tobacco cell cultures. The effect of RepA was mitigated by over-expression of ZmRb1. RepA increased transformation frequency and callus growth rate of high type II maize germplasm. RepA-containing transgenic maize calli remained embryogenic, were readily regenerable, and produced fertile plants that transmitted transgene expression in a Mendelian fashion. In high type II, transformation frequency increased with the strength of the promoter driving RepA expression. When a construct in which RepA was expressed behind its native LIR promoter was used, primary transformation frequencies did not improve for two elite Pioneer maize inbreds. However, when LIR:RepA-containing transgenic embryos were used in subsequent rounds of transformation, frequencies were higher in the RepA+ embryos. These data demonstrate that RepA can stimulate cell division and callus growth in culture, and improve maize transformation.

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Figures

Figure 1

Figure 1

Effect of RepA expression on plant cell division and callus growth. (a) Recent cell division in a GFP-expressing BY-2 cell culture clearly showing two daughter cells separated by a newly formed transverse wall. (b) GFP expression 2 weeks after particle delivery of Ubi:moPAT∼moGFP:pinII DNA into Hi-II immature embryos; only single cells expressing GFP were observed. (c) GFP expression 2 weeks after particle delivery of Ubi:moPAT∼moGFP:pinII and Ubi:RepA; multiple GFP-expressing multicellular colonies were observed in addition to single cells expressing GFP. (Scale markers in b and_c_ = 500 μm.)

References

    1. Gutierrez C. EMBO J. 2000;19:792–799. - PMC - PubMed
    1. Gutierrez C. Plant Mol Biol. 2000;43:763–772. - PubMed
    1. Palmer K E, Rybicki E P. Adv Virus Res. 1998;50:183–234. - PubMed
    1. Schalk H J, Matzeit V, Schiller B, Schell J, Gronenborn B. EMBO J. 1989;8:359–364. - PMC - PubMed
    1. Xie Q, Sanz-Burgos A P, Guo H, Garcia J A, Gutierrez C. Plant Mol Biol. 1999;39:647–656. - PubMed

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