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Abstract: Insertion mutagenesis has become one of the most popular methods for gene functions analysis. Here we report a two-element ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice. The excision of Ds element was examined by PCR amplification. The excision frequency of Ds element varied from 0% to 40% among 20 F2 populations derived from 11 different Ds parents. Southern blot analysis revealed that more than 70% of excised Ds elements reinserted into rice genome and above 70% of the reinserted Ds elements were located at different positions of the chromosome in rice. The result of histochemical GUS analysis indicated that 28% of enhancer trap and 22% of gene trap tagging plants displayed GUS activity in leaves, roots, flowers or seeds. The GUS positive lines will be useful for identifying gene function in rice.

[1] Altmann, T., Felix, G., Jessop, A., Kauschmann, A., Uwer, U., Pena-Cortes, H., Willmitzer, L., 1995.Ac/Dstransposon mutagenesis in Arabidopsis thaliana: mutant spectrum and frequency of Ds insertion mutants.Mol. Gen. Genet.,247:646-652.

[2] Balcells, L., Coupland, G., 1994. The presence of enhancers adjacent to theAcpromoter increases the abundance of transposase mRNA and alters the timing ofDsexcision in Arabidopsis.Plant Mol. Biol.,24:789-798.

[3] Bancroft, I., Dean, C., 1993. Factors affecting the excision frequency of the maize transposable elementDsin Arabidopsis thaliana.Mol. Gen. Genet.,240:65-72.

[4] Campisi, L., Yang, Y., Yi, Y., Heilig, E., Herman, B., Cassista, A.J., Allen, D.W., Xiang, H., Jack, T., 1999. Generation of enhancer trap lines in Arabidopsis and characterization of expression patterns in the inflorescence.Plant J.,17:699-707.

[5] Chin, H.G., Choe, M.S., Lee, S.H., Park, S.H., Koo, J.C., Kim, N.Y., Lee, J.J., Oh, B.G., Yi, G.H., Kim, S.C., Choi, H.C., Cho, M.J., Han, C.D., 1999. Molecular analysis of rice plants harboring anAc/Dstransposable element-mediated gene trapping system.Plant J.,19:615-623.

[6] Christensen, A.H., Quail, P.H., 1996. Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants.Transgenic Res.,5:213-218.

[7] Enoki, H., Izawa, T., Kawahara, M., Komatsu, M., Koh, S., Kyozuka, J., Shimamoto, K., 1999.Acas a tool for the functional genomics of rice.Plant J.,19:605-613.

[8] Greco, R., Ouwerkerk, P.B., Sallaud, C., Kohli, A., Colombo, L., Puigdomenech, P., Guiderdoni, E., Christou, P., Hoge, J.H., Pereira, A., 2001. Transposon insertional mutagenesis in rice.Plant Physiol.,125:1175-1177.

[9] Hiei, Y., Ohta, S., Komari, T., Kumashiro, T., 1994. Efficient transformation of rice (Oryza sativaL.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.Plant J.,6:271-282.

[10] Hood, E.E., Gelvin, S.B., Melchers, L.S., Hoekema, A., 1993. New Agrobacterium helper plasmids for gene transfer to plants.Transgen. Res.,2:208-218.

[11] Ito, T., Seki, M., Hayashida, N., Shibata, D., Shinozaki, K., 1999. Regional insertional mutagenesis of genes on Arabidopsis thaliana chromosome V using theAc/Dstransposon in combination with a cDNA scanning method.Plant J.,17:433-444.

[12] Izawa, T., Ohnishi, T., Nakano, T., Ishida, N., Enoki, H., Hashimoto, H., Itoh, K., Terada, R., Wu, C., Miyazaki, C., Endo, T., Iida, S., Shimamoto, K., 1997. Transposon tagging in rice.Plant Mol. Biol.,35:219-229.

[13] Jeon, J.S., Lee, S., Jung, K.H., Jun, S.H., Jeong, D.H., Lee, J., Kim, C., Jang, S., Yang, K., Nam, J., An, K., Han, M.J., Sung, R.J., Choi, H.S., Yu, J.H., Choi, J.H., Cho, S.Y., Cha, S.S., Kim, S.I., An, G., 2000. T-DNA insertional mutagenesis for functional genomics in rice.Plant J.,22:561-570.

[14] Koprek, T., McElroy, D., Louwerse, J., Williams-Carrier, R., Lemaux, P.G., 2000. An efficient method for dispersingDselements in the barley genome as a tool for determining gene function.Plant J.,24:253-263.

[15] Long, D., Martin, M., Sundberg, E., Swinburne, J., Puangsomlee, P., Coupland, G., 1993a. The maize transposable element systemAc/Dsas a mutagen in Arabidopsis: identification of an albino mutation induced byDsinsertion.Proc. Natl. Acad. Sci. U.S.A.,90:10370-10374.

[16] Long, D., Swinburne, J., Martin, M., Wilson, K., Sundberg, E., Lee, K., Coupland, G., 1993b. Analysis of the frequency of inheritance of transposedDselements in Arabidopsis after activation by a CaMV 35S promoter fusion to theActransposase gene.Mol. Gen. Genet.,241:627-636.

[17] Nakagawa, Y., Machida, C., Machida, Y., Toriyama, K., 2000. Frequency and pattern of transposition of the maize transposable elementDsin transgenic rice plants.Plant Cell Physiol.,41:733-742.

[18] Rinehart, T.A., Dean, C., Weil, C.F., 1997. Comparative analysis of non-random DNA repair following Ac transposon excision in maize and Arabidopsis.Plant J.,12:1419-1427.

[19] Smith, D., Yanai, Y., Liu, Y.G., Ishiguro, S., Okada, K., Shibata, D., Whittier, R.F., Fedoroff, N.V., 1996. Characterization and mapping of Ds-GUS-T-DNA lines for targeted insertional mutagenesis.Plant J.,10:721-732.

[20] Springer, P.S., McCombie, W.R., Sundaresan, V., Martienssen, R.A., 1995. Gene trap tagging of PROLIFERA, an essential MCM2-3-5-like gene in Arabidopsis.Science,268:877-880.

[21] Sundaresan, V., Springer, P., Volpe, T., Haward, S., Jones, J.D., Dean, C., Ma, H., Martienssen, R., 1995. Patterns of gene action in plant development revealed by enhancer trap and gene trap transposable elements.Genes Dev.,9:1797-1810.

[22] Suzuki, Y., Uemura, S., Saito, Y., Murofushi, N., Schmitz, G., Theres, K., Yamaguchi, I., 2001. A novel transposon tagging element for obtaining gain-of-function mutants based on a self-stabilizingAcderivative.Plant Mol. Biol.,45:123-131.