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Chemically inducible gene expression in seeds before testa rupture

  • Mariko Nonogaki (a1), Taira Sekine (a2) and Hiroyuki Nonogaki (a1)


Impermeability of the testa hinders efficient penetration of some small chemicals, such as transcriptional inhibitors, through the endosperm and the embryo during seed experiments. In Arabidopsis seeds, 5-bromo-4-chloro-3-indolyl β-d-glucuronic acid, a substrate for β-glucuronidase, did not permeate through the endosperm and embryo efficiently at the stages before testa rupture. The Arabidopsis testa also limited efficient entry of methoxyfenozide, a chemical ligand that was used for inducible gene expression experiments, into seeds. While the detection of a reporter gene at the early imbibitional stages could be replaced by reverse transcription-polymerase chain reaction (RT-PCR), the interference of entry of the chemical ligand into seeds by the testa was still problematic to gene induction experiments. To develop an efficient inducible expression system for gene function analysis in seeds, an inducible expression system with nitrate, which is a testa-permeable ligand, was examined. The vector containing the 2.1-kb upstream sequence of NITRITE REDUCTASE 1 was able to cause expression of a test gene (long non-coding RNA) in imbibed seeds at the stage before testa rupture in a nitrate-dependent manner. This system can be used not only for characterization of genes associated with seed dormancy and germination in basic research, but also for the development of germination recovery or enhancement technologies for agricultural applications.


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Aoyama, T., Dong, C.H., Wu, Y., Carabelli, M., Sessa, G., Ruberti, I., Morelli, G. and Chua, N.H. (1995) Ectopic expression of the Arabidopsis transcriptional activator Athb-1 alters leaf cell fate in tobacco. Plant Cell 7, 17731785.
Böhner, S., Lenk, I., Rieping, M., Herold, M. and Gatz, C. (1999) Transcriptional activator TGV mediates dexamethasone-inducible and tetracycline-inactivatable gene expression. Plant Journal 19, 8795.
Bruce, W., Folkerts, O., Garnaat, C., Crasta, O., Roth, B. and Bowen, B. (2000) Expression profiling of the maize flavonoid pathway genes controlled by estradiol-inducible transcription factors CRC and P. Plant Cell 12, 6580.
Caddick, M.X., Greenland, A.J., Jepson, I., Krause, K.P., Qu, N., Riddell, K.V., Salter, M.G., Schuch, W., Sonnewald, U. and Tomsett, A.B. (1998) An ethanol inducible gene switch for plants used to manipulate carbon metabolism. Nature Biotechnology 16, 177180.
Debeaujon, I., Leon-Kloosterziel, K.M. and Koornneef, M. (2000) Influence of the testa on seed dormancy, germination, and longevity in Arabidopsis. Plant Physiology 122, 403414.
De Veylder, L., Beeckman, T., Van Montagu, M. and Inzé, D. (2000) Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene. Journal of Experimental Botany 51, 16471653.
Gatz, C. (1997) Chemical control of gene expression. Annual Review of Plant Physiology and Plant Molecular Biology 48, 89108.
Konishi, M. and Yanagisawa, S. (2010) Identification of a nitrate-responsive cis-element in the Arabidopsis NIR1 promoter defines the presence of multiple cis-regulatory elements for nitrogen response. Plant Journal 63, 269282.
Koo, J.C., Asurmendi, S., Bick, J., Woodford-Thomas, T. and Beachy, R.N. (2004) Ecdysone agonist-inducible expression of a coat protein gene from tobacco mosaic virus confers viral resistance in transgenic Arabidopsis . Plant Journal 37, 439448.
Liu, P.P., Koizuka, N., Martin, R.C. and Nonogaki, H. (2005a) The BME3 (Blue Micropylar End 3) GATA zinc finger transcription factor is a positive regulator of Arabidopsis seed germination. Plant Journal 44, 960971.
Liu, P.P., Koizuka, N., Homrichhausen, T.M., Hewitt, J.R., Martin, R.C. and Nonogaki, H. (2005b) Large-scale screening of Arabidopsis enhancer-trap lines for seed germination-associated genes. Plant Journal 41, 936944.
Lloyd, A.M., Schena, M., Walbot, V. and Davis, R.W. (1994) Epidermal cell fate determination in Arabidopsis: patterns defined by a steroid-inducible regulator. Science 266, 436439.
Love, J., Scott, A.C. and Thompson, W.F. (2000) Stringent control of transgene expression in Arabidopsis thaliana using the Top10 promoter system. Plant Journal 21, 579588.
Martinez-Andujar, C., Ordiz, M.I., Huang, Z., Nonogaki, M., Beachy, R.N. and Nonogaki, H. (2011) Induction of 9-cis-epoxycarotenoid dioxygenase in Arabidopsis thaliana seeds enhances seed dormancy. Proceedings of the National Academy of Sciences USA 108, 1722517229.
Matakiadis, T., Alboresi, A., Jikumaru, Y., Tatematsu, K., Pichon, O., Renou, J.-P., Kamiya, Y., Nambara, E. and Truong, H.-N. (2009) The Arabidopsis abscisic acid catabolic gene CYP707A2 plays a key role in nitrate control of seed dormancy. Plant Physiology 149, 949960.
Mosallanejad, H., Badisco, L., Swevers, L., Soin, T., Knapen, D., Vanden Broeck, J. and Smagghe, G. (2010) Ecdysone signaling and transcript signature in Drosophila cells resistant against methoxyfenozide. Journal of Insect Physiology 56, 19731985.
Nesi, N., Jond, C., Debeaujon, I., Caboche, M. and Lepiniec, L. (2001) The Arabidopsis TT2 gene encodes an R2R3 MYB domain protein that acts as a key determinant for proanthocyanidin accumulation in developing seed. The Plant Cell 13, 20992114.
Nonogaki, H. (2014) Seed dormancy and germination-emerging mechanisms and new hypotheses. Frontiers in Plant Science 5, 233.
Nonogaki, M., Sall, K., Nambara, E. and Nonogaki, H. (2014) Amplification of ABA biosynthesis and signaling through a positive feedback mechanism in seeds. Plant Journal 78, 527539.
Padidam, M. (2003) Chemically regulated gene expression in plants. Current Opinion in Plant Biology 6, 169177.
Piskurewicz, U., Jikumaru, Y., Kinoshita, N., Nambara, E., Kamiya, Y. and Lopez-Molina, L. (2008) The gibberellic acid signaling repressor RGL2 inhibits Arabidopsis seed germination by stimulating abscisic acid synthesis and ABI5 activity. The Plant Cell 20, 27292745.
Rajjou, L., Gallardo, K., Debeaujon, I., Vandekerckhove, J., Job, C. and Job, D. (2004) The effect of alpha-amanitin on the Arabidopsis seed proteome highlights the distinct roles of stored and neosynthesized mRNAs during germination. Plant Physiology 134, 15981613.
Roslan, H.A., Salter, M.G., Wood, C.D., White, M.R.H., Croft, K.P., Robson, F., Coupland, G., Doonan, J., Laufs, P., Tomsett, A.B. and Caddick, M.X. (2001) Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana . Plant Journal 28, 225235.
Tavva, V.S., Dinkins, R.D., Palli, S.R. and Collins, G.B. (2006) Development of a methoxyfenozide-responsive gene switch for applications in plants. Plant Journal 45, 457469.
Zuo, J., Niu, Q.-W., Frugis, G. and Chua, N.-H. (2002) The WUSCHEL gene promotes vegetative-to-embryonic transition in Arabidopsis . Plant Journal 30, 349359.


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Chemically inducible gene expression in seeds before testa rupture

  • Mariko Nonogaki (a1), Taira Sekine (a2) and Hiroyuki Nonogaki (a1)


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