Abstract
Transient GUS (β-glucuronidase) expression was visualized in cell suspensions of Triticum aestivum, Zea mays, Pennisetum glaucum, Saccharum officinarum, Pennisetum purpureum and Panicum maximum after microprojectile bombardment with pBARGUS and pAHC25 plasmid DNAs. pBARGUS contains the GUS (UidA) gene coding region driven by the Adh1 promoter and the Adh1 intron 1, as well as the BAR gene coding region driven by the CaMV 35S promoter and the Adh1 intron 1. pAHC25 contains the GUS and BAR gene coding regions driven by the maize ubiquitin promoter, first exon and first intron (Ubi1). The effectiveness of the constructs was first compared in cell suspension cultures by counting blue expression units (b.e.u.). The expression of construct pAHC25 ranged from 3 to 50 fold greater than pBARGUS in different species. In addition, the two plasmids were quantitatively compared in Triticum aestivum and Zea mays by using the more sensitive GUS fluorometric assay to determine the amount of methylumbellyferride (MU) produced. There was more than a 30 fold increase in MU production with pAHC25 than with pBARGUS in the wheat suspension, while the maize suspension showed only a 2.5 fold increase with the pAHC25 construct. Transient GUS expression was also visualized in immature embryos of Pennisetum glaucum following bombardment with pBARGUS and pAHC25 DNA. Expression of plasmid pAHC25 was twice as high as pBARGUS. A comparison of two DNA/gold preparation methods, as well as repeated sonications of the DNA/gold mixture, had no effect on the number of b.e.u.
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References
Bowers R, Birch RG (1992) The Plant J 2:409–416
Callis J, Fromm M, Walbot V (1987) Genes Dev 1:1183–1200
Christensen AH, Sharrock RA, Quail PH (1992) Plant Mol Biol 18:675–689
Christou P, Ford TL, Kofron M (1991) Bio/Technology 9:957–962
Freeling M (1973) Molec Gen Genet 127:215–227
Fromm M, Morrish F, Armstrong C, Williams R, Thomas J, Klein TM (1990) Bio/Technology 8:833–839
Fromm M, Taylor LP, Walbot V (1985) Proc Nat Acad Sci USA 82:5824–5828
Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brien JV, Chambers SA, Adams WR Jr., Willets NG, Rice TB, Mackey CJ, Krueger RW, Kausch AP, Lemaux PG (1990) The Plant Cell 2:603–618.
Hauptmann RM, Ozias-Akins P, Vasil V, Tabaeizadeh Z, Rogers SG, Horsch RB, Vasil IK, Fraley RT (1987) Plant Cell Rep 6:265–270
Ho WJ, Vasil IK (1983) Ann Bot 51:719–726
Jefferson RA, Kavanagh TA, Bevan MW (1987) EMBO J 6:3901–3907
Karlsson SB, Vasil IK (1984) J Plant Physiol 123:211–227
Klein TM, Gradziel T, Fromm ME, Sanford JC (1988) Bio/Technology 6:559–563
Kyozuka J, Izawa T, Nakajima M, Shimamoto K (1990) Maydica 35:353–357
Last DI, Brettell RIS, Chamberlain DA, Chaudhury AM, Larkin PJ Marsh EL, Peacock WJ, Dennis ES (1991) Theor Appl Genet 81:581–588
Lee BT, Murdoch K, Topping J, Both MTJ de, Wu QS, Karp A, Steele gnS, Symonds C, Kreis M, Jones MGK (1987) In: Puite KJ, Dons JJM, Huizing HJ, Kool AJ, Koornneef M, Krens FA (eds) Progress in Plant Protoplast Research. Proc 7th Int Protoplast Symp, Wageningen, The Netherlands, December 6–11, 1987, pp 343–346
Lu C, Vasil IK (1981) Ann Bot 47:543–548
Luehrson KR, Walbot V (1991) Molec Gen Genet 225:81–93
Maas C, Laufs J, Grant S, Korfhage C, Werr W (1991) Plant Mol Biol 16:199–207
McElroy D, Zhang W, Cao J, Wu R (1990) The Plant Cell 2:163–171
Murashige T, Skoog F (1962) Physiol Plant 15:473–497
Oard JH, Paige DF, Simmonds JA, Gradziel TM (1990) Plant Physiol 92:334–339
Paul A-L, Ferl RJ (1991) Maydica 36:129–134
Redway FA, Vasil V, Vasil IK (1990) Plant Cell Rep 8:714–717
Reggiardo MI, Arana JL, Orsaria LM, Permingeat HR, Spitteler MA, Vallejos RH (1991) Plant Science 75:237–243
Russell JA, Mihir KR, Sanford JC (1992) In Vitro Cell Dev Biol 28P:97–105
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY
Sanford JC, Devit MJ, Russell JA, Smith FD, Harpending PR, Roy MK, Johnston SA (1991) Technique 3:3–16
Taylor MG, Vasil IK (1991) Plant Cell Rep 10:120–125
Vasil V, Castillo A, Fromm ME, Vasil IK (1992) Bio/Technology 6:667–672
Vasil V, Clancy M, Ferl RJ, Vasil IK, Hannah LC (1989) Plant Physiol 91:1575–1579
Vasil V, Vasil IK (1981) Ann Bot 47:669–678
Vasil V, Vasil IK (1986) J Plant Physiol 124:399–408
Zhang W, Wu R (1988) Theor Appl Genet 76:835–840
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Communicated by R. N. Beachy
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Taylor, M.G., Vasil, V. & Vasil, I.K. Enhanced GUS gene expression in cereal/grass cell suspensions and immature embryos using the maize uhiquitin-based plasmid pAHC25. Plant Cell Reports 12, 491–495 (1993). https://doi.org/10.1007/BF00236093
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DOI: https://doi.org/10.1007/BF00236093