Abstract
An aluminum borate whiskers-mediated transformation system for calluses of tobacco (Nicotiana tabacum, cv. SR-1) has been developed. A total of 50 small pieces of calluses were vigorously agitated in a liquid medium containing aluminum borate whiskers, pBI221 plasmid carrying the β-glucuronidase (GUS) gene, and pBI222 plasmid carrying the hygromycin phosphotransferase (HPT) gene. After treatment, calluses were cultured to select for hygromycin resistance, and three resistant calluses were obtained. Adventitious shoots were produced from each hygromycin-resistant callus and were transferred to rooting medium. A total of three plantlets obtained from each hygromycin-resistant callus were acclimatized and established in soil. Polymerase chain reaction analysis revealed that all the plantlets were cotransformed with both the GUS and HPT genes. Detached leaves of transgenic individuals showed clear hygromycin resistance when cultured in liquid medium. Histochemical assay for GUS revealed that one of these transgenic plants expressed the GUS gene, indicating coexpression of foreign genes.
Similar content being viewed by others
Abbreviations
- ABW:
-
aluminum borate whiskers
- BA:
-
6-benzyladenine
- CaMV:
-
cauliflower mosaic virus
- GUS:
-
β-glucuronidase
- HPT:
-
hygromycin phosphotransferase
- NAA:
-
1-naphthaleneacetic acid
- PCR:
-
polymerase chain reaction
- SCW:
-
silicon carbide whiskers
References
Y Asano Y Otsuki M Ugaki (1991) ArticleTitleElectroporation-mediated and silicon carbide fiber-mediated DNA delivery in Agrostis alba L. (Red top). Plant Sci. 79 247–252
EA Brisibe A Gajdosova A Olesen SB Andersen (2000) ArticleTitleCytodifferentiation and transformation of embryogenic callus lines derived from anther culture of wheat J. Exp. Bot. 51 187–196
SJ Dalton AJE Bettany E Timms P Morris (1998) ArticleTitleTransgenic plants of Lolium multiflorum, Lolium perenne, Festuca arundinacea and Agrostis stolonifera by silicon carbide fibre-mediated transformation of cell suspension cultures Plant Sci. 132 31–43
BR Frame PR Drayton SV Bagnall CJ Lewnau WP Bullock HM Wilson JM Dunwell JA Thompson K Wang (1994) ArticleTitleProduction of fertile transgenic maize plants by silicon carbide whisker-mediated transformation Plant J. 6 941–948
GB Gharti-Chhetri W Cherdshewasart J Dewulf J Paszkowski M Jacobs I Negrutiu (1990) ArticleTitleHybrid genes in the analysis of transformation conditions. 3. Temporal/spatial fate of NPT II gene integration, its inheritance and factors affecting these processes in Nicotiana plumbaginifolia. Plant Mol. Biol. 14 687–696
R Hunold R Bronner G Hahne (1994) ArticleTitleEarly events in microprojectile bombardment: cell viability and particle location Plant J. 5 593–604
RA Jefferson TA Kavanagh MW Bevan (1987) ArticleTitleGUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants EMBO J. 6 3901–3907
HF Kaeppler W Gu DA Somers HW Rines AF Cockburn (1990) ArticleTitleSilicon carbide fiber-mediated DNA delivery into plant cells Plant Cell Rep. 9 415–418
HF Kaeppler DA Somers HW Rines AF Cockburn (1992) ArticleTitleSilicon carbide fiber-mediated stable transformation of plant cells Theor. Appl. Genet. 84 560–566
S Kartzke H Saedler P Meyer (1990) ArticleTitleMolecular analysis of transgenic plants derived from transformations of protoplasts at various stages of the cell cycle Plant Sci. 67 63–72
EL Kemper Silva MJ da P Arruda (1996) ArticleTitleEffect of microprojectile bombardment parameters and osmotic treatment on particle penetration and tissue damage in transiently transformed cultured immature maize (Zea mays L.) embryos Plant Sci. 121 85–93
S Kosugi Y Ohashi K Nakajima Y Arai (1990) ArticleTitleAn improved assay for β-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous β-glucuronidase activity Plant Sci. 70 133–140
J Matsushita M Otani Y Wakita O Tanaka T Shimada (1999) ArticleTitleTransgenic plant regeneration through silicon carbide whisker-mediated transformation of rice (Oryza sativa L.) Breed. Sci. 49 21–26
K Mizuno W Takahashi T Ohyama T Shimada O Tanaka (2004) ArticleTitleImprovement of the aluminum borate whisker-mediated method of DNA delivery into rice callus Plant Prod. Sci. 7 45–49
T Murashige F Skoog (1962) ArticleTitleA revised medium for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant. 15 473–497
N Nagatani H Honda T Shimada T Kobayashi (1997) ArticleTitleDNA delivery into rice cells and transformation using silicon carbide whiskers Biotechnol. Tech. 11 471–473
S Omirulleh A Ismagulva M Karabaev T Meshi M Iwabuchi (1996) ArticleTitleSilicon carbide fiber-mediated DNA delivery into cells of wheat (Triticum aestivum L.) mature embryos Plant Cell Rep. 16 133–136
JF Petolino NF Hopkins BD Kosegi M Skokut (2000) ArticleTitleWhisker-mediated transformation of embryogenic callus of maize Plant Cell Rep. 19 781–786
SO Rogers AJ Bendich (1985) ArticleTitleExtraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues Plant Mol. Biol. 5 69–76
I Svensson E Artursson P Leanderson R Berglind F Lindgren (1997) ArticleTitleToxicity in vitro of some silicon carbides and silicon nitrides: whiskers and powders Am. J. Ind. Med. 31 335–343
W Takahashi T Shimada J Matsushita O Tanaka (2000) ArticleTitleAluminium borate whisker-mediated DNA delivery into callus of rice and production of transgenic rice plant Plant Prod. Sci. 3 219–224
W Takahashi H Oishi M Ebina T Takamizo T Komatsu (2002) ArticleTitleProduction of transgenic Italian ryegrass (Lolium multiflorum Lam.) via microprojectile bombardment of embryogenic calli Plant Biotechnol. 19 241–249
P Vain MD McMullen JJ Finer (1993) ArticleTitleOsmotic treatment enhances particle bombardment-mediated transient and stable transformation of maize Plant Cell Rep. 12 84–88
GL Vaughan J Jordan S Karr (1991) ArticleTitleThe toxicity, in vitro, of silicon carbide whiskers Environ. Res. 56 57–67
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mizuno, K., Takahashi, W., Beppu, T. et al. Aluminum borate whisker-mediated production of transgenic tobacco plants. Plant Cell Tiss Organ Cult 80, 163–169 (2005). https://doi.org/10.1007/s11240-004-9542-9
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11240-004-9542-9