Summary
Lepidopteran insects are major defoliating pests of soybean in the southeastern United States. Soybean plants transgenic for a nativecryIA(b) gene fromBacillus thuringiensis var.kurstaki HD-1 were obtained. Embryogenic cultures were induced by plating cotyledons on a Murashige and Skoog-based medium supplemented with 40 mg/liter of 2,4-dichlorophenoxyacetic acid (2,4-D). The embryogenic cultures were maintained in liquid medium containing 5 mg/liter 2,4-D. These cultures were subjected to microprojectile bombardment, followed by selection on 50 mg/liter hygromycin. Resistant embryogenic cell lines were transferred to growth regulator-free medium to permit recovery of mature somatic embryos. After a desiccation period, the somatic embryos were returned to growth regulator-free medium for conversion into plants. Southern hybridization analysis verified transformation. Feeding assays of T1 plants from one cell line deterred feeding, development, and survival of velvetbean caterpillar at a level comparable to that of GatIR81-296, a soybean breeding line with a high level of insect resistance. Reduced feeding on T1 plants correlated with the presence of the transgene.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Adang, M. J.; Brody, M. S.; Cardineau, G., et al. The construction and expression of aBacillus thuringiensis cryIIIA gene in protoplasts and potato plants. Plant Mol. Bol. 21:1131–1145; 1993.
Adang, M. J.; DeBoer, D.; Endres, J., et al. Manipulation ofBacillus thuringiensis genes for pest control. In: Roberts, D. W.; Granandos, R. R., eds. Biotechnology, biological pesticides, and novel plantpest resistance for insect pest management. Ithaca, NY: Cornell University; 1988:31–37.
Adang, M. J.; Idler, K. F.; Rocheleau, A. Structural and antigenic relationship among three crystal proteins ofBacillus thuringiensis subsp.kurstaki. In: Maramorosch, K., ed. Biotechnology in invertebrate pathology and cell culture. New York: Academic Press, Inc.; 1987.
Bailey, M. A.; Boerma, H. R.; Parrott, W. A. Genotype effects on proliferative embryogenesis and plant regeneration of soybean. In Vitro Cell. Dev. Biol. 29P:102–108; 1993.
Barton, K. A.; Whitely, H. R.; Yang, N.-S.Bacillus thuringiensis deltaendotoxin expressed in transgenicNicotiana tabacum provides resistance to lepidopteran insects. Plant Physiol. 85:1103–1109; 1987.
Beach, R. M.; Todd, J. W. Resistance of the soybean breeding line GatIR 81-296 to foliar feeding by threeSpodoptera sp. J. Agric. Entomol. 4:193–199; 1987.
Beach, R. M.; Todd, J. W. Foliage consumption and developmental parameters of the soybean looper and the velvetbean caterpillar (Lepidoptera: Noctuidae) reared on susceptible and resistant soybean genotypes. J. Econ. Entomol. 81:310–316; 1988.
Benedict, J. H.; Altman, D. W.; Umbeck, P. F., et al. Behavior, growth, survival, and plant injury byHeliothis virescens (F.) (Lepidoptera: Noctuidae) on transgenicBt cottons. J. Econ. Entomol. 85:589–593; 1992.
Benedict, J. H.; Sachs, E. S.; Altman, D. W., et al. Impact of δ-endotoxin-producing transgenic cotton on insect-plant interactions withHeliothis virescens andHelicoverpa zea (Lepidoptera: Noctuidae). Environ. Entomol. 22:1–9; 1993.
Brattsten, L. B. Bioengineering of crop plants and resistant biotype evolution in insects: counteracting coevolution. Arch. Ins. Bioch. Physiol. 17:353–267; 1991.
Breyne, P.; Van Montagu, M.; Depicker, A., et al. Characterization of a plant scaffold attachment region in a DNA fragment that normalizes transgene expression. Plant Cell 4:463–471; 1992.
Carozzi, N. B.; Warren, G. W.; Desai, N., et al. Expression of a chimeric CaMV 35SBacillus thuringiensis insecticidal protein gene in transgenic tobacco. Plant Mol. Biol. 20:539–548; 1992.
Chang, H.-H.; Chan, M.-T.Agrobacterium tumefaciens-mediated transformation of soybean (Glycine max (L.) Merr.) is promoted by the inclusion of potato suspension culture. Bot. Bull. Acad. Sin. 32:171–178; 1991.
Chee, P. P.; Fober, K. A.; Slightom, J. L. Transformation of soybean (Glycine max) by infecting germinating seeds withAgrobacterium tumefaciens. Plant Physiol. 91:1212–1218; 1989.
Cheng, J.; Bolyard, M. G.; Saxena, R. C., et al. Production of insect resistant potato by genetic transformation with a δ-endotoxin gene fromBacillus thuringiensis varkurstaki. Plant Sci. 81:83–91; 1992.
Christou, P. Morphological description of transgenic soybean chimeras created by the delivery, integration and expression of foreign DNA using electric discharge particle acceleration. Ann. Bot. 66:379–386; 1990.
Christou, P. Soybean transformation by electric discharge particle acceleration. Physiol. Plant 79:210–212; 1990.
Christou, P.; McCabe, E. Prediction of germ-line transformation events in chimeric R0 transgenic soybean plantlets using tissue-specific expression patterns. Plant J. 2:283–290; 1992.
Christou, P.; McCabe, D. E.; Martinell, B. J., et al. Soybean genetic engineering—commercial production of transgenic plants. Trends Biotech. 8:145–151; 1990.
Christou, P.; Swain, W. F.; Yang, N. S., et al. Inheritance and expression of foreign genes in transgenic soybean plants. Proc. Natl. Acad. Sci. USA 86:7500–7504; 1989.
Delannay, X.; LaVallee, J.; Proksch, K., et al. Field performance of transgenic tomato plants expressing theBacillus thuringiensis var.kurstaki insect control protein. Bio/Technology. 7:1265–1269; 1989.
Feiltelson, J. S.; Payne, J.; Kim, L.Bacillus thuringiensis: insects and beyond. Bio/Technology 10:271–276; 1992.
Finer, J. J.; McMullen, M. D. Transformation of soybean via particle bombardment of embryogenic suspension culture tissue. In Vitro Cell. Dev. Biol. 27P:175–182; 1991.
Finer, J. J.; Nagasawa, A. Development of an embryogenic suspension culture of soybean (Glycine max Merrill). Plant Cell Tissue Organ Cult. 15:125–136; 1988.
Firoozabady, E.; DeBoer, D. L.; Merlo, D. J., et al. Transformation of cotton (Gossypium hirsutum L.) byAgrobacterium tumefaciens and regeneration of transgenic plants. Plant Mol. Biol. 10:105–116; 1987.
Fischoff, D. A.; Bowdish, K. S.; Perlak, F. J., et al. Insect tolerant transgenic tomato plants. Bio/Technology 5:807–813; 1987.
Gamborg, O. L.; Miller, R. A.; Ojima, K. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50:150–158; 1968.
Gordon-Kamm, W. J.; Spencer, T. M.; Mangano, M. L., et al. Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell 2:603–618; 1990.
Hinchee, M. A. W.; Connor-Ward, D. V.; Newell, C. A., et al. Production of transgenic soybean plants usingAgrobacterium-mediated gene transfer. Bio/Technology 6:915–922; 1988.
Hoffmann, M. P.; Zalom, F. G.; Wilson, L. T., et al. Field evaluation of transgenic tobacco containing genes encodingBacillus thuringiensis δ-endotoxin or cowpea trypsin inhibitor: Efficacy againstHelicoverpa zea (Lepidoptera: Noctuidae). J. Econ. Entomol. 85:2517–2522; 1992.
Hudson, R. D.; Jones, D. C.; McPherson, R. M. Soybean insects. In: Douce, G. K.; McPherson, R. M., eds. Special publication 70.: Georgia Agricultural Experiment Stations; 1991.
Jenkins, J. N.; Parrott, W. L.; McCarty, J. C., Jr., et al. Growth and survival ofHeliothis virescens *Lepidoptera: Noctuidae) on transgenic cotton containing a truncated form of the delta endotoxin gene fromBacillus thuringiensis. J. Econ. Entomol. 86:181–185; 1993.
Kao, K. N. Staining methods for plant protoplasts and cells. In: Wetter, L. R.; Constabel, F., eds. Plant tissue culture methods. Saskatoon, Saskatchewan: Prairie Regional Laboratory of the National Research Council of Canada; 1982:67–71.
Keim, P.; Olson, T. C.; Shoemaker, R. C. A rapid protocol for isolating soybean DNA. Soybean Genet. Newslett. 15:150–152; 1988.
Kilby, N. J.; Leyser, H. M. O.; Furner, I. J. Promoter methylation and progressive transgene inactivation inArabidopsis. Plant Mol. Biol. 20:103–112; 1992.
Komatsuda, T.; Ohyama, K. Genotypes of high competence for somatic embryogenesis and plant regeneration in soybeanGlycine max. Theor. Appl. Genet. 75:695–700; 1988.
Koziel, M. G.; Beland, G. L.; Bowman, C., et al. Field performance of elite transgenic maize plants expressing an insecticidal protein derived fromBacillus thuringiensis. Bio/Technology 11:194–200; 1993.
Liu, B.-L.; Yue, S.-X.; Hu, N.-B., et al. Transfer of atrazine resistant gene from nightshade to soybean chloroplast genome and its expression in transgenic plants. Ch. Sci. Bull. 34:1670–1672; 1989.
Liu, B.-L.; Yue, S.-X.; Hu, N.-B., et al. Transfer of the atrazine-resistant gene of black nightshade to soybean chloroplast genome and its expression in transgenic plants. Sci. Chi. Ser. B. 33:444–452; 1990.
Luthy, P.; Ebersold, H. R.Bacillus thuringiensis delta endotoxin: Histopathology and molecular mode of action. In: Davison, E. W., ed. Pathogenesis of invertebrate microbial Diseases. Toronto: Allenheld; 1981:235–237.
Maniatis, T.; Fritsch, E. F.; Sambrook, J. Molecular cloning. A laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1982.
McCabe, D. E.; Swain, W. F.; Martinell, B. J., et al. Stable transformation of soybean (Glycine max) by particle acceleration. Bio/Technology 6:923–926; 1988.
McCouch, S. R.; Kochert, G.; Yu, Z. H., et al. Molecular mapping of rice chromosomes. Theor. Appl. Genet. 76:815–829; 1988.
McGaughey, W. H.; Whalon, M. E. Managing insect resistance toBacillus thuringiensis toxins. Science 258:1451–1455; 1992.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Murray, E. E.; Rochleau, T. R.; Eberle, M., et al. Analysis of unstable RNA transcripts of insecticidal crystal protein genes ofBacillus thuringiensis in transgenic plants and electroporated protoplasts. Plant Mol. Biol. 16:1035–1060; 1991.
Perlak, F. J.; Deaton, R. W.; Armstrong, T. A., et al. Insect resistant cotton plants. Bio/Technology 8:939–943; 1990.
Portillo, H. E.; Pitre, H. N. Effect of four soybean genotypes on the development and fecundity ofHeliothis virescens andPseudoplusia includens (Lepidoptera: Noctuidae). Fl. Entomol. 75:386–390; 1992.
Renckens, S.; De Greve, H.; Van Montagu, M., et al.Petunia plants escape from negative selection against a transgene by silencing the foreign DNA via methylation. Mol. Gen. Genet. 233:53–64; 1992.
Rufener, G. K., II.; St. Martin, S. K.; Cooper, R. L., et al. Genetics of antibiosis resistance to Mexican bean beetle in soybean. Crop Sci. 29:618–622; 1989.
Stewart, C. N., Jr.; Via, L. E. A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. Bio-Techniques 14:748–751; 1993.
Vaeck, M.; Reynaerts, A.; Höfte, H., et al. Transgenic plants protected from insect attack. Nature 238:33–37; 1987.
Van Rie, J. Insect control with transgenic plants: resistance proof? Tibtech 9:177–179; 1991.
Warren, G. W.; Carozzi, N. D.; Desai, N., et al. Field evaluation of transgenic tobacco containing aBacillus thuringiensis insecticidal protein gene. J. Econ. Entomol. 85:1651–1659; 1992.
Wilson, F. D.; Flint, H. M.; Deaton, R. W., et al. Resistance of cotton lines containing aBacillus thuringiensis toxin to pink bollworm (Lepidoptera: Gelechiidae) and other insects. J. Econ. Entomol. 85:1516–1521; 1992.
Yang, Y.-S.; Wada, K.; Goto, M., et al.In vitro formation of nodular calli in soybean (Glycine max L.) induced by cocultivatedPseudomonas maltophilia. Jpn. J. Breed. 41:595–604; 1991.
Zhou, J. H.; Atherly, A. G. In situ detection of transposition of the maize controlling element (Ac) in transgenic soybean tissues. Plant Cell Rep. 8:542–545; 1990.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Parrott, W.A., All, J.N., Adang, M.J. et al. Recovery and evaluation of soybean plants transgenic for aBacillus thuringiensis var.Kurstaki insecticidal gene. In Vitro Cell Dev Biol - Plant 30, 144–149 (1994). https://doi.org/10.1007/BF02632204
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02632204