PROJECT PLAN TITLE:
Genetic engineering of floral bulb crops for virus and nematode resistance
INTRODUCTION:
Floral bulb crops are propagated vegetatively making it difficult to eliminate viruses that cause stunting of the plant and streaking on the flowers that make the flowers unmarketable. Virus resistance by genetic engineering has been achieved in a variety of crops but not the flower bulb crops. Cucumber mosaic virus is one of the main viruses in gladiolus and infects numerous agricultural crops as well. Both gladiolus and lilies will be developed that contain an antiviral gene for cucumber mosaic virus, and these plants will be analyzed for their resistance to the virus. The root lesion nematode is the number one pest of Easter lilies grown in the field, and it ranks third for the damage that it causes to agricultural crops. Genes that affect the development of the nematode will be isolated and used to develop Easter lilies with these genes for possible resistance against nematodes.
PROGRESS:
1. Developed the technology for genetic engineering of gladiolus that is reproducible and applicable to many cultivars. Applied the technology to develop gladiolus plants with antiviral genes and found that the plants had short-term, but not long-term, resistance to Bean yellow mosaic virus. Developed gladiolus plants with short-term resistance to Cucumber mosaic virus. 2. Have developed a system for genetic engineering of Easter lilies that is currently being optimized. 3. Isolated a polyubiquitin promoter from gladiolus that has been found to direct high levels of gene expression in gladiolus. This promoter will be useful for genetic engineering when high levels of a gene are required for effective resistance.
PLANS:
1. Develop both gladiolus and lilies with a double stranded RNA for cucumber mosaic virus resistance. 2. Identify genes that affect the development of the root lesion nematode and use these genes in lilies to develop resistance to nematodes.
RECENT PUBLICATIONS:
2000. Kamo, K., Blowers, A., and McElroy, D. Effect of the cauliflower mosaic virus 35S, actin, and ubiquitin promoters on uidA expression from a bar-uidA fusion gene in transgenic Gladiolus plants. In Vitro Cell. Dev. Biol.-Plant 36:13-20.
2000. Kamo, K., McElroy, D., and Chamberlain, D. Transforming embryogenic cells lines of Gladiolus with either a bar-uidA fusion gene or cobombardment. In Vitro Cell. Dev. Biol.-Plant 36:182-187.
2000. De Villiers, S. M., Kamo, K., Thomson, J. A., Bornman, C. H. and Berger, D. K. Biolistic transformation of chincherinchee (Ornithogalum) and regeneration of transgenic plants. Physiol. Plant. 109: 450-455.
2001. Kamo, K., Roh, M., Blowers, A., Smith, F. and Van Eck, J. Transgenic Gladiolus. In: Y.P.S. Bajaj (Editor) Biotechnology and Agriculture and Forestry 48. Transgenic Crops III, Springer Verlag, Berlin, pp. 155-169.
2001. Joung, Y. H., Roh, M. S., Kamo, K. and Song, J. S. Agrobacterium-mediated transformation of Campanula glomerata. Plant Cell Rep. 20:289-295.
2001. Kamo, K. Expression of the bar and uidA genes by Gladiolus following three seasons of dormancy. Acta Hort. 560:165-167.
2002. Castillon, J. and Kamo, K. Maturation and conversion of somatic embryos of three genetically diverse rose cultivars. HortScience 37:973-977.
2002. Joung, Y. H., Liao, M. S., Roh, M. S., Kamo, K. and Song, J. S. In vitro propagation of Campanula glomerata, ‘Acaulis’ from leaf blades. Scientia Hort. 92:137-146.
2002. Lipsky, A., Cohen, A., Gaba, V., Kamo, K., Gera, A., Watad, A. Transformation of Lilium longiflorum plants for cucumber mosaic virus resistance by particle bombardment. Acta Hort. 568:209-214.
2003. Kamo, K. K. Long-term expression of the uidA gene in Gladiolus plants under control of either the ubiquitin, rolD, mannopine synthase, or cauliflower mosaic virus promoters following three seasons of dormancy. Plant Cell Rep. 21:797-803.
2004. Kamo, K., Jones, B., Castillon, J., Bolar, J. and Smith, F. Dispersal and filtration of embryogenic callus increases the frequency of embryo maturation and conversion for hybrid tea roses. Plant Cell Rep. 22:787-792.
2004. Kim, C. K., Chung, J. D., Park, M. Kamo, K. and Byrne, D. H. Agrobacterium tumefaciens-mediated transformation of Rosa hybrida using the green fluorescent protein (GFP) gene. Plant Cell Tiss. Org. Cult. 78:107-111.
2004. Ahn, J., Joung, Y.H., and Kamo, K. Transformation of Easter lilies. Transgenic plants of Easter lily (Lilium longiflorum) with phosphinothricin resistance. J. Plant Biotech. 6:9-13.
2005. Kamo, K., Gera, A., Cohen, J., Hammond, J., Blowers, A., Smith, F. and Van Eck, J. Transgenic Gladiolus plants transformed with the Bean yellow mosaic virus coat protein gene in either sense or antisense orientations. Plant Cell Rep. 23:654-663.
2005. Aebig, J. A., Kamo, K. and Hsu, H.T. Biolistic inoculation of gladiolus with cucumber mosaic cucumovirus. J. Virol. Meth. 123:89-94.
2005. Kamo, K., Jones, B., Bolar, J. and Smith, F. Regeneration from long-term embryogenic callus of the Rosa hybrida cultivar Kardinal. In Vitro Cell. Dev. Biol.-Plant 41:32-36.
2006. Joung, Y. H. and Kamo, K. Expression of a polyubiquitin promoter isolated from Gladiolus. Plant Cell Rep. 25:1081-1088.
2007. Joung, H. Y., Cantor, M., Ellis, D. and Kamo, K. Vitrification of Gladiolus shoot tips from cormels. J. Korean Society Horticultural Science 48:251-255.
2007. Kamo, K. and Joung, Y. H. A polyubiquitin promoter isolated from Gladiolus and its expression in Gladiolus plants. Acta Hort. 763:251-258.
2007. Joung, H. Y., Cantor, M., Kamo, K. and Ellis, D. Cryopreservation of Gladiolus cultivars. Acta Hort. 760:225-232.
2008. Kamo, K. Transgene expression for Gladiolus plants grown outdoors and in the greenhouse. Scientia Horticulturae 117:275-280.
2008. Kamo, K. and Han, B. H. Biolistic-mediated transformation of Lilium longiflorum cv. Nellie White. HortScience 43:1-6.
2009. Kamo, K. and Joung, H.Y. Long-term gus expression from Gladiolus callus lines containing either a bar-uidA fusion gene or bar and uidA delivered on separate plasmids. Plant Cell Tiss. Organ Cult. 98:263-272.
2009. Kamo, K., Young, Y.H., and Green, K. GUS expression in Gladiolus plants controlled by two Gladiolus ubiquitin promoters. Floriculture and Ornamental Biotechnology 3:10-14.
2010. Kamo, K., Jordan, R., Guaragna, M.A., Hsu, H.-t., and Ueng, P. Resistance to Cucumber mosaic virus in Gladiolus plants transformed with either a defective replicase or coat protein subgroup II gene from Cucumber mosaic virus. Plant Cell Rep. 29:695-704.
2011. Kamo, K. Inherited transgene expression of the uidA and bar genes in Lilium longiflorum cv. ‘Nellie White’. Floriculture and Ornamental Biotechnology 5:35-39.
2012. Lakshman, D.K., Pandey, R., Kamo, K. Bauchan, G., and Mitra, A. Genetic transformation of Fusarium oxysporum f.sp. gladioli with Agrobacterium to study pathogenesis in Gladiolus. Eur. J. Plant Pathol. DOI 10.1007/s10658-012-9953-0. Online 2012.
2012. Kamo, K., Aebig, J., Guaragna, M.A., James, C., Hsu, H.-t., and Jordan, R. Gladiolus plants transformed with single-chain variable fragment antibodies to Cucumber mosaic virus. Plant Cell Tiss. Organ Cult. DOI 10.1007/s11240-012-0124-y. Online 2012.
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