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Title: DEVELOPMENT OF A GENETIC TRANSFORMATION AND REGENERATION SYSTEM FOR TARO

Author
item HE, X. - UNIVERSITY OF HAWAII
item MIYASAKA, S. - UNIVERSITY OF HAWAII
item Fitch, Maureen
item ZHU, Y. - HARC
item Moore, Paul

Submitted to: CTAHR Student Research Symposium
Publication Type: Proceedings
Publication Acceptance Date: 3/15/2003
Publication Date: 4/24/2003
Citation: He, X., Miyasaka, S.C., Fitch, M.M., Zhu, Y.J., Moore, P.H. 2003. Development of a genetic transformation and regeneration system for taro. 15th Annual CTAHR Student Research Symposium (Abstracts) p.21. 2003.

Interpretive Summary: abstract only

Technical Abstract: Taro (Colocasia esculenta) is an important staple crop in the Pacific Islands. To improve resistance of taro to fungal pathogens, a genetic transformation and regeneration system has been developed. Highly regenerative taro calli of cv. Bun Long were obtained in a Murashige and Skoog (MS) medium with 2 mg L-1 benzyladenine (BA) and 1 mg L-1 napthaleneacetic acid (NAA). Alternating between MS with 2 mg L-1 BA plus 1 mg L-1 NAA and MS with 4 mg L-1 BA every two weeks induced shoot regeneration from callus. The rice chitinase gene, CHI11, was introduced into taro calli using high-velocity particle bombardment with the plasmid, pBI121. The candidate transformant calli were screened by polymerase chain reaction (PCR) amplification using a primer for the CHI11 gene and three positive lines were obtained, a transformation efficiency rate of 1%. Transformation of calli were conducted using the Agrobacterium tumefaciens strain, EHA 105, and the plasmid, pCNL65. The plasmid carried the reporter gene, ß-glucoronidase (GUS) driven by the cauliflower mosaic virus (CaMV) 35S promoter. After 1 week of selection on antibiotic plates, 80 calli samples showed a strong blue color upon exposure to 5-bromo-4-chloro-3-indolyl-beta-D-glucurononide, or a transformation efficiency rate of 90%. Also, transformation of shoots of cvs. Bun Long and Maui Lehua were conducted using Agrobacterium strain EHA105. After 1 week of selection on antibiotic plates, approximately 50% of the shoot samples stained positive for GUS activity.