1. In Vitro Regeneration and Genetic Transformation of Cucumis metuliferus through Cotyledon Organogenesis
- Author
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Hsiu-Yin Ho, Chien-Hung Chung, Hsin-Mei Ku, Chia-Wei Lin, Fuh-Jyh Jan, Yu-Tsung Lin, Shi-Dong Yeh, and Mei-Hsiu Su
- Subjects
food.ingredient ,Somatic embryogenesis ,Agrobacterium ,fungi ,food and beverages ,Organogenesis ,Horticulture ,Biology ,biology.organism_classification ,food.food ,food ,Shoot ,Botany ,Primordium ,Cucumis metuliferus ,Cotyledon ,Explant culture - Abstract
This study was undertaken to establish for the first time an efficient regeneration and transformation system for Cucumis metuliferus line PI292190, which is the source of a well-defined resistant gene, Wmv, that provides resistance against Papaya ringspot virus type P (PRSV-P) and PRSV-W (formerly known as Watermelon mosaic virus 1, WMV-1). Different combinations of growth regulators were evaluated for the regeneration of cotyledon explants. Adventitious buds or shoot primordia were obtained within 3 to 4 weeks on regeneration medium. After shoot development, adventitious buds or shoot primordia were transferred to elongation medium for 3 to 4 weeks and these shoots were subcultured onto rooting medium for another 1 to 2 weeks. Under optimal culture conditions, a total of 7 to 10 weeks was necessary to obtain C. metuliferus plantlets from cotyledons. Furthermore, transgenic plants were successfully obtained using an Agrobacterium tumefaciens-mediated transformation method as shown by polymerase chain reaction analysis and histochemical β-glucuronidase (GUS) assay. A total of nine transgenic plants were developed from 360 cotyledon explants, giving a transformation frequency of 2.5%.
- Published
- 2011
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