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Stable transformation of papaya via microprojectile bombardment

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Summary

Stable transformation of papaya (Carica papaya L.) has been achieved following DNA delivery via high velocity microprojectiles. Three types of embryogenic tissues, including immature zygotic embryos, freshly explanted hypocotyl sections, and somatic embryos derived from both, were bombarded with tungsten particles carrying chimeric NPTII and GUS genes. All tissue types were cultured prior to and following bombardment on half-strength MS medium supplemented with 10 mg 1−1 2,4-D, 400 mg 1−1 glutamine, and 6% sucrose. Upon transfer to 2,4-D-free medium containing 150 mg 1−1 kanamycin sulfate, ten putative transgenic isolates produced somatic embryos and five regenerated leafy shoots. Leafy shoots were produced six to nine months following bombardment. Tissues from 13 of these isolates were assayed for NPTII activity, and 10 were positive. Six out of 15 isolates assayed for GUS expression were positive. Three isolates were positive for both NPTII and GUS,

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Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

GUS:

β -glucuronidase

X-gluc:

5-Br-4-Cl-3-indolyl-β-D-glucuronic acid

CaMV:

cauliflower mosaic virus

NOS:

nopaline synthase

NPTII:

neomycin phosphotransferase II

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Communicated by C. Quiros

Journal Series no. 3448 of the Hawaii Institute of Tropical Agriculture and Human Resources

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Fitch, M.M.M., Manshardt, R.M., Gonsalves, D. et al. Stable transformation of papaya via microprojectile bombardment. Plant Cell Reports 9, 189–194 (1990). https://doi.org/10.1007/BF00232177

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  • DOI: https://doi.org/10.1007/BF00232177

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