Articles
AGROBACTERIUM-MEDIATED TRANSFORMATION OF GRAPEFRUIT (CITRUS PARADISI MACF.) WITH GENES FROM CITRUS TRISTEZA VIRUS
Article number
535_29
Pages
237 – 244
Language
Abstract
A protocol for the Agrobacterium-mediated transformation of epicotyl segments of etiolated Duncan grapefruit seedlings has been developed and will be described.
Transformation frequencies are low in terms of number of solidly transgenic shoots produced per number of epicotyl segments cultured.
However, selection is very stringent and relatively few shoots must be screened.
Furthermore, unlike previous species tested, regenerated transgenic grapefruit shoots root readily.
Transgenic plants have been produced that contain the coat protein (CP) gene and a portion of the gene encoding the RNA-dependent RNA polymerase from Florida severe citrus tristeza virus (CTV) isolate T36. Buds from these plants have been grafted onto T36-infected mandarin rootstock to determine whether the transgenic scions show resistance to the virus.
Experiments are also underway with other CTV constructs, including CP genes from Florida mild isolate T30 and Venezuelan stem pitting isolate B249, p27 (the diverged CP from T36), and p20, an expressed gene of T36 of unknown function.
Transformation frequencies are low in terms of number of solidly transgenic shoots produced per number of epicotyl segments cultured.
However, selection is very stringent and relatively few shoots must be screened.
Furthermore, unlike previous species tested, regenerated transgenic grapefruit shoots root readily.
Transgenic plants have been produced that contain the coat protein (CP) gene and a portion of the gene encoding the RNA-dependent RNA polymerase from Florida severe citrus tristeza virus (CTV) isolate T36. Buds from these plants have been grafted onto T36-infected mandarin rootstock to determine whether the transgenic scions show resistance to the virus.
Experiments are also underway with other CTV constructs, including CP genes from Florida mild isolate T30 and Venezuelan stem pitting isolate B249, p27 (the diverged CP from T36), and p20, an expressed gene of T36 of unknown function.
Authors
G.A. Moore, V.J. Febres, C.L. Niblett, D. Luth, M. McCaffery, S.M. Garnsey
Keywords
citrus, genetic transformation, Agrobacterium tumefaciens, plant improvement
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