Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Patent
1996-06-05
1998-08-11
Fox, David T.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
800DIG52, 800DIG65, 435 691, 435 693, 435 694, 435 6951, 435 6952, 435 696, 435 697, 4351723, 4352522, 435418, A01H 500, C12N 1518, C12N 1524, C12N 1582
Patent
active
057929351
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to methods for transforming Musa species such as bananas or plantains using Agrobacterium tumefaciens.
2. Description of the Prior Art
Bananas and plantains are perennial giant herbs belonging to the Genus Musa; their fruits are the fourth most important food in the developing world. Approximately 10% of the world's production of bananas (over 9 million tons in 1990, at a value of over U.S. $4 billion) enters the export market to generate an important source of income for tropical and subtropical regions. In light of the fact that Musa spp. make such an enormous contribution to food security and also provide export revenue in developing countries, it is a remarkable paradox that these crops have never benefited from traditional crop breeding. Production around the world is entirely dependent on unimproved clones that were often collected from nature, domesticated and maintained by clonal propagation. In terms of important crops for which biotechnology offers the possibility for dramatic genetic improvement, there are few opportunities as open for novel approaches as Musa spp.
There is general agreement that edible bananas originated from the two species, M. accuminata and M. balbisiana. Edible bananas have three different ploidy levels: 2N=22, 3N=33 and 4N=44. They have been propagated vegetatively for hundreds of years, with somatic mutations providing variability. Triploids are the most numerous and widely utilized cultivars (including dessert bananas used in world export trade). Efforts to breed Musa using conventional methods are fraught with obstacles, including low fertility, levels of ploidy, and lack of genetic variability. Since almost all accepted cultivars are seedless, sterile, clonally-propagated plants, conventional breeding efforts must begin with unimproved material that has been poorly characterized for genetic traits. It would be a major advantage if it were to be possible to make genetic improvements in the currently accepted seedless cultivars that are in production as food crops; this manuscript describes a system for reproducible and rapid genetic transformation of Musa that will make this possible.
Although traditional breeding has been slow for Musa spp., application of the tools of plant tissue culture have been significant value to crop improvement. Embryo rescue, cell suspension cultures, and related cell culture techniques have been used in research activities to overcome limitations in crop breeding, with resultant selection of genetic variants with new phenotypes. Plant cells grown in unorganized (callus, cells and protoplasts) cultures undergo ubiquitous genetic change or somaclonal variation. While this genetic variability is useful in creating new germplasm, it is a distinct negative feature in the clonal propagation of desired cultivars for which genetic uniformity is necessary. This has led to development of shoot-tip culture protocols which are now widely in use in developed and developing countries for Musa multiplication. Characteristically, these micropropagation procedures require only short periods of exposure of the plant tissues to growth in an undifferentiated state, and have a very low level of somaclonal variation among progeny.
In the development of the transformation system described below, we have attempted to devise a system that would mimic the widely used shoot micropropagation procedures now in use commercially. Our goal was to make available a system that would be useful for targeted genetic modification of existing, valuable cultivars with the least possible probability of introducing unanticipated somaclonal variability.
Virulent strains of the gram negative soil bacterium Agrobacterium tumefaciens are known to infect dicotyledonous plants and certain monocotyledonous plants. The tumor-inducing agent in the A. tumefaciens is a plasmid that functions by transferring some of its DNA into its host plan's cells. This plasmid (the Ti plasmid), and the virulence of the various str
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Recomendations from the INIBAP Workshop on "Biotechnology for Banana and Plantain" San Jose, C
Arntzen Charles J.
May Gregory D.
Fox David T.
Remenick James
Texas A&M University
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