PAG-II Plant Genome II Conference

Town & Country Conference Center, San Diego, CA, January, 1994.


PG-II: RAPD DETECTION AND CHARACTERIZATION OF A DNA POLYMORPHISM ASSOCIATED WITH DWARF BANANA SOMACLONAL VARIANTS

RAPD DETECTION AND CHARACTERIZATION OF A DNA POLYMORPHISM ASSOCIATED WITH DWARF BANANA SOMACLONAL VARIANTS

Olivia P. Damasco 1, Ian D. Godwin 1, Robert J. Henry 2, Stephen W. Adkins 1 and Mike K. Smith 3, 1 Department of Agriculture, and 2 Qld Agricultural Biotechnology Centre, The University of Queensland, QLD 4072, 3 DPI Maroochy Horticultural Research Centre, Nambour, QLD 4560, Australia.


Meristem culture is an ideal source of disease-free planting material in vegetatively reproduced species such as banana. In many instances however, somaclonal variation occurs, with resultant plants having deleterious phenotypes, most frequent of which is dwarfism. Dwarfs are rarely detected in vitro, and are commonly not apparent until field planting has taken place. Our experiments demonstrate that the dwarfs have a reduced sensitivity to GA3. We have applied RAPD marker techniques to assist with early detection of dwarfism. "Normal" and dwarf in vitro plants of cv. New Guinea Cavendish were screened with 66 random decamer primers, of which 19 revealed polymorphisms. These 19 primers were used to screen a range of known "normal" and dwarf plants growing in vitro, in the glasshouse and the field, all representing different somaclonal variation events. In all, total genomic DNA of 80 plants of two cultivars was analysed, including cv. New Guinea Cavendish (18 "normal", 21 dwarf) and cv. Williams (23 "normal", 18 dwarf). One primer, OPJ-04 (Operon Tech., 5'-CCGAACACGG-3') consistently amplified a band of approximately 1.5 kb in "normal" plants which was absent in all dwarfs tested. This primer was then shown to consistently amplify a band of similar size in "normal" chloroplast DNA which was always absent in dwarf chloroplast DNA. This is preliminary evidence that at least one component of this dwarf-ism may be controlled by a highly mutable locus in the chloroplast genome. Sequencing of the amplified fragment is in progress to characterise the putative chloroplast DNA.


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