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Data/hora: 08/06/2017 22:23:16
Biblioteca(s): Embrapa Mandioca e Fruticultura.
Data corrente: 15/04/2004
Data da última atualização: 15/04/2004
Autoria: MBA, R.; TOHME, J.; ROCA, V.; FREGENE, M.
Título: Lowering the cost of biotechnologies for national cassava programs: microsatellite
markers to facilitate use of the cassava molecular genetic map and provide new genetic
information.
Ano de publicação: 1998
Fonte/Imprenta: Revista Brasileira de Mandioca, Cruz das Almas, v. 17, p. 25, nov., 1998. Suplemento.
Idioma: Inglês
Conteúdo: Abstract: The applicabillity of the majority of molecular markers, currently available on
the molecular genetic map of cassava, in marker-assisted studies of complex traits and
marker-assisted improvement of cassava has been questioned by collaborators in the
NARs considering the low technology status of most cassava breeding program. The
speed and low cost of PCR markers. with high levels of allelic diversity. easy to assay.
and codominant make them the markers of choice for transfer of gene tags for important
cassava traits to collaborators in the National Programs. PCR technology requires a
relatively small capital out-lay for its integration into day-to-day plant breeding
operations and it eliminates labor costs involved in the tedious Southern analysis of RFLP
markers. More microsatellite markers can be assayed for a large number of plants, in a
given time, for a given budgest, compared to RFLP markers. A DNA library, enriched for
microsatellite markers. was developed by the affinity capture method and cloned into
pUC 18 plasmid. The library was transformed into DH 10B competent cells. A total of
18.000 colonies from the enriched library was screened for microsatellite markers by
hybridization with microsatellite oligonucleotides and by anchored primer PCR. A total
of 120 positive clones were identified and sequenced on an AB1377 automated
sequencer. Primers. 20-mers long. were designed from regions flanking microsatellite
markers and were used in PCR amplification of the parental genotype of the mapping
population. Polymorphic microsatellite markers were scored in the progeny and mapped
onto the existing map of cassava using the linkage analysis software MAPMAKER 3.0
Results reveal a rather low efficiency of enrichment using the affinitv capture method,
though different di and tri-nucleotide microsatellite markers could be obtained.
Approximately 200 mapped microsatellite markers is the targest of this ongoing project.
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