INVESTIGADORES
TALIA Paola Monica
artículos
Título:
Detection of single copy sequences using BAC-FISH and C-PRINS techniques in sunflower chromosomes
Autor/es:
PAOLA TALIA; EDUARDO J. GREIZERSTEIN; H. ESTEBAN HOPP; NORMA PANIEGO; LIDIA POGGIO; RUTH A. HEINZ
Revista:
BIOCELL
Editorial:
INST HISTOL EMBRIOL-CONICET
Referencias:
Lugar: Mendoza; Año: 2011 vol. 35 p. 19 - 28
ISSN:
0327-9545
Resumen:
ABSTRACT: Bacterial artificial chromosome - fluorescence in situ hybridization (BAC-FISH) and cyclingprimed
in
situ
labeling (C-PRINS) techniques were
evaluated
for integration
of physical
and genetic maps of
sunflower
(Helianthus
annuus
annuus
L.). Single-site SSR markers
were
selected from three linkage groups
of a
high-density
sunflower
genetic map. This
selection was
based on previously
identified
QTL associated to S.
sclerotiorum.
sclerotiorum.
These
markers
were
used to select BACs
contaning single copy
sequences for BAC-FISH
aplication.
Blocking of highly
dispersed repetitive
sunflower
sequences reduced unspecific
hybridization,
and
allowed
the detection of specific
signals for BACs
containing SSR markers
HA4222 and HA2600, anchored
to LG 16 and LG 10, respectively.
Single-site FISH signal detection was
optimized by
adjusting the
relative
quantity and quality of unlabelled repetitive
sequences present in the blocking
DNA.
The
SSR marker
ORS1247
anchored to the LG 17 was
detected by
C-PRINS, which
yielded fluorescence signals that were
specific
and intense. This
progress
in localizing single-copy
sequences using BAC-FISH
and indirect CPRINS
strategies
in sunflower
will facilitate
the integration
of genetic and physical
maps, allowing
the identification
of chromosomes containing key
genes and/or QTL associated to agronomic
important
traits in
sunflower.Bacterial artificial chromosome - fluorescence in situ hybridization (BAC-FISH) and cyclingprimed
in
situ
labeling (C-PRINS) techniques were
evaluated
for integration
of physical
and genetic maps of
sunflower
(Helianthus
annuus
annuus
L.). Single-site SSR markers
were
selected from three linkage groups
of a
high-density
sunflower
genetic map. This
selection was
based on previously
identified
QTL associated to S.
sclerotiorum.
sclerotiorum.
These
markers
were
used to select BACs
contaning single copy
sequences for BAC-FISH
aplication.
Blocking of highly
dispersed repetitive
sunflower
sequences reduced unspecific
hybridization,
and
allowed
the detection of specific
signals for BACs
containing SSR markers
HA4222 and HA2600, anchored
to LG 16 and LG 10, respectively.
Single-site FISH signal detection was
optimized by
adjusting the
relative
quantity and quality of unlabelled repetitive
sequences present in the blocking
DNA.
The
SSR marker
ORS1247
anchored to the LG 17 was
detected by
C-PRINS, which
yielded fluorescence signals that were
specific
and intense. This
progress
in localizing single-copy
sequences using BAC-FISH
and indirect CPRINS
strategies
in sunflower
will facilitate
the integration
of genetic and physical
maps, allowing
the identification
of chromosomes containing key
genes and/or QTL associated to agronomic
important
traits in
sunflower.