PMID- 24244840
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20131118
LR  - 20211021
IS  - 2041-2851 (Print)
IS  - 2041-2851 (Electronic)
VI  - 5
DP  - 2013
TI  - Mapping pachytene chromosomes of coffee using a modified protocol for 
      fluorescence in situ hybridization.
PG  - plt040
LID - 10.1093/aobpla/plt040 [doi]
LID - plt040
AB  - Fluorescence in situ hybridization (FISH) is the most direct method for 
      physically mapping DNA sequences on chromosomes. Fluorescence in situ 
      hybridization mapping of meiotic chromosomes during the pachytene stage is an 
      important tool in plant cytogenetics, because it provides high-resolution 
      measurements of physical distances. Fluorescence in situ hybridization mapping of 
      coffee pachytene chromosomes offers significant advantages compared with FISH 
      mapping of somatic chromosomes, because pachytene chromosomes are 30 times longer 
      and provide additional cytological markers. However, the application of this 
      technique to pachytene chromosomes has been complicated by problems in making 
      preparations of meiotic chromosomes and by difficulties in the application of 
      standard FISH protocols. We have been able to overcome most of these obstacles in 
      applying the FISH technique to the pachytene chromosomes of coffee plants. 
      Digesting the external callose layer surrounding the pollen mother cells (PMCs) 
      in conjunction with other procedures permitted suitable pachytene chromosomes to 
      be obtained by increasing cell permeability, which allowed the probe sequences to 
      enter the cells. For the first time, hybridization signals were registered on 
      coffee pachytene chromosomes using the FISH technique with a repetitive sequence 
      as a probe. We obtained slides on which 80 % of the PMCs had hybridization 
      signals, resulting in FISH labelling with high efficiency. The procedure does not 
      seem to be dependent on the genotype, because hybridization signals were detected 
      in genetically different coffee plants. These findings enhance the possibilities 
      for high-resolution physical mapping of coffee chromosomes.
FAU - Iacia, Ana Amelia Sanchez
AU  - Iacia AA
AD  - Centro de Pesquisa e Desenvolvimento de Recursos Geneticos Vegetais , Instituto 
      Agronomico de Campinas , CxP. 028, Campinas, Sao Paulo 13012-970 , Brazil.
FAU - Pinto-Maglio, Cecilia A F
AU  - Pinto-Maglio CA
LA  - eng
PT  - Journal Article
DEP - 20131107
PL  - England
TA  - AoB Plants
JT  - AoB PLANTS
JID - 101539425
PMC - PMC3828664
OTO - NOTNLM
OT  - Coffea
OT  - FISH
OT  - meiotic chromosomes
OT  - molecular cytogenetics
OT  - physical mapping
OT  - rDNA.
EDAT- 2013/11/19 06:00
MHDA- 2013/11/19 06:01
PMCR- 2013/01/01
CRDT- 2013/11/19 06:00
PHST- 2013/03/11 00:00 [received]
PHST- 2013/08/12 00:00 [accepted]
PHST- 2013/11/19 06:00 [entrez]
PHST- 2013/11/19 06:00 [pubmed]
PHST- 2013/11/19 06:01 [medline]
PHST- 2013/01/01 00:00 [pmc-release]
AID - plt040 [pii]
AID - 10.1093/aobpla/plt040 [doi]
PST - ppublish
SO  - AoB Plants. 2013;5:plt040. doi: 10.1093/aobpla/plt040. Epub 2013 Nov 7.