Paternity determination of interspecific rhododendron hybrids by genomic in situ hybridization (GISH).
Genomic in situ hybridization (GISH) has been proved to be the most effective and accurate technique for confirmation of hybrid character. The objective of our study was to adapt and optimize a GISH protocol for identification of donor chromatin in hybrids obtained by interspecific crosses between five Rhododendron taxa (R. aureum, R. brachycarpum, R. catawbiense 'Catharine van Tol', R. catawbiense 'Nova Zembla', and R. yakushimanum 'Koichiro Wada'). Positive results were obtained only when we used mitotic chromosome spreads prepared from anthers. The best differentiation of maternal and paternal chromosomes in hybrid genomes was obtained when 50 ng of probe was applied together with blocking DNA at a concentration of 3.0 microg/microL. The results demonstrate that GISH is a practical tool for detection of alien genomes and analysis of the constitution of the chromosomes in rhododendron hybrids.
Czernicka
Małgorzata
M
Department of Genetics, Plant Breeding and Seed Science, Faculty of Horticulture, University of Agriculture in Krakow, Al. 29 Listopada 54, 31-425 Krakow, Poland. czernickam@ogr.ar.krakow.pl
Mścichowska
Anna
A
Klein
Maria
M
Muras
Piotr
P
Grzebelus
Ewa
E
D032461:Chromosomes, Plant / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D018745:Genome, Plant; D006824:Hybridization, Genetic; D017403:In Situ Hybridization / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D015203:Reproducibility of Results; D029793:Rhododendron / Q000145:classification / Q000235:genetics*; D013045:Species Specificity
D015342:DNA Probes; D018744:DNA, Plant