Gypsophila is a genus of about 100 species of flowering plants in the family Caryophyllaceae, native to Europe, Asia and North Africa. The flowers, often used to fill flower bouquets, are characteristic, small in diameter (3-10 mm) with five white or pink petals and produced in large inflorescences. The most commonly cultivated species is G. paniculata. The identification of cultivars and accessions using molecular markers is a crucial aim of modern horticulture, with applications in breeding programs and in germplasm collection management. In this work we carried out the analysis of the genetic variability of different Gypsophila cultivars with the aim to develop a panel of molecular markers useful in breeding programs. Up to now, with the exception of RAPD characterization (Rady, 2006. Biol Plantarum, 50 (4): 507-513) no molecular data are available despite the economic importance of the species. To better analyze the degree of variability among cultivated varieties, the molecular markers used in this work were ISSR (Inter Simple Sequences Repeats) and TRAP (Targeted Region Amplified Polymorphism) markers, which generate a rich pattern of fragments. ISSRs were visualized on agarose gels, while TRAPs (Hu & Vick, 2003 Pl. Mol. Biol. Reporter 21:289–294) based on expressed sequence information to generate polymorphic markers around targeted candidate genome regions, were analyzed by an automated fluorescent Sequencer (ABI PRISM 310). We designed primers anchored on interstitial telomers. The PCR amplifications were based on a fixed (targeted) primer and on an arbitrary labelled primer. Both the markers used were capable to evidence polymorphisms among the 12 commercial accessions examined. In particular the two methodologies gave rise to a 0-1 patterns of data processed by means of NtSys software (Rohlf, F.J. 2002. NTSYS pc: Numerical Taxonomy System, Version 2.1. Exeter Publishing, Setauket, NY.) to obtain UPGMA dendrograms with SM similarity coefficient determined using the binary matrix. Results separated the cultivars in two main clusters in accordance to breeders information. In particular the cultivars of the Perfecta group were placed together in the same cluster. Further analysis on species and varieties will be necessary to confirm these preliminary results in order to clarify the cultivars genetic relationships and to plane a molecular markers assisted breeding program.
MOLECULAR CHARACTERIZATION OF GYPSOPHILA CULTIVARS WITH ISSR AND TRAP MARKERS / INTRIERI M.C.; CALISTRI E.; NIEDDU F.; PASQUALETTO P.; BUIATTI M.; BOGANI P.. - ELETTRONICO. - (2010), pp. 1-1. (Intervento presentato al convegno 54th Italian Society of Agricultural Genetics Annual Congress tenutosi a Matera, Italy nel 27/30 September).
MOLECULAR CHARACTERIZATION OF GYPSOPHILA CULTIVARS WITH ISSR AND TRAP MARKERS
INTRIERI, MARIA CARMELA;BUIATTI, MARCELLO;BOGANI, PATRIZIA
2010
Abstract
Gypsophila is a genus of about 100 species of flowering plants in the family Caryophyllaceae, native to Europe, Asia and North Africa. The flowers, often used to fill flower bouquets, are characteristic, small in diameter (3-10 mm) with five white or pink petals and produced in large inflorescences. The most commonly cultivated species is G. paniculata. The identification of cultivars and accessions using molecular markers is a crucial aim of modern horticulture, with applications in breeding programs and in germplasm collection management. In this work we carried out the analysis of the genetic variability of different Gypsophila cultivars with the aim to develop a panel of molecular markers useful in breeding programs. Up to now, with the exception of RAPD characterization (Rady, 2006. Biol Plantarum, 50 (4): 507-513) no molecular data are available despite the economic importance of the species. To better analyze the degree of variability among cultivated varieties, the molecular markers used in this work were ISSR (Inter Simple Sequences Repeats) and TRAP (Targeted Region Amplified Polymorphism) markers, which generate a rich pattern of fragments. ISSRs were visualized on agarose gels, while TRAPs (Hu & Vick, 2003 Pl. Mol. Biol. Reporter 21:289–294) based on expressed sequence information to generate polymorphic markers around targeted candidate genome regions, were analyzed by an automated fluorescent Sequencer (ABI PRISM 310). We designed primers anchored on interstitial telomers. The PCR amplifications were based on a fixed (targeted) primer and on an arbitrary labelled primer. Both the markers used were capable to evidence polymorphisms among the 12 commercial accessions examined. In particular the two methodologies gave rise to a 0-1 patterns of data processed by means of NtSys software (Rohlf, F.J. 2002. NTSYS pc: Numerical Taxonomy System, Version 2.1. Exeter Publishing, Setauket, NY.) to obtain UPGMA dendrograms with SM similarity coefficient determined using the binary matrix. Results separated the cultivars in two main clusters in accordance to breeders information. In particular the cultivars of the Perfecta group were placed together in the same cluster. Further analysis on species and varieties will be necessary to confirm these preliminary results in order to clarify the cultivars genetic relationships and to plane a molecular markers assisted breeding program.
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