Prins and C-PRINS: Promising tools for the physical mapping of the lupin genome

Abstract

Two molecular cytogenetics methods, PRINS (primed in situ DNA labeling) and C-PRINS (cycling PRINS), were optimized for the physical mapping of several types of DNA sequences on the mitotic chromosomes of the narrow-leafed lupin (Lupinus angustifolius L.). The fragment of the FokI element from Vicia faba was localised by indirect PRINS reaction. Two other sequences, fragments of the coding sequences of L. luteus and of L. angustifolius, were localised by indirect C-PRINS. These techniques are faster and more sensitive than FISH, and they allowed the mapping of short DNA fragments. The data obtained shows that both types of PRINS are valuable tools for chromosome identification in lupin.

Karyotyping of the narrow-leafed lupin (Lupinus angustifolius L.) by using FISH, PRINS and computer measurements of chromosomes

“…In contrast, several cytogenetic markers from PRINS (primed in situ DNA labelling) might be of use for L. angustifolius karyotyping. Oligonucleotide sequences (fragment of the FokI repeat element from Vicia faba, sequences from L. luteus and from L. angustifolius) have been localised by our group in chromosomes by PRINS and its variant C-PRINS, allowing the identification of several chromosome pairs (Kaczmarek et al 2007).…”

Physical localization of molecular markers and assignment of the 15th linkage group to chromosome 11 of the karyotype in cassava (Manihot esculenta Crantz) by primed in situ labeling

“…PRINS can be used to detect relatively small chromosomal regions, DNA sequences, and molecular marker loci that cannot be observed using the FISH technique (Zhu et al, 1995;Shi et al, 1996;Macas et al, 2000;Kubaláková et al, 2001;Tatum and Rayburn, 2006;Kaczmarek et al, 2007). In this study, we demonstrated the feasibility of determining the physical location of molecular markers and assigning a linkage group to an individual chromosome of the karyotype in cassava using 2 markers by PRINS.…”

“…Fluorescence in situ hybridization (FISH) has been conventionally used to integrate genetic and chromosomal maps in several plant species (Jiang et al, 1995;Dong et al, 2000;Sadder et al, 2000;Cheng et al, 2001a,b;Kulikova et al, 2001;Howell et al, 2002;Islam-Faridi et al, 2002;Kim et al, 2002;Pedrosa et al, 2002;Koumbaris and Bass, 2003;Zhang et al, 2005;Wang et al, 2008). Because of the combination of accuracy and sensitivity of polymerase chain reaction (PCR) with FISH, primed in situ labeling (PRINS) has been found to be a more powerful tool for localizing DNA sequences, single-copy genes (Macas et al, 2000;Kubaláková et al, 2001;Tatum and Rayburn, 2006;Kaczmarek et al, 2007;Gao et al, 2011), and restriction fragment length polymorphism loci (Zhu et al, 1995;Shi et al, 1996) in some plants (Birchler and Danilova, 2012).…”

The Development of a Primed <i>In Situ</i> Hybridization Technique for Chromosome Labeling in Cultivated Strawberry (<i>Fragaria</i>×<i>ananassa</i>)

“…Since then, the PRINS and C-PRINS techniques have been used to detect the localization of the DNA sequence on the chromosomes in various plants as summarized by Kubaláková et al (2001). In particular, some studies have detected the localization of single copy sequence in situ using C-PRINS (Zhu et al 1995, Mukai and Appels 1996, Kubaláková and Doležel 1998, Kaczmarek et al 2007, Tui and Roy 2008, Talia et al 2011. However, PRINS has not been used with the exception of the study by Abbo et al (1993b).…”