Early Identification of Intra- and Intergeneric Hybrids among Colchicaceous Ornamentals, <i>Gloriosa</i> spp., <i>Littonia modesta</i> Hook. and <i>Sandersonia aurantiaca</i> Hook., by Flow Cytometry and Random Amplified Polymorphic DNA Analyses

Abstract

Plantlets derived from ovule culture following intra-or intergeneric crosses among Colchicaceous ornamentals, Gloriosa spp., Littonia modesta Hook. and Sandersonia aurantiaca Hook., were subjected to flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses in order to verify their hybridity. For crosses between plants with apparently distinct relative fluorescence intensity (RFI) of nuclei, i.e., intrageneric crosses between Gloriosa genotypes with different ploidy levels, and most intergeneric crosses, detection of hybrid plantlets could be readily accomplished by FCM analysis. The results of hybrid identification by RAPD analysis supported those of FCM analysis. In addition, RAPD analysis allowed the verification of the hybridity of intra-or intergeneric cross-derived plantlets, which could not be identified as hybrids by FCM analysis due to the similarity of RFI of the parents or appearance of the RFI peak in an unexpected position. Totally, 110 independent hybrid plantlets (60 intrageneric and 50 intergeneric hybrids) have so far been identified by FCM and/or RAPD analyses. Thus, FCM in combination with RAPD analyses offer simple and rapid means for the early detection of intrageneric and intergeneric hybrids in Colchicaceous ornamentals.

Cross-compatibility in Interspecific Hybridization of Lychnis (Caryophyllaceae) and Characterization of Interspecific Hybrids between L. fulgens and L. sieboldii

“…Isolation of total genomic DNA from leaves and RAPD analysis were performed according to Amano et al (2007) and Godo et al (2009). For RAPD analysis, five DNA oligomer (12) sets, C3, C43, G4, G5, and G64 (Wako Pure Chemical Industries, Ltd., Osaka, Japan), were used.…”

Identification of Polyploidy of in vitro Anther-derived Shoots of Asparagus officinalis L. by Flow Cytometric Analysis and Measurement of Stomatal Length

“…Recently, a rapid and exact method to determine nuclear DNA contents by flow cytometry (FCM) has been developed for various plant species, such as cloudberry (Thiem and Sliwinska, 2003), hop (Koutoulis et al, 2005), lily (Nakano et al, 2006), Gloriosa spp. (Amano et al, 2007), and Diospyros kaki Thunb. (Yamada and Tao, 2007).…”

Interspecific hybridization between triploid Senno (Lychnis senno Siebold et Zucc., Caryophyllaceae) and allied taxa of the genus Lychnis

“…Isolation of total genomic DNA from leaves and RAPD analysis were performed according to Amano et al (2007). For RAPD analysis, three sets of DNA Oligomer (12) Set, C43, G64, and G81 (Wako Pure Chemical Industries, Ltd., Osaka, Japan) were tested.…”

“…Embryo, ovule, and immature seed cultures have been successfully applied to produce interspecific or intergeneric hybrids in various ornamental plants such as Lilium species (Asano 1978(Asano , 1980, Cyclamen species (Ishizaka 1997), Primula species (Kato and Mii 2000;Amano et al 2006;Hayashi et al 2007a, b), Cosmos species (Oku et al 2008), and Colchicaceous plants (Amano et al 2007). Although all the Japanese Senno strains previously found are triploid, they had relatively high pollen fertility (about 70%) as assessed with acetocarmine staining and sometimes produced a few seeds by artificial self-pollination (Godo et al 2004b).…”