Breeding Aspects of Selected Ornamental Bulbous Crops

This article provides an overview of the origin, genetic diversity and methods and trends in breeding of selected ornamental geophytes (Lilium, Tulipa, Narcissus and Hippeastrum). The role of interspecific hybridisation and polyploidisation in assortment development is reviewed. A great variety of cultivars with traits of interest have been generated over the last century by using classical breeding. Geophyte breeders have been interested in a diversity of traits, including resistance to diseases, flower colour and shape, long lasting flowering and a long vase life. Shortening the long breeding process of many geophytes by reducing the juvenile phase and using in vitro techniques are reviewed. Currently, the breeding process has been enhanced by using modern molecular cytogenetic techniques. Genomic in situ hybridisation is frequently used, among other techniques, for genome differentiation in interspecific hybrids, and for assessment of the extent of intergenomic recombination in backcross progenies. Furthermore, several molecular marker techniques are used for verification of hybrid status, identification of genetic diversity, confirmation of the genetic fidelity of in vitro propagated plants and construction of high-density linkage maps. Recently, a myriad of new plant breeding technologies, such as cisgenetics and genome editing technologies have been used to improve the traits of ornamental geophytes, an endeavour that is discussed here. Breeding trends, cultivar novelties as well a new cultivars registered by international authorities during the last five years are presented in detail.

Euchromatic Supernumerary Chromosomal Segments—Remnants of Ongoing Karyotype Restructuring in the Prospero autumnale Complex?

Supernumerary chromosomal segments (SCSs) represent additional chromosomal material that, unlike B chromosomes, is attached to the standard chromosome complement. The Prospero autumnale complex (Hyacinthaceae) is polymorphic for euchromatic large terminal SCSs located on the short arm of chromosome 1 in diploid cytotypes AA and B7B7, and tetraploid AAB7B7 and B6B6B7B7, in addition to on the short arm of chromosome 4 in polyploid B7B7B7B7 and B7B7B7B7B7B7 cytotypes. The genomic composition and evolutionary relationships among these SCSs have been assessed using fluorescence in situ hybridisation (FISH) with 5S and 35S ribosomal DNAs (rDNAs), satellite DNA PaB6, and a vertebrate-type telomeric repeat TTAGGG. Neither of the rDNA repeats were detected in SCSs, but most contained PaB6 and telomeric repeats, although these never spanned whole SCSs. Genomic in situ hybridisation (GISH) using A, B6, and B7 diploid genomic parental DNAs as probes revealed the consistently higher genomic affinity of SCSs in diploid hybrid B6B7 and allopolyploids AAB7B7 and B6B6B7B7 to genomic DNA of the B7 diploid cytotype. GISH results suggest a possible early origin of SCSs, especially that on chromosome 1, as by-products of the extensive genome restructuring within a putative ancestral P. autumnale B7 genome, predating the complex diversification at the diploid level and perhaps linked to B-chromosome evolution.

Genome differentiation, natural hybridisation and taxonomic relationships among Eleocharis viridans, E. niederleinii and E. ramboana (Cyperaceae)

The role of natural hybridisation and genome changes in the differentiation and speciation of Eleocharis (Cyperaceae) was addressed through the study of the following three closely related species of the polyphyletic series Tenuissimae: Eleocharis viridans Kük. ex Osten, E. ramboana R.Trevis & Boldrini and E. niederleinii Boech., which often reproduce asexually. Molecular and cytogenetic data were used to understand the genomic and karyotypic relationships in the group. Genomes were compared using internal transcribed spacer–cleaved amplified polymorphic sequence (ITS-CAPS) marker and confirmed with random amplified polymorphic DNA, which allowed identification of different genetic groups, with clear evidence of natural hybrids. Karyotype analysis showed numerical variation from 2n = 20–42, with occurrence of chromosome heteromorphisms and polymorphisms, including variability in 35S rDNA site numbers. Meiotic studies demonstrated irregular pairing in some samples, which is associated with hybridisation and asexual reproduction. Genomic in situ hybridisation (GISH) reactions were conducted using two well defined genetic groups as probes, with 2n = 20 and normal meiosis. Probes were tested against each one of the genetic groups and showed positive, partial and negative GISH results, which supported the molecular analysis data. The results indicated that the three studied species are undergoing an intense process of genomic and karyotypic re-arrangement, which results in overlapping of morphological and genomic characteristics. The present study has exemplified the value of an integrative taxonomic approach to solve conflicts in species delimitation in groups undergoing hybridisation.

Introgression of yellow flower colour in Buddleja davidii by means of polyploidisation and interspecific hybridisation

To introduce yellow colour in the commercial Buddleja davidii (2n = 4x = 76) assortment, an interspecific breeding programme with B. globosa (2n = 2x = 38) was started. The first step was to perform chromosome doubling in B. globosa. Two of the obtained tetraploid B. globosa plants were subsequently used as male parent in interspecific crosses with the white flowering B. davidii cv. Nanhoensis Alba. In total 182 interspecific crosses were made and 18 F1 hybrids were obtained. Genome size measurements, chromosome counts and genomic in situ hybridisation (GISH) analysis proved the hybrid nature of most of the F1 hybrids. Plant morphology also expressed hybrid characteristics. F1 seedlings inherited the yellowish flower colour from B. globosa. As for many other woody ornamentals, the creation of hybrids through interspecific hybridisation along with polyploidisation offers new opportunities for breeding in Buddleja.

Hybridism in the Kunzea ericoides complex (Myrtaceae): an analysis of artificial crosses

Observations of wild plants and herbarium specimens suggest that hybridism is a feature of the Australasian Kunzea ericoides (Myrtaceae) complex. In this study 73 artificial cross combinations were attempted with New Zealand material of Leptospermum scoparium, five Kunzea species, two varieties and six informally recognised entities within the New Zealand K. ericoides complex. The results of these crosses are documented, and for five hybrids spanning the intergeneric, interspecific and intraspecific crosses attempted, we provide a more detailed assessment based on morphology, molecular (nrDNA and chloroplast) sequence variation, and genomic in situ hybridisation (GISH). This is the first time GISH has been used in the Myrtaceae. Hybrids were easily generated between the New Zealand members of the K. ericoides complex, but not between them and the Australian K. ericoides complex. We were unable to produce hybrids between the New Zealand K. ericoides complex and two more distantly related Australian species, K. baxteri and K. parvifolia. Intergeneric crosses between New Zealand plants of Leptospermum scoparium, Kunzea sinclairii and an informally recognised variant K. aff. ericoides (b) were successfully produced, but failed to flower. The molecular evidence and observations after GISH show that even when low levels of sequence divergence exist, genome differentiation, to different extents, can be observed. The results confirm some suspected New Zealand hybrid complexes. However, while hybrids were easily generated artificially, natural instances of hybridism appear to be largely confined to those habitats significantly disturbed since European settlement of both countries.

Interspecific chromosomal rearrangements in monosomic addition lines of Allium

Monosomic alien addition lines (MAALs) are useful for assigning linkage groups to chromosomes. We examined whether the chromosomal rearrangements following the introduction of a single onion (Allium cepa) chromosome into the Allium fistulosum genome were produced by homeologous crossing over or by a nonreciprocal conversion event. Among the monosomic lines available, 17 were studied by fluorescent genomic in situ hybridisation, using total A. cepa genomic DNA as the probe and total A. fistulosum genomic DNA as the competitor. In this way, rearrangements such as chromosomal translocations between A. cepa and A. fistulosum were identified as terminal regions consisting of tandem DNA repeats. Homeologous crossing over between the two closely related genomes occurred in 4 of the 17 lines, suggesting that such events are not rare. On the basis of a detailed molecular cytogenetic characterisation, we identified true monosomic alien addition lines for A. cepa chromosomes 3, 4, 5, 7, and 8 that can reliably be used in genetic studies.Key words: chromatin transfer, genomic in situ hybridisation, GISH, monosomic alien addition lines, MAALs, Allium.

Chromosome structure of Triticum timopheevii relative to T. turgidum

The chromosome structure of four different wild populations and a cultivated line of Triticum timopheevii (2n = 28, AtAtGG) relative to Triticum turgidum (2n = 28, AABB) was studied, using genomic in situ hybridisation (GISH) and C-banding analysis of meiotic configurations in interspecific hybrids. Two wild accessions and the cultivated line showed the standard C-banding karyotype. The other two accessions are homozygous for translocation 5At/3G and translocations 1G/2G and 5G/6G. GISH analysis revealed that all the T. timopheevii accessions carry intergenome translocations 6At/1G and 1G/4G and identified the position of the breakpoint in translocation 5At/3G. C-banding analysis of pairing at metaphase I in the hybrids with T. turgidum provides evidence that four species-specific translocations (6AtS/1GS, 1GS/4GS, 4GS/4AtL, and 4AtL/3AtL) exist in T. timopheevii, and that T. timopheevii and T. turgidum differ in the pericentric inversion of chromosome 4A. Bridge plus acentric fragment configurations involving 4AL and 4AtL were identified in cells at anaphase I. This result suggests that the paracentric inversion of 4AL from T. turgidum does not exist in T. timopheevii. Both tetraploid species have undergone independent and distinct evolutionary chromosomal rearrangements. The position, intercalary or subdistal, of the breakpoints in species-specific translocations and inversions contrasts with the position, at or close to the centromere, of intraspecific translocations. Different mechanisms for intraspecific and species-specific chromosome rearrangements are suggested.Key words: Triticum timopheevii, chromosome pairing, translocation, evolution, C-banding, GISH.