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BackgroundBulbous flowers such as lily and tulip (Liliaceae family) are monocot perennial herbs that are economically very important ornamental plants worldwide. However, there are hardly any genetic studies performed and genomic resources are lacking. To build genomic resources and develop tools to speed up the breeding in both crops, next generation sequencing was implemented. We sequenced and assembled transcriptomes of four lily and five tulip genotypes using 454 pyro-sequencing technology.ResultsSuccessfully, we developed the first set of 81,791 contigs with an average length of 514 bp for tulip, and enriched the very limited number of 3,329 available ESTs (Expressed Sequence Tags) for lily with 52,172 contigs with an average length of 555 bp. The contigs together with singletons covered on average 37% of lily and 39% of tulip estimated transcriptome. Mining lily and tulip sequence data for SSRs (Simple Sequence Repeats) showed that di-nucleotide repeats were twice more abundant in UTRs (UnTranslated Regions) compared to coding regions, while tri-nucleotide repeats were equally spread over coding and UTR regions. Two sets of single nucleotide polymorphism (SNP) markers suitable for high throughput genotyping were developed. In the first set, no SNPs flanking the target SNP (50 bp on either side) were allowed. In the second set, one SNP in the flanking regions was allowed, which resulted in a 2 to 3 fold increase in SNP marker numbers compared with the first set. Orthologous groups between the two flower bulbs: lily and tulip (12,017 groups) and among the three monocot species: lily, tulip, and rice (6,900 groups) were determined using OrthoMCL. Orthologous groups were screened for common SNP markers and EST-SSRs to study synteny between lily and tulip, which resulted in 113 common SNP markers and 292 common EST-SSR. Lily and tulip contigs generated were annotated and described according to Gene Ontology terminology.ConclusionsTwo transcriptome sets were built that are valuable resources for marker development, comparative genomic studies and candidate gene approaches. Next generation sequencing of leaf transcriptome is very effective; however, deeper sequencing and using more tissues and stages is advisable for extended comparative studies.

Interspecific hybridization of Lilium longiflorum (L) with Asiatic (A) lily hybrids results in so-called LA-hybrids. Some of these hybrids produce 2n-pollen, which were used to perform crosses on Asiatic and Oriental (O) hybrids, resulting in ALA- and OLA-hybrids. Recombination between homoeologous chromosomes (introgression) and the mechanism of 2n-pollen formation in these hybrids were studied using genomic in situ hybridization (GISH). A clear differentiation between the chromosomes of L. longiflorum, Asiatic, and Oriental hybrids was observed in four ALA- and one OLA-hybrid using GISH. Two ALA-hybrids showed 3 and 5 recombinant chromosomes with a total of 5 and 10 crossover sites per hybrid, respectively. These occurred at random positions on the chromosomes. The number and the location of the rDNA-sites were determined using in situ hybridization and provided a tool, the FISH-marker, for identifying the NOR-bearing chromosomes in the lily hybrids. Evidence for the occurrence of the FDR-mechanism (first division restitution) of 2n-pollen formation in the LA-hybrids was obtained on the basis of absence of homologous chromosomes of L. longiflorum in the ALA- and OLA-hybrids.Key words: Lilium longiflorum, introgression, FDR, interspecific hybridization, FISH.

Sixteen Oriental and 12 Asiatic cultivars were crossed in 158 different combinations. A total of 708 F1 hybrids were obtained from 86 of the different combinations of 15 Oriental and 11 Asiatic cultivars. Because the Lilium cultivars (2n=2x=24) used for the production of these hybrids belong to two different taxonomic sections-Archelirion (0) and Sinomartagon (A), respectively-the F1 hybrids (OA) could be obtained only through embryo, embryo sac rescue, ovary slice or ovule culture. Most of the F, hybrids were highly sterile (did not produce viable n gametes) due to the failure of chromosome pairing. However, in a few cases F1 plants were found that produced viable 2n pollen at variable frequencies. These 2n pollen grains were successfully used for the production of backcross progenies. Using genomic in situ hybridization we found intergenomic recombinant chromosomes in the sexual polyploid progenies. These results indicate that there are effective prospects for combining important horticultural traits from the two main groups of cultivars of lilies through sexual polyploidization.