Erycina pusilla is considered a potential model organism for orchids, because of several advantageous features, such as short juvenile period, low chromosome number and all year round blooming. Two different chromosome numbers (2n = 10 and 2n = 12) are reported for E. pusilla, which suggests two cytotypes. To reveal the genome homology between these two cytotypes, we generated hybrids from the intercytotypic reciprocal crosses (2n = 10 9 2n = 12 and 2n = 12 9 2n = 10), and applied 4 0 ,6-diamidino-2-phenylindole staining, genomic in situ hybridization and flow cytometry for genomic and cytogenetic analysis. The parental genomes showed high similarity both in genomic composition and content. The hybrids displayed a chromosome number of 2n = 11 in mitotic cells. Moreover, five bivalents and one univalent were observed at meiotic metaphase I stage. We observed meiotic synaptic behavior and found homeologous pairing with unpaired loops between parental chromosome pairing segments. These results demonstrated that chromosome rearrangement events have occurred between parental cytotypes during evolution. This study also illustrated the genome homology and homeologous pairing at pachytene phase, indicating that the chromosome number variation of two cytotypes mainly resulted from chromosome rearrangements, not changes in genomic constitution.

Each species is characterized by a specific set of chromosomes, which is described as the chromosome portrait or karyotype. In general, such a karyotype is the same for all individuals in the population. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Here, we examined the karyotypes of the two cytotypes and found differences in arm ratios and heterochromatin patterns as well as in the presence of satellite chromosomes and in the number and location of rDNA and telomeric repeat sites. These differences are extensive and would have required multiple chromosome rearrangements to generate the differences between the two karyotypes. We also found that F1 hybrids between the parents with the two different chromosome numbers resulted in sterile offspring, in accordance with our previous findings. The combination of hybrid sterility and extensively rearranged chromosomes supports the hypothesis that these two reported cytotypes are, in fact, two different species.

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