Genetic diversity of Cosmos species revealed by RAPD and ISSR markers.

The genus Cosmos is native of America and is constituted by 34 species; 28 of them are endemic of Mexico. The cosmos are used as a nematicide, antimalarial, and antioxidative agent. The aim of this study was to estimate the genetic diversity among 7 cosmos species based on random amplified polymorphic DNA (RAPD) and inter-simple sequences repeats (ISSR) markers. With RAPD markers, the obtained polymorphism was 91.7 % and the genetic diversity was 0.33, whereas these values were 65.6%, and 0.22 from ISSR markers, respectively, indicating the presence of high genetic diversity among the Cosmos species that were analyzed. The unweighted pair group method with arithmetic mean dendrograms that were obtained with both markers were notably similar, revealing 2 clusters and indicating a clear genetic differentiation among the Cosmos species that were assessed. The first cluster comprised the species Cosmos sulphureus, Cosmos pacificus, and Cosmos diversifolius, while the second cluster included the species Cosmos purpureus, Cosmos crithmifolius, Cosmos bipinnatus, and Cosmos parviflorus. Besides this, the Cosmos species were clustered according to their collection sites. The Mantel test corroborates the correlation between the genetic distance and the geographic altitude of each Cosmos species. The results suggest that it is necessary to preserve the Cosmos species in their natural habitat in addition to the germoplasm collection for ex situ conservation.

Interspecific hybridization between Tigridia pavonia and T. augusta through ovary slice culture.

Tigridia pavonia is the most popular species in the Tigridia genus, and is currently marketed in Europe, Asia, and Australia as a landscape plant. Although it is native to Mexico, there are no breeding programs for it. In this study, we attempted to increase its flower color spectrum and growth habit by interspecific hybridization with T. augusta. Interspecific hybrids between T. pavonia and T. augusta were successfully obtained for the first time using the cut-style pollination and ovary slice culture techniques. On the contrary, no hybrids were obtained from a reciprocal cross. At three, four, and five days after pollination (DAP) ovaries were sliced and cultured on Murashige and Skoog medium without growth regulators and ammonium nitrate, but were supplemented with 6% sucrose, 50 mg/L yeast extract, and 0.25% Gelrite. After 80 days of culture initiation, the germination of only 10 embryos was observed in ovary slices cultured at three DAP. After transfer to identical fresh medium, six hybrid embryos developed into seedlings. All obtained hybrid seedlings were transplanted successfully to soil, and grew normally. The progenies investigated were identified as true hybrids based on randomly amplified polymorphic DNA analysis.

Efficient callus induction and plant regeneration from filaments with anther in lily (Lilium longiflorum Thunb.).

Bulb scale propagation makes it difficult to obtain a large number of bulblets from disease-free stocks in a short time. The establishment of improved micropropagation procedures by in vitro culture is therefore desirable. Easter lily (Lilium longiflorum Thunb.) filaments with and without anther were excised and cultured in vitro with different media and culture conditions. In cultures of filaments with anther, callus developed and led to bulb, shoot, and root formation, whereas in cultures of filaments lacking anther, callus development did not occur. Among the various media tested, the B5 medium combined with darkness and the N6 medium combined with darkness or light, both supplemented with 9% sucrose, proved to be superior. A total of 1260 plants were regenerated from callus, acclimatized under a mist, and transferred to the greenhouse with a 100% success rate. No morphological abnormalities were observed among plants regenerated from filament-derived callus and all plants displayed isozyme banding patterns identical to the original cultivar. Chromosome observations revealed that all callus-regenerated plantlets tested were diploid (2n=24). The results suggest that in vitro culture of filaments with anther can be cultured for mass propagation.