Abstract
The present study was an attempt to develop an in vitro colchicine chromosome doubling protocol to restore the fertility of an F1 interspecific hybrid in Lilium. Basal scale segments of Lilium × formolongi × Oriental hybrid (FO) bulblets were pre-cultured for three durations (6, 15 and 25 days) and soaked in three colchicine concentrations (0.00, 1.25 and 2.50 mM) for 18, 24 and 36 h. To separate mixoploids, three cycles of adventitious bud induction were performed. The ploidy levels of the surviving plantlets were detected by flow cytometry at 30–31 weeks after induction and were confirmed by chromosome counts. The results indicated that the pre-culture duration, colchicine concentration and exposure time all had significant impacts on the tetraploid induction rate. The preferred procedure was to pre-culture the segments for 6 days and then treat them with 1.25 mM colchicine for 24 h. The morphological traits of rosette leaves were significantly different between the tetraploid and diploid plants. The adult tetraploid plants had considerably longer and wider leaves, larger flowers, and delayed flowering time (8 days later) than did diploid plants. Pollen viability tests and backcross trials of FO hybrids demonstrated fertility restoration at the tetraploid level. This protocol provides a feasible method for inducing fertile tetraploid FO hybrids for further breeding.
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