Morphological Characterization of Tetraploids of Limonium sinuatum (L.) Mill. Produced by Oryzalin Treatment of Seeds

Limonium sinuatum (L.) Mill. (2n = 2x = 16) is a popular ornamental plant with dimorphism of pollen grains (type A and type B) and stigmas (papilla and cob-like). We applied polyploidy breeding to this species in order to introduce desirable traits. Tetraploid and mixoploid L. sinuatum plants were successfully obtained with oryzalin treatment of L. sinuatum ‘Early Blue’ seeds. All three tetraploids had increased leaf width, stomatal size, flower length, and pollen width compared to those of the diploid, and tetraploids had four germinal pores of pollen grains, whereas the diploid had three. All tetraploids had type A pollen grains and cob-like stigmas. Furthermore, the growth of cultivated tetraploid plants was slow, with later bolting and flowering times. Mixoploids Mixo-1 and Mixo-3 were estimated to be polyploidy periclinal chimeric plants consisting of a tetraploid L1 layer and diploid L2 layer, and Mixo-2 was estimated to be a polyploidy periclinal chimeric plant consisting of the diploid L1 layer and tetraploid L2 layer. Mixo-4 had tetraploid L1 and L2 layers. Mixoploids, except Mixo-4, had type A pollen grains and cob-like stigmas, whereas Mixo-4 had type B pollen grains and papilla stigmas. These polyploids will be useful as polyploidy breeding materials.

Induction of Tetraploids from Petiole Explants through Colchicine Treatments inEchinacea purpureaL.

Petiole explants were obtained from in vitro grown diploid (2x=22)Echinacea purpureaplantlets. Shoots were regenerated by culturing the explants on MS basal medium containing 0.3 mg/L benzyladenine (BA), 0.01 mg/L naphthaleneacetic acid (NAA) and four concentrations (30, 60, 120, and 240 mg/L) of colchicine for 30 days, or 120 mg/L of colchicine for various durations (7, 14, 21, and 28 days). The regenerated shoots were induced to root on MS basal medium with 0.01 mg/L NAA, and then the root-tips of the regenerated shoots were sampled for count of chromosome number. It was found that a treatment duration of >7 days was necessary for induction of tetraploid (4x=44) shoots, and treatment with 120 mg/L colchicine for 28 days was the most efficient for induction of tetraploids, yielding 23.5% of tetraploids among all the regenerated shoots. Chimeras were observed in almost all the treatments. However, the ratio of tetraploid to diploid cells in a chimeric plant was usually low. In comparison with diploid plants, tetraploid plants in vitro had larger stomata and thicker roots with more root branches, and had prominently shorter inflorescence stalk when mature.