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.

MicroRNA and Putative Target Discoveries in Chrysanthemum Polyploidy Breeding

MicroRNAs (miRNAs), around 22 nucleotides (nt) in length, are a class of endogenous and noncoding RNA molecule that play an essential role in plant development, either by suppressing the transcription of target genes at a transcriptional level or inhibiting translation at a posttranscriptional level. To understand the roles of miRNAs and their target genes in chrysanthemum polyploidy breeding, three sRNA libraries of normal and abnormal embryos after hybridization were performed by RNA-Seq. As a result, a total of 170 miRNAs were identified and there are 41 special miRNAs in cross of paternal chromosome doubling, such as miR169b, miR440, and miR528-5p. miR164c and miR159a were highly expressed in a normal embryo at 18 days after pollination, suggesting the regulatory role at the late stage of embryonic development. miR172c was only detected in the normal embryo at 18 days after pollination, which means that miR172c mainly mediates gene expression in postembryonic development and these genes may promote embryo maturation. Other miRNAs, including miR414, miR2661, and miR5021, may regulate the genes participated in pathways of auxin response and energy metabolism; then they regulate the complex embryonic development together.