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Hsuan Chen, Lan Xue, Tong Li and Ryan N. Contreras

Hibiscus syriacus is a woody shrub in the Malvaceae family that is common in landscapes due to its broad adaptability and variable ornamental characteristics. Interspecific hybridization has been used to improve Hibiscus by building novel floral traits, hybrid vigor, and hybrid infertility. A few interspecific hybrid Hibiscus cultivars (H. syriacus × H. paramutabilis), such as Lohengrin and Resi, are notable because of their vigorous vegetative growth, female infertility, and large flowers. However, little is known about the male fertility and breeding potential of these hybrid cultivars, which could increase flower size by backcrossing to H. syriacus. In this study, we estimated male fertility of the two hybrid cultivars by acetocarmine staining and in vivo pollination and assessed selection methods for floral traits, specifically flower size and petal number. A BC1F1 population of 294 individuals was developed by crossing hybrid cultivars Lohengrin or Resi with a variety of double-flowered H. syriacus cultivars. A negative correlation between petal number and petal area was detected by quantile regression, which is a method that circumvents the problem of simple linear regression, which violates statistical assumptions. Quantile regression was used to build simultaneous selection thresholds for different levels of required stringency. As expected, the female fertility of hybrid cultivars was extremely low or zero; however, the male fertility of hybrid cultivars was not reduced compared with H. syriacus cultivars. A negative linear correlation between the petal number and petal area of the BC1F1 individuals was observed. In addition, quantile regression was recommended to set a single selection threshold to be applied to the selection of two negatively correlated traits, which was more effective than independent selection of petal numbers and petal areas among progeny.

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Jason D. Lattier, Hsuan Chen and Ryan N. Contreras

Chromosome numbers are an important botanical character for multiple fields of plant sciences, from plant breeding and genetics to systematics and taxonomy. Accurate chromosome counts in root tips of woody plants are often limited by their small, friable roots with numerous, small chromosomes. Current hydrolysis and enzyme digestion techniques require handling of roots before the root squash. However, optimum chromosome spread occurs when the cell walls have degraded past the point of easy handling. Here, we present a new enzyme digestion protocol that is fast, efficient, and flexible. This protocol reduces handling of the roots allowing for long-duration enzyme digestion. Digestions are performed on a microscope slide, eliminating the need for handling digested cells with forceps or pipettes. To illustrate the flexibility of this method across woody plant taxa, we performed chromosome counts on five angiosperms and one gymnosperm. Ploidy levels included diploids, triploids, and tetraploids with chromosome numbers ranging from 2n = 16 to 2n = 80. The range of holoploid 2C genome sizes spanned 1.54–24.71 pg. This protocol will provide a useful technique for plant cytologists working with taxa that exhibit a wide range of genome size and ploidy levels.