Assessing Genetic Diversity and Population Structure of Kalmia latifolia L. in the Eastern United States: An Essential Step towards Breeding for Adaptability to Southeastern Environmental Conditions

Kalmia latifolia L. (mountain laurel), an attractive flowering shrub, is considered to be a high-value ornamental plant for the eastern United States. Limited information on the genetic diversity and structure of K. latifolia is available, which obstructs efficient germplasm utilization and breeding for adaptability to southeastern environmental conditions. In this study, the genetic diversity of 48 wild K. latifolia plants sampled from eight populations in the eastern U.S. was assessed using eight inter simple sequence repeat (ISSR) markers. A total of 116 bands were amplified, 90.52% of which (105) were polymorphic. A high level of genetic diversity at the species level was determined by Nei’s gene diversity (0.3089) and Shannon’s information index (0.4654), indicating that K. latifolia was able to adapt to environmental changes and thus was able to distribute over a wide latitudinal range. In terms of the distribution of genetic diversity, Nei’s genetic differentiation and analysis of molecular variance (AMOVA) showed 38.09% and 29.54% of diversity existed among populations, respectively, elucidating a low-to-moderate level of among-population genetic differentiation. Although a relatively large proportion of diversity was attributed to within-population variation, low diversity within populations (mean genetic diversity within populations (HS) = 0.19) was observed. Both STRUCTURE and unweighted pair group method with arithmetic mean (UPGMA) dendrograms exhibited the clustering of populations that inhabit the same geographic region, and four clusters correlated with four geographic regions, which might be attributed to insect pollination, small population size, and environmental conditions in different habitats. These results function as an essential step towards better conserving and utilizing wild K. latifolia resources, and hence promoting its genetic improvement and breeding for adaptability to southeastern environmental conditions.

Pygmaeomycetaceae, a new root associated family in Mucoromycotina from the pygmy pine plains

ABSTRACTA new genus, Pygmaeomyces, and two new species are described based on phylogenetic analyses, phenotypic and ecological characters. The species delimitation was based on concordance of gene genealogies. The Pygmaeomyces cultures were isolated from the roots of mountain laurel (Kalmia latifolia) and pitch pine (Pinus rigida) from the acidic and oligotrophic New Jersey Pygmy Pine Plains; however, they likely have a broader distribution because their internal transcribed spacer (ITS) sequences have high similarity with a number of environmental sequences from multiple independent studies. Based on the phylogeny and phenotypical characters, a new family Pygmaeomycetaceae is proposed to accommodate this new lineage in Mucoromycotina. Pygmaeomycetaceae corresponds to Clade GS23, which was identified based on a sequence-only soil fungal survey and was believed to be a distinct new class. Compared to the culture-based methods, we observed that sequence-only analyses tend to over-estimate the taxonomic level. Results from this work will facilitate ecological and evolutionary studies on root-associated fungi.

Implementation of Soil Moisture Sensor Based Automated Irrigation in Woody Ornamental Production1

A soil moisture sensor-based automated irrigation system was trialed in a commercial ornamental nursery over the 2014-2015 growing seasons. In both years, use of the sensor-based system resulted in an approximate 50% reduction in irrigation application (volume) when compared to grower-managed irrigation. No differences in growth and equivalent or slightly reduced crop losses were noted when comparing the sensor-based irrigation system to grower-managed irrigation in production of Pieris japonica, Hydrangea quercifolia, and Kalmia latifolia. In 2014, Rhododendron catawbiense had equivalent canopy size and reduced mortality when comparing sensor-based irrigation to grower-managed irrigation. However, in 2015 irrigation control with the sensor-based system resulted in significant (>50%) Rhododendron losses. High mortality was thought to have resulted from use of averaged (across crop species) soil moisture readings to establish irrigation set points. Canopy structure of Rhododendron obstructed water capture to a greater degree than the other three species due to canopy architecture. This effect, combined with precision irrigation applications, resulted in persistent drought conditions within the Rhododendron block. Soil moisture sensor-based automated irrigation can be an effective means of automating irrigation. Support from crop consultants is highly desirable to minimize disruption and maximize adoption during implementation. Index words: Pieris Japonica D. Don ex G. Don ‘Prelude', Hydrangea quercifolia W. Bartram ‘Jet Stream', Rhododendron catawbiense Michx. ‘Roseum Elegans', Kalmia latifolia L. ‘Sarah', irrigation groupings, automation, canopy structure, irrigation capture, technology transfer, outreach, education, precision irrigation Species used in this study: Japanese andromeda ‘Prelude' (Pieris Japonica D. Don ex G. Don); Oakleaf hydrangea ‘Jet Stream' (Hydrangea quercifolia W. Bartram); Rhododendron ‘Roseum Elegans' (Rhododendron catawbiense Michx.); Mountain laurel ‘Sarah' (Kalmia latifolia L.)

Comparing Insect Pollinator Visitation for Six Native Shrub Species and Their Cultivars

Interest in native landscape plants to support pollinators has increased. Most native plants sold by nurseries are cultivars, and some consumer and conservation groups question the suitability of native cultivars to support pollinators. In 2017 and 2018, insect pollinator visitation was quantified for six native shrub species and one or more cultivars of each species (Aronia melanocarpa, A. melanocarpa ‘UCONNAM012’ Ground Hog®, A. melanocarpa ‘UCONNAM165’ Low Scape Mound®, Clethra alnifolia, C. alnifolia ‘Hummingbird’, C. alnifolia ‘Ruby Spice’, Dasiphora fruticosa, D. fruticosa ‘Goldfinger’, D. fruticosa ‘Pink Beauty’, Hydrangea arborescens, H. arborescens ‘Annabelle’, Kalmia latifolia, K. latifolia ‘Sarah’, Physocarpus opulifolius, and P. opulifolius ‘Monlo’ Diabolo®). Insects were identified into 12 categories (Apis mellifera, Bombus spp., Andrenidae, Halictidae, Megachilidae, other bees, Lepidoptera, Syrphidae, other flies, wasps, Coleoptera, and other insects). The number of inflorescences and insect visitation was similar for C. alnifolia and its cultivars, and the compact cultivar Hummingbird had the greatest floral density. A. melanocarpa had more total visitors of Andrenidae than both of its compact cultivars because it was larger and produced more inflorescences. Compact Aronia cultivars and the straight species were mostly similar for Andrenidae visitation when compared on a per-inflorescence basis. D. fruticosa had more visitors of Bombus spp. and Megachilidae than both of its cultivars. These insects may have been less attracted to ‘Pink Beauty’ because of its pink flower color and ‘Goldfinger’ because of its wider flowers, which result from it being a tetraploid. H. arborescens ‘Annabelle’ had one-third the number of Bombus spp. visitors as H. arborescens because ‘Annabelle’ produces >50% fewer fertile florets. P. opulifolius ‘Monlo’ attracted more syrphids than P. opulifolius possibly because flowers contrasted more strongly with the reddish purple foliage of ‘Monlo’ than with the green foliage of the straight species. Insect visitation was similar for K. latifolia and K. latifolia ‘Sarah’. Based on this work, we determined that native shrub cultivars are not universally less or more attractive to pollinators and must be evaluated on a case-by-case basis.

In Vitro Seed Germination of Kalmia latifolia L. Hybrids: A Means for Improving Germination and Speeding Up Breeding Cycle

Kalmia latifolia L. (mountain laurel), a member of Ericaceae, is a beautiful ornamental shrub native to the eastern United States. The plant is not common in the southeastern United States landscapes because of the limited heat tolerance of most commercial cultivars. Breeding of heat-tolerant cultivars can be achieved by cross hybridization, but is often challenged by low germination percentage, long germination time, and potential abortion of cross-hybridized seeds. We used in vitro seed germination to enhance germination and shorten germination time and investigated the appropriate collecting time, optimal basal medium, and pH for this approach. Collecting time affected in vitro seed germination, with more mature hybrid seeds [collected 4–5 months after pollination (MAP)] having higher germination rate (90% in 4 weeks) than the less mature seeds collected in 2 MAP (20% in 7 weeks). Seedlings from the mature seeds also produced two true leaves on average after 8 weeks of culture, whereas seedlings from the less mature seeds had no true leaves. Woody Plant Medium (WPM) better enhanced in vitro seed germination compared with Murashige and Skoog (MS) or Gamborg’s B5 (B5) medium. WPM yielded higher germination (98%) than MS (90%) and significantly greater total leaf area per seedling (67 mm3) than MS (50 mm3) and B5 (52 mm3) for seeds of ‘Firecracker’ × ‘Snowdrift’. Similar effects had been observed on seeds from ‘Little Linda’ × ‘Starburst’ and ‘Pristine’ × ‘Peppermint’. The pH ranging from 4.2 to 5.4 did not affect seed germination and seedling development of mountain laurel hybrids. Our protocol enabled early collection of mountain laurel hybrid seeds 1 month before their full maturation and permitted seeds to germinate in 4 weeks on WPM, which shortened the period from crossing to the seedling stage from up to 15 to 6 months and enhanced germination percentage from 30% to more than 90% compared with traditional seed germination. This protocol should be applied to promote the breeding and selection of new mountain laurel cultivars for the southeastern United States landscapes.

Genome Sizes and Ploidy Levels in the Genus Kalmia

Kalmia is a highly ornamental genus of shrubs native to North America and Cuba and grown as a valuable nursery crop throughout much of the temperate world. Although most species of Kalmia have previously been found to be diploid with 2n = 2x = 24, one species, Kalmia polifolia, has been found to be tetraploid. However, sampling within the genus has been limited, and information on the ploidy levels of specific cultivars is lacking. The objective of this study was to determine the relative genome sizes and ploidy levels of species, hybrids, and cultivars of Kalmia. Flow cytometry was used to determine the relative genome sizes of 67 accessions representing species, interspecific hybrids, cultivars, and chemically induced polyploids. Traditional cytology was used to calibrate genome sizes with ploidy levels. Results showed that relative genome sizes were conserved with 1Cx values ranging from 0.57 pg for Kalmia carolina to 0.70 pg for Kalmia latifolia. Most species of Kalmia were diploid including K. buxifolia (Leiophyllum buxifolium), K. carolina, K. cuneata, K. hirsuta, K. latifolia, and K. microphylla. Although plants of K. carolina (Kalmia angustifolia var. carolina) were uniformly diploid, the closely related, but more northerly distributed, K. angustifolia was primarily tetraploid, providing additional justification for treating these as separate species. An unusual triploid of K. angustifolia f. candida was also documented. Kalmia polifolia included both tetraploid and potentially pentaploid individuals, indicating a ploidy series within this species. Kalmia latifolia cultivars also included one triploid, two cytochimeras, and two chemically induced tetraploids. Overall, polyploidy was more prevalent in Kalmia than previously reported and varied both within and among species. This broader survey of relative genome sizes and ploidy levels in Kalmia provides valuable information for plant breeders and new insights into the systematics and cytogenetics of the genus.

Impacts of repeated wildfire on long-unburned plant communities of the southern Appalachian Mountains

The infrequent occurrence of large wildfires in the southern Appalachian Mountains over the last several decades has offered few opportunities to study their impacts. From 2000 to 2008, five wildfires burned a large portion of the area in and surrounding the Linville Gorge Wilderness in North Carolina. Areas were burned either once or twice. The response of acid cove and thermic oak plant communities (structure, cover, richness, diversity) was measured in 78 vegetation monitoring plots, established in 1992 and remeasured in 2010–11. Fire altered forest structure in both communities, resulting in the mortality of larger trees and increases in the abundance of smaller (<5 cm diameter at breast height (DBH)) stems. Burning twice decreased stem counts for mountain laurel (Kalmia latifolia) in both communities, whereas oaks (Quercus spp.) responded positively to burning twice in the thermic oak community. Table Mountain pine stem counts increased in acid cove and thermic oak communities burned once. Fire appears to promote princesstree (Paulownia tomentosa) invasion. Herbaceous species cover responded positively to fire (once or twice; both communities), with concurrent increases in woody species richness and diversity. Tree species composition in acid cove plots was not affected by burning, although some slight changes occurred in thermic oak plots burned twice.