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Tissue proliferation (TP) is characterized primarily by the formation of galls or tumors at the crown of container-grown rhododendrons propagated in vitro. However, TP of Rhododendron `Montego' is observed initially in in vitro shoot cultures and it is characterized by the formation of multiple shoots with small leaves and nodal tumors. The formation of shoots in `Montego' TP (TP+) shoot cultures occurs without the presence of exogenous cytokinin in the medium, unlike normal `Montego' (TP–) shoot cultures, which require cytokinin for shoot growth. Structural studies have shown that tumors are composed of many adventitious buds and parenchyma cells, suggesting that TP is a result of abnormal cytokinin regulation that is controlling tumor and shoot formation. Two approaches are being used to determine if differences in cytokinin concentration and/or metabolism exist between TP+ and TP– shoot cultures. In the first approach, shoot cultures are grown in vitro for 1 week in the presence of tritiated isopentenyladenine (iP). Cytokinin uptake and metabolism are analyzed using HPLC and other analytical methods. Experiments suggest that extensive degradation and N-glucoside conjugation occur in TP+ and TP– shoots, resulting in the removal of most of the exogenous iP. In the second approach, the levels of endogenous cytokinins such as iP, isopentenyladenosine, zeatin, and zeatin riboside, are being measured in TP+ tumors and shoots and in TP– shoots by an ELISA method.

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An interspecific hybridization program involving five species of Impatiens was initiated to delineate incompatibility barriers. With the exception of one cross, no viable hybrid seed was recovered. Fluorescence microscopy revealed foreign pollen tubes to reach ovules in all crosses, although not all ovules were approached. A histological study involving I. auricoma Baill. and I. walleriana Hook f. ensued to confirm the presence of hybrid embryos. Developing I. walleriana × I. auricoma and reciprocal hybrid embryos were compared to self embryos. Development of hybrid embryos was delayed as early as five days post-pollination. I. walleriana × I. auricoma embryos continued to develop for 8 days post-pollination, but did not reach a size greater than a 5-day self embryo. Excessive endosperm was observed in the hybrid. I. auricoma × I. walleriana embryos continued to enlarge up to ovary abortion but did not reach a size greater than a 7-day self embryo and little to no endosperm developed. Disintegration of ovules included disorganization and collapse of the endosperm, and vacuolization and loss of turgidity of the embryo.

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The genus Aronia Medik., also known as chokeberry, is a group of deciduous shrubs in the Rosaceae family, subtribe Malinae. The two commonly accepted black-fruited Aronia species are black chokeberry [Aronia melanocarpa (Michx.) Elliott] and aroniaberry [Aronia mitschurinii (A.K. Skvortsov & Maitul)]. The geographic range of wild A. melanocarpa is the Great Lakes region and the northeastern United States, with a southerly extension into the higher elevations of the Appalachian Mountains. Wild A. melanocarpa found in New England are diploids, whereas plants throughout the rest of the range are tetraploids. A. mitschurinii is a cultivated hybrid between ×Sorbaronia fallax (C.K.Schneid.) C.K.Schneid. and A. melanocarpa and exists as a tetraploid. There is currently limited diversity of Aronia genotypes in the ornamental and fruit industries, and many of the current cultivars are not adapted to the southern United States and similar environs with limited chilling to break winter dormancy. The goal of this study was to determine 1) the chilling requirements for A. mitschurinii ‘Viking’ and 2) the range of chilling requirements for wild A. melanocarpa genotypes from different geographic origins. Two experiments were conducted in which plants were subjected to various chilling accumulation treatments and then moved to a greenhouse for observation of budbreak and subsequent growth. Expt. 1 was conducted at the University of Maryland at Wye, MD, and focused solely on the commercial cultivar A. mitschurinii ‘Viking’. Outdoor, ambient fall and winter temperatures were used to achieve the chilling treatments. In Expt. 1, we determined the optimal chilling requirements for A. mitschurinii ‘Viking’ to be greater than 900 h using the single temperature model. Expt. 2 was conducted at the University of Connecticut and focused on wild genotypes, plus A. mitschurinii ‘Viking’. A fixed temperature cold room was used to achieve chilling treatments. In Expt. 2, we found A. melanocarpa genotypes from southern regions in the United States required chilling accumulation of 600 h (single temperature model), compared with genotypes from northern regions that required more than 900 h of chilling accumulation. Tetraploid A. melanocarpa required 900 h of chilling to break bud, but diploid A. melanocarpa required 1200 h of chilling to break bud. Expt. 2 confirmed the 900-h chilling requirement for A. mitschurinii ‘Viking’. For both experiments, the rate of budbreak and shoot growth was positively correlated with increasing amounts of chilling.

Open Access

Comptonia peregrina is a desirable native ornamental plant for challenging landscapes, but it cannot be produced using conventional softwood stem cuttings. We demonstrate that C. peregrina can be successfully propagated using young shoots (6 to 8 cm in length) recently emerged from rhizomes taken as cuttings. Although significantly more cuttings rooted using intermittent mist (99%) than propagation dome (70%), cuttings under propagation domes had greater shoot counts. Due to the drier and warmer conditions under propagation domes, cutting shoot tips were killed, which relieved apical dominance and stimulated lateral budbreak. Cuttings rooted under propagation domes produced plants having greater height, width, and size after 90 d than cuttings rooted under intermittent mist. Treatment of cuttings with talc-based rooting hormone at 3000 and 8000 ppm indole-3-butyric acid (IBA) significantly improved rooting percentage and shoot count over untreated cuttings. Cuttings treated with 8000 ppm IBA produced the most roots. Container plants grown from cuttings and pruned to 7 cm in height produced twice as many shoots as unpruned plants. Using cuttings taken from young shoots (6 to 8 cm) produced from rhizomes, 3000 or 8000 ppm IBA, and intermittent mist nursery growers can achieve rooting percentages for C. peregrina above the 80% benchmark preferred for commercial plant production.

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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.

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Hyperhydricity of shoots initiated in vitro, poor shoot extension, inability of shoot cultures to maintain good growth over an extended time, and unsuccessful ex vitro rooting have limited the development of a commercial scale micropropagation system for hemp (Cannabis sativa). We present a culture initiation method that prevents shoot hyperhydricity using vented-lid vessels with 0.2-µm pores and medium containing agar at 1% (w/v). To optimize shoot multiplication in vitro, a control medium (medium A) and four treatment media (medium B, C, D, and E), with varying inorganic nutrients and vitamins were tested. Control medium A consisted of 1× Murashige and Skoog (MS) with vitamins plus 3% (w/v) sucrose, 0.5 mg·L−1 metatopolin, 0.1 mg·L−1 gibberellic acid, and 0.8% agar (w/v) at pH 5.7. The four treatment media differed from the control medium as follows: medium B, 2.5× MS with vitamins; medium C, 1× MS with vitamins plus added mesos [calcium chloride (anhydrous), magnesium sulfate (anhydrous), and potassium phosphate (monobasic) nutrients]; medium D, 1× MS with vitamins plus added vitamins; and medium E, 1× MS with vitamins plus added mesos and vitamins. Medium C and medium E produced more microcuttings than the control at 6 weeks after the initial subculture with shoot multiplication media and all other treatments at 9 and 12 weeks. Shoots grown on these two media displayed optimal extension and leaf lamina development; however, they exhibited slight chlorosis by 12 weeks after subculture with shoot multiplication media. In a separate experiment, medium E was supplemented with ammonium nitrate at 0, 500, 1000, or 1500 mg·L−1, and cultures grown with 500 mg·L−1 produced the most microcuttings and exhibited the best combination of shoot extension and leaf lamina development. We provide a method of prerooting microshoots in vitro that has resulted in 75% to 100% rooting ex vitro in rockwool. Using 10 recently micropropagated plants, ≈300 retip cuttings (cuttings taken from new shoots from recently micropropagated plants) were harvested over 10 weeks. The average weekly rooting was more than 90%. Retipping can produce nine-times as many plants in a similar amount of floor space as stem cuttings derived from traditional stock mother plants. The micropropagation/retipping method proposed can be a more efficient way to generate clonal liner plants for commercial-scale production.

Open Access

Feminized hemp seed producers often use selfing to maintain a strain name; however, selfing may lead to inferior plants for cannabidiol (CBD) production. Using three different hemp strains as parents [Candida (CD-1), Dinamed CBD, and Abacus], two outcrosses [Candida (CD-1) × Abacus and Dinamed CBD × Candida (CD-1)] and one self-cross [Candida (CD-1) × Candida (CD-1)] were conducted to produce feminized seed. Progeny from the self-cross were significantly smaller and had less yield than outcrossed progeny. Selfed progeny were variegated and highly variable for total dry weight and floral dry weight. Discriminant analysis of principal components (DAPC) using amplified fragment length polymorphism (AFLP) separated the three progeny populations and showed that outcrossed populations clustered closer to the maternal parent, possibly the result of a maternal effect. Analysis of molecular variance (AMOVA) indicated that most variation (74.5%) was within populations, because the progeny from all three populations are half-siblings of each other. The selfed progeny population had lower expected heterozygosity (He = 0.085) than each of the outcrossed progeny populations (He ≈ 0.10). These results suggest that selfed progeny may demonstrate inbreeding depression resulting from enhanced expression of homozygous recessive traits. It may be beneficial for feminized seed producers to use outcrossing instead of selfing to generate feminized seed for CBD production.

Open Access

There is demand for micropropagated Cannabis sativa liner plants, because they are uniform, vigorous, and pathogen free; however, availability is limited because of challenges with in vitro culture decline and ex vitro rooting. Ex vitro rooting success of microcuttings was evaluated for ‘Abacus’ and ‘Wife’ when cultures were 6, 9, 12, 15, and 18 weeks old from initiation. Microcuttings of ‘Wife’ harvested from 6, 9, and 12-week-old cultures rooted at or above 80%, but rooting declined to 50% and 30% for 15- and 18-week-old cultures, respectively. Rooting for ‘Abacus’ remained relatively constant between 47% and 70% for microcuttings harvested from 6- to 18-week-old cultures. ‘Wife’ plants grown from microcuttings, stem cuttings, and retip cuttings (cuttings taken from new shoots on recently micropropagated plants) had equivalent total shoot length, number of shoots, and flower dry weight, whereas micropropagated ‘Abacus’ plants had less shoot length and flower dry weight than plants from stem cuttings. However, when micropropagated ‘Abacus’ plants were provided an extra week of vegetative growth to reach an initial size equivalent to stem and retip plants, all plants performed the same. Propagation method did not change cannabinoid content for both ‘Abacus’ and ‘Wife’. Retip cuttings of ‘Abacus’ and ‘Wife’ rooted at 76% to 81% without rooting hormone, which is comparable to rates reported for stem cuttings of C. sativa treated with rooting hormone. Propagators should consider retipping to expand their liner production, because retips root well and possess the same desirable attributes as micropropagated plants.

Open Access

Aronia Medik., commonly known as chokeberry, is a genus of deciduous, multistemmed, rosaceous shrubs native to eastern North America. Three species of chokeberry are commonly accepted, A. arbutifolia (L.) Pers., red chokeberry, A. melanocarpa (Michx.) Elliott, black chokeberry, and A. prunifolia (Marshall) Rehder, or purple chokeberry. In Europe, a fourth species of human origin is recognized as Aronia mitschurinii A.K.Skvortsov & Maitul. In North America this type of Aronia is described as cultivars of A. melanocarpa, including ‘Viking’, ‘Nero’, and ‘Aron’. This species is characterized by near homogeneity of the population, tetraploidy, and a distinct morphology with more robust stems, wider leaf blades, and larger fruits than wild populations of A. melanocarpa. It has been proposed that this genotype originated from Russian pomologist Ivan Michurin’s early 20th century experiments involving Aronia × Sorbus hybridization. In this study we used amplified fragment length polymorphic (AFLP) markers to elucidate the relationships of A. mitschurinii to wild North American Aronia, ×Sorbaronia C.K. Schneid, Sorbus L., and six additional genera from subtribe Pyrinae (Rosaceae). Data from seven primer combinations were interpreted by the NTSYSpc software package into a similarity matrix using Jaccard’s coefficient. Clustering of AFLP similarity data using the unweighted pair group method with arithmetic mean (UPGMA) identified A. mitschurinii as distinct from wild Aronia, grouping it close to ×Sorbaronia fallax C. K. Schneid. and ×Sorbaronia ‘Ivan’s Beauty’. Non-metric multidimensional scaling (nMDS) also demonstrated a relationship between A. mitschurinii, ×Sorbaronia fallax, a ×Sorbaronia × Aronia backcross and compound-leaved Sorbus.

Free access

We investigated the role of an ornamental, purple-leaved specimen of Japanese barberry in a local invasion using parentage analysis. We focused on a landscape plant of B. thunbergii var. atropurpurea in Willington, CT, that was first established at least 30 years ago. We genotyped every barberry plant found within a 92-m radius of this individual. In contrast to feral populations that are distant from residential or commercial plantings, 14% of the 43 feral plants in our sample had purple foliage and 30% were found growing within 16.5 m of the focal individual. Parentage analysis identified seven plants (five purple-leaved and two green-leaved) as descendants of the focal individual. Five of these descendants are likely first-generation offspring and two are likely second-generation seedlings. In addition, one plant was identified as a backcross between the focal plant and one of its offspring. Our results show that purple-leaved Japanese barberry used in residential landscapes can contribute to plant invasions, at least under some circumstances.

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