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  • Author or Editor: Donglin Zhang x
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Cephalotaxus species are needle evergreens offering the aesthetic qualities of Taxus, yew, yet are heat- and drought-tolerant, sun- and shade-adaptable, and resist deer browsing. They are adaptable to nursery and garden cultivation in USDA hardiness zones (5)6–9. Unfortunately, the various species are frequently confused in the American nursery trade due to their extreme similarity in morphology. Recently, molecular data have been widely applied in the taxonomic studies, especially DNA sequencing. The chloroplast gene rbcL of Cephalotaxus has been sequenced for determining species relationships. The preliminary results show that C. oliveri Mast. has 10 base changes from C. drupacea Sieb. et Zucc., while only one base difference occurred between C. drupacea and C. harringtonia (Forbes) Koch. There are between one and 10 base substitutions among C. fortunei Hooker, C. koreana Nakai, and C. sinensis (Rehd. et Wils.) Li. Compared with other closely related conifers, Cephalotaxus has a substantial number of differences among species except between C. drupacea and C. harringtonia, which may not be distinct species. Detailed data relative to gene sequencing, growth morphology, and horticultural characteristics should lead to correct identification of species and great horticultural uses. Furthermore, the method of rbcL sequence can be applied to distinguish other morphologically homogeneous ornamental plants.

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The identity of heath-leaved cypress is controversial. In this study nucleotide sequences of nuclear ribosomal DNA were used to identify heath-leaved cypress (Chamaecyparis `Ericoides') species. Sixteen individuals were sampled representing the five species of Chamaecyparis, `Ericoides', and four other genera of Cupressaceae (Cupressus, Fokienia, Juniperus, and Thuja). The results placed `Ericoides' unequivocally to Chamaecyparis thyoides, supporting a conclusion derived from wood anatomy. This study supports the usefulness and integrity of using molecular data to identify the genetic affinity of cultivars that are morphologically different from the parent species.

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The correct identification of horticultural taxa becomes more and more important for intellectual property protection and economic reasons. Traditionally, morphological characteristics have been used to differentiate among the horticultural taxa. However, the morphological characteristics may vary with plant age, cultural conditions, and climate. Modern technologies, such as DNA markers, are now employed in the identification of horticultural taxa. Currently, technologies of DNA sequencing (gene sequences) and DNA fingerprinting (RAPD, RFLP, SSR, and AFLP) are available for distinguishing among horticultural taxa. The literature and our personal experience indicate that the application of each technique depends on the taxon and ultimate goal for the research. DNA sequencing of a variety of nuclear or chloroplast encoded genes or intergenic spacers (rbcL, ndhF, matK, ITS) can be applied to distinguish different species. All DNA fingerprinting technologies can be used to classify infraspecies taxa. AFLP (the most modern technique) is the better and more-reliable to identify taxa subordinate to the species, while RAPDs can be employed in clonal or individual identification. Techniques of RFLP and SSR lie between AFLP and RAPD in their effectiveness to delineate taxa. Mechanics, laboratory procedures, and inherent difficulties of each technique will be briefly discussed. Application of the above technologies to the classification of Cephalo taxus will be discussed in concert with the morphological and horticultural characteristics. Future classification and identification of horticultural taxa should combine DNA technology and standard morphological markers.

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To shorten Ilex seed germination time and speed up breeding cycles, immature embryos of Ilex crenata ‘Sky Pencil’ seedlings were removed from fruits at their heart-shape stage and cultured in vitro on Murashige and Skoog (MS) medium or Woody Plant Medium (WPM) with 3% sucrose and 0.65% agar. Cultures were incubated at 27 °C for 2 weeks in darkness and subsequently moved to a growth chamber with 14-hour photoperiod (115 μmol⋅m−2⋅s–1). Embryos began to germinate 2–3 weeks after culture. The highest germination rate was 91.67% under 1/4 MS medium. Embryos cultured on MS medium also had high germination rates and produced the longest seedlings to 8.02 mm. Nodal segments with one axillary bud taken from embryo germination seedlings were cultured on MS medium with various concentrations of cytokinins and auxins for micropropagation. Zeatin (ZT; 4-hydroxy-3-methyl-trans-2-butenylaminopurine) increased the number of shoots and shoot lengths significantly more than 6-benzylaminopurine (6-BA). The recommended ZT concentration should be 2.28 µM. Rooting induction could be established on 1/4 MS medium with various concentrations of indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA). IBA at 4.14 µM produced the best rooting percentage (91.67%) and good-root quality. All rooted plantlets were transplanted into a mixture of peatmoss and perlite (1:1 v/v) and acclimatized in a mist system. The average survival rate was 88.8%. The rapid embryo germination protocol for Ilex crenata could save Ilex breeders at least 2 years compared with traditional seed germination.

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The influence of red and blue light wavelengths was tested to improve the initial in vitro multiplication of apple (Malus × domestica) rootstock cultivars Budagovsky 9 (B.9), Geneva 30 (G.30), and Geneva 41 (G.41). Single-node segments were established in semisolid Murashige and Skoog media and then transferred to proliferation media and cultured 40 days under white, red, or blue light irradiance. In a second experiment, G.30 was cultured under red, blue, or white light with and without gibberellic acid (GA3). The three rootstocks responded similarly under white light in terms of shoot number, length of the longest shoot, and the number of elongated shoots. Red light increased the number of shoots, length of the longest shoot, and the number of elongated shoots of B.9 and G.30 when compared with white or blue light. Red light increased the number of elongated B.9 and G.30 shoots to five per explant compared with one per explant under white light. In contrast, shoot growth of G.41 showed no difference under the three light quality treatments, and the number of elongated shoots per explant was less than one. When compared with an absence of GA3, a concentration of GA3 at 0.5 mg·L−1 promoted in vitro shoot growth of G.30 under red and blue light.

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These studies were conducted to determine the most effective methods for increasing shoot elongation during the initial proliferation stage of micropropagation in two dwarfing apple, Malus ×domestica (Borkh.), rootstock cultivars. Several experiments were conducted to compare explant collection date, exposure to chilling (5 ± 1 °C) temperatures, and varying concentrations of plant growth regulators in Murashige and Skoog (MS) media. Microshoot growth of ‘Geneva 41’ (‘G.41’) was very low and unaffected by chilling duration from 0 to 8 weeks or by gibberellic acid (GA3) concentration from 0 to 1.0 mg·L−1, but was improved by an additional subculture which increased shoot length from 1 to 15 mm. In ‘Geneva 30’ (‘G.30’), shoot elongation was most affected by date, chilling explants, and by optimizing cytokinin concentration and type. Explant collection date in April increased shoot growth compared with August or November. Microshoot growth of ‘G.30’ was increased by chilling nodal explants for 4 and 6 weeks when explants were collected in August and November, but not in April. Eight weeks chilling was detrimental for explants collected in April, and generally had little or no effect with August and November. The cytokinin 6-benzylaminopurine (BA) increased shoot number to a greater extent than thidiazuron (TDZ) or zeatin (ZT), and was also more effective for increasing shoot elongation with concentrations of 0 to 2.0 mg·L−1. In ‘G.30’, GA3 increased shoot growth at the optimum concentration of BA, but not with lower concentrations. ‘G.30’ microshoots were fewer and shorter with 24-epi-brassinolide (EBR) at concentrations of 0.1 and 1.0 mg·L−1. Chemical names: N-phenyl-N’-(1,2,3-thiadiazol-5-yl)urea (TDZ), 6-(4-hydroxy-3-methylbut-2-enylamino)purine (ZT).

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We documented a successful embryo rescue (ER) protocol for butterfly weed (Asclepias tuberosa), a member of the milkweed family (Asclepiadaceae). Milkweed (Asclepias sp.) includes more than 100 species native to the United States, is an important pollinator plant, and has many commercially desirable traits. However, there is little commercial production outside of native plant nurseries because milkweed species are typically seed-grown and suffer from low seed set during pollination, late-term abortion of seed pods, and nonuniform germination. This project determined the optimal growing media (study one) and embryo maturity (study two) to recover mature seedlings from excised embryos and compared the results to those of traditional methods of seed germination (in soilless substrate). Study one investigated three different media: Murashige and Skoog (MS) medium at full strength and half strength and woody plant medium. MS medium at half strength was optimal for butterfly weed germination and maturation, with greater root and shoot lengths at the time of harvest. In study two, the effects of MS medium at half strength on embryo maturation 90, 60, and 30 days after pollination (DAP) were investigated. The optimal time to harvest embryos was 60 DAP; embryos at 30 DAP were capable of germination but not maturation. A mean germination rate of 97.4% was observed when using embryo rescue, but it was 72.3% with mature seed germinated in soilless substrate typical of commercial production. A similar increase in germination rates was observed for all embryo maturities when compared with seed germinated using soilless substrate. The protocol developed for this study should help to standardize production, reduce propagation time, and improve the commercial acceptance and profitability of milkweed.

Open Access

Cephalotaxus Sieb. and Zucc. (plum yew) species and cultivars have become popular because of their sun and shade tolerance, resistance to deer browsing, disease and insect tolerance, and cold and heat adaptability. Unfortunately, the nomenclature and classification in the literature and nursery trade are confusing due to their extreme similarity in morphology. In this study, amplified fragment-length polymorphism (AFLP) markers were used to discriminate taxa and evaluate genetic differences among 90 Cephalotaxus accessions. A total of 403 useful markers between 75 and 500 base pairs (bps) was generated from three primer-pair combinations. Cluster analysis showed that the 90 accessions can be classified as four species, C. oliveri Mast., C. fortunei Hooker, C. harringtonia (Forbes) Koch., and C. ×sinensis (a hybrid species); four varieties, C. fortunei var. alpina Li, C. harringtonia var. koreana (Nakai) Rehd., C. harringtonia var. nana (Nakai) Hornibr., and C. harringtonia var. wilsoniana (Hayata) Kitamura; and eight cultivars. Suggested names are provided for mislabeled or misidentified taxa. The Cephalotaxus AFLP data serve as a guide to researchers and growers for identification and genetic differences of a taxon, and a model to establish a cultivar library against which later introductions or problematic collections can be cross-referenced.

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