Hydrangea Plants Research Articles

The Influence of Combined Pruning and the Use of Root Application of Two Biostimulants and Foliar Nutrition on the Growth and Flowering of Panicle Hydrangea Plants

The aim of this experiment was to assess how the interaction of two treatments influenced the growth and flowering of two varieties of Panicle hydrangea shrubs. The first treatment was plant pruning. Simultaneously, the plants received one of the three following treatments: root application of Trichoderma atroviride, root application of BlackJak biostimulant, or foliar application of a multi-component fertilizer. Simultaneous pruning and inoculation of the plants with the Trichoderma atroviride mycelium improved the length of hydrangea shoots the most, as compared with the control plants (18%). These two treatments also increased the number of flowers (16–47%, depending on the variety) and the fresh weight of plants (10–28%) compared with the control plants. T. atroviride alone improved the number of flowers in both varieties (19–24%) and the diameter of inflorescences in the ‘Silver Dollar’ one (17%). The foliar nutrition increased the fresh weight of plants by 7–57%, depending on the cultivar and pruning. It also increased the diameter and number of inflorescences in one of the varieties. Together with pruning, it intensified the growth of shoots in both cultivars (5–10%). The BlackJak biostimulant treatment gave ambiguous results. In combination with pruning, it improved the length of shoots (15%) in one cultivar and the fresh weight in the other (18%). Without pruning, the treatment increased the number of flowers (16%) and the diameter of inflorescences (9%) in one cultivar. It increased the fresh weight of plants in both cultivars (19–21%). Regardless of the other treatments, pruning increased the length of the shoots and the fresh weight of the plants. On the other hand, it reduced the number of flowers and their diameter. In most cases, the biostimulant treatment and foliar fertilization improved the growth and flowering of the plants. In combination with pruning, they improved the growth of the hydrangea shrubs but reduced the number and diameter of flowers. The simultaneous Ta treatment and pruning were the most beneficial for the growth and flowering of the panicled hydrangea plants grown in containers in a nursery.

In vitro and in vivo biocontrol studies against Botrytis cinerea, the causal agent of grey mould in Hydrangea macrophylla

ABSTRACT Necroses on aerial parts of Hydrangea macrophylla were regularly observed in plant storage chamber at 2°C in Hortensia France company. A total of 34 Botrytis isolates were collected from Hydrangea plants and their environment. To develop biocontrol methods against Botrytis, eight Trichoderma isolates were collected from Hydrangea or the plant soil substrate in the same cold room. Two Trichoderma isolates (E10SM, E014) from this study and one reference isolate (M03) exhibited a high degree of in vitro parasitism against B. cinerea, by comparison with Trichoderma C04 reference isolate (no parasitism). Culture filtrates of isolates E014 and M03 also showed significant in vitro antibiotic properties against B. cinerea growth. A pathosystem H. macrophylla – B. cinerea was set up in the greenhouse, with the inoculation of leaves using mycelial explants selected as the best infection mode. Using this pathosystem, a biocontrol assay showed a significant effect of M03 isolate towards B. cinerea, using Trichoderma conidia applied to leaves 5 h prior pathogen inoculation. After elicitor addition (B. cinerea grinded mycelia) in Trichoderma cultures, antagonistic potentialities of isolate C04 towards the pathogen was highly enhanced, reaching the inhibition level assessed with isolate M03 grown without elicitor. Optimisation of Trichoderma culture filtrates for antagonistic abilities was investigated, and specific cultural conditions were selected for M03 (12 g L−1 PDB, pH 4.2, 22°C, 22 days) and C04 (2.4 g L−1 PDB, pH 6.2, 13°C, 7 days). Overall, our results suggest great properties of several Trichoderma isolates for future development as biocontrol products.

Powdery Mildew of Bigleaf Hydrangea: Biology, Control, and Breeding Strategies for Resistance

Hydrangea macrophylla, commonly known as bigleaf, garden, French, or florist hydrangea, is the most economically important member of the Hydrangea genus, with a breeding history spanning hundreds of years. Bigleaf hydrangea breeding improvement has largely focused on aesthetic traits and there are few varieties tolerant or resistant to major diseases such as powdery mildew. Powdery mildew is an obligate biotrophic Ascomycete in the order Erysiphales represented by approximately 900 species worldwide. The disease-causing agent in hydrangeas is Golovinomyces orontii (formerly Erysiphe polygoni DC), which tarnishes the beauty, growth, and salability of bigleaf hydrangea plants, especially those packed closely in production environments. Chemical or biological control is commonly used in production. A recently published haplotype-resolved genome of bigleaf hydrangea enables targeted analyses and breeding techniques for powdery mildew resistance. Analyzing transcriptomes of tolerant and susceptible hydrangeas through RNA sequencing will lead to the identification of differentially expressed genes and/or pathways. Concurrent application of marker-assisted selection, genetic transformation, and gene editing will contribute to the development of powdery-mildew-resistant varieties of bigleaf hydrangea. The aim of this review is to give a general overview of powdery mildew, its impact on bigleaf hydrangea, current control methods, molecular mechanisms, and breeding prospects for powdery mildew resistance in bigleaf hydrangea.

Growing media is the major source of damaging population of Systena frontalis (Coleoptera: Chrysomelidae) in ornamental plant nurseries.

Systena frontalis (F.) (Coleoptera: Chrysomelidae) is a serious pest of ornamental shrubs in containerized ornamental plant nurseries in the central and eastern United States. Adult S. frontalis cause numerous shot holes on foliage, rendering ornamental plants unmarketable. Growing media in plant containers is an overwintering site of S. frontalis, but the extent to which adults emerging from the growing media can damage the plants is unclear. Experiments were conducted on panicled hydrangea (Hydrangea paniculata Siebold) in Georgia, North Carolina, and Virginia nurseries in the spring of 2021 and 2022 to answer this question. The treatments were (i) canopy caged, (ii) whole-plant caged, and (iii) noncaged hydrangea plants. In all 3 states, beetle abundance and feeding damage found on caged (whole plant) and noncaged plants were significantly greater than those on plants where only the canopy was caged. In most sites and years, beetle abundance and feeding damage were not significantly different between the noncaged plants and those where the canopy and containers were caged, suggesting that the majority of S. frontalis emerged from the growing media and the majority of damage suffered by the hydrangea plants were caused by beetles emerging from the containers. Because growing media contributed to a significant proportion of the S. frontalis population in a nursery, treatment targeting larvae in the growing media should be a critical component of a holistic management plan against S. frontalis.

Assessment of Fresh Miscanthus Straw as Growing Media Amendment in Nursery Production of Sedum spectabile 'Stardust' and Hydrangea arborescens 'Annabelle'.

The aim of this research was to assess the influence of fresh miscanthus straw shreds as a component of growing media in nursery production of perennial Sedum spectabile 'Stardust' and woody shrub Hydrangea arborescens 'Annabelle'. A total of five substrate mixes composed of peatmoss and miscanthus straw were used: 100%P, 70%P:30%M, 50%P:50%M, 70%P:30%M, 100%M. Each substrate was subjected to three fertilizer treatments: Basacote, Basacote + YaraMila, and YaraMila. The growth response of both tested species was very similar. In general, plants performed best in 100%P, and the quality decreased with increasing miscanthus straw amendment; however, differences in height and dry weight at the level of ~9% suggest that Sedum plants obtained market value with up to 50% miscanthus amendment and Hydrangea plants with up to 30% miscanthus mixed in media. The most favorable effect on the tested parameters was a combination of Basacote + YaraMila, which delivered more soluble salts, and in higher rates than Basacote and YaraMila used separately. Decrease in EC and nutrients in the substrate with increase in miscanthus straw amendment suggest that uniform irrigation among all the treatments contributed to nutrients leaching from miscanthus media due to its lower water holding capacity.

First Report of Leaf Spot Caused by Alternaria tenuissima on Panicle Hydrangea (Hydrangea paniculata) in China.

Panicle hydrangea (Hydrangea paniculata), belonging to the Saxifragaceae family (Wu et al. 2001), is an ornamental flowering plant which is native to China and Japan. In July 2021, brown leaf spots (diameter ranged from 3 to 5 mm) were observed on panicle hydrangea plants in a 0.1 ha field of Northeast Agriculture University (126.72°E, 45.74°N), Heilongjiang Province. The incidence was approximately 30%. Small circular or irregular brown spots initially appeared on the older leaves, and these lesions gradually expanded with time. In some serious cases, lesions joint together and caused leaf wilting. To identify the causal agent, ten symptomatic leaves from different panicle hydrangea plants were collected, and surface sterilized with 70% ethanol for 30 s, followed by 0.5% NaClO treatment for 4 min, and then rinsed with sterile water three times. Tissues between healthy and necrotic area were cut into 5×5 mm pieces after air drying on sterile filter paper. The pieces were placed on potato dextrose agar (PDA) amended with streptomycin sulfate (50 mg/liter) and incubated at 25°C for 5 days. Fifteen pure isolates were obtained using the hyphal tip technique and cultured on PDA for 7 days at 25°C for morphological and molecular identification. Colonies of all isolates were dark olivaceous with white margin. Conidiophores were septate, singly arising, and light brown, with a size range of 10.2-60.1 × 1.4-5.6 µm (n=50). Conidia were obclavate to obpyriform, brown to dark brown, and in a size range of 15.0-25.2 × 5.1-15.3 µm (n=50). To further identify these isolates, four different genomic DNA regions including ribosomal DNA internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), RNA polymerase II second largest subunit (rpb2), and histone 3 (HIS3) were amplified and sequenced with primers ITS1/ITS4 (White et al. 1990), gpd1/gpd2 (Berbee et al. 1999), RPB2-5F2/RPB2-7R (Sung et al. 2007), and H3-1a/H3-1b (Xu et al. 2022) for three representative isolates (DXQ2-2, DXQ2-3, and DXQ2-4), respectively. The sequences of ITS, GADPH, rpb2, and HIS3 for these three isolates were identical and only sequences of DXQ2-2 were deposited in GenBank (GenBank accession nos. OL305828, OL333601, OL333602, and OL436242). These sequences showed 100%, 100%, 100%, and 99.61% identity with A. tenuissima BJ-CX-1 (GenBank accession nos. MK683974, MK683784, MK684069, and MK683879), respectively. Based on morphological features and DNA sequences analyses, these isolates were identified as Alternaria tenuissima (Simmons 2007). To fulfill Koch's postulates, ten healthy and surface disinfected leaves of one panicle hydrangea plant grown in pot were sprayed with a conidial suspension (1×106 conidia/mL) of isolate DXQ2-2. Meanwhile, ten surface disinfected leaves of another panicle hydrangea plant grown in pot sprayed with sterilize water served as the control. All plants were maintained in a greenhouse at 25℃ and 70% relative humidity. Five days after inoculation, leaves inoculated with conidial suspension showed leaf spot symptoms that were similar to those observed in the field, whereas no symptom was observed on the control leaves. The experiment was conducted twice. The Alternaria tenuissima isolate was successfully re-isolated from the symptomatic leaves and confirmed based on above morphological and molecular methods. To our knowledge, this is the first report of leaf spot on panicle hydrangea caused by A. tenuissima. Leaf spot has a negative effect on the aesthetic value of panicle hydrangea, so further investigation and management is needed to control this disease.

Comparative Transcriptome Analysis Unveils the Molecular Mechanism Underlying Sepal Colour Changes under Acidic pH Substratum in Hydrangea macrophylla

The hydrangea (Hydrangea macrophylla (Thunb). Ser.), an ornamental plant, has good marketing potential and is known for its capacity to change the colour of its inflorescence depending on the pH of the cultivation media. The molecular mechanisms causing these changes are still uncertain. In the present study, transcriptome and targeted metabolic profiling were used to identify molecular changes in the RNAome of hydrangea plants cultured at two different pH levels. De novo assembly yielded 186,477 unigenes. Transcriptomic datasets provided a comprehensive and systemic overview of the dynamic networks of the gene expression underlying flower colour formation in hydrangeas. Weighted analyses of gene co-expression network identified candidate genes and hub genes from the modules linked closely to the hyper accumulation of Al3+ during different stages of flower development. F3'5'H, ANS, FLS, CHS, UA3GT, CHI, DFR, and F3H were enhanced significantly in the modules. In addition, MYB, bHLH, PAL6, PAL9, and WD40 were identified as hub genes. Thus, a hypothesis elucidating the colour change in the flowers of Al3+-treated plants was established. This study identified many potential key regulators of flower pigmentation, providing novel insights into the molecular networks in hydrangea flowers.

A New Ilarvirus Found in French Hydrangea.

In this study, a new virus was identified in French hydrangea plants, exhibiting chlorotic vein banding and necrotic ring spots on older leaves. The virus was mechanically transmitted to herbaceous hosts, in which it induced local and systemic or only local symptoms. The genome of the new virus was characterized and consisted of three RNA sequences that were 3422 (RNA 1), 2905 (RNA 2) and 2299 (RNA 3) nucleotides long, with five predicted open reading frames; RNA2 was bicistronic and contained conserved domains and motifs typical of ilarviruses. The phylogenetic analysis of the predicted proteins—p1, p2a, p3a and p3b—revealed its close relationship to recognized members of subgroup 2 within the genus Ilarvirus. Homologous antiserum was effective in the detection of the virus in plant extracts and no cross reactions with two other distinct members of subgroup 2 were observed. Overall, the biological features, phylogenetic relationships and serological data suggest that this virus is a new member of the genus, for which we propose the name hydrangea vein banding virus (HdVBV).

First Report of Leaf Spot Caused by Cladosporium tenuissimum on Panicle Hydrangea (Hydrangea paniculate) in China.

Panicle Hydrangea (Hydrangea paniculate) is an ornamental flowering plant native to China and Japan. In August 2019, leaf spot symptoms with about 30% disease incidence were observed on panicle hydrangea in two grower fields (about 0.1 ha in total) of Northeast Agriculture University, China (126.72°E, 45.74°N). Symptoms initially appeared on the lower and older leaves and showed small subcircular brown spots with dark-brown edges on both sides. As the disease progressed, the necrotic spots enlarged, became irregular, coalesced, and the infected leaf blighted in approximately 2 weeks. Panicle hydrangea leaf samples (n=15) from different plants that showed spot symptoms were collected and surface sterilized with 70% ethanol for 10 s, followed by 0.5% NaClO treatment for 4 min, and rinsed in sterile water 3 times. Thereafter, leaf samples were placed on potato dextrose agar (PDA) and incubated at 25°C for 7 days. Fifteen hyphal-tipped pure cultures were obtained. Colonies growing on PDA for 7 days were olive green to dark green, exhibited a velvet-like texture and sometimes were radially furrowed and wrinkled. Margins varied from white gray to dark green without prominent exudates. The back of the plate showed dark green to black. Conidiophores were up to 180 to 600 µm long, 2.8 to 4.5 µm wide (n=50), subcylindrical-filiform, straight, septate, and unbranched or rarely branched. Ramoconidia were 0 to 1 septate, cylindrical to clavate, smooth-walled, 8 to 22 μm long (n=50). Conidia were single-celled, lemon-shaped, smooth-walled and 2.0 to 5.0 µm (diameter) (n=50). To confirm the identity, three genomic DNA regions, internal transcribed spacer (ITS), partial translation elongation factor-1 alpha (EF), and actin (ACT) of the representative isolate BAI-1 were amplified with primer pairs ITS1/4, EF1-728F/986R, and ACT-512F/783R, respectively (Bensch et al. 2012; Jo et al. 2018). DNA sequences of the isolate from ITS, EF, and ACT showed 99.81% (514/515 bp), 99.10% (219/221 bp), and 99.54% (216/217 bp) nucleotide identity with those of C. tenuissimum CBS 125995, respectively (GenBank accession nos. HM148197, HM148442, and HM148687). The sequences of isolate BAI-1 were deposited in GenBank (accession nos. MW045455, MW052465, and MW052466). To fulfill Koch's postulates, five healthy 2-year-old panicle hydrangea plants grown in pots were surface sterilized with 70% ethanol, washed twice with sterile distilled water, and sprayed with a conidial suspension of strain BAI-1 (adjusted to 1×106 conidia/ml using a hemocytometer), maintained in a greenhouse at 25°C and 85% relative humidity. Five plants sprayed with sterilized water served as controls. The inoculated plants showed leaf spot symptoms that were similar to those previously observed in the fields after 7 days, whereas control leaves remained healthy. The fungus was reisolated from symptomatic leaves and its identity was confirmed by morphological and molecular method. These experiments were repeated twice. So far, C. tenuissimum was reported to cause leaf spot of alfalfa (Han et al. 2019) and castor (Liu et al. 2019). To our knowledge, this is the first report of leaf spot disease in panicle hydrangea caused by C. tenuissimum in China. Leaf spot has a negative effect on the aesthetic value of panicle hydrangea, and this report will assist with monitoring distribution of the disease as well as developing management recommendations.

Comparative Performance of Reduced-risk Fungicides and Biorational Products in Management of Postharvest Botrytis Blight on Bigleaf Hydrangea Cut Flowers

Botrytis cinerea is one of the problematic and notorious postharvest pathogens of bigleaf hydrangea (Hydrangea macrophylla) cut flowers. It causes flower blight, leaf blight, and stem rot, reducing the ornamental value (such as longevity, color, and texture) of flowers, ultimately making them unsalable. The objective of this study was to identify effective conventional fungicides and biorational products for botrytis blight management on bigleaf hydrangea cut flowers that can be easily and readily adopted by growers of ornamentals. Preventive preharvest whole-plant spray and postharvest dip treatment applications were used in this study. For the whole-plant spray applications, bigleaf hydrangea plants were sprayed with treatment solution 3 days before harvesting flowers. For the dip applications, cut flowers were dipped in treatment solutions after harvest. For both application types, flowers were inoculated with B. cinerea spores once treatment solutions dried. Flowers were stored in cold storage for 3 days and then displayed in conditions similar to retail stores. Botrytis blight disease severity, marketability of flower (postharvest vase life), phytotoxicity, and application residue were assessed in the study. Treatments showed variable efficacy in managing postharvest B. cinerea infection in bigleaf hydrangea cut flowers. Preventive preharvest whole-plant spray and postharvest dip applications of isofetamid and fluxapyroxad + pyraclostrobin significantly reduced the postharvest botrytis blight disease severity and area under disease progress curve (AUDPC) compared with the positive control (nontreated, inoculated with B. cinerea). When applied as a postharvest dip, the fungicide fludioxonil and biofungicide Aureobasidium pullulans strains DSM 14940 and DSM 14941 effectively lowered the disease severity and disease progress (AUDPC). These effective treatments also maintained a significantly longer postharvest vase life of bigleaf hydrangea cut flowers compared with the nontreated, inoculated control. The longer vase life may be attributed to lowered botrytis blight disease severity and the resultant proper physiological functioning of flowers.

A Novel Method for Estimating Nitrogen Stress in Plants Using Smartphones

For profits in crop production, it is important to ensure that plants are not subjected to nitrogen stress (NS). Methods to detect NS in plants are either time-consuming (e.g., laboratory analysis) or require expensive equipment (e.g., a chlorophyll meter). In this study, a smartphone-based index was developed for detecting NS in plants. The index can be measured in real time by capturing images and processing them on a smartphone with network connectivity. The index is calculated as the ratio of blue reflectance to the combined reflectance of blue, green, and red wavelengths. Our results indicated that the index was specific to NS and decreased with increasing stress exposure in plants. Further, the index was related to photosynthesis based on the path analysis of several physiological traits. Our results further indicate that index decreased in the NS treatment due to increase in reflectance of red and green (or yellow) wavelengths, thus it is likely related to loss of chlorophyll in plants. The index response was further validated in strawberry and hydrangea plants, with contrasting plant architecture and N requirement than petunia.

Salt Tolerance of Hydrangea Plants Varied among Species and Cultivar within a Species

A greenhouse study was conducted to assess the relative salt tolerance of 11 cultivars of hydrangea: Hydrangea macrophylla ‘Ayesha’, ‘Emotion’, ‘Mathilda Gutges’, ‘Merritt’s Supreme’ and ‘Passion’; H. paniculata ‘Interhydia’ and ‘Bulk’; H. quercifolia ‘Snowflake’; H. serrata ‘Preciosa’; and H. serrata × macrophylla ‘Sabrina’ and ‘Selina’. Plants were treated with a nutrient solution at an electrical conductivity (EC) of 1.0 dS·m−1, and nutrient solution-based saline solutions at an EC of 5.0 dS·m−1 (EC 5) or 10 dS·m−1 (EC 10). The study was repeated in time (Experiments 1 and 2). In both experiments, by the fourth week after treatment, ‘Bulk’ plants in EC 10 exhibited severe salt damage with most of them dead. ‘Interhydia’ was also sensitive, showing severe salt damage in EC 10 with a high mortality rate by the end of the experiment. The leaf area and total shoot dry weight (DW) of all cultivars in EC 5 and EC 10 treatments were significantly reduced compared to the control. Leaf sodium (Na+) and chloride (Cl−) concentrations were negatively correlated with visual quality, leaf area and shoot DW. The salt-sensitive cultivars ‘Bulk’, ‘Interhydia’ and ‘Snowflake’ had inherently low leaf Na+ and Cl− concentrations in both control and salt-treated plants compared to other cultivars. Salt tolerance varied among species and cultivars within H. macrophylla. Among the 11 cultivars, H. macrophylla ‘Ayesha’ and two hybrids, ‘Sabrina’ and ‘Selina’, were relatively salt-tolerant. H. macrophylla ‘Merritt’s Supreme’ and ‘Mathilda’ were moderately tolerant. H. paniculata ‘Bulk’ was the most sensitive, followed by H. paniculata ‘Interhydia’, and then by H. serrata ‘Preciosa’ and H. macrophylla ‘Passion’, as evidenced by high mortality and severe salt damage symptoms. H. quercifolia ‘Snowflake’ and H. macrophylla ‘Emotion’ were moderately salt-sensitive.

First report of stem rot on hydrangea caused by Phytophthora hedraiandra in Japan

A stem rot disease appeared on hydrangea plants in Tochigi Prefecture, Japan. Four Phytophthora-like isolates were obtained, and a representative isolate was selected for analysis. Hydrangea plants inoculated with the isolate developed similar symptoms. Molecular phylogenetic analyses using the rDNA ITS region and cox1, EF1-α, and β-tubulin genes indicated that the isolate was Phytophthora hedraiandra. The sporangia were papillate, laterally sessile, and caducous. Antheridia were paragynous or amphygynous, and laterally sessile, and the oospores were aplerotic. Based on these characteristics, the isolate was identified as P. hedraiandra. We suggest naming this hydrangea disease “Ajisai kukieki byo”.

Nitrogen Fertilization, Container Type, and Irrigation Frequency Affect Mineral Nutrient Uptake of Hydrangea

Mineral nutrient uptake of Hydrangea macrophylla ‘Merritt’s Supreme’ affected by nitrogen (N) fertilization rate, container type, and irrigation frequency was investigated. Rooted liners of hydrangea plants were fertilized twice weekly with a N-free fertilizer plus five N rates including 0, 5, 10, 15, or 20 mM N from ammonium nitrate (NH4NO3), irrigated once or twice daily with the same total irrigation volume, and grown in two types of one-gallon containers: a black plastic container and a biodegradable container (biocontainer), made from recycled paper. Concentrations of calcium (Ca) and magnesium (Mg) averaged in the entire plant, and iron (Fe) and manganese (Mn) in roots had increasing trends with increasing N rate. By comparison, increasing N rate had a dilution effect on root phosphorus (P), stem and root potassium (K), stem Ca and Mg, and leaf boron (B) concentrations. In general, nutrient content of each tested element increased with increasing N rate in each structure, or total in the plant. When there was a significant container type effect, plastic containers consistently had increased nutrient concentrations and content compared to biocontainers. One irrigation per day was beneficial in increasing nutrient concentrations of P, Ca, and zinc (Zn) in different plant structures.

Hydrangea DNA Methylation Caused by pH Substrate Changes to Modify Sepal Colour is Detected by MSAP and ISSR Markers

The hydrangea (Hydrangea macrophylla (Thunb). Ser.) is an ornamental species with great market potential. It is known for its ability to change the colour of its inflorescence, according to the pH of the culture substrate. The molecular mechanisms that underlie these changes are still unclear. It is known that epigenetic mechanisms, such as DNA methylation, play an important role in genetic expression, so they could be responsible for this phenomenon in hydrangea. In the present study, the molecular markers ISSR (Inter Simple Sequence Repeat) and MSAP (Methyl-Sensitive Amplification Polymorphism) were used to detect molecular changes in the genome of hydrangea plants that were cultivated under different pH levels to modify the colour of the sepals. The results showed a correspondence between the methylation signal measured with MSAP and amplification ISSR patterns when compared before and after the modification of pH culture substrates. These results suggest that DNA methylation might be involved as a molecular mechanism underlying the colour change of hydrangea sepals in response to a differential pH in the substrate. In addition, the results pave the way to study the relationship between DNA methylation and ISSR marker profiles.

Nitrogen Fertilization and Irrigation Frequency Affect Hydrangea Growth and Nutrient Uptake in Two Container Types

Plant growth, water use, photosynthetic performance, and nitrogen (N) uptake of ‘Merritt’s Supreme’ hydrangea (Hydrangea macrophylla) were investigated. Plants were fertilized with one of five N rates (0, 5, 10, 15, or 20 mm from NH4NO3), irrigated once or twice per day with the same total daily amount of water, and grown in either a paper biodegradable container or a traditional plastic container. Greater N rate generally increased plant growth index (PGI) in both plastic and biocontainers. Leaf and total plant dry weight (DW) increased with increasing N rate from 0 to 20 mm and stem and root DW were greatest when fertilized with 15 mm and 20 mm N. Plants fertilized with 20 mm N had the greatest leaf area and chlorophyll content in terms of SPAD reading. Container type had no influence on DW accumulation or leaf area. N concentrations (%) in leaves, roots, and the entire plant increased with increasing N rate. N concentrations in roots and in the entire plant were lower in biocontainers compared with plastic containers. Greater N rate generally increased daily water use (DWU), and biocontainers had greater DWU than plastic containers. The 20 mm N rate resulted in the highest net photosynthetic rate measured on 11 Sept. and 22 Sept. (65 and 76 days after treatment). Net photosynthetic rate (measured on 8 Oct.) and stomatal conductance (gS) (measured on 27 Aug., 22 Sept., and 8 Oct.) were lower in biocontainers compared with plastic containers. Two irrigations per day resulted in higher substrate moisture at 5-cm depth than one irrigation per day, and slightly increased PGI on 19 Aug. However, irrigation frequency did not affect photosynthetic rate, gS, or N uptake of hydrangea plants except in stems. Considering the increased water use of hydrangea plants when grown in the paper biocontainer and lower plant photosynthesis and N uptake, the tested paper biocontainer may not serve as a satisfactory sustainable alternative to traditional plastic containers.

Occurrence of hydrangea ringspot virus and hydrangea chlorotic mottle virus in hydrangea plants in South Korea

Approximately 25 species of Hydrangea, often grown for cut flowers and landscaping, are widely distributed in Asia, the Americas, Europe, and South Africa and can be affected by multiple viruses. To identify any viruses affecting hydrangea populations in South Korea, we analyzed symptomatic samples from three different regions in South Korea using RT-PCR and sequenced the amplified products. From the 211 hydrangea plants sampled with virus-like symptoms such as chlorotic mottle, chlorotic and brown spots on the leaves and distortion of leaves, hydrangea ringspot virus (HdRSV) was found in 33 plants, and hydrangea chlorotic mottle virus (HdCMV) was detected in 10; two plants were co-infected with HdRSV and HdCMV. The coat proteins of HdRSV and HdCMV had the greatest amino acid sequence homology to HdRSV Hyd-CZ (98.7 % identity, GenBank accession GQ265901) and HdCMV-US (97.9 % identity, GenBank accession DQ412999), respectively. To our knowledge, this study is the first report of HdRSV and HdCMV infection in hydrangea plants in South Korea.

Complete Genome Sequences of Two Hydrangea Ringspot Virus Isolates from Japan.

Hydrangea ringspot virus (HdRSV) is a plant RNA virus, naturally infecting Hydrangea macrophylla. Here, we report the first genomic sequences of two HdRSV isolates from hydrangea plants in Japan. The overall nucleotide sequences of these Japanese isolates were 96.0 to 96.3% identical to those of known European isolates.

Foliar Application of Dikegulac Sodium Increases Branching of ‘Merritt’s Supreme’ Bigleaf Hydrangea

The goal of this experiment was to evaluate the efficiency of foliar application of dikegulac sodium on increasing the lateral branching of ‘Merritt’s Supreme’ bigleaf hydrangea (Hydrangea macrophylla). Plants were grown in greenhouses at two locations including El Paso, TX and Kosciusko, MS. Two weeks before application of dikegulac sodium, half of plants were hand-pinched leaving two nodes. Foliar spray of dikegulac sodium at 400, 800, or 1600 mg·L−1 was then applied to pinched and unpinched plants. There were two additional control treatments: pinched or unpinched without application of dikegulac sodium. Data were collected at 2 weeks, 6 weeks, 80 days, and 10 months after treatments. Bigleaf hydrangea plants exhibited severe phytotoxicity including interveinal chlorosis or bleaching of new growth at 2 weeks after application of dikegulac sodium with more pronounced symptoms at higher dikegulac sodium concentrations. The severity of phytotoxicity symptoms became less significant at 6 weeks after treatment. The effect of dikegulac sodium on bigleaf hydrangea plant growth, number of branches, and number of flowers depended on both locations and dosages. In El Paso, TX, dikegulac sodium at 800 or 1600 mg·L−1 inhibited bigleaf hydrangea plant growth at 6 weeks and 80 days after treatment, and this effect disappeared at 10 months after treatment. Dikegulac sodium at all tested dosages doubled or tripled the number of branches of pinched or unpinched bigleaf hydrangea, respectively, at 80 days after treatment. At 10 months after treatment, the number of branches and flowers of bigleaf hydrangea plants tended to increase, but was insignificant. In Kosciusko, MS, dikegulac sodium at 1600 mg·L−1 reduced the plant growth at 6 weeks after treatment. This treatment increased the number of branches and flowers of unpinched plants by 196% and 95% and pinched plants by 53% and 31%, respectively, at 10 months after treatment. Dikegulac sodium application could be used to increase number of branches and flowers and produce compact ‘Merritt’s Supreme’ bigleaf hydrangea. However, the efficacy varied with environmental conditions.

First Report of Alfalfa mosaic virus Occurrence in Hydrangea in the United States.

In spring of 2012, a previously unrecorded virus-like disease characterized by conspicuous yellow leaf blotching (calico symptoms) was observed in plants of Hydrangea macrophylla in a single location in Southampton, NY. Bacilliform and spherical particles resembling those of Alfalfa mosaic virus (AMV) were observed by transmission electron microscopy (TEM) in partially purified extracts from symptomatic leaf tissue. The identity of the virus was confirmed by immunosorbent electron microscopy (ISEM) (4) using antiserum to AMV (ATCC PVAS 92) that both trapped and decorated the virions. Three primer pairs designed from available AMV RNA 1, RNA 2, and RNA 3 genomic sequences were used to generate amplicons from the hydrangea AMV isolate. Reverse-transcription (RT)-PCR was done using total RNA extracted from symptomatic hydrangea leaf tissue with a Qiagen RNeasy kit, and Ready-to-Go RT-PCR beads (GE Healthcare). Amplicons of 1,049, 1,013, and 658 bp were obtained using the primer pairs AMV1F (5'-ATCCACCGATGCCAGCCTTA)/AMV1R (5'-TTCCGCCTCACTGCTGTCTG), AMV2F (5'-GATCGCCGGAAGTGATCCAG)/AMV2R (5'-TCACCGGAAGCAACAACGAA), and AMV3F (5'-GCCGGTTCTCCAAAGGGTCT)/AMV3R (5'-CGCGTCGAAGTCCAGACAGA), respectively. The PCR products were cloned using a TOPO TA cloning kit (Invitrogen) and three clones of each were sequenced. The sequences obtained from the hydrangea AMV RNA 1 (JX154090), RNA 2 (JX154091), and RNA 3 (JX154092) had 95 to 98% nucleotide sequence identity to homologous genomic sequences of known AMV isolates. To our knowledge, this is the first report of AMV occurrence in H. macrophylla in the United States. This virus has been reported to occur in H. macrophylla in British Columbia (3), but in a previous survey its presence was not detected in hydrangeas in the United States (1). A report of possible AMV infection in H. macrophylla in Italy (2) was based solely on symptomatology and cross-protection tests and therefore cannot be verified. The AMV-infected hydrangea plants were found by ISEM to also contain low concentrations of Hydrangea ringspot virus (HRSV) and Hydrangea chlorotic mottle virus (HdCMV). However, based on previous evidence of single and mixed infections (3), it is unlikely that the calico symptoms observed were influenced by the presence of HRSV and HdCMV. This report is of interest both because AMV, unlike HRSV and HdCMV, causes foliar symptoms that would render hydrangea plant unmarketable, and because the disease can be spread by a number of common aphid species that transmit AMV. It will also serve to alert growers and diagnosticians to the potential threat posed by AMV infection.