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Development of bioactive functions in hydrangeae dulcis folium. III. On the antiallergic and antimicrobial principles of hydrangeae dulcis folium. (1). Thunberginols A, B, and F.

From the less polar fraction of Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla Seringe var. thunbergii Makino, Eight antiallergic and antimicrobial principles were isolated together with several known compounds. Among the newly isolated bioactive constituents, the chemical structures of thunberginols A, B, and F have been determined on the basis of chemical and physicochemical evidence. Thunberginols A, B, and F were found to exhibit more potent antiallergic activity than phyllodulcin, hydrangenol, disodium cromoglycate (DSCG), and tranilast. In addition, these thunberginols showed antimicrobial activity against oral bacteria.

Animals↗

Development of bioactive functions in hydrangeae dulcis folium. V. On the antiallergic and antimicrobial principles of hydrangeae dulcis folium. (2). Thunberginols C, D, and E, thunberginol G 3'-O-glucoside, (-)-hydrangenol 4'-o-glucoside, and (+)-hydrangenol 4'-O-glucoside.

Following the characterization of thunberginols A, B, and F, six bioactive principles, thunberginols C, D, and E, thunberginol G 3'-O-glucoside, (-)-hydrangenol 4'-O-glucoside, and (+)-hydrangenol 4'-O-glucoside, were isolated from Hydrangeae Dulcis Folium, the processed leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO, together with four kaempferol and quercetin oligoglycosides. Their chemical structures have been determined on the basis of chemical and physicochemical evidence. Thunberginols C, D, E, and G and (-)-hydrangenol 4'-O-glucoside showed antiallergic activity in the in vitro bioassay using the Schultz-Dale reaction. These components also exhibited inhibitory activities on the histamine release from rat mast cells and on the histamine-induced contraction in isolated guinea pig tracheal chain. In addition, thunberginols C, D, E, and G showed antimicrobial activities against oral bacteria.

Animals↗

[Development of bioactive functions in hydrangeae dulcis folium. II. Antiulcer, antiallergy, and cholagoic effects of the extract from hydrangeae dulcis folium].

In order to develop new bioactive functions of Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla Seringe var. thunbergii Makino, effects of the methanolic extract from the crude drug on antiucler, antiallergic, cholagoic, and various pharmacological actions were investigated. Consequently, the methanolic extract was found to exhibit potent antiulcer, antiallergic, and cholagoic activities. By monitoring with these activities, it was found that the active constituents were contained in the lipophilic portion of the methanolic extract. Furthermore, the known lipophilic constituents such as phyllodulcin and hydrangenol were found to show little antiulcer and cholagoic activities, while it was also found that they showed antiallergic activity on Schultz-Dale reactions.

Animals↗

Genetic diversity, phylogenetic relationships, and marker development between Hydrangea serrata and H. macrophylla based on plastome and 45S nrDNA.

Ornamental hydrangeas (genus Hydrangea) are cultivated worldwide for their diverse flower colors and attractive morphology. Here, we assembled the complete plastid genome (plastome) and 45S nuclear ribosomal DNA (45S nrDNA) sequences of 22 individuals representing H. serrata, H. macrophylla, and related species (H. arborescens, H. paniculata, H. petiolaris, and H. hydrangeoides). The plastomes contained up to 2,344 single-nucleotide polymorphisms (SNPs) and 367 insertions/deletions (InDels) within the genus, whereas the assembled 45S nrDNA sequences showed 119 SNPs and 10 InDels. Phylogenetic analyses based on plastome and 45S nrDNA sequences clearly separated H. serrata and H. macrophylla from the other Hydrangea species. In the plastome-based tree, H. petiolaris was placed in the same clade as H. arborescens, whereas in the 45S nrDNA-based tree it showed a close relationship to H. hydrangeoides. The H. serrata and H. macrophylla samples were not always separated according to their species boundaries, as observed in samples Hse8-Hse12. Notably, one H. serrata sample (Hse8), collected from a wild mountainous region of Japan, exhibited a closer genetic relationship to H. macrophylla samples, indicating that cultivated hydrangeas may have originated from a specific wild lineage of H. serrata adapted to mountainous habitats. Using plastome-derived molecular markers, 66 Hydrangea samples were further classified into five groups, with Group II comprising both cultivated H. macrophylla and a subset of wild H. serrata samples, suggesting a close genetic affinity between this group and the ancestral gene pool of cultivated H. macrophylla. Based on these genomic resources, eight plastome-derived molecular markers were developed to differentiate cultivated hydrangeas from wild genotypes and to assess genetic diversity within H. serrata and H. macrophylla, providing practical tools for germplasm identification, breeding, and genetic resource management of Hydrangea species.

hydrangea↗

Genome-wide AP2/ERF analysis identifies HmaERF87 as a positive regulator of Hydrangea macrophylla leaf spot resistance.

A total of 164 APETALA2/ethylene-responsive factor (AP2/ERF) genes were identified in Hydrangea macrophylla, and HmaERF87 positively contributes to leaf spot resistance. The APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor family plays important roles in plant stress responses, but its contribution to disease resistance in Hydrangea macrophylla (hydrangea) remains poorly understood. In this study, 164 AP2/ERF genes were identified in the H. macrophylla genome and classified into APETALA2 (AP2), ethylene-responsive factor (ERF), dehydration-responsive element-binding (DREB), and related to ABI3/VP1 (RAV) subfamilies. Their chromosomal distribution, conserved motifs, gene structures, and duplication patterns were analyzed. A total of 46 pathogen-responsive H. macrophylla AP2/ERF (HmaERF) genes were identified from the RNA sequencing (RNA-seq) dataset of resistant and susceptible cultivar leaves collected before and after Corynespora cassiicola inoculation. Promoter analysis revealed that the HmaERF genes with upregulated expression post-C. cassiicola infection showed a higher frequency and copy number of jasmonate-responsive cis-regulatory elements, suggesting their possible involvement in hormone-mediated defense responses. Three infection-induced candidate genes, including HmaERF56, HmaERF87, and HmaERF129, were selected for functional validation using virus-induced gene silencing (VIGS) in hydrangea leaf discs. Silencing of HmaERF87 expression via VIGS significantly increased lesion development after C. cassiicola inoculation, whereas the transient overexpression of HmaERF87 reduced the area of leaf disc lesions. Subcellular localization showed that the HmaERF87 protein was localized in the nucleus, and yeast assays indicated that its transcriptional activation activity was mainly associated with the C-terminal region of the protein. These results support a role for HmaERF87 as a positive regulator of H. macrophylla resistance to leaf spot disease and provide a candidate gene for further studies of disease resistance in hydrangea.

Plant Proteins↗

Population dynamics of Tetranychus kanzawai (Acari: Tetranychidae) on hydrangea.

The seasonal occurrence of Tetranychus kanzawai Kishida populations on hydrangea (Hydrangea macrophylla) was studied at two different localities in Ibaraki, Japan, during a three-year period. There were two types of seasonal population trends: one with a population peak from May to June, and the other with the spring peak in June and the autumn peak in September-October. Each year the populations on hydrangea plants abruptly declined just after the spring peak. Predators showed a delayed density-dependent numerical response. The population crashed even in the absence of predators, suggesting that the predators had nothing to do with the June decline. Furthermore, the rate of development from larva to adult and the fecundity in adult females on detached hydrangea leaves decreased markedly just prior to the abrupt decline in density in June. Consequently, seasonal changes in plant quality (perhaps influenced by secondary compounds) seem to contribute to the drastic decline of T. kanzawai density on hydrangea in June.

Animals↗

Allergic contact dermatitis from hydrangea--is it so rare?

8 cases of allergic contact dermatitis from hydrangea seen in Angers, France, during the last 15 years are reported and compared to other cases found in the literature. In this review, allergic contact dermatitis from hydrangeas appears to be an occupational dermatosis among nursery workers, presenting as an eczema involving the hands and especially the first 3 fingers, with chronic features of fissuring and scaling and with a chronic course. Differential diagnosis from irritant contact dermatitis may be difficult. Patch tests with the stem as well as the leaf of hydrangeas gave strong positive reactions in all patients, and hydrangenol, the allergen of hydrangea, when tested, always also gave a positive reaction. Sensitization seems to occur after close and prolonged contact with the plant, which could explain the relative frequency in Angers because it provides almost 90% of hydrangea seedling production in France.

Adult↗

Internal Detoxification Mechanism of Al in Hydrangea (Identification of Al Form in the Leaves).

An internal detoxification mechanism for Al was investigated in an Al-accumulating plant, hydrangea (Hydrangea macrophylla), focusing on Al forms present in the cells. The leaves of hydrangea contained as much as 15.7 mmol Al kg-1 fresh weight, and more than two-thirds of the Al was found in the cell sap. Using 27Al- nuclear magnetic resonance, the dominant peak of Al was observed at a chemical shift of 11 to 12 parts per million in both intact leaves and the extracted cell sap, which is in good accordance with the chemical shift for the 1:1 Al-citrate complex. Purification of cell sap by molecular sieve chromatography (Sephadex G-10) combined with ion-exclusion chromatography indicated that Al in fractions with the same retention time as citric acid contributed to the observed 27Al peak in the intact leaves. The molar ratio of Al to citric acid in the crude and purified cell sap approximated 1. The structure of the ligand chelated with Al was identified to be citric acid. Bioassay experiments showed that the purified Al complex from the cell sap did not inhibit root elongation of corn (Zea mays L.) and the viability of cells on the root tip surface was also not affected. These observations indicate that Al is bound to citric acid in the cells of hydrangea leaves.

Journal Article↗

Anti-malarial activity of leaf-extract of hydrangea macrophylla, a common Japanese plant.

To find a new anti-malarial medicine derived from natural resources, we examined the leaves of 13 common Japanese plants in vitro. Among them, a leaf-extract of Hydrangea macrophylla, a common Japanese flower, inhibited the parasitic growth of Plasmodium falciparum. The IC50 of Hydrangea macrophylla leaf extract to Plasmodium falciparum was 0.18 microg/ml. The IC50 to NIH 3T3-3 cells, from a normal mouse cell line, was 7.2 microg/ml. Thus, selective toxicity was 40. For the in vivo test, we inoculated Plasmodium berghei, a rodent malaria parasite, to ddY mice and administered the leaf-extract of Hydrangea macrophylla (3.6 mg/0.2 ml) orally 3 times a day for 3 days. Malaria parasites did not appear in the blood of in the treated mice, but they did appear in the control group on day 3 or 4 after inoculation with the parasites. When leaf extract was administered to 5 mice 2 times a day for 3 days, malaria parasites did not appear in 4 of the mice but did appear in 1 mouse. In addition, the leaf-extract was administered orally 3 times a day for 3 days to Plasmodium berghei infected mice with a parasitemia of 2.7%. In the latter group, malaria parasites disappeared on day 3 after initiating the treatment, but they appeared again after day 5 or 6. Although we could not cure the mice entirely, we confirmed that the Hydrangea macrophylla leaf extract did contain an anti-malarial substance that can be administered orally.

Animals↗

Synonymy between two spider mite species, Tetranychus kanzawai and T. hydrangeae (Acari: Tetranychidae), shown by ribosomal ITS2 sequences and cross-breeding experiments.

Amplification of the second internal transcribed spacer (ITS2) of ribosomal DNA was used to compare seven samples of the Tetranychus kanzawai Kishida-- T. hydrangeae Pritchard & Baker mite complex from five different countries: Australia, the Congo, Indonesia, Japan and the USA. No morphological differences were detected between these mites and their ITS2 sequences displayed strong similarity except for a small nucleotide divergence of 0.2% in specimens from Australia and Indonesia. Reciprocal crosses and backcrosses between mites assumed to be T. kanzawai and T. hydrangeae respectively showed reproductive compatibility. Fertile hybrid females were obtained in all cases, indicating conspecificity of the mites tested. It is concluded that T. hydrangeae is a synonym of T. kanzawai. The evidence suggests that T. kanzawai originated in South-east Asia and probably spread throughout the world on Hydrangea spp. cuttings.

Animals↗

Occupational contact dermatitis to hydrangea.

Two female commercial hydrangea growers, from separate nurseries, presented with similar hand and facial dermatitis. Both had a hand dermatitis affecting particularly the first three fingers and backs of both hands and complained of a recurrent facial dermatitis affecting the forehead, around both the eyes and bridge of nose. They related their dermatitis to their work. Patch tests confirmed allergy to all components of hydrangeas including petal, leaf and stem. Avoidance resulted in resolution of their dermatoses. Allergy to hydrangeas has been reported previously although infrequently.

Adult↗

Sepal color variation of Hydrangea macrophylla and vacuolar pH measured with a proton-selective microelectrode.

Sepal color of hydrangea varies with the environmental conditions. Although chemical and biological studies on this color variation have a long history, little correct knowledge has been generated about color development. All colored sepals contain the same anthocyanin, delphinidin 3-glucoside. Thus, there must be some other system for developing the wide variety of colors. In hydrangea sepals the cells of the epidermis are colorless and only the second layer of cells contain pigment. We prepared protoplasts without any color change during enzyme treatment of sepals and measured the vacuolar pH of each of the colored cells. We could correlate the color of a single hydrangea cell with its vacuolar pH using a combination of micro-spectrophotometry and a proton-selective microelectrode. Values for the vacuolar pH of blue (lambda vismax: 589 nm) and red cells (lambda vismax: 537 nm) were 4.1 and 3.3, respectively, the vacuolar pH of blue cells being significantly higher.

Anthocyanins↗

[Quantitative analysis of dihydroisocoumarin constituents of Hydrangeae Dulcis Folium by means of high performance liquid chromatography. Chemical characterization of the processing, distribution in plant, and seasonal fluctuation].

In order to characterize the chemical change of the constituents during the processing of Hydrangeae Dulcis Folium, quantitative analyses of phyllodulcin, hydrangenol, and their 8-O-glucosides were developed by means of high performance liquid chromatography. As an application of this HPLC method, the distribution of those dihydroisocoumarins in different parts of Hydrangea macrophylla var. thunbergii was investigated. It was found that these dihydroisocoumarins were contained at the highest concentration in the leaves. Furthermore, the seasonal fluctuation of these compounds in the leaves, together with the height of the plant and total dry weight of the leaves, were clarified and so that the suitable period for the harvest of Hydrangea macrophylla var. thunbergii was deduced to be from Oct. to Nov.

Benzopyrans↗

Suppression by Hydrangeae Dulcis Folium of D-galactosamine-induced liver injury in vitro and in vivo.

Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla SER. var. thunbergii MAKINO, suppressed D-galactosamine-induced liver injury by 85.2% when added to the diet at 1% and fed to rats for fifteen days. The hepatoprotective effect is more potent than that of a milk thistle extract and turmeric powder. Some fractionated extracts showed hepatoprotective activity in the D-galactosamine-induced in vitro liver injury model.

Alanine Transaminase↗

Immunomodulatory activity of thunberginol A and related compounds isolated from Hydrangeae Dulcis Folium on splenocyte proliferation activated by mitogens.

We investigated the immunomodulatory effects of antiallergic constituents from Hydrangeae Dulcis Folium, the processed leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO, on splenocyte proliferation in mice. Thunberginol A and hydrangenol significantly suppressed T lymphocyte proliferation induced by concanavalin A. Thunberginol A also suppressed B lymphocyte proliferation induced by lipopolysaccharide, but other constituents induced significant increases. These inhibitory effects of thunberginol A on splenocyte proliferation seemed to contribute to the suppressive effect on type IV allergy.

Adjuvants, Immunologic↗

Terpenoids of Salvia hydrangea: two new, rearranged 20-norabietanes and the effect of oleanolic acid on erythrocyte membranes.

Four abietane-type terpenoids, including two known royleanones and two new, rearranged 20-norabietanes, were isolated from the roots of the Iranian medicinal plant Salvia hydrangea DC. ex Bentham (Lamiaceae), which is used as an anthelmintic and antileishmanial remedy. Their structures were established using COSY, NOESY, HSQC, and HMBC spectral data. The possible identity of one of the 20-norabietanes with demethylmulticauline, previously reported from a different Salvia species, is discussed. A moderate in vitro antiplasmodial effect of the extract of S. hydrangea flowers was found to be associated with the presence of large amounts of pentacyclic triterpenes, mainly oleanolic acid. The observed antiplasmodial activity of oleanolic acid is apparently due to its incorporation into the erythrocyte membrane, which adversely affects the growth of Plasmodium falciparum parasites. Thus, oleanolic acid caused transformation of erythrocytes into stomatocytes in the concentration range where the in vitro antiplasmodial activity was observed.

Abietanes↗

'Candidatus phytoplasma japonicum', a new phytoplasma taxon associated with Japanese Hydrangea phyllody.

A phytoplasma was discovered in diseased specimens of field-grown hortensia (Hydrangea spp.) exhibiting typical phyllody symptoms. PCR amplification of DNA using phytoplasma specific primers detected phytoplasma DNA in all of the diseased plants examined. No phytoplasma DNA was found in healthy hortensia seedlings. RFLP patterns of amplified 16S rDNA differed from the patterns previously described for other phytoplasmas including six isolates of foreign hortensia phytoplasmas. Based on the RFLP, the Japanese Hydrangea phyllody (JHP) phytoplasma was classified as a representative of a new subgroup in the phytoplasma 16S rRNA group I (aster yellows, onion yellows, all of the previously reported hortensia phytoplasmas, and related phytoplasmas). A phylogenetic analysis of 16S rRNA gene sequences from this and other group I phytoplasmas identified the JHP phytoplasma as a member of a distinct sub-group (sub-group Id) in the phytoplasma clade of the class Mollicutes. The phylogenetic tree constructed from 16S rRNA gene sequences was consistent with the hypothesis that the JHP phytoplasma and its closest known relatives, the Australian grapevine yellows (AUSGY), Phormium yellow leaf (PYL), Stolbur of Capsicum annuum (STOL) and Vergilbungskrankheit of grapevine (VK) share a common ancestor. The unique properties of the DNA from the JHP phytoplasma clearly establish that it represents a new taxon, 'Candidatus Phytoplasma japonicum'.

DNA, Bacterial↗