Dendrobium Flowers Research Articles

Peppermint essential oil enhances the vase life of Dendrobium orchids

To extend the vase life of cut flowers, there is now a trend of using plant essential oils in place of synthetic chemicals, as they are fully biodegradable, more eco-friendly, and safer. The objective of this study was to examine the possible application and postharvest quality effects of three plant essential oils namely, ginger (Zingiber officinale Roscoe), peppermint (Mentha piperita L.), and citronella (Cymbopogon nardus Rendle), as natural vase solution for cut Dendrobium flowers. Peppermint essential oil showed promise as a holding solution for extending the vase life of Dendrobium orchids. To confirm vase life extension, emulsions containing peppermint essential oil at concentrations of 50 and 100 μg mL−1 combined with 4 % glucose to formulate holding solutions applied to Dendrobium orchids. Vase life, some biochemical changes, electrolyte leakage, total microbial count in the holding solution, and physical condition via scanning electron microscopy (SEM) were evaluated over a period of 25 days. The three major compounds in peppermint essential oil were identified as menthol (33.24 %), 1-menthone (18.91 %) and menthofuran (14.85 %). The essential oil was applied in emulsion form as a holding solution. Treatment with 4 % glucose and either 50 or 100 μg mL−1 peppermint essential oil prolonged the vase life of Dendrobium orchids to up to 28 days. Scanning electron microscopy on Day 7 showed that the xylem vessels of treated orchids remained clear, suggesting reduced microbial plugging at the stalk end. Similarly, on Day 20, a reduced microbial cell count was observed for treated orchids (<1 log CFU mL−1) in comparison with controls (7.20 ± 0.04 log CFU mL−1). Finally, the essential oil improved flower quality by helping preserve petal membrane stability and petal anthocyanin content. Our results suggest the application of peppermint essential oil as a novel alternative to chemicals used in holding solutions for extending the vase life of Dendrobium orchids.

Analysis of metabolic differences between Jiaosu fermented from dendrobium flowers and stems based on untargeted metabolomics

Dendrobium officinale is an important traditional Chinese medicinal herb containing bioactive polysaccharides and alkaloids. This study characterized metabolite differences between jiaosu (fermented plant product) from Dendrobium flowers versus stems using untargeted metabolomics. The jiaosu was fermented by mixed fermentation of Saccharomyces cerevisiae, Lactobacillus bulgaricus and Streptococcus thermophilus. Liquid chromatography-mass spectrometry analysis identified 476 differentially expressed metabolites between the two Jiaosu products. Key results showed downregulation of flavonoid metabolism in Dendrobium Stems Edible Plant Jiaosu (SEP) but increased flavonoid synthesis in Dendrobium Flowers Edible Plant Jiaosu (FEP), likely an antioxidant response. SEP displayed upregulation of lignin metabolites with potential antioxidant properties. The findings demonstrate significant metabolite profile differences between SEP and FEP, providing the basis for developing functional jiaosu products targeting specific health benefits.

Transcriptomic and Metabolomic Analyses Reveal Differences in Flavonoid Pathway Gene Expression Profiles between Two Dendrobium Varieties during Vernalization.

Dendrobium (Orchidaceae, Epidendoideae) plants have flowers with a wide variety of colors that persist for a long period throughout the year. The yellow coloration of Dendrobium flowers is mainly determined by the flavonol pathway and the flavone pathway, but the relevant biosynthesis mechanisms during vernalization remain unclear. To explore the similarities and differences in flavonoid biosynthesis in different tissues during vernalization, we selected two species of Dendrobium for a flower color study: Dendrobium capillipes Rchb (which has yellow flowers) and Dendrobium nobile Lindl (which has white flowers). We collected a total of 36 samples from six tissue types and both Dendrobium species during vernalization and subjected the samples to metabolic profiling and transcriptome sequencing. A total of 31,504 differentially expressed genes (DEGs) were identified between different tissues of the two Dendrobium species by transcriptomic analysis. However, many differentially accumulated metabolites (DAMs) and DEGs were enriched not only in the general pathway of "flavonoid biosynthesis" but also in multiple subpathways of "flavone and flavonol biosynthesis". According to a combined transcriptome and metabolome analysis, Putrescine hydroxycinnamoyl transferase 1 (LOC110093422) may be the main gene responsible for the differences in flavonoid accumulation during vernalization, which is closely associated with yellow flowers. Taken together, the results of our study preliminarily revealed the metabolites responsible for and the key genes regulating flavonoid biosynthesis during vernalization. These results provide a basis for the further study of the molecular mechanism of flavonoid synthesis during vernalization.

1-MCP prevents ultrastructural changes in the organelles of Dendrobium petals that are induced by exogenous ethylene

Ethylene is a plant hormone that causes flower senescence. Dendrobium flowers are sensitive to ethylene and ethylene can induce premature senescence depending on the cultivar and the ethylene concentration. Dendrobium ‘Lucky Duan’ is one of the most sensitive cultivars to ethylene exposure. Open florets of ‘Lucky Duan’ were subjected to ethylene, 1-MCP, or 1-MCP plus ethylene treatments and compared with an untreated control. Ethylene induced earlier development of color fading, drooping and venation in petals, whereas 1-MCP pre-treatment counteracted these changes. Under light microscopy, epidermal cells and mesophyll parenchyma tissue around the vascular bundles of petals treated with ethylene showed collapsed cells whereas 1-MCP pre-treatment counteracted this collapse. An scanning electron microscopy (SEM) study confirmed clearly that ethylene treatment caused the collapse of mesophyll parenchyma tissue around vascular bundles. Ultrastructural changes were also studied using transmission electron microscopy (TEM) and showed that ethylene treatment induced morphological changes in conjunction with disorganization of the plasma membrane, the nuclei, chromatin, the nucleoli, myelin bodies, multivesicular bodies, and mitochondria including changes in size and number, breakages of membranes, enlargement of intercellular spaces and disintegration. 1-MCP pre-treatment was observed to counter these changes that were induced by ethylene. The role of ethylene-induced ultrastructural changes in the different organelles was apparently associated with membrane damage.

The Dendrobium Orchid: Botany, horticulture, and utilization

AbstractDendrobium, one of the largest genera of tropical orchids, is grown commercially on a large scale with an important economic impact in several ASEAN countries.Dendrobiumorchids can be grown in a wide range of climates, but flower production is best under tropical conditions. Due to being the most popular orchid for cut flower production, many hybrids ofDendrobiumhave been produced with colorful and attractive blooms. Cut flowers ofDendrobiumare popular worldwide. In addition, manyDendrobiumcultivars can be grown as flowering potted plants and for other purposes such as in medicinal applications and for use in decorative products. Postharvest physiology and technology are important in maintaining the quality of both cut flowers and potted plants from production sites to the marketplace. Plants of theDendrobiumgenus also contain a wide diversity of bioactive compounds that can be developed into medicinal and cosmetic products. This review describes the botany, cultivation, postharvest physiology, handling and utilization ofDendrobium.

Complementary water and nutrient utilization of perianth structural units help maintain long floral lifespan in Dendrobium.

Most orchids have high ornamental value with long-lived flowers. However, the mechanisms by which orchids maintain floral longevity are poorly understood. Here, we hypothesized that floral longevity in Dendrobium is maintained by high resource investment and complementary water and nutrient utilization in different structural units of the perianth. To test this hypothesis, we determined which water- and nutrient-related traits are correlated with flower longevity in 23 Dendrobium species or cultivars, and examined variations of the related traits during flower development of one long-lived cultivar. We found that floral longevity was correlated with dry mass per unit area of perianths and total flower biomass, which indicates that maintaining floral longevity requires increased resource investment. During development of long-lived flowers, labella showed a high capacity for water storage and nutrient reutilization, which could partly remedy high water demand and biomass investment. Sepals and petals, in contrast, had stronger desiccation avoidance and higher metabolic activity with lower biomass investment. These findings indicate that Dendrobium flowers maintain longevity by complementary water and nutrient utilization strategies in the sepals, petals and labella, with labella consuming more water and nutrients to extend flower display, and sepals and petals using a more conservative strategy.

Differential expression of ethylene biosynthetic and receptor genes in pollination-induced senescence of Dendrobium flowers

Following successful pollination, Dendrobium orchid flowers rapidly undergo senescence. In Dendrobium cv. Khao Chaimongkol, compatible pollination resulted in faster ethylene production and more rapid development of senescence symptoms, such as drooping, epinasty, venation and yellowing, compared with non-pollinated controls or pollination with incompatible pollinia. The DenACS1 and DenACO1 genes in the perianth of florets that had been pollinated with compatible pollinia were expressed more highly than those in non-pollinated open florets. Incompatible pollinia reduced the expression of DenACS1 and DenACO1 genes in the perianth. Transcript levels of the ethylene receptor gene DenERS1 and signaling genes DenEIL1 and DenERF1 showed differential spatial regulation with greater expression in the perianth than in the column plus ovary following compatible pollination. Compatible pollinia increased ethylene production concomitant with premature senescence and the increased expression of the DenACS1 and DenACO1 genes, and suppressed the ethylene receptor gene DenERS1, whereas incompatible pollinia did not stimulate ethylene production nor induce premature senescence but induced higher expression of DenERS1 both in the perianth and in the column plus ovary. These results suggest that the increased ethylene production in open florets pollinated with compatible pollen was partially due to an increase in the expression of DenACS1 and DenACO1 genes. The compatible pollinia induced a negative regulation of DenERS1 which may play an important role in ethylene perception and in modulating ethylene signaling transduction during pollinia-induced flower senescence.

1-Methylcyclopropene inhibits ethylene production and post-pollination changes of Dendrobium flowers

1-Methylcyclopropene inhibits ethylene production and post-pollination changes of Dendrobium flowers

Influence of sucrose feeding on sugar metabolism and invertase activity in cut Dendrobium flowers

Influence of sucrose feeding on sugar metabolism and invertase activity in cut Dendrobium flowers

High-quality RNA isolation from pigment-rich Dendrobium flowers.

Isolation of high-quality RNA from Dendrobium flowers is challenging because of the high levels of pigment, polysaccharides, and polyphenols. In the present study, an efficient CTAB method for RNA extraction from the pigment-rich flowers of Dendrobium was optimised. The optimised method yielded high quantities of RNA (10.1-12.9µg/g). Spectrophotometric values of A260/280 in the range of 2.2 to 2.4 and A260/230 values of 2.0 suggested that the isolated RNA was free of polyphenols, polysaccharides, and protein contaminants. RNA integrity numbers determined by microfluidics were in the range of 7.9-8.9 indicative of intact RNA. In the improved method, the addition of 3M NaCl and 3% PVP-10 in the extraction buffer, followed by an incubation period of 45min at 65°C, eliminated most of the polysaccharides, polyphenolic compounds, and denatured protein. Extraction with phenol:chloroform:isoamyl alcohol (125:24:1) effectively removed pigments from the aqueous phase, while the precipitation of RNA with lithium chloride minimised the co-precipitation of protein, DNA, and polysaccharide and resulted in the extraction of high quality of RNA. The suitability of the RNA for downstream processing was confirmed via RT-PCR amplification of Chalcone synthase gene from cDNA prepared from RNA isolated from different developmental stages of the flower of a Dendrobium hybrid. The present method will be highly useful for the isolation of RNA from pigment, polyphenol, and polysaccharide-rich plant tissues.

Increased hydrolase gene expression and hydrolase activity in the abscission zone involved in chilling-induced abscission of Dendrobium flowers

After 4days exposure to15°C and 5°C temperature, cut cv. Ear Sakul Dendrobium inflorescences were returned to 25°C.The exposure of open flowers and floral buds to 5°C induced more abscission than exposure to 15°C after transfer to 25°C. Ethylene production rates of floral buds and open flowers with and without 1-MCP treatment were comparatively low during exposure to to15°C and 5°C. Flower buds had a greater increase of ethylene production than open flowers after upon transfer to 25°C and flower buds stored at 5°C had a greater increase than that of flower buds stored at 15°C. 1-MCP treatment reduced significantly more ethylene production rates of floral buds stored at 5°C than those stored at 15°C compared with open flowers upon transfer to 25°C on the last 3days. The abscission zone (AZ) activities of pectin methylesterase (PME), polygalacturonase (PG), β-1,4 glucanase (cellulase, CEL) and β-galactosidase (β-GAL) increased after exposure to5°C, while β-GAL also increased after 15°C exposure. Treatment with 100nlL−1 1-MCP for 4h at 25°C, lower temperature exposure reduced abscission in both open flowers and floral buds. 1-MCP also prevented an increase in the activities of PME, PG, CEL and β-GAL in the AZ. Genes encoding a PME, a CEL, and a β-GAL were isolated from the AZ. In the absence of 1-MCP an early increase was found in the expression of the β-GAL and PME genes in the AZ, that was correlated with the abscission rates. An increase in the expression of the CEL gene was found later in abscission. 1-MCP prevented the increase in expression of the β-GAL, PME and CEL genes in the AZ. It was concluded that chilling-induced abscission was mediated by ethylene, that induced gene expression and increased the activity of cell wall hydrolases. The high correlation between abscission and gene expression of β-GAL and CEL suggests that they were involved relatively early and again in late stages of abscission. The role of PME and PG was less clear, but they were possibly involved in the abscission after the 5°C treatment.

Vesicles between plasma membrane and cell wall prior to visible senescence of Iris and Dendrobium flowers

Cut Iris flowers (Iris x hollandica, cv. Blue Magic) show visible senescence about two days after full opening. Epidermal cells of the outer tepals collapse due to programmed cell death (PCD). Transmission electron microscopy (TEM) showed irregular swelling of the cell walls, starting prior to cell collapse. Compared to cells in flowers that had just opened, wall thickness increased up to tenfold prior to cell death. Fibrils were visible in the swollen walls. After cell death very little of the cell wall remained. Prior to and during visible wall swelling, vesicles (paramural bodies) were observed between the plasma membrane and the cell walls. The vesicles were also found in groups and were accompanied by amorphous substance. They usually showed a single membrane, and had a variety of diameters and electron densities. Cut Dendrobium hybrid cv. Lucky Duan flowers exhibited visible senescence about 14 days after full flower opening. Paramural bodies were also found in Dendrobium tepal epidermis and mesophyll cells, related to wall swelling and degradation. Although alternative explanations are well possible, it is hypothesized that paramural bodies carry enzymes involved in cell wall breakdown. The literature has not yet reported such bodies in association with senescence/PCD.

Pollinia-borne chemicals that induce early postpollination effects in Dendrobium flowers move rapidly into agar blocks and include ACC and compounds with auxin activity

The early visible effects of pollination in orchids are likely due to pollinia-borne chemicals. In Dendrobium we tested whether such compounds were water soluble and would diffuse in solid-aqueous phase, and determined both 1-aminocyclopropane-1-carboxylic acid (ACC) concentrations and auxin activity. Following pollination, the flower peduncle showed epinastic movement, followed by yellowing of the flower lip, flower senescence and ovary growth. Placing pollinia on agar blocks for 3, 6, 9 or 12h, prior to transferring them to the stigma, increased the time to these early postpollination effects or prevented them. Placing agar blocks that had been used for contact with the pollinia on the stigma also induced the early postpollination effects. The concentrations of ACC, the direct precursor of ethylene, in pollinia was lower the longer the pollinia had been in contact with the agar blocks, whilst the ACC content in the agar blocks increased with the period of contact. The auxin activity of the agar blocks also increased with the time of contact with pollinia. It is concluded that chemicals in the pollinia are responsible for the early visible postpollination effects, and that these (a) rapidly diffuse in aqueous media, and (b) comprise at least ACC and compounds with auxin activity. The idea is discussed that ACC plus auxin is adequate for the production of the early postpollination effects.

Population ecology of Thrips palmi (Thysanoptera:Thripidae) in orchid farms in Thailand

The population dynamics and in-farm distribution of Thrips palmi Karny were evaluated in two orchid farms, one in the Bangkok metropolitan area and the other in Ratchaburi Province, Thailand. Yellow sticky traps were hung above the orchid canopy (three traps/220 m2), and the trapped adults were counted weekly from November 2009 to October 2010 at the first location and from January to December of 2012 at the second location. T. palmi nymphs and adults were both counted weekly on 300 orchid panicles in an area of 2,200 m2 at both locations. We found that the T. palmi population dynamics at the first location was represented by a W-shaped curve, with the distribution clumped throughout the year, whereas the curve was J-shaped at the second location, with the distribution mostly clumped. Qualitative surveys of thrips species were conducted on ten different orchid genera in ten locations and on 20 different Dendrobium hybrids in 20 locations. Five random sampling plots (1 × 4 m2) were examined on each farm. The results indicated that only T. palmi was found on every farm. Moreover, quantitative comparison among five orchid types revealed that Dendrobium flowers with darker color are more attractive to T. palmi. However, this phenomenon does not coincide with Oncidium sp. and Mokara sp. because of their unfavorable morphology for thrips behavior.

Role of stigmatic fluid in ovary growth and senescence of pollinated Dendrobium flowers

Pollination induced an increase in ethylene production concomitantly with ovary growth, epinasty, drooping, changes in venation andfading of the colour of the flowers of Dendrobium ‘Pompadour’. Removal of stigmatic fluid prior to pollination significantly reduced ovarygrowth, while ethylene production, epinasty, drooping, venation and colour of pollinated flowers were not significantly affected. Removalof stigma fluid had no effect on ovary growth of non-pollinated flowers treated with 1-aminocyclopropane-1-carboxylic acid (ACC ), butsignificantly reduced the ovary growth of non-pollinated flowers induced by the application of napthaleneacetic acid (NAA). The effect ofstigmatic fluid on ovary growth of pollinated flowers is discussed.

Compatible and incompatible pollination and the senescence and ovary growth of Dendrobium flowers

The pollinia from five cultivars of Dendrobium (‘Miss Teen’, ‘Karen’, ‘Sakura’, ‘Willie’, and ‘Pompadour’) were placed on the stigma of openflowers of Dendrobium ‘Miss Teen’ (or ‘Kenny’). The ethylene production and post-pollination development of cross-pollinated flowerswere monitored. The pollinated flowers showed two different development patterns depending upon pollination compatibility. The firstgroup, which was pollinated with incompatible pollen from flowers of ‘Karen’ and ‘Miss Teen’, did not exhibit post-pollination symptomsand ovary growth. The second group, which was pollinated with compatible pollen from flowers of ‘Pompadour’, ‘Sakura’ and ‘Willie’,showed premature perianth senescence and induced ovary growth within a day. Compatible pollination of ‘Miss Teen’ flowers resulted inan immediate burst in the production of ethylene and a rapid ovary growth followed by an increase in pollen tube length. In contrast,when ‘Miss Teen’ flowers were self-pollinated or pollinated with ‘Karen’ pollinia, these changes did not occur. In the pollinia of ‘Pompadour’,‘Sakura’ and ‘Willie’ there were higher levels of auxin and 1-aminocyclopropane-1-carboxylic acicd (ACC ) than in those of ‘Miss Teen’and ‘Karen’. Compatible pollination also resulted in higher ACC synthase and ACC oxidase activities within 12 h after pollination than afterpollination with incompatible pollen. There was a low level of ACC oxidase activity in non-pollinated flowers and a higher ACC oxidaseactivity in the column plus pedicel than petal plus sepal and lip.

Isolation of an actin promoter for strong expression of transgenes in the orchid genus Dendrobium

We isolated and functionally characterized a Dendrobium Actin1 (DmACT1) promoter that drives strong gene expression in the orchid genus Dendrobium. A genomic fragment containing the region 3227 bp upstream of the coding region of DmACT1 was obtained by inverse PCR. Detailed comparison of the full-length cDNA and genomic sequences revealed that DmACT1 has a 1374 bp first intron in the 5' UTR. However, the 5' flanking sequences upstream of the coding region showed no obvious sequence similarities compared to those of known promoters, including plant actin promoters. Serial deletion constructs of the 5' flanking region from the translation initiation codon were fused to the coding sequence of a GUS/luciferase fusion reporter to identify the regulatory elements necessary for promoter activity. Transient assays in the flowers of Dendrobium revealed that the 5' UTR-intron greatly enhanced promoter activity. Moreover, the DmACT1 promoter with its 5' UTR-intron yielded approximately 10-fold higher reporter activity than the rice Act1 promoter-intron. Our data suggest that the DmACT1 promoter with its 5' UTR-intron is a useful tool for strong expression of transgenes in Dendrobium orchids.

Effect of wetting period on infection of orchid flowers by Alternaria alternata and Curvularia eragrostidis

The effect of different durations of the incubation wetting period on the infection of Dendrobium flowers by two serious pathogens—Alternaria alternata (Fr.) Keissl. 1912 and Curvularia eragrostidis (Henn.) J.A. Mey. 1959—was investigated. Cut orchid flowers of the genus Dendrobium were used for the inoculation. The courses of symptom development and mycelium growth were determined. The results indicate that the infection and the subsequent development of disease on the flowers occur rapidly even if the conditions favourable for pathogen germination on the plant tissues persist for only 2 h. However, the proportion of infected flowers was significantly higher after a longer incubation period (6 h). The mycelium growth of the pathogen was also significantly influenced by the length of the incubation period.

Sucrose feeding of Cut Dendrobium inflorescences promotes bud opening, inhibits abscission of open flowers, and delays tepal senescence

Inflorescences of Dendrobium cv. Khao Sanan were held in distilled water (controls) or in solutions containing two antimicrobial compounds (silver nitrate and 8-hydroxyquinoline sulfate) with or without 4gL−1 sucrose or 4gL−1 glucose. Sugar+antimicrobial compounds promoted bud opening and largely prevented abscission of open flowers. It also resulted in a delay of tepal senescence. No clear effect was found of sucrose feeding on the concentrations of sucrose, glucose and fructose in the tepals of open flowers, but an increase in sucrose, glucose, and fructose concentrations was found in the column+labellum. It is concluded that the effect of sugar feeding on abscission and tepal senescence might relate to a reduction of ethylene sensitivity. It was previously concluded that tepal senescence in Dendrobium flowers was not regulated in the tepals but in the column. It is not clear how the increased in sugar levels in the column+labellum relates to the delay of tepal senescence after sugar feeding.

Ethylene-regulated hastening of perianth senescence after pollination in Dendrobium flowers is not due to an increase in perianth ethylene production

Flowers of Dendrobium cv. Kenny were hand-pollinated using pollinia from cv. Sakura. This resulted in a large increase in flower ethylene emission and rapid perianth (tepal) senescence. The increase in flower ethylene emission was correlated in time with an increase in ethylene emitted by the column (the fused stigma, style and stamens) plus the ovary. No ethylene emission was observed from perianth parts that were isolated at various periods after pollination. The increased ethylene emission by the column plus ovary was correlated with an increase in ACC synthase and ACC oxidase activity in these flower parts. The perianth parts, in contrast, only showed an increase in ACC oxidase activity, following pollination. The data show that pollination-induced early perianth senescence in Dendrobium is mediated by increased ethylene biosynthesis by the column + ovary, and not due to increased ethylene biosynthesis in the perianth parts. Apparently, ethylene synthesized in the gynoecium diffuses to the perianth parts where it induces senescence. The data are very similar to those found previously in pollinated Phaleanopsis orchids and in emasculated Cymbidium orchids, with the exception that ethylene was emitted from the tepals of these two orchids and not from Dendrobium.