Orchid Research Articles

Application of DNA barcoding tool for authentication of traditionally important medicinal Dendrobium (Orchidaceae) species from Sikkim Himalaya

Dendrobium, the second largest genus of the Orchidaceae family, is equally important for ornamental value and traditional medicine. Sikkim Himalaya region is rich in traditional knowledge of medicinal plants and their diversity. However, these regions were least explored through modern molecular tools. Reference databases association with DNA Barcoding technique is a new tool for species authentication. Here, we tested the efficiency of DNA barcoding loci (ITS, matK, rbcL, trnH-psbA) based on similarity search, characters based and phylogenetic methods to authenticate the medicinally important Dendrobium species. DNA from young leaves of studied species were isolated, and ITS (∼700 bp), rbcL(∼650 bp), matK (∼ 800 bp), trnH-psbA (∼ 450 bp) were amplified and sequenced for species-level identification. ITS, rbcL and trnH-psbA regions showed easy amplification and sequencing. Among the 54 sequences, 9 sequences were first time submitted to GenBank. In case of similarity search, ITS showed most efficient to identify all Dendrobium species except two species Dendrobium thyrsiflorum and Dendrobium densiflorum. However, the characters-based method easily differentiated between Dendrobium thyrsiflorum and Dendrobium densiflorum. The similarity-based methods and ITS sequence information together may help correct species identification for medicinal plants of Dendrobium.

Comparative and phylogenetic analyses of six Kenya Polystachya (Orchidaceae) species based on the complete chloroplast genome sequences

BackgroundPolystachya Hook. is a large pantropical orchid genus (c. 240 species) distributed in Africa, southern Asia and the Americas, with the center of diversity in Africa. Previous studies on species of this genus have not obtained the complete chloroplast genomes, structures and variations. Additionally, the phylogenetic position of the genus in the Orchidaceae is still controversial and uncertain. Therefore, in this study, we sequenced the complete plastomes of six Kenya Polystachya species based on genome skimming, subjected them to comparative genomic analysis, and reconstructed the phylogenetic relationships with other Orchidaceae species.ResultsThe results exhibited that the chloroplast genomes had a typical quadripartite structure with conserved genome arrangement and moderate divergence. The plastomes of the six Polystachya species ranged from 145,484 bp to 149,274 bp in length and had an almost similar GC content of 36.9–37.0%. Gene annotation revealed 106–109 single-copy genes. In addition, 19 genes are duplicated in the inverted regions, and 16 genes each possessd one or more introns. Although no large structural variations were observed among the Polystachya plastomes, about 1 kb inversion was found in Polystachya modesta and all 11 ndh genes in the Polystachya plastomes were lost or pseudogenized. Comparative analysis of the overall sequence identity among six complete chloroplast genomes confirmed that for both coding and non-coding regions in Polystachya, SC regions exhibit higher sequence variation than IRs. Furthermore, there were various amplifications in the IR regions among the six Polystachya species. Most of the protein-coding genes of these species had a high degree of codon preference. We screened out SSRs and found seven relatively highly variable loci. Moreover, 13 genes were discovered with significant positive selection. Phylogenetic analysis showed that the six Polystachya species formed a monophyletic clade and were more closely related to the tribe Vandeae. Phylogenetic relationships of the family Orchidaceae inferred from the 85 chloroplast genome sequences were generally consistent with previous studies and robust.ConclusionsOur study is the initial report of the complete chloroplast genomes of the six Polystachya species, elucidates the structural characteristics of the chloroplast genome of Polystachya, and filters out highly variable sequences that can contribute to the development of DNA markers for use in the study of genetic variability and evolutionary studies in Polystachya. In addition, the phylogenetic results strongly support that the genus of Polystachya is a part of the tribe Vandeae.

First Report of Leaf Anthracnose Caused by Colletotrichum karstii of Piper nigrum in Yunnan Province, China.

Pepper (Piper nigrum L.) is one of the economically important spice crops of China and mainly grown in the Hainan and Yunnan provinces. In January 2021, the classic anthracnose lesions were observed on pepper leaves at a plantation (24°57’50"N, 98°53’00"E) in Baoshan city, Yunnan, China. Most of the diseased spots occurred at the tips and margins of the old pepper leaves. Lesions were grayish brown or pale white with a slight yellow halo, concentric whorl black dots or scattered black dots were observed on the leaves spots sometimes (Fig. 1). Five symptomatic leaves from different parts of the field were sampled for pathogen isolation. Lesion tissues removed from the border between symptomatic and healthy tissue were surface sterilized in 75% ethanol, then air-dried, plated on potato dextrose agar medium plates (PDA), and incubated in a 12-h photoperiod at 28℃. Similar fungal colonies developed from all plated tissues after 5 days. And five isolates from different leaves (one isolate per leaf) were sub-cultured using the single-spore method. The colonies appeared white, cottony, aerial mycelium dense and slow-growing (mean 1.01 mm day–1) on PDA plates in 6 days. Conidia were short-cylindric, straight, sometimes slightly constricted near the center, ends broadly rounded, measuring 11.05 to 14.43 × 3.78 to 6.08 µm (average = 12.03 × 5.48 µm, n=200). Appressoria were single, subglobose to elliptic, light brown to dark black. Among them, genomic DNA of two isolates (21HJ0301-1 and 21HJ0301-2) were extracted from mycelium and used as a template for molecular identification. The internal transcribed spacer (ITS) region of ribosomal DNA, and partial sequence of chitin synthase (CHS-1), actin (ACT) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene regions were amplified with primer pairs ITS1/ITS4, CHS-79F/CHS-354R, ACT-512F/ACT-783R, GDF/GDR, respectively (Weir et al. 2012). These four gene sequences were deposited in GenBank (Accession No. MZ725047 and MZ725048 for ITS, MZ733415 and MZ733416 for GAPDH, MZ733408 and MZ733409 for ACT, MZ733422 and MZ733423 for CHS-1). A multilocus phylogenetic analysis performed with the reference sequences revealed that both 21HJ0301-1 and 21HJ0301-2 isolates clustered with C. karstii (Fig.2). Based on morphology and molecular results, isolates were confirmed to be C. karstii. Pathogenicity tests were carried out on potted seedlings in the greenhouse, six healthy leaves per isolate were inoculated with six-day-old cultures of C. karstii mycelial discs of 5 mm in diameter after being wounded with a needle or non-wounded. Control leaves were inoculated with PDA agar. Inoculated plants were incubated under high relative humidity at room temperature. Anthracnose symptoms appeared within 5 days using non-wounded or wounded inoculation methods. All control leaves remained asymptomatic. The fungus was re-isolated from inoculated leaves fulfilling Koch’s postulates, but not on controls. C. karstii has a wide range of hosts, such as rubber tree, tea-oil tree, chili, and some other plants belonging to the family Orchidaceae in China (Cai et al. 2016; Jiang and Li 2018; Diao et al. 2017; Yang et al. 2011). To the best of our knowledge, this is the first report of C. karstii on Piper nigrum in China. This report will help us to recognize the anthracnose disease of Piper nigrum and establish a foundation for future studies on C.karstii to address effective management strategies.

Interaction between Cymbidium aloifolium and Apis cerana: Incidence of an outlier in modular pollination network of oil flowers.

So far, oil‐rewarding flowers are known to be pollinated only by oil‐collecting bees, which gather and use lipids for larval feed and nest building. As honeybees do not have oil‐collecting appendages on their legs, they have not been associated with pollination of such flowers. In a predominantly Apis pollinated and food deceptive clade of wild Cymbidiums, we investigated the reproductive strategy of Cymbidium aloifolium, hitherto unknown for its floral oil reward. Our study demonstrates the requisites for establishment of mutualistic interaction between the oil flower and Apis cerana indica, a corbiculate bee. Success in pollination requires learning by honeybees to access the food reward, thereby displaying cognitive ability of the pollinator to access the customized reward. Morphometric matching between orchid flowers and the pollinator, and that between pollinia and stigmatic cavity also appear to be essential in the pollination success. Absence of pollinator competition and prolonged flower‐handling time are suggested to promote floral constancy. The present study highlights the need to explore the spectrum of pollination rewards pursued by honeybees, which may include unconventional composition of floral resources.

Over a century later and 400 kilometers apart: rediscovery of Bulbophyllum barbatum (Orchidaceae) in Diamantina, Minas Gerais, Brazil

Fieldwork focused on the Orchidaceae family from the Diamantina Plateau of the Espinhaço Range in Minas Gerais, Southeast Brazil, resulted in the rediscovery of Bulbophyllum barbatum almost 140 years after its original description, in 1882. The species, known only from the original illustration by Barbosa Rodrigues, was originally described from the region known as Zona da Mata Mineira, and its rediscovery is reported from over 400 km to the northwest of that region. The new record allowed a more detailed study of its morphology, the first photographs of the species, as well as the preliminary assessment of its conservation status as Critically Endangered. Such unexpected rediscovery highlights the importance of conservation actions in the Brazilian campos rupestres given its high diversity and endemism, the deficient knowledge on the taxonomy and distribution of its species, and the increasing pressure of human activities.

Estimation of metabolic activity of cut flowers during a study to enhance vase life of Cymbidium hybrid cut-flowers

In this study, the metabolic activity (MA) of orchid cut flowers was estimated by the modified alkali trap method for the first time, when an approach was made to increase the vase life of Cymbidium miniature hybrid and Cymbidium hybrid ‘H. C. Aurora’ cut flowers. Metabolic inhibitors like-systemic fungicides (Carbendazim and Metalaxyl) and darkness were used to increase the vase life of the cut flowers. Cut flowers kept with metabolic inhibitors had significantly higher relative fresh weight (RFW), rate of vase solution uptake (RVSU) and significantly lower MA compared to control, which consequently increased the vase life of the cut flowers. The vase life was maximum under Carbendazim + darkness treatment (C200 + D), followed by Carbendazim (C200)> Metalaxyl +Darkness (M200 + D)> Metalaxyl (M200)> Distilled water + Darkness (D)> Control, whereas the MA was maximum under the Control treatment, followed by D > M200 > M200 + D > C200 > C200 + D treatments for both the cut flowers. The C200 + D treatment increased the vase life by ∼31 and 27% for Cymbidium miniature hybrid and Cymbidium hybrid ‘H. C. Aurora’, respectively, compared to the control treatment. It can be inferred that Carbendazim+ darkness treatment can increase the vase life of orchid cut flowers and the modified alkali trap method would be effective for estimation of MA of the cut flowers.

The Genetic and Hormonal Inducers of Continuous Flowering in Orchids: An Emerging View.

Orchids are the flowers of magnetic beauty. Vivid and attractive flowers with magnificent shapes make them the king of the floriculture industry. However, the long-awaited flowering is a drawback to their market success, and therefore, flowering time regulation is the key to studies about orchid flower development. Although there are some rare orchids with a continuous flowering pattern, the molecular regulatory mechanisms are yet to be elucidated to find applicable solutions to other orchid species. Multiple regulatory pathways, such as photoperiod, vernalization, circadian clock, temperature and hormonal pathways are thought to signalize flower timing using a group of floral integrators. This mini review, thus, organizes the current knowledge of floral time regulators to suggest future perspectives on the continuous flowering mechanism that may help to plan functional studies to induce flowering revolution in precious orchid species.

Seed viability testing for research and conservation of epiphytic and terrestrial orchids

BackgroundSeed viability testing is essential in plant conservation and research. Seed viability testing determines the success of ex-situ conservation efforts, such as seed banking but commonly testing protocols of orchids lack consistency and accuracy, therefore, there is a need to select an appropriate and reliable viability test, especially when conducting comparative studies. Here, we evaluated the suitability of three seed viability tests, Evans blue test (EB), Fluorescein diacetate test (FDA) and Tetrazolium test (TTC), with and without sterilization, on seeds of 20 orchid species, which included five epiphytes and fifteen terrestrials, using both fresh seeds and seeds stored at − 18 ºC for 6 to 8 years.ResultsWe found that sterilization and lifeform of seeds affected seed viability across all tests but the storage time was not an influential factor. Sterilization negatively affected seed viability under EB and FDA test conditions but increased the detection of viable seeds in the TTC test in both epiphytic and terrestrial species. The EB test, when administered without sterilization provided the highest viability results. Being non-enzymatic unlike TTC and FDA tests, as expected, the EB test was the most reliable with similar results between sterilized and not sterilized seeds for most epiphytic and terrestrial species as well as when compared between groups.ConclusionsThe lifeform of the species and seed sterilization prior to testing are important influential factors in orchid seed viability testing. Since EB test was found to be reliable we recommend the EB test for seed viability assessment in orchids rather than the less reliable but commonly used TTC test, or the FDA test, which require more expensive and sophisticated instrumentation. Since storage time was not an influential factor in orchid seed viability testing, the recommendations of this study can be used for both fresh as well as long-term stored orchid seeds. This is helpful for research and especially for conservation measures such as seed banking. However, due to the species specificity of the bio-physiology of orchids, we call for comprehensive viability test assessment in the hyper diverse orchid family to be extended to a greater number of species to facilitate efficient conservation and research.

<i>Eulophia dabia</i> - A Lesser Known Critically Endangered Terrestrial Orchid

Eulophia dabia (D.Don) Hochr is an endemic and critically endangered, species of the family Orchidaceae. It prefers to grow on sandy soil near water embankments. The tubers of this species are highly medicinal and used by the local healer for the treatment of various ailments. The present paper provides insight of its medicinal utility and their mode of propagation.

Calcium Channel Blockade Mediates the Vasorelaxant Activity of Dichloromethane Extract from Roots of Oncidium cebolleta on Isolated Rat Aorta

Oncidium cebolleta belongs to the Orchidaceae family, and it is used in preparations in Mexican traditional medicine to treat several diseases. This research aimed to study the vasorelaxant activity of dichloromethane extract from the roots of Oncidium cebolleta (DEROc) on endothelium-intact and -removed rat aorta segments to determine its mode of vasorelaxant action. Several extracts (hexane, dichloromethane, and methanol) obtained from leaves and roots were evaluated on isolated rat aorta rings with and without endothelium to determine their vasorelaxant effect. Except for methanolic extract from leaves, all extracts induced a concentration-dependent vasorelaxant effect on endothelium-intact rat aorta rings pre-contracted with noradrenaline (NA, 0.1 µM). Dichloromethane extract from roots (DEROc) was the most potent and efficient, and the effect was endothelium-independent. Also, DEROc could relax KCl-induced contraction (80 mM) and inhibit the contraction induced by CaCl2-cumulative concentration on endothelium-denuded rings. However, DEROc was less potent than control nifedipine (L-type calcium channel blocker), but its effectiveness was similar. Contractions induced by NA and 5-HT [0.001 µM to 0.1 µM] were significantly reduced by DEROc. However, DEROc vasorelaxation was not reduced by TEA (potassium channel blocker, 10 mM) nor by ODQ (soluble guanylyl cyclase inhibitor, 0.1 µM) in endothelium-denuded aortic rings. Our functional results suggest that DEROc induces its relaxant effect by an endothelium-independent mechanism due to a Ca2+ channels blockade in rat aortic rings.

First Report of Fusarium commune Causing Leaf Spot Disease on Bletilla striata in China.

Bletilla striata (Thunb.) Rchb. f. (Orchidaceae family, known as Baiji in Chinese) is a perennial herb and has been traditionally used for hemostasis and detumescence in China. In April of 2020, a leaf spot disease on B. striata was observed in plant nurseries (∼0.2 h) in Guilin, Guangxi Province, China. Approximately 20% of the plants were symptomatic, of which 150 plants were randomly selected for investigation. Initial symptoms include the appearance of small, circular or irregular light brown spots, randomly scattered on the edges and surfaces of the leaves, which progressively expand into large, suborbicular or irregular-shaped dark brown, necrotic areas. At the severe stage, the lesions coalesced into large necrotic areas and ultimately resulted in leaf abscission. To isolate the pathogen, three representative plants exhibiting symptoms were collected from the nurseries. Leaf tissues (5 × 5 mm) were cut from the margin of necrotic lesions (n = 18), surface-disinfected in 1% sodium hypochlorite (NaOCl) solution for 2 min, then rinsed three times in sterile water before isolation. The tissues were plated on potato dextrose agar (PDA) medium, and incubated at 28°C (12-h photoperiod) for 3 days. Hyphal tips from recently germinated spores were transferred to PDA to obtain pure cultures. Nine fungal isolates with similar morphological characteristics were obtained. Three single-spore isolates, BJ23.1, BJ55.1, and BJ91.3, were subjected to further morphological and molecular characterisation. Colonies on PDA plates were villose, had a dense growth of aerial mycelia and appeared white (1A1) to yellowish white (3A2). Macroconidia were smooth, hyaline, straight to slightly curved, usually contained three or five septa, and measuring 23.3 to 42.1 × 3.0 to 6.2 μm (mean ± SD: 31.2 ± 5.1 × 4.2 ± 0.6 μm, n = 50). Microconidia were generally cylindrical, straight to slightly curved, aseptate, and measuring 7.2 to 18.8 × 2.5 to 4.3 μm (mean ± SD: 12.1 ± 2.8 × 3.3 ± 0.5 μm, n = 62). Morphological characteristics are similar to those of F. commune (Skovgaard et al. 2003). For molecular identification, the genomic DNA of the isolates BJ23.1, BJ55.1, and BJ91.3 were extracted using the CTAB method (Guo et al. 2000). The internal transcribed spacer (ITS) region of rDNA, partial translation elongation factor-1 alpha (TEF-1α), RNA polymerase second largest subunit (RPB2), and the mitochondrial small subunit rDNA (mtSSU) genes were amplified using primer pairs [ITS1/ITS4 (White et al. 1990), EF-1/EF-2 (O'Donnell et al. 1998), and 5f2/11ar (Liu et al. 1999, Reeb et al. 2004), MS1/MS2 (Li et al. 1994), respectively]. The obtained sequences were deposited in NCBI GenBank under the following accession numbers: ITS (MZ424697 to MZ424699), TEF-1α (MZ513467 to MZ513469), RPB2 (MZ513473 to MZ513475), and mtSSU (MZ513470 to MZ513472). BLAST® analysis of the deposited sequences showed 99 to 100% identity with those of F. commune present in GenBank (Accession numbers: DQ016205, MH582348, MH582181, AF077383). In addition, a phylogenetic analysis using concatenated sequences of ITS, TEF-1α, mtSSU genes showed that BJ23.1, BJ55.1, and BJ91.3 located on the same clade with strains of F. commune. Therefore, based on morphological and molecular characteristics, the isolates were identified as F. commune (Skovgaard et al. 2003, Stewart et al. 2006). Pathogenicity was tested using 1.5-year-old B. striata plants. Healthy leaves on plants were inoculated with 5 × 5 mm mycelial discs of strains BJ23.1, BJ55.1, and BJ91.3 from 3-day-old PDA cultures, each isolate was inoculated onto three plants; three other plants inoculated with sterile PDA discs served as controls. All plants were enclosed in transparent plastic bags and incubated in a greenhouse at 28°C for 14 days (12-h photoperiod). Three days post-inoculation, leaf spot symptoms appeared on the inoculated leaves. No symptoms were detected on control plants. Experiments were replicated three times with similar results. To fulfill Koch's postulates, F. commune was consistently re-isolated from symptomatic tissue and confirmed by morphology and sequencing, whereas no fungus was isolated from the control plants. F. commune has been reported to cause diseases on some plants, including sugarcane (Wang et al. 2018), maize (Xi et al. 2019) and Wax Gourd (Zeng et al. 2020). To our knowledge, this is the first report of F. commune causing leaf spot disease on B. striata in China. Identification of this pathogen provides the information for further studies to develop management strategies to control the disease.

Organ-Specific Gene Expression Reveals the Role of the Cymbidium ensifolium-miR396/Growth-Regulating Factors Module in Flower Development of the Orchid Plant Cymbidium ensifolium.

Orchids are some of the most popular ornamental plants worldwide. Orchid floral morphology has increasingly attracted horticultural and commercial attention. Although multiple genes have been shown to be involved in the formation of the orchid flower, the underlying multi-level regulatory networks are largely unknown. In this study, we analyzed the ontogeny of flower development in Cymbidium ensifolium, a traditional orchid in the tropical and subtropical regions of Asia, by performing deep sequencing of the transcriptome of individual flower organs to discover organ-specific genes potentially involved in their growth. We identified 3,017 differentially-expressed genes (DEGs) during the development of various flower organs, and observed over-representation of GROWTH-REGULATING FACTORS (GRFs) specific to flower column (gynostemium). Eleven C. ensifolium GRFs (CeGRFs) from our transcriptome data clustered into five phylogenetic subgroups. Ten of these GRFs shared a region complementary to C. ensifolium microRNA396 (Ce-miR396), and degradome sequencing confirmed the cleavage of transcripts derived from seven CeGRFs. We cloned Ce-miR396 and used a protoplast-based transient expression system to overexpress it in Cymbidium protoplasts. We observed a significant decrease in the transcripts of several CeGRFs in flowers and leaves, indicating a potential role for miR396–GRF module in organ development through the cleavage of distinct CeGRFs. Temporal and spatial expression analysis indicated that most CeGRF transcripts accumulated in flower buds and column tissues, where Ce-miR396 expression was the lowest. Expression dynamics in wild type and floral-defective mutants further confirmed a strong correlation between Ce-miR396, CeGRFs, and flower organ development and column specification. Moreover, overexpression of Ce-miR396 in Nicotiana tabacum resulted in curved pistils and reduced fertility, implying that the conserved role of Ce-miR396 in floral development. These results provide tools to better understand the biological roles of GRFs in orchid development, and open new avenues for the diversification of orchid floral patterns.

Advances in Breeding, Bioprospecting, and In Vitro Culture of Laelia Orchid Species

Orchids (Orchidaceae) are plants that are highly appreciated by their beautiful flowers worldwide. Moreover, they represent a source of metabolites with applications in medicine and biotechnology. Within the Orchidaceae family, the Laelia genus is a group of orchid species from the Neotropics and is probably one of the most representative genera of America. Laelia orchids are cultivated by their splendid flowers and are widely used in orchid breeding. Here, we revise the use of the Laelia genus in orchid breeding and metabolite bioprospecting. We also analyze the use of plant tissue culture (PTC) as an alternative to conventional propagation and as a strategy for the recovery of those Laelia species threatened with extinction. We summarize and discuss the recent advances in the application of different PTC techniques for mass multiplication based on asymbiotic germination, organogenesis, protocorm-like bodies development, and somatic embryogenesis, and the advances of in vitro conservation by cryoconservation and the use of slow-growth promoting hormones. Finally, we suggest future directions and venues in research for Laelia species.

Chromosome-level genome assembly of the fully mycoheterotrophic orchid Gastrodia elata.

Gastrodia elata, an obligate mycoheterotrophic orchid, requires complete carbon and mineral nutrient supplementation from mycorrhizal fungi during its entire life cycle. Although full mycoheterotrophy occurs most often in the Orchidaceae family, no chromosome-level reference genome from this group has been assembled to date. Here, we report a high-quality chromosome-level genome assembly of G. elata, using Illumina and PacBio sequencing methods with Hi-C technique. The assembled genome size was found to be 1045 Mb, with an N50 of 50.6 Mb and 488 scaffolds. A total of 935 complete (64.9%) matches to the 1440 embryophyte Benchmarking Universal Single-Copy Orthologs were identified in this genome assembly. Hi-C scaffolding of the assembled genome resulted in 18 pseudochromosomes, 1008 Mb in size and containing 96.5% of the scaffolds. A total of 18,844 protein-coding sequences (CDSs) were predicted in the G. elata genome, of which 15,619 CDSs (82.89%) were functionally annotated. In addition, 74.92% of the assembled genome was found to be composed of transposable elements. Phylogenetic analysis indicated a significant contraction of genes involved in various biosynthetic processes and cellular components and an expansion of genes for novel metabolic processes and mycorrhizal association. This result suggests an evolutionary adaptation of G. elata to a mycoheterotrophic lifestyle. In summary, the genomic resources generated in this study will provide a valuable reference genome for investigating the molecular mechanisms of G. elata biological functions. Furthermore, the complete G. elata genome will greatly improve our understanding of the genetics of Orchidaceae and its mycoheterotrophic evolution.

Paphiopedilum charlesworthii var. lannaense, a new slipper orchid from Northern Thailand identified by morphological and AFLP analyses

A new endemic slipper orchid in the family Orchidaceae from Northern Thailand was described according to morphological and molecular analyses. Morphological information specifies the new slipper orchid Paphiopedilum charlesworthii var. lannaense to be similar to Paphiopedilum charlesworthii (Rolfe) Pfitzer 1895, with the exception of its staminode being obovate-obcordate with yellow color, glittery and rough surface. Molecular analysis by AFLP technique indicates that Paphiopedilum charlesworthii var. lannaense can be included as a member of section Paphiopedilum and is closely related to Paphiopedilum coccineum Perner, H. & Herrmann, R. (2000) (syn. Paphiopedilum barbigerum var. coccineum), from which it differs morphologically by dorsal sepal and petal characters. The morphological and molecular evidence supported that Paphiopedilum charlesworthii var. lannaense is a new slipper orchid in the genus Paphiopedilum (Cypripedioideae, Orchidaceae).

Research Advances in Multi-Omics on the Traditional Chinese Herb Dendrobium officinale

Dendrobium officinale Kimura et Migo is an important epiphytic plant, belonging to the Orchidaceae family. There are various bioactive components in D. officinale plants, mainly including polysaccharides, alkaloids, and phenolic compounds. These compounds have been demonstrated to possess multiple functions, such as anti-oxidation, immune regulation, and anti-cancer. Due to serious shortages of wild resources, deterioration of cultivated germplasm and the unstable quality of D. officinale, the study has been focused on the biosynthetic pathway and regulation mechanisms of bioactive compounds. In recent years, with rapid developments in detection technologies and analysis tools, omics research including genomics, transcriptomics, proteomics and metabolomics have all been widely applied in various medicinal plants, including D. officinale. Many important advances have been achieved in D. officinale research, such as chromosome-level reference genome assembly and the identification of key genes involved in the biosynthesis of active components. In this review, we summarize the latest research advances in D. officinale based on multiple omics studies. At the same time, we discuss limitations of the current research. Finally, we put forward prospective topics in need of further study on D. officinale.

Dactylorhiza hatagirea (D. Doon) Soo, an important medicinal herb of the Himalaya and urgent need for its conservation- A review

Dactylorhiza hatagirea (D. Doon) Soo (Family Orchidaceae) is a terrestrial orchid, native to alpine and sub-alpine zones of Himalaya, well known for medicinal uses because of its immense pharmacological properties and therapeutic potential. Leaves and tubers of the herb finds enormous uses in traditional Indian medicinal system like Ayurveda, Siddha, Unani and Sowa-rigpa for treatment of different ailments like dysentery, diarrhoea, chronic fever, cough, wounds, burns, fractures, stomach-ache, etc. The present review analyses the distribution, medicinal uses, phytochemistry, pharmacology, and conservation status of D. hatagirea. The review is based on literature available on this orchid with an objective to compile and understand diverse medicinal uses among the indigenous Himalayan communities and phytochemical potential in pharmacology. This review also highlights the potential threats faced by this orchid and suggests future conservation prospects and strategies.

Orchidaceae of the municipality of Igarapé-Miri, Pará, Brazilian Amazon

Abstract The municipality of Igarapé-Miri is located in the Endemism Center of Belém, which corresponds to the most deforested portion of the Brazilian Amazon with only 38% of its native forest remaining. The family Orchidaceae has several endemic taxa and expressive diversity in the Amazon phytogeographic domain, including in the state of Pará where 97 genera and 395 species have been reported. Considering the advanced state of fragmentation and the loss of diversity in the region, together with the lack of information about the flora of the municipality of Igarapé-Miri (Pará, Brazil), a floristic-taxonomic study was undertaken of the species of Orchidaceae in the municipality, including an identification key, morphological descriptions, taxonomic comments, illustrations and information on habitat, phenology and the geographic distribution of species. Fifty-eight species distributed in 38 genera were recorded for the municipality of Igarapé-Miri. The most diverse genera were Epidendrum (7 spp.) and Catasetum (4 spp.), followed by Maxillaria, Scaphyglottis and Stelis (3 spp. each). Most species occur as epiphytes and in Terra Firme Forest, with 31 species being exclusive to this type of vegetation. The occurrence of Anathallis fastigiata, Catasetum albovirens, Dichaea brachyphylla and Palmorchis triquilhada, species restricted to the Brazilian Amazon, stands out. The distribution of Stelis palmeiraensis is expanded to the Amazon phytogeographic domain and Stelis ciliaris and Uleiorchis ulei are documented in the state of Pará for the first time.

Optimising conservation translocations of threatened

Context Conservation translocations are increasingly being used in the management of rare plants, yet have low success in maintaining populations through recruitment. Aims We investigated whether the survival of translocated plants, recruitment and, therefore, cost effectiveness, can be improved by selecting optimal microsites for both adults and seedlings. Methods Caladenia colorata plants propagated symbiotically with Serendipita australiana (n = 735) were introduced to four sites where the pollinator was present and vegetation matched wild populations. Plant demography was monitored over 6 years. The relationship between microsite variables and measures of orchid survival, re-emergence, flowering and recruitment were analysed with generalised linear mixed-effects models. We then estimated potential improvement in emergence and recruitment, if microsite selection was optimised. Key results A total of 77% of plants survived translocation, and populations grew by 84% through recruitment (n = 615). Survival was positively associated with cover of leaf litter, graminoids and cryptogams. Recruitment was positively correlated with soil moisture. The majority of recruitment was within 5 cm of adult C. colorata plants. The potential improvement by selecting favourable microsites increased adult survival by up to 8% and recruitment by 10–40%. Conclusions Incorporating both the germination niche and adult plant niche within plant translocations more broadly could significantly improve long-term population persistence and the utilisation of conservation funding. Implications Our results are directly applicable to 58 endangered Caladenia species in the subgenus Calonema, owing to their shared mycorrhizal association with S. australiana. Furthermore, our results are applicable to all plant translocations as understanding germination niche and microhabitat requirements is likely to improve success overall.

Identification of predominant bacteria and controlling of microbial population in vase solution of cut orchid flowers

Identification of predominant bacteria and controlling of microbial population in vase solution of cut orchid flowers