INTRODUCTION:

India has abundant knowledge of herbs base treatment call as ayurveda. in India ayurveda practiced from more than 5000 years. Ayurveda, which is the oldest India system of medicine, reported approximately 2000 medicinal plant species for the treatment of various diseases. Dactylorhiza hatagirea (D.Don) Soo is a member of family Orchidaceae⁽¹⁾. Dactylorhiza hatagirea (D.Don) Soo is native from the Indian Himalayan region and Its distribution outstretch to Pakistan, Afghanistan, Nepal, Tibet and Bhutan¹. but in India, it is originate from are of Jammu and Kashmir, Sikkim, Arunachal Pradesh Uttarakhand, and Himachal Pradesh. Dactylorhiza hatagirea (D.Don) Soo is commonly known as Hatajari in Uttarakhand and Salampanja in Kasmir².

Dactylorhiza hatagirea (D.Don) Soo is a terrestrial, erect herb, up to 60-70cm high, along with palmate divided tuberous. Leaves are broadly lanceolate or oblong-lingulate or elliptic. Flowers purplish-lilac, or rarely white, in many-flowered densely cylindrical inflorescence³. The main characteristics of this plant is that, it relics erect in excessive snowstorm. Tubers of Dactylorhiza hatagirea (D.Don) Soo are sweet, cooling, emollient, astringent, aphrodisiac, demulcent, rejuvenating and nervine tonic⁴. The tubers of Dactylorhiza hatagirea (D.Don) Soo contain a glycoside, starch, mucilage, albumen, a trace of volatile oil and ash. Chemically, dactylorhins A-E, dactyloses A and B and lipids etc. are found as major constituents^5,6. Dactylorhiza hatagirea (D.Don) Soo conventionally it used to cure various diseases like dysentery, diarrhoea, chronic fever, cough, stomach-ache, wounds, cuts, burns, fractures, general weakness and aphrodisiac but few activities had been established scientifically.

Classification and Morphology:

Dactylorhiza hatagirea (D. don) belongs to the Orchidaceae family. The orchids have attracted the admiration of scientists, horticulturists, herbalists, and laypeople alike, and deserve the pride of place in any discussion on flowering plants among the ornamentals. Orchidaceae is believed to be the second-largest family of flowering plants after Asteraceae, with between 21,950 and 26,049 currently accepted species, grouped into 880 genera (World Checklist of Selected Plant Families, 2013). Nepal harbours 451 species of orchid belonging to 104 genera (Rajbhandari, 2015). Central Nepal harbours the highest number of orchid species (69 species) followed by east Nepal (58 species) and west Nepal (33 species). Among them, D. hatagirea is a terrestrial-ground-dwelling perennial herb with erect, hollow and obtuse stem, bears palmately lobed rhizome, and lanceolate leaves with sheathing leaf-base The cylindrical and terminal spike bears rosy-purple flowers with green bracts. The inflorescence consists of a compact raceme with 25 to 50 flowers developed from axillary buds. Flowers are 1.7-1.9 cm long with a curved spur, and the dark purple-spotted lip of the flower is rounded and lobed. The structure of orchid flowers is unique among floral plants. The orchid flower typically has an outer whorl of three sepals, an inner whorl of three petals, two identical and one modified (Lip), and a single massive column (the gynostemium, composed of the male stamens attached to the female pistil) in the centre. The plants store a large amount of water in their palmately lobed tuberous roots to survive in arid conditions.⁷

Distribution:

The distribution of Dactylorhiza covers most of Europe, temperate Asia, North Africa, Japan, Aleutian Islands, and northern parts of North America with the highest species richness in north-western Europe, though only nine species of Dactylorhiza are endemic. D. hatagirea is distributed in Nepal, India, Bhutan, China, Mongolia, Pakistan, and Russia⁶. This Himalayan endemic medicinal orchid is also documented in Hindu Kush Himalaya range in terrestrial habitat from Western Himalaya (Afghanistan, Pakistan), India (Jammu and Kashmir, Himachal Pradesh, Uttarakhand, Sikkim), Bhutan, South-East China and Nepal (Hulma, Dolpa, Doti, Kaski, Gorkha, Rasuwa, Sindhupalchok, Dolakha) (Flora of China Editorial Committee, 2009). Its habitat is in grassland slopes in the sub-alpine and the alpine zones between 2800m and 4200m altitudes above the mean sea level (IUCN, 2004). The altitudinal range of habitat distribution of D. hatagirea in Annapurna Conservation Area is reported as 3200 to 3600 meters above sea level with abundance in the slope from 30° to 60° at north-east aspect and maximum species richness of total medicinal orchid is observed at an elevation of 1700m above sea level.⁷

Figure 1: Morphology of D. hatagirea(a) aerial part of flowering plants

Figure 2: Morphology of D. hatagirea and name of each part of a flower

Figure 3: Morphology of D. hatagirea rhizomes.

Figure 5: Distribution of Dactylorhizahatagirea Phytochemical Study

D. hatagirea is one of the highest valued orchids extensively used in traditional medicine as a farinaceous food and nerve tonic for sick, stomachache, headache, typhoid, effective in chronic diarrhoea, fever, general debility, in treating weakness in children and women, root powder is spread on wounds to control bleeding⁶. It is also considered an alternative source of salep used very commonly in Europe and is considered an essential aphrodisiac plant in Ayurveda, Siddha, Unani literature. Therefore, it is employed to enhance performance and to increase vigour and vitality.

Antipyretic Activity:

The Hydroalcoholic Extract of Dactylorhiza Hatagirea Roots and Lavandula Stoechas Flowers showed a significant (P<0.01) dose dependent antipyretic effect in yeast induced elevation of body temperature in experimental rats. The antipyretic effect taking placed as from the first hour and the effect was remained for 4 h, after administration of the extract. The results obtained from both standards and extracts treated groups were compared with the control group. A significant reduction in the yeast elevated rectal temperature was observed in the test drug⁷.

Anti-Inflammatory Activity:

The hydroethanolic extract of Dactylorhiza hatagirea possesses a strong anti-inflammatory activity and may be considered an appealing source of effective anti-inflammatory compounds. The Hydroalcoholic extract of D. hatagirea showed potent anti- inflammatory activity on carrageenan induced paw oedema in rats. The anti-inflammatory activity of root extract of D. hatagirea confirmed and dose-dependent response and were comparable to diclofenac (standard drug). The extracts were most active at the dose level of 200 mg/kg body weight in the fourth hour of treatment. The extracts of D. hatagirea could be an alternative phyto active resource for generating anti-inflammatory agents⁸.

Sedative and Hypnotics Activity:

The hypnotics activity activities of D. hatagirea roots and L. stoechas flower were examined by thiopental sodium induced sleeping time in mice at the doses of 100, 200 and 300mg/kg p.o body weight. The extracts exhibited significant reduction of onset and potentiated duration of sleep in thiopental sodium induced sleeping time test⁹. Altogether, results suggested that hydroalcoholic extracts of D. hatagirea roots and L. stoechas flower holds potent hypnotic properties. To elucidate the precise mechanism action and bioactive compounds which responsible for the hypnotics activity of D. hatagirea is not establish¹⁰.

Anti‑diabetic Activities:

3T3 L1 cells were treated with diverse concentrations (25μg – 400μg/mL) of D. hatagirea extract and were investigate for their cytotoxic effect. The extract of D. hatagirea had not elicit any cytotoxic effect on the cells. Percentage inhibition of Apha amylase activity of D. hatagirea leaf extract was estimated with acarbose as positive control. The extract elicit 27.13±0.3% inhibition of Apha amylase activity at 31.25μg/mL and 74.53±0.5 % inhibition at 500μg/mL concentration. the inhibition of Alpha Glucosidase activity by D. hatagirea leaf extract was estimated with acarbose as positive control. Methanolic extract of D. hatagirea leaves showed significant inhibition of α Glucosidase activity⁸. Methanolic extract of D. hatagirea leaves also elevated GLUT4 8). The results also revealed that the quantity of 2(N (7 Nitrobenz 2 oxa 1,3 diazol 4 yl) Amino) 2 Deoxyglucose taken up by 3T3-L1 cells treated with D. hatagirea leaf extract and metformin is higher than that of untreated cells⁸. So that D. hatagirea leaves showed potential antidiabetic activity¹¹.

Aphrodisiac Activity:

The aqueous extract of D. hatagirea causes significant anabolic effect which is comparable to testosterone treatment. D. hatagirea causes also caused Attraction toward the female rats.

The indulgence of D. hatagirea treated animals in copulation increased and the number of bouts increased significantly as well. D. hatagirea also increase endogenous NO and T level as compared to the control group. In another study it also reveal that extract of D. hatagirea Improved of penile erection, sperm count and seminal fructose levels^12-14.

Genetic Diversity:

Effective conservation, management, recovery, and utilization of rare and endangered species can be achieved through variability analysis. Genetic diversity assessment provides the basis for in situ conservation and sustainable utilization of the plant genetic resources as the marked and robust differentiation among natural populations. Variation in the morphological characters is possible due to variations in topography, elevation, soil fertility, rainfall, and other climatic conditions, which are very specific to Himalaya. Genetic diversity inherent in plants plays a significant role in the ability of a population to respond adaptively to environmental changes. Morphological variability in plants, along with biochemical differences in protein and sugar levels, are essential parameters to study natural populations of rare plants, reflecting an adaptive feature of the species. Molecular markers are the essential tools to study the genetic diversity among different populations, which shows actual genetic differences existing between different populations and can overcome the environmental effects.

Plant Propagation:

D. hatagirea is temperate to alpine, monocotyledonous, perennial, and terrestrial orchid valued for its ornamental and medicinal use. It is a habitat-specific and inherently slow-growing species in nature and poorly regenerating species because seeds are microscopic and non-endospermous with undifferentiated embryos posing a prime concern for long-term survival in a natural condition. It has a prolonged rate of vegetative propagation and very poor seed germination in nature that is 0.2 to 0.3%. The in vitro propagation of D. hatagirea is very difficult and slow as the process of seed germination to plantlets formation is very lengthy (>1 yr), and very few numbers of in vitro raised plantlets can be obtained using green pod culture in M.S. medium. The in vitro seed germination is slow not only in D. hatagirea but also in the other species of Dactylorhiza. In D. ruthei, and D. praetermissa seeds started germinating after four months of culture, and only 20.0 to 25.0% germination was achieved in Norstog medium after eight months of inoculation.^8-10

There are very few reports on reintroducing in vitro propagated species of medicinal orchids to natural habitat or their cultivation is always collected from the wild for trade. Symbiotic germination has practical merit for both conservation and horticulture, but it remained an underutilized tool for orchids in our context^11-16. There is a a successful reintroduction of D. hatagirea seedlings grown from mycorrhizal fungi inoculated seeds, which resulted in 100% germination within ten days of sowing, and healthy protocorms were obtained after 40 days, seedlings with well-developed roots, tubers and leaves were obtained after three months. The fungi isolated from the mature plants were characterized using molecular techniques. To substitute for the habitat protection and species recovery, it is crucial to identify and reintroduce over-exposed species through its multiplication using tissue culture.^17-21

CONCLUSION:

D. hatagirea has been categorized as endangered and prioritized for its research and conservation by the Government of Nepal. However, there are fewer efforts made to exploit the modern biotechnological tools for its research, conservation, and utilization. In-vitro mass propagation of this orchid and their reintroduction in the natural condition can be a good initiative for its in-situ conservation. At the same time development of successful agro-technology for its cultivation in commercial-scale under ex-situ conditions can fulfil both the medicinal and floricultural sectors. The use of molecular marker technology can give a good idea on the genetic diversity of D. hatagirea found in Nepal, and conservation efforts can be directed accordingly. Biomolecules present in the plant make it of high value. Therefore, intensive research on the biochemical constitution of the medicinal plants deserves special attention. There is very little documented information regarding genes related to essential metabolites and their expression. The development of trait-specific molecular markers will help in the identification of diversity in populations. Latest biotechnological tools like transcriptomics and metabolomics can be utilized to understand the biosynthetic pathwaysfor essential metabolites from this orchid. This will help in better utilization for medicinal and therapeutic purposes. If research is focused on this sector, panchaule can be one of the promising high-value products shortly for people living in the mountains of Nepal.

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Received on 26.06.2021 Modified on 10.10.2021

Asian J. Pharm. Tech. 2022; 12(2):141-145.

DOI: 10.52711/2231-5713.2022.00024