Cultivation and population status of Ephedra (Ephedra gerardiana Wall. Ex Stapf) critically endangered medicinal plant for the conservation in cold desert of Ladakh, India


 Ephedra gerardiana is critically endangered and one of the most important medicinal plant of Trans-Himalaya of Ladakh. It is majorly used medicinal plant of Ayurveda, Sowa-Rigpa, Unani, Homeopathy, Siddha, Allopathy and other traditional medical systems, hence it is very important to understand its density, and cultivation technique for effective sustainable collection, conservation and utilization. The three valleys of Ladakh viz., Indus, Suru and Nubra valleys were analysed for the population census where the density of the species were ranges between 0.2-3.85 ind/m2 revealed the degree of threat of the species. Germplasm collected from different locations were studied for cultivation packages and practices in three conditions such as polycarbonate greenhouse, trench greenhouse and open field as controlled. We observed that the highest germination percentage (82.67±3.79) and survivability percentage (75.67±4.04) in polycarbonate greenhouse followed by trench greenhouse (58.00±2.00) (50.67±1.53) after 30 minutes of hot water treatment, sown in farm yard manure (FYM), clean sand, clay soil and sandy soil in the ration of 2:1:1:1. Morphological characters such as plant height (7.00±1.00), number of leaves (31.67±1.53) and root length (18.33±1.53) were also observed highest in polycarbonate greenhouse. Owing to its potential uses in the region and other part of the world make the species under threat. Results provide the comprehensive information about this valuable species for cultivation, ethnobotany, and population census for conservation of E. gerardiana critically endangered medicinal plant of Himalaya.


Introduction
The distribution pattern of E. gerardiana in the Himalayan region considered as critically endangered using IUCN criteria (Samant and Pant 2006) and endangered in Trans-Himalayan cold desert of Ladakh and Lahaul-Spiti (Chaurasia and Gurmet 2003). It is locally known as Tsepat, and mTshe-lDum in Sowa-Rigpa (Gurmet and Rath 2020) and popular trade name is Somlata. Himalayan region medicinal plants have occupied an important values spiritually, culturally, and health bene ts of the peoples. More than 34% of total plant wealth have known to have medicinal value (Ved 2008). The genus Ephedra belonging to member of Ephedraceae, is a group of plants that inhabit temperate regions in Asia, Eurasia, Northern Africa, South-western North America, and western South America. Due to its xerophytic characters, it is a drought and frost resistant. It is represented by 50-65 species of shrubs, rarely small trees (Stevenson 1993; Sharma and Uniyal 2008;Sharma et al. 2010). Three species were reported from Ladakh including E. gerardiana, E. intermedia and E. regeliana (Dvorsky et al. 2018). In India, its distribution extends from Sikkim in Eastern Himalaya to North-West in Uttarakhand, Ladakh, Himachal Pradesh, and Jammu & Kashmir. Genus Ephedra have eleven species that distributed in the Indian sub-continent, mostly in higher elevations of Himalaya preferring alkaline soils. Different species that occur in various regions of country are markedly different from each other in term of their habitat preferences that promote diversity within the genus (Sharma and Uniyal 2008; Sharma et al. 2010). The drug Ephedrine and many active compounds such as ephedrine, pseudoephedrine, norephedrine, norpseudoephedrine, methyl ephedrine, methyl pseudoephedrine, alkaloids, phenoles, terpenoids are obtained from the E. gerardiana and is one such important endangered medicinal plant. In trans-Himalaya of Ladakh the E. gerardiana is being used by local healears (Amchis) and ethnic people to cure various diseases such as chronic fever, wounds, tumours, discomfort in breathing, cough, sweating, urine obstructions (Gurmet and Stobgais 2016). The fruit and shoot is also used for the treatment of asthma, rheumatism and heart stimulant in Sowa-Rigpa by local healears (Amchis) of High altitude of Ladakh (Kunzes et al. 2012). The decoction of areal part is used against bronchial problems and liver disorders. It is also cure menstrual irregularities (Navchoo and Buth 1992). E. gerardiana is also used in Ayurveda, Unani, Siddha, Homeopathy Chines medicines, folk medicine and other traditional m edicines. Due to its resinous smell and taste it is used with tobacco after drying and making powder called Sotak, a pinch of which local people place under the tongue in Ladakh. Twigs used as a tooth brush (Bhattacharyya 1991). The E. gerardiana has been also used for other purposes by ethnic communities, where fruit is eaten in high altitude areas, whole plant used for fuel, aerial part is used for washing utensils and also used for religious purposes in cold desert (Samant and Lal 2019). Owing to its many fold medicinal properties and their folk uses, E. gerardiana is in high demand by pharmaceutical industries and traditional healers. Lack of suitable and effective conservation management, over exploitation, habitat degradation the genus is under brink of extension. For long-term conservation management of this genus, a better and comprehensive understanding of the species remains a top priority, for which detailed studies on its habitat ecology, biology, and agro-technology are not done so far (Samant and Lal 2019). E. gerardiana is an erect shrub of varying sizes, bears cylindrical, striated, often curved branches arising in whorls, dark green in colour. The internodes of branchlet measure 1-5 cm in length and the fruits are 1-3 mm in width are red in colour, sweet in taste and edible. The ovoid fruits contain 1-3 seeds covered by bracts. The rhizomes have large knobs. It is found scattered in the drier regions of temperate and alpine Himalayas from Kashmir to Sikkim at altitude range 2100-4800 m amsl (meter above mean sea level) and similar heights at Pangi (Chamba), Lahaul and Spiti, Chini and Kilba Kailash ranges of Kanawar (Kinnaur), Shali hills (Shimla), Kashmir and Ladakh. The habitat preference of the plant are sandy slopes, rocky slopes, sandy places and driers area of Himalayas. In this present study we are focusing on population census, ethnobotany, agro-technology for in-situ and ex-situ conservation in Ladakh region.

Materials And Methods
Temperature and lux intensity . Semi-structured questionnaires, interviews and participatory reviews were used to elucidate information from the experienced persons (Amchis) using standard methods (Martin 1995). Information on local names of plants, and plant parts used for curing various ailments were recorded. Field visits were also planned with the elders and Amchis to ascertain the plants and also to obtain rst-hand information on their distribution. The plants is collected for identi cation and herbarium preparation following standard methods (Jain and Rao 1976). The voucher specimens are submitted to National Institute of Sowa-Rigpa Leh, Ladakh for future reference.  Table 1.
Survey, sampling, identi cation and analysis of data The eld surveys and samplings were carried out in 2018-2019 within the selected sites. In each site a plot of 20x20m was laid. Species were sampled by randomly placed 20 quadrats of 1x1 m in each plot. The size and number of quadrate was determined following Misra (1968) and Kersaw (1973). For the collection of data from these quadrats and data analysis were done following standard ecological methods (Grieg-Smith 1957; Kersaw, 1973 In the questionnaire survey of the study area, we collected the information from 75% and 25% male and female respectively. Out of total informants, only 41% males and 31% females were well versed with the use of E. gerardiana and they also know the potential sites of the targeted species where they used to visit along with the researchers and tourists. According to the respondents, it was clears that the target species facing grave threat due to the over exploitation and illegal collection made by the local inhabitants and also by the outsiders. In optimized cultivation packages and practices in Ladakh condition, we observed that the species can be cultivated by raising the nurseries with freshly harvested seeds treating with hot water for 30 minute and sown in the mixture of farm yard manure (FYM), clean sand, clay soil and sandy soil in different conditions. In polycarbonate greenhouse average annual temperature is maximum 32.

Discussion
Based on present study E. gerardiana is an important plant species for local dwellers and Amchies who take care of health of the inhabitants of the region, and other part of the world. Many important active compounds are obtained from the species having high demand globally. Owing to its many important medicinal and folk importance leads to over exploitation and habitat degradation, due to which it may extinct from the wild of cold desert of Ladakh, study also con rm that the species has very low density individual/m 2 in the region, study conducted by Samant and Pant (2006) using IUCN criteria in the Himalayan region also con rmed that it is on brink of extinction on distribution of E. gerardiana therefore, it's very important to conserve the species. On the basis of density of the species in the region and ethno-botanical study, optimization of cultivation techniques was made, for standardization of the packages and practices we opt for the simple seeds treatments with hot water with different timing and used high cost and low cost greenhouses to raise the nurseries so that distinguish farmers can afford for the cultivation of the E. gerardiana. The study revealed that freshly harvested seeds can be used to raising the nursery which required quite moist and warm temperature, therefore different protective conditions is recommended coupled with hot water treatment is highly

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