RESEARCH ARTICLE Exploration, Conservation, and Cultivation of japonica (Thunb.) Miers Rama Shankar1, Shazia Neyaz2, Gyati Anku3, Late Wngsu Rungsung4

Abstract​ Aims and objectives: Stephania japonica (Thunb.) Miers belonging to family and is a highly demanded medicinal used in the name of Rajpatha for treatment of fever, malaria, jaundice, and various stomach disorders by the local traditional healers and pharmaceuticals for the preparation of medicines for related diseases. It is widely distributed in Northeastern , i.e., Arunachal Pradesh, Assam, Manipur, Meghalaya, Mizoram, Nagaland and Tripura. Because of acute pressure on wild sources due to rapid exploitation and changing forest areas into agriculture land and various other developmental activities in a very short time, there will be the shortage of drug in the wild and needs to be cultivated this along with several others. Accordingly, a study has been carried out with the aim to develop easily accessible agro- technique for undertaking cultivation by the farmers in an easily accessible manner. Outcome of the study: This plant is a highly demanded medicinal plant distributed in North East India. Its cultivation is through tuberous root. Seeds also act as a tool for giving rise to new plants initiating from the mature viable seeds fallen on the ground. Seed germination is more accessible for this plant. The authors has developed various cultivation parameters in Regional Ayurveda Research Institute, Itanagar (Arunachal Pradesh) through seed germination and roots. Conclusion: Seeking out the practices of traditional use of tuberous root of S. japonica (Thunb.) Miers, there is an urgent need of cultivation. Accordingly, easily acceptable techniques for its cultivation is described in the present communication and cultivation can be made by establishing plant nurseries. Keywords: Agrotechnique, Cultivation, Stephania japonica. Journal of Drug Research in Ayurvedic Sciences (2020): 10.5005/jdras-10059-0098

Introduction​ 1Regional Ayurveda Research Institute, Jhansi, Uttar Pradesh, India The plant of Stephania japonica (Thunb.) Miers belonging to family 2,3Regional Ayurveda Research Institute, Itanagar, Arunachal Pradesh, Menispermaceae and is commonly known as Rajapatha (locally India Tape ). Occasionally distributed in foot hills of Arunachal 4Herbal Ayurveda Research Centre, Nagaland University, Lumami, Pradesh, Assam, Manipur, Meghalaya, Mizoram and Nagaland, Nagaland, India it has been recorded to be used in the cure of various ailments, Corresponding Author: Rama Shankar, Regional Ayurveda Research namely, asthma, tuberculosis, dysentery, hyperglycemia, malaria, Institute, Jhansi, Uttar Pradesh, India, Phone: +91 9436898754, e-mail: cancer, and fever.1–4 [email protected] Over 150 alkaloids together with flavonoids, lignans, steroids, How to cite this article: Shankar R, Neyaz S, Anku G, et al. Exploration, terpenoids, and coumarins have been identified in the genus Conservation, and Cultivation of Stephania japonica (Thunb.) Miers. Stephania, and many of these have been evaluated for biological J Drug Res Ayurvedic Sci 2020;5(3):139–142. activity. Protostephanine, an antihypertensive alkaloid, is present in Source of support: Nil 5–7 the species under stud. Flowering takes place during the month Conflict of interest: None of May with the appearance of of around 5–6.5 cm. This plant is of great commercial value in all the states and are being exploited from natural habitat without any concept of developing suitable agro-technique for cultivation. Accordingly, the future conservational aspects to fulfill the need of pharmaceutical authors have made exhaustive exploration and also made attempt firms in all parts of the country. The use of roots of this plant to develop suitable agro-technique for the cultivation of this plant in malarial fever, stomach and cardiac disorders, and diabetes under which following initiative steps were taken: develops better scope for its cultivation through easily accessible agrotechnique and being recommended for in situ conservation • An attempt had been made to transplant naturally occurring in this communication. plant from the natural habitat to the herbal garden of Regional Ayurveda Research Institute, Itanagar. After getting successful luxuriant growth, the attempt had been made for plantation Aims and Objectives​ through tuberous root with terminal buds of this plant. The With S. japonica (Thunb.) Miers being a most valuable medicinal bulbils showed the development of new plants. After obtaining plant distributed in most parts of Northeastern states and Western experiences on growth and development of new plants, a large- Himalayas, and being taken under trade and exploited from wild scale plantation was made. For in situ conservation, studies were sources due to an acute pressure in the crude drug market, there is regularly conducted during various field excursions from time an urgent need to cultivate this plant in the Northeastern states by to time in areas where the naturally occurring plants were found

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with respect to fruiting and seed formation. Field observations were on viability of seed germination in natural wild field and found that the mature seeds fallen on ground in the vicinity of the plants get germinated and young seedlings also developed. • Recorded various positive and negative aspects for development of new plants in field and places of plantation. Germination of seeds in the field including viability period and other precautionary measures were also recorded. • Observations on the growth and development of seedlings under nursery plantation plots. • Records were maintained on gestation period for the study.

Materials and Methods​ During various explorations of medicinal plants, voucher specimens were collected from the field along with collection of germ plasm, Fig. 1: Photographs of different stages of cultivation of Stephania i.e., young plants from field. The newly germinated plants that japonica were collected from the field were planted into earthen pots as well as in the garden of the institute for first-hand plantation. After successful growth of plant in the garden with support for climbing growth in comparison to other months (Fig. 1). For transplantation, the germ plasm, young tubers of this plant were planted in the the individual planting material is the whole piece of tuberous field, and nursery bags were filled with manure and garden soil at root with active buds of 2.5–3 cm size. These root pieces are different monthly intervals. Young plants in natural field showed transplanted in field directly. Maximum plantation has been made that seeds of this plant are the basic germ plasm. Organic manure during premonsoon, i.e., in month of April–June. Delay in sowing made from litter, cow dung, and decayed plant litter was used for causes poor initiation of shoot, particularly during extreme rains healthy growth. On completion of sowing, growth of the planted due to precipitation of organic manures and minerals from the soil. seedlings was observed at different intervals. Thus, in situ conservation is a suitable practice for its sustainability. Land for transplantation was prepared by digging pits of 30 × 30 Germinated young plants in nature can be transplanted to other × 30 cm and filled with a mixture of sand, clay, and organic manure protected places also. Rooting does not initiate in stem cuttings in equal proportion. For propagation through seeds, attempts to develop into new plants. Fertilizers used for this plant is organic were made to study the germination of seed in the nursery. In situ nitrogen-rich manure and wormy compost. Nitrogen acts as strong conservational aspects were also considered. The soil conditions fertilizer for the growth and development of tuberous plants. of the natural fields were also recorded. Watering is done twice daily, as the plant is moist and shade loving. Due to heavy rains for a longer period, weeds affect the growth of Observations​ the plant. Tuberous roots act as a weed-suppressing crop, and one or two light weeding in the first and second month of planting is The plants of S. japonica (Thunb.) Miers is distributed in Arunachal sufficient to minimize the weed population. Removal of old and dry Pradesh in foot hills adjoining Assam up to West Kameng, East and sheath is beneficial for root elongation. Temporary barbed and West Siang, Mizoram in Kolasib, Lunglei, Serchip and Mamit wire fencing round the cultivation area supports the growth and forest divisions, in Manipur in Senapati and Ukhrul, in Nagaland development of the plant in the field. Weeding in the vicinity of in Dimapur and Mokokchung forest divisions, and certain parts plants is helpful in seed germination fallen on ground. Sandy loam, of Assam and Meghalaya. Attempt has been made for developing red laterite, humus-rich soil is the best for cultivation. A well-drained easy and accessible cultivation methodology of this plant through soil rich in organic matter is very good for the growth of this plant. seed germination, stem cuttings, and tuberous root cuttings. The plant being very hard can be grown in tropical and subtropical Seed germination and developing seedlings through seeds were climatic conditions. successful both in nursery and the natural fields. Being a dioeciously New seedlings attaining the length of 2–3 m are safe for vine/climber, fruiting is minimum in nature. Natural climatic transplantation with support for further growth. The stem conditions were selected for plantation of the two plants located cuttings planted during heavy rains or winter remain as such in foot hills having climbing support. Sandy and sandy loam to in the polythene bags till the arrival of next favorable season, clay loam with organic litter was found suitable at geographical and initiation of tender vine takes place from node during the positioning given as under: months of March, April, September, or October. It shows 90% N 27°06.138; E 093°38.883 results in the growth and development of new plants. The soil of the plantation site is acidic with pH value of can be harvested after 1 year and root during third year from the 4.63 and moisture content 21.7%. Soil sample contains sodium, side without damaging the plant which is confined to only one or potassium, calcium, and nitrogen at 0.56%, 1.17%, 10.48%, and two adjacent places in the tuber. Results of the study is supported 0.37%, respectively, whereas the polythene bags were filled with with photographs (Fig. 2). garden soil mixed with organic manure and sand. Seed germination in nursery bags as well as in natural habitat is a viable practice. Germinated seedlings having entire tuberous root is used as Conclusion​ propagules. Roots were collected during January and February Stephania japonica (Thunb.) Miers being a climbing plant after attaining a tuber size of 5 cm. The diameter shows better grows very well in the slopes and showed good results on seed

140 Journal of Drug Research in Ayurvedic Sciences, Volume 5 Issue 3 (July–September 2020) Exploration, Conservation, and Cultivation of Stephania japonica (Thunb.) Miers

Figs 2A to D: Photographs showing various stages of developments germination and plantation of its tuberous root. Tuberous roots Pharmacognosy is also thankfully acknowledged. Thanks are have sufficient nutrient, water content, and chemical constituents also due to Shri DJ Payeng, Field Attendant for various assistance for self-defense and survival but the growth is slow. In case of the rendered in field and garden of Regional Ayurveda Research young seedlings with leaves, i.e., 3-months-old seedlings, they Institute Itanagar during course of studies. are field planted in 30× 30 × 30 cm pit at 3 m spacing with the supports of bamboo sticks below the shade of trees like Bauhinia eferences variegata, Melia azedarach, Schima wallichii, Vitex negundo, etc., R for the support. 1. Anonymous. The Wealth of India, Row Materials. New Delhi, India: In situ conservation is best suitable practice for sustainability. Publication. and Information Directorate CSIR; 1989. p. 41. 2. Anonymous. Indian Medicinal Plants. Orient Longman, India: Arya Removal of old and dry leaf and sheath is beneficial for root Vaidya Sala; 1994. p. 280. elongation. New seedlings attaining the length of 2–3 m are safe 3. Chopra RN, Chopra IC, Handa KL, et al. Chopra’s Indigenous Drugs of for transplantation with support for further growth. The stem India. 2nd edn, Calcutta, India: Char UN and Sons, Ltd.; 1958. p. 412. cuttings planted during heavy rains or winter remain as such in 4. Chopra RN, Nayar SL, Chopra IC. Supplementary to Glossary of Indian the polythene bags till the arrival of next favorable season, and Medicinal Plants. New Delhi, India: PID; 1956. initiation of tender vine takes place from node during the months 5. Nishanthi M, Artharthanarieswaran P, Devdas G, et al. Pharma­ March, April, or September–October. It shows 90% results in the cognostical studies on leaves of stephania japonica var. timoriensis. growth and developing. Int J Pharm Sci 2011;2(1):139–141. 6. Kumar SD, Ruchi B, Ravindra S, et al. The genus Stephania (Menispermaceae): chemical and pharmacological perspectives. cknowledgments J Ethnopharmacol 2010;132:369–383. DOI: 10.1016/j.jep.2010.08.047. A 7. Habibur Rahman M, Badrul Alam M, Chowdhury NS, et al. Antioxidant, Authors are thankful to the Director General, CCRAS for financial analgesic and toxic potentiality of Stephania japonica (Thunb.) Miers. assistance through project. Programme officer and Asstt. Director leaf. Internat. J Pharmacology 2011;7(2):257–262.

Journal of Drug Research in Ayurvedic Sciences, Volume 5 Issue 3 (July–September 2020) 141 Exploration, Conservation, and Cultivation of Stephania japonica (Thunb.) Miers

142 Journal of Drug Research in Ayurvedic Sciences, Volume 5 Issue 3 (July–September 2020)