Indian J. of Agroforestry Vol. 12 No. 1 :71-76 (2010) ” Effect of tree-crop combinations and nitrogen levels on herbage yield of sacred basil (Ocimum sanctum L.) grown in agrihorti-silvi-pasture system in mid hill Himalayas K.S. Verma’, N.S. Thakur’ and R.C. Rana’ "Department of Environmental Sciences, Dr. YS Parmar University of Horticulture and Forestry, Nauni, Solan-173 230 (Himachal Pradesh) The Energy and Resources Institute, Darbari Seth Block, IHC Complex, Lodhi Road, New Delhi-110 003 Department of Forest Products, Dr. YS Parmar University of Horticulture and Forestry, Nauni, Solan-173 230 (Himachal Pradesh) ABSTRACT : Investigations on effect of tree-crop combinations and nitrogen levels on growth and herbage yield of Ocimum sanctum L. were carried out during 2004-2005 and 2005-2006, in experimental farm of Department of Silviculture and Agroforestry, Or. Y S Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh (india), 0. sanctum was grown in association with Prunus persica (Peach), Grewia optiva, Morus alba and Setaria sphaceleta. The distinctive tree-crop combinations were, Peach + Grewia + Setaria + O. sanctum, Peach + Morus * Setaria + O. sanctum, Peach + Setaria + O. sanctum, Grewia + Setaria + O. sanctum, Morus + Setaria + O. sanctum and ©. sanctum as sole crop. Three nitrogen doses applied to O. sanctum were 40, 80 and 120 kg/ha. Maximum plant height (90.70 cm) and leaf area (14.62cmi‘lleaf) of O. sanctum was recorded under tree-crop combination Peach+Morus+Setaria. However, leaf area index (LAI) was higher (2.77) for the plants grown under Peach+Grewia+ Setaria, Nitogen dose of 120 kg/ha produced tallest plants (90.96em) with leaf area of 15.84 cmleafand LAL 3.16. The total herbage yield was not affected by tree-crop combinations however, it was mayimum (5.56tha) under Peach+Norus+Setera, Plant growth attributesi.e. plant height, leaf area LAl along with herbage yield increased significantly with the increase in N dose. Key words: Ocimum sanctum, tulsi, agroforestry nitrogen, herbage yield 1. INTRODUCTION Ocimum sanctum L.. (Family Lamiaceae) “Sacred Basil or Holy basil’, Basil, the "Royal Herb" in India, is popularly known as Tulsi. Tulsi has been described as early as the vedic period. Itis the most sacred and holy plant of India dedicated to Hindu culture and ceremonies. Its leaves are used in the worship of gods, goddesses and part taken as Prasad (Holy food). Apart from having religious importance in India, it is an important medicinal plant used to treat common cold, asthama, bronchitis and fever. All plant parts have medicinal value (Charak Samhita, 1997). It is used in numerous formulations by more than 7,000 pharmacies of Ayurveda, Sidha, Unani ‘and Homeopathy in India. O. sanctum has also been included in the list of species prioritized by National Medicinal Plants Board (Herbals in India, NABARD publication). Singh and Parabia (2003) have estimated 30.5 tonnes annual consumption of leaves of holy basil by 34 per cent pharmacies in Gujrat state alone, which come from cultivated land. The demand and supply gap for O. sanctum was estimated to the tune of 925.1 to 3038.4 tonnes from year 2001-2002 to 2004-2005 (Anon.., 2002). National Medicinal Plants Board of India (New Delhi, India) has estimated annual demand of holy basil 3296.8 tonnes in 2001-2002, which at an annual growth rate of 17.9 per cent, might have increased manifolds till now. It is very essential to conserve and cultivate medicinal plants on ‘commercial scale to meet the growing demand which our depleting wild stock can not cater. It is well established fact that medicinal plants grow in association with trees and shrubs in nature. This association could be replicated successfully on farms and in the agricultural landscape. About 75 percent of Himalayan population depends directly or indirectly on agroforestry products and services in rural and urban areas as a result of changing forest management policies and laws which are conservation centric and debars the people to use the natural wealth. However, the increased ‘awareness of the benefit of medicinal and aromatic plants, their market demand coupled with restrictions on collection from natural wild habitats have offered an opportunity to the farmers to cultivate these medicinal herbs and diversify the existing agricultural land use systems. Therefore, the present investigations were carried out to find out the growth and yield of O. sanctum in association with fruit and fodder trees and grass species to evolve suitable tree-crop combinations and to find out the response of O. sanctum to varying nitrogen levels. 2. MATERIALS AND METHODS 2.1 Study area The investigations were carried out at experimental farm of Dr. Y.S. Parmar University cf Horticulture and Forestry, Nauni Solan, Himachal Pradesh, India, located within 30°51 N latitude and 76°11" E longitude (survey of India Toposheet No. 55 F/1) at an elevation of 1250m above mean sea level. Climate of the area is transitional between subtropical to sub-temperate with maximum temperature rising up to 37.8°C during summer. ‘The mean annual temperature is 19.8°C. May and June are the hottest months, whereas December and January are the coldest. The annual rain fall2 ect of teorp combinatons and nitrogen levels on herbage yield of sacred bas (Ocimum sanctum L.) ranges between 800-1300 mm of which 75 percent is received during mid June to mid September. The Parent material consists of sand stone, conglomerate, boulders, dolomite and is calcarious. The soil type belongs to order inceptisols and typic entrochrepts with gravely sandy loam texture. The surface soil was high in organic carbon (1.01%), medium in nitrogen (457.70 kg/ha), high in available phosphorus (31.50 kg/ha) and potassium (354.20 kg/ha) with a PHof7.0. 2.2 Tree-crop combinations The structural components of agroforestry system were Prunus persica (fruit), Morus alba and Grewia optiva (fodder) and Setaria sphacelata (grass), planted in row planting geometry, in East to West direction during 1992, 2001 and 2002, respectively The O. sanctum was cultivated between the tree and grass component. The tree-crop combinations, formed were: Peach + Grewia + Setaria + O. sanctum (T,), Peach + Morus + Setaria + O. sanctum (T,), Peach + Setaria + O. sanctum (T,), Grewia + Setaria + 0. sanctum (T.), Morus + Setaria + O. sanctum (T,), O. sanctum sole (T,). Three nitrogen doses 40 (Nj), 80 (Ny:) and 120 (Ni) kg/ha, were applied to find out the effect on growth and yield of O. sanctum. The experiment was laid out in randomized block design replicated thrice. Details of area occupancy and magnitude of different components under different tree-crop combinations have been given in Table 1. Field preparation involved two ploughings followed by harrowing and planking with tractor. Plots were prepared as per the treatment details. Well rotten farm yard manure @20 tonnes ha" was applied before transplanting. Recommended basal dose of phosphorus and potassium for O. sanctum (40 kg P,O, and 40 kg K,O/ha) were also applied. With the commencement of monsoon in last week of June, healthy seedlings of O. sanctum were transplanted at a spacing of 40 x 40 cm. Light irrigation was given after transplanting to facilitate better establishment of seedlings. Nitrogen was applied in two split doses i.e. half dose of N in the form of urea along with full basal doses of P and K at transplanting. The remaining dose was given 30- 40 days after transplanting. Hand weeding was done as and when required. 2.3 Growth and herbage yield of 0. sanctum Plant height was measured in centimetres with the help of measuring tape from randomly selected Plants. The leaf area of fully extended leaf was measured using pre-calibrated portable leaf area meter (Cl-203, INC. USA) and is expressed in cm’ per leaf. Leaf area index (LAI) was measured with the help of pre-calibrated, pre programmed LAI- 2000 plant canopy analyzer (LICOR-USA). The above and belowground biomass was taken at full bloom stage. Fresh weight was recorded immediately after harvesting. Dry herbage yield (leaves, stems and roots) was taken after drying in shade to a constant weight. 2.4 Growth and yield of fruitand fodder trees The growth parameters (tree height, diameter, basal area and crown area) were measured following standard mansurational techniques and formulae. The silvological characteristics (average values) of fruit and fodder trees have been presented in Table 2. Yield of fodder and fuel wood was taken when the lower leaves tuned yellow. The fodder and fuel wood yield was calculated on fresh as well as dry weight basis. Fuel wood yield for peach was calculated by taking all the prunes from individual tree. Likewise, fruit yield for peach was taken at the time of harvesting. The data generated were analysed with the procedure outlined by Gomez and Gomez (1984). 3. RESULTS AND DISCUSSION 3.4 Plantgrowth of 0. sanctum It is clear from Table 3 that plant height of ©. sanctum had significant influence of different tree- crop combinations and N levels. It was maximum (90.70 cm) in T, (Peach + Morus + Setaria) and minimum (82.23 cm) in T, (Morus + Setaria) Nitrogen level of 120 kg/ha produced the tallest Plants (90.96 cm) and 40 kg N/ha shortest (81.97 cm). In general, increase in the N level increased plant height, in all the tree-crop combinations. The interaction effect between different tree-crop combinations and nitrogen levels was although non-significant, the highest value of 93.24 cm was recorded by T,N,,.. Asintercrop, O. sanctum plants had either equal or significantly higher height compared to sole crop. The results thus indicated that intercropping of O. sanctum with woody perennials in agroforestry will have enhancing effect on the plant height. However, association of certain woody perennials like Morus [T,(Morus + Setaria + 0. sanctum)] may depress height growth Investigation carried out by Dutt (2004) under Poplar based agroforestry systems, reported adverse effect on plant height of O. sanctum but at closer spacing. Thus, it can be inferred that if suitable woody species planted at optimum spacing, plant height of O sanctum can be enhanced under various agroforestry systems. Further, @ significant increase in height was ‘observed due to N application. The N dose of 120 kgiha produced tallest plants. Nitrogen bei responsible for stimulating vegetative growth in Plants, the above results are hence in accordance with its role in plants. A significant increase in plantIndian J. of Agroforestry Vol. 12 No.1 : 71-76 (2010) 3 Table 1. Details of area occupancy and magnitude of different components (per hectare) of various tree-crop combinations ‘Tree-crop combinations Spacing No-of tangas Landes treesiha under ‘grasses trees inlercropping (mi) MAPS (mr) T, (Peach + Grewia + Setaria + 0. sanctum) Peach Grewia Setaria T, (Peach +Monus + Setaia + 0. sanctum) Peach Moris Setaia Peach Setaria Grewia Setaria Morus Setaria Sx10m 5x10 m T,, (Peach + Setaria + 0, sanctum) 5x10 T,. (Grewia + Stara + O, sanctum) 1x10 (Morus + Setara + 0, sanctum) axiom T, (0. sanctum sole) 2002000 ~=—=«OOO ‘x10 m (3 trees in between 2trees of peach) 600 Planted in blocks (400 m2) on stip for trees 200 © 2000» 8000 ‘x10 m (8 trees in between two tres of peach) 800 Planted in blocks ($00 m2) on stip for trees 200 = 2000000 Planted i blocks (1200 m2) on strip for tees 1000 20008000 Planted in blocks (1000 m2) on stip for tees 1000 2000-8000 Planted in blocks (1000 m2) on stip for tees Ni NL 8000 Table 2. Silvological characteristics (average values) of fruit and fodder tree under Ocimum sanctum based tree-crep combinations Teezepconbinatons Peach Crone cpa Tra aba ‘Voor’ Oameter “Sewn, Magee “Saree “eraee Agaee Messe Aone (m) (cm) _area(m’) (im) (cm) area (mm) (m) (cm) area (m’) T, (P+G+S+0. sanctum) 419 975 10.71 8.86 4.76 287 . . . T, (P#M#S+0, sanctum) 3.34 1061 647 - - - 422 632 6.09 T.(P*8+0 sanctum) 31879885 - is i : : T.(G+S+O. sanctum) : : : T.(M#S#0. sanctum) : T.(O. sanctum) “ u 7 301378248 2 7 : : - 7 380-551 4.28 "Som above the graft union N= Nitrogen, P = Peach, G = Grewia optiva, M = Morus alba, S = Setaria sphacelata, O = Ocimum height at higher level of nitrogen application may also have occurred owing to medium status of soil nitrogen. Similar results were observed by Pareek et al. (1980) for O. sanctum and Ajimoddin (2005) for O. basilicum. Positive effect of nitrogen application on plant height of other medicinal plants like Melisa offinalis (Sood. et al,1996) and Salvia sclaria (Rastogi etal, 1997) have been reported. Leaf area was influenced significantly due to different tree-crop combinations. Plants grown in association with tree-crop combination T, (Peach + Morus + Setaria) had maximum leaf area (14.62 cm‘leaf). However, it was statistically at par to the leaf area (14.00 cm*/leaf) of plants grown as sole crop. The plants grown in T, (Peach + Grewia + Setaria) attained the minimum leaf area (13.370m‘lleaf). Leaf area of the plants in treatments T,, T,, Tand T. was statistically at par at P<0.5 (Table 3). Among N levels, the highest dose (120 kg/ha) produced leaves with significantly greater leaf area (15.84 cmilleaf), whereas 40 kg Nina produced plants with lesser leaf area (12.25 cm‘lleaf). The interaction between tree-crop combinations x N levels was observed to be statistically non-significant. All tree-crop combinations significantly influenced the leaf area index (LAI). The maximum LAI (2.77) was found in T, which was statistically at par with LAI at T, and T,. Minimum LAI (2.67) was for plants grown in, and'T,, however the values were at parT, ‘Le. Morus + Setaria + O. sanctum (Table 3). Varying N levels influenced the LAI significantly. The maximum LAI (3.16) was at level of 120 kg/ha which ‘was statistically higher over the remaining two levels viz. 40 kg (2.31) and 80 kg/ha (2.31). The interaction effect between tree-crop combinations and N levels™ rect oftree.crop combinations and nitrogen levels on herbage eld of sacred bas (Ocimum sanctum L.) \was statistically significant at P0.05. The maximum LAI (3.38) was recorded at T, with 120 kg N/ha whereas, minimum (2.21) was also at T, but with application of 40 kg Niha (Table 3). Leafarea index of . sanctum was also affected significantly due to tree-crop combinations, which was significantly less at T, over T,. The remaining combinations including sole crop did not differ significantly among themselves. Application of N_ progressively increased the leaf area (from 40, 80 to 120 kg Niha), with maximum at 120 kg Niha. Plants grown in association with Peach + Grewia + Setaria (T,), followed by Peach + Setaria (T,) had significantly higher leaf area index as compared to sole crop. The values of LAI obtained in present investigation are in conformity to those reported by Dutt (2004) under poplar based tree-crop combinations. He reported that plants grown under closely spaced poplar plantations showed lesser LAI whereas higher leaf area under wider spacing compared to sole crop of O sanctum. It is amply clear from the findings that varying N levels significantly increased the LAI (Table 4) with maximum at 120 kg N/ha. This is attributed to the fact that higher dose of N resulted in tallest plants with large leaf size and dense crop. 3.2 Herbage yield O. sanctum Different tree-crop combinations did not affect the aboveground herb yield significantly but nitrogen levels of 40, 80 and 120 kg/ha influenced it, significantly. The N dose of 120 kg/ha gave maximum aboveground herb yield (5.65 tha) and lowest dose (40 kg/ha) gave minimum (4.05 ha). The interaction effect of tree-crop combinations and N levels (Table 4) was non-significant. Belowground biomass yield varied significantly due to tree-crop combinations (Table 4). The maximum yield (0.45 tha) was in T, (Grewia + Setaria + O. sanctum). But it was at parwith T,, T,, T,, and T,, Minimum yield (0.39 t/ha) was obtained under tree-crop combination T, (Peach + Morus + Setaria + O. sanctum) which was at par with T, (Peach + Grewia + Setaria + O. sanctum). The varying nitrogen levels had significant effect on belowground biomass yield. The nitrogen dose (120 kg/ha) produced maximum root yield (0.49 Uha) and the lowest dose i.e. 40 kg N per hectare produced the minimum (0.37 tha) root biomass. The belowground biomass owing to interaction between tree-crop combinations and Nlevels bear no significant effect The total herbage yield showed non-significant effect on its yield due to different tree-crop combinations (Table 4). On the other hand different N levels influenced the total herbage yield (above and belowground) significantly. The highest dose of N (120 kg/ha) gave the maximum yield (6.15 tha) and the lowest dose 40 kg Niha yielded minimum. The total herbage yield owing to interaction between tree-crop combination and N doses was non-significant. The highest aboveground and total herbage yield was achieved under tree-crop combination comprising Peach + Morus + Setaria + O, sanctum (T,). The above findings revealed that O. sanctum could be boosted if {grown with suitable woody perennials. Herbage yield obtained in present study was less compared to that reported by Pareek and Gupta (1984) for sole crop. This may be attributed to variation in climatic and soil conditions which were different under both cases. Furthermore, the varying levels of N (40, 80 and 120 Table 3. Effect of tree-crop combinations and nitrogen levels on plant height, leaf area and leaf area index (LAI) of Ocimum sanctum grown under rainfed conditions Nievel kglnall Piantheight(om) Leatarea(enTeat) TAL Tree-rop combinations Ne Ne Ne Mean Ne Ne Na Mean No Nw Nw Mean T.(P#G#S+0. sanctum) 140 6452 9024 8699 1142 1349 1851 1337 221 271 338 277 T.(P#M+S+0, sanctum) 8601 9089 9521 9070 1918 1468 1599 1462 294 269 312 272 T,(P+8+0. sanctum) 8207 84.19 9094 STE 1224 1345 1553 1364 233 279 918 278 T.(G+S+0, sanctum) 8153 8225 9099 8492 1260 1359 1658 1393 233 260 918 267 T,(M#S+0, sanctum) 7123 8127 8819 2.29 1195 1363 1692 1383 298 262 304 270 T,(O. sanctum) 8155 B01 8817 8492 1211 1396 1652 1400 224 265 309 267 Mean 8197 8482 90.96, 1225 1360 1584 231 267 316 CDi T 188 068 0.96 N 133 048 0.04 TN Ns NS. ost NeNirogen, P=Peach, G= Grewia optva, M= Morus alba, S= Selariaphacelate O=OcmumIndian J. of Agroforestry Vol. 12 No.1 : 71-76 (2010) kg N/ha) progressively increased the biomass yield and higher biomass yield was achieved with the application of 120 kg Niha. Pareek et al,, (1980) reported that increasing levels of nitrogen from 0 to 40 and 80 kg N/ha progressively increased herbage yield. Pandey et al, (1978) reported that application 0f 120 kg N, 105kg each of P and K per hectare gave maximum herbage yield of O. sanctum. Increased herbage yield at varying N doses have also been reported for O. basilicum (Ajimoddin et al., 2005), Salivia sclaria (Rastogi eft al, 1997) and Melissa officinalis (Sood et al, 1996). 3.3 Yield attributes of Peach, Grewia, Morus and Setaria Yield attributes of tree and grass species forming different tree-crop combinations have been presented in Table 5. Results obtained have been described componentwise as under: 75 Peach : Fuel wood yield (prune wood) was maximum (8.06 q/ha) from tree-crop combination ‘ofeach + Grewia + Setaria (T,) followed by T, and T, (Table 5). Fruit yield was highest (31.90q/ha) for treatment T, (Peach + Grewia + Setaria) followed byT, and, (30.50 q/ha). Grewia optiva : Tree-crop combination (T,) Grewia + Setaria gave maximum (9.20 q/ha) average dry fuel wood yield followed by T, (6.57q/ha). Average fodder yield expressed both as fresh as well as dry weight basis, was maximum (7.40 q/ha fresh: 3.50 q/ha dry) in T, followed by T, Morus alba : Fuel wood yield was highest (13.30 qjha) for T, (Morus + Setaria) followed byT, (8.89 qlha). Tree-crop combinations with components Morus + Setaria provided maximum (18.90q/ha fresh ; 7.10 qiha dry) fodder, followed by T, (12.68 qhha;7.73 qiha dry) Table 4. Effect of tree-crop combinations and nitrogen levels on herbage yield of Ocimum sanctum grown under rainfed conditions Wievel (kg Herbage yield (ha) Tree-crop combinations Above ground Below ground Total No Ne Ns Mean Ne Na Nw Mean Ny Na Na Mean T(PsG+S+0 sanctum) —“418—«496—=—«565 «MSCS «(Odd (MTA <5Ad 609536 T,(PAM*S+O, sanctum) 4405.17 5845.14 035 Ot O44) 099475 558 635 5.56 T,(P#8+0, sanctum) 379 471 561 470 037 048 049 04d 416 5.13 tT 54 T.(G+S#0. sanctum) 390 «479-562-477 038 048 «053045428524 BIS 522. T.(M#S+0, sanctum) 397 479-557 478 036 046 084 «04S 433 525 609 5.23 T.(0. sanctum) 407 482-558-482 038 047 081 «045 445 529 609 5.28 Mean 405 488 5.65 037 045 049 442 533 648 Dee T Ns 0.04 NS. N 020 0.03 021 THN Ns NS Ns N= Nitrogen, P = Peach, G = Grewia optva, M= Morus alba, S= Sefaria sphacelata, O= Ocimum Table 5. Yield attributes of peach, Grewia, Morus and Setaria under Ocimum sanctum based tree-crop combinations Tree-cop combinations Peach Grows optiva Morisalba ‘Solara Fal Te Fal Fiera) Fuad Fodierio(aha) gas fed (ha) yield Fresh = s(Dy yield Fresh Dry (ha) (ha) (aha) _weight__weight__(@iha)_—_—_weight weight T.(P*G¥S+O. sanctum 806 3190 —~=«SST SCC - - 854 T.(P#M#S+0.sanctum) 5.76 3050 : : : 8.89 1268 7739.88 T.(P#8+0. sanctum) 72 3050 : . ‘ 5 23.78 T.(G+8+0, sanctum) - - 920 7403.50 2 : : 21.55 T.(M#S+0, sanctum) : : : 13.30 1890 710-2330 T.(O. sanctum) : N=Nitrogen, P=Peach, G= Grewia otiva, M= Morus alba, S= Setaria sphacelat, O= ocimum76 Efecto tree-cop combinations andnirogen lvls on herbage yield o sacred bas (Ocimum sanctum L.) Setaria sphacelata : Setaria dry grass yield was maximum (28.78 q/ha) with T, (Peach + Setaria) followed byT,,T,,T,andT, (Table 5). 4. CONCLUSION The present study have indicated that plant height, leaf area, leaf area index (LAl) aboveground, belowground and total herbage yield of O. sanctum was maximum in tree-crop combination Peach + Morus + Setaria + 0. sanctum. The nitrogen dose of 120 kg/ha resulted in pronounced growth and herb yield over the two N levels i.e. 40 kg and 80 kg Niha. Tree-crop combination Peach + Morus + Setaria gave maximum herbage yield, nonetheless, the tree-crop combinations Peach’ + Grewia + Setaria + O. sanctum would be equally important in meeting the diverse needs of farmers for fruit, fodder, fuel wood and also fulfil the escalating demand for tulsi herb. Thus, medicinal plants based agroforestry systems will ensure socio- economic growth, health care, availabilty of fodder round the year ‘as well as supplementing the demand of pharmaceutical industry. REFERENCES Ajimodii, 1, Vasundhara,M., Radhaksishna 0, Bradar, SL. ‘and Rad, G.G.E. 2005. integrated nutrient management studies in sweet basil (Ocimum basicum L.). Indian Pertumer 49(1), 95-101 Anon. 2002. Ministry of Health and Family Welfare. GOL Charak Samhita, 1997. Chikitsa Sthana. Chowkhamaba Publishers, 36 (English Editon) Dutt, V. 2004. Screening of medicinal and aromatic plants for popular (Populus doltoides Marsh) based agroforestry system under sub-Himalayan conditions. Ph.D. Thesis, Dr. Y.S, Parmar University of Horticulture and Forestry, Nauni, Solan (H.P.)-Ingia, Gomez, K.A. and Gomez, AA. 1984. Statistical Procedures for ‘Agficuitural Research, John Willey and Sons, New York. pp 89-107 sha, K.K. and Gupta, C. 1991. Interecopping of medicinal plants with poplar and their phenology. Indian Forester. 117:535- 5a Pandey, M.B., Sharma, M.L. and Austhana, J.R. 1978. Indian Perfumer.22(2): 100-103, Pareek. $.K. and Gupta, R. 1988. Exploratory studies on yield ‘and comparative economics of medicinal plant based Cropping systems in North-Western plains of India, Annals of Agricultural Research. §(1-2):169-177, Pareek, SK, Maheshwari, M.L. and Gupta, R. 1980. Domestication studies on Ocimum sanctum for high oll and feugenol content. indian Perfumer.24(2): 93-100. Rastogi J M., Sood, M., Srivastava, LJ. and Romesh, C. 1997. Response of ciary sage (Salvia scalarea L.) to nitogen levels androw spacing. Indian Perfumer. 41(3): 119-116. Singh, A.P.and Minoo Prabia. 2003. Status of medicinal plants ‘consumption by the pharmaceutical industries in Gujrat state. Indian Forester. 123 (2):198-212. Sood, M., Singh, J.M, Srivastava, LJ. and Rana, R.C. 1996, ‘Studies on the response Melisa officinalis L. to nitrogen levels androw spacings. Indian Perfumer40(4): 107-109,