Effect of Some Trace Elements on Growth, Yield and Chemical Constituents of Trachyspermum ammi L (AJOWAN) Plants under Sinai Conditions

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1 Research Journal of Agriculture and Biological Sciences, 4(6): , , INSInet Publication Effect of Some Trace Elements on Growth, Yield and Chemical Constituents of Trachyspermum ammi L (AJOWAN) Plants under Sinai Conditions Abd El- Wahab, Mohamed A. Dept. of Medicinal and Aromatic Plants,Desert Research Center, El- Matareya, Cairo. Abstract: spraying of iron, zinc and manganese as each alone at 50 or 100 ppm or in triple combination of them at 50 ppm for each on plant grown under sandy soil conditions resulted in significant enhancement of the vegetative characters of Trachyspermum ammi L. in addition to essential oil percentage, essential oil yield/plant, oil components,npk, Fe, Zn, Mn and total carbohydrate content in the herb. The triple combination of them surpassed the solo application of each. The major oil component was thymol giving 55.51% with the triple combination. Ñ-cymene came in the second order of thymol giving 31.42% and ã-terpinene in the third order by 20.66% and followed by á-pinene in the 2.62% for the same treatment. Key words: Trachyspermum ammi L., volatile oil, Thymol, trace elements INTRODUCTION Ajowan fruit (Trachyspermum ammi L) (Carum copticum.benth and Hook) Fam. Apiaceace (Umbelliferae) is cultivated around the Mediterranean Sea and in south West Asia extending from Iraq to India. In India the essential oil and its main component (thymol) used as medicine, particularly for cholera. Although it grows also in Afghanistan and Egypt, [1] ajowan is produced chiefly in India Lawless. Micronutrients are required in very small quantities and are often supplied as impurities in commonly used fertilizers and in clippings, liming materials, topdressing, certain pesticides, and irrigation water. Sandiness increases the possibility for micronutrient deficiencies; however, most sands used for soil modification are not pure and are usually modified some with soil and organic matter. The intensive publications on iron (Fe), manganese (Mn) and zinc (Zn) reflect the important roles of these elements in plant growth and yield of aromatic and medicinal plants. The main functions of Fe concern growth, respiration, chlorophyll synthesis and photosynthesis. Iron is the micronutrient most often found to be deficient in turf. Its deficiency is characterized by an interveinal chlorosis (yellowing) of turf grass leaves and an eventual thinning of the turf. Deficiencyes of iron is a serious problem under conditions of high ph (above 7.5) and high soil phosphorus contents. The problem can be corrected by spraying every 2 weeks with 1 to 2 ounces of ferrous sulfate per 1,000 square feet of turf. Manganese is considered as an activator in enzyme systems of the carboxylic acid cycle and carbohydrate metabolites Zinc is an essential element for tryptophan as precursor to form indoles. For some dehydrogenase; it functions as a part or cofactor for lot of enzymes and it is also necessary for RNA and protein synthesis. Deficiencies of other micronutrients are not common in the field of turf, but high ph is also known to induce deficiencies of manganese, zinc, or copper in other crops. High soil phosphorus in conjunction with high ph further [2] [3] aggravates a zinc deficiency Hewitt Hopkins, [4] Marschner. Many investigators confirmed the role of these elements on growth and yield of many aromatic and [5] medicinal plants, El-Khateeb on Ruta graveolens L., [6] [7] El-Ghadban on Mentha viridis L., Nagiub et al. on [8] [9] dill, Swaefy on Mentha piperita L., Abd El-latif on [10] Pimpinella anisum L., Abd El-Salam on Foeniculum [11] [12] vulgare Mill., Mohamed on geranium, Mohamed on Coriandrum sativum L. andcarum carvi L., [13] Kassem on Rosemary, Swaefy on Trachyspermum ammi L. As the plants in sandy soil mostly suffer from deficiency of trace elements, this work was conducted aiming to investigate the effect of iron, manganese and zinc on the productivity of Trachyspermum ammi L. plants in sand soil at El- Maghara location, Middle Sinai Corresponding Author: Abd El- Wahab, Mohamed A., Dept. of Medicinal and Aromatic Plants,Desert Research Center, El- Matareya, Cairo. 717

2 MATERIALS AND METHODS A Field Experiment Was Carried out at Elmaghara Research Station Desert Research Center at 100 km South El-Arish City in the Middle of Sinai during the two successive seasons of 2003/2004 and 2004/2005, Seeds were sown on 15 th November directly in rows. The distance between rows was 75 cm and 50 cm between hills. In complete randomized block design containing 11 treatments with three replicates. After two weeks from germination, the plants were thinned at two plants per hill.drip irrigation system was used in this experiment with drippers (4.0 l/h) in the whole period of both seasons for only one hour every two days. The fertilization treatments included the following: Nitrogen: at the rate of 450 kg / fed. Ammonium sulphate (20.5 % N) Phosphorus at the rate of 200 kg /fed. calcium super phosphate (16 % P2O 5) Potassium at the rate of 100 kg / fed. potassium sulfate (48 % K2O) -. Fertilization with calcium super phosphate and 3 Poultry manure at 10 m /fed. was conducted immediately before planting in each season in one dose. As for nitrogen and potassium fertilizers, they were applied in five equal doses in both seasons. The first was added 30 days from appearance of the real leaves then every two weeks. From December to first May, the plants were monthly sprayed with an aqueous solution of trace elements, using 0.5 ml/l Bio-film as a wetting agent. The concentrations and source of elements were as follows: Control plant, were sprayed with tap water. Iron at 50 or100 ppm, as a foliar spray of the chelated Fe-EDTA ( Fe 6% ) Zinc at 50 or100 ppm, as a foliar spray of the chelated Zn-EDTA (Zn14%) Manganese at 50 or100 ppm, as a foliar spray of the chelated Mn-EDTA (Mn 14%). Iron (50 ppm) + zinc (50 ppm). Zinc (50 ppm) + Mn (50 ppm). Iron (50 ppm) + Mn (50 ppm). Iron (50 ppm) + Zn (50 ppm) + Mn (50 ppm). The analysis of poultry manure, soil and water is shown in Tables A,B,C and D Statistical Analysis: The complete randomized block design was used in the experiments (11 treatments) with 3 replicates. Every replicate contained 10 hills. The statistical analysis was carried out according to [15] Costat Statistical Software. L.S.D.test was used to compare the means of treatments. Harvesting: Harvesting was carried out on 15 th May in the first and second seasons. Determination of dry matter : according to [16] A.O.A.C. Determination of oil percentage: according to [17] British Pharmacopoeia. GC/ Mass analysis of volatile oil, GC: Trace [18] Gc200 according to Robert Adams. N, P and K elements were determined in the acid digested solution, which was prepared according to [19] Hach et al.. The results were calculated from a standard curve made on ammonium sulfate. Nitrogen content was determined by modified micro kjeldahl [16] method as described by A.O.A.C.. For phosphorus determination, the ammonium molybdate method [20] according to Murphy and Riley. was used. As for potassium, Iron, Zinc, Manganese it was estimated using atomic absorption apparatus according to [21] Cottenie et al.. Total carbohydrates percentages in [22] the herb were determined according to Herbert et al,. RESULTS AND DISCUSSION Effect on Vegetative Growth: Data presented in Table (5) Show the effect of some trace elements (Fe, Mn and Zn) on vegetative growth parameters of Trachyspermum ammi plant. The results obtained can be discussed as follows: Plant Height: The effective treatments on increasing plant height were zinc at 50 or 100 ppm without significant differences between them. However the triple application of iron, zinc and manganese was more effective in this regard for the highest maximum plant height (111.7, cm during the two seasons, respectively). These results are in harmony with El- [5] [8] Kateeb on Ruta graveolens L., Swaefy on Mentha piperita L. and Swaefy on Trachyspermum ammi L. However the obtained increase in plant height can be attributed to the role of zinc on enhancing synthesis of tryptophan as a precorsure of indole, synthesis which enhances the apical dominance, consequently plant height increase. 718

3 Table 1: ** The chemical properties of Poultry manure ( PM ). Soluble cations meq/l. Soluble anions meq/l ph E.C mmhos Ca Mg Na K CO3 HCO3 Cl SO Humidity Ash O.M N% P% K% - 55% 6% 40% 1.92% 0.88% 1.17% Table 2: ** The chemical properties of soil in ppm (water extract 1:2.5) Soluble cations mg/l Soluble anions mg/l ph E.C mmhos Ca Mg Na K CO3 HCO3 Cl SO Total dissolved TOC,% Total nitrogen, % Phosphate, ìg/l solids (TDS), mg/l Table 3: ** Water analysis of the irrigation water Soluble cations mg/l Soluble anions mg/l ph E.C mmhos Ca Mg Na K CO3 HCO3 Cl SO Total dissolved TOC,% Nitrate, mg/l Phosphate, ìg/l solids (TDS), mg/l N.D N.D N.D Table 4: **Analysis of available trace elements of soil and irrigation water: Parameter Water Soil Iron, mg/l Manganese, mg/l Zinc, mg/l ** analyzed in the Desert Research Center laboratories according to Chapman, H. D. and P. F. Pratt (15) Number of Branches per Plant: Data in Table (5) show that the number of branches/plant was significantly increased with zinc or manganese application at 50 or 100 ppm without significant differences between the two concentrations. However the effective treatment on enhancing number of branches was the application of iron + zinc + manganese each at 50 ppm, which resulted in the highest number of branches/plant (20.33and branches/plant) during the two seasons, respectively. [7] Similar findings were reported by Naguib on dill [12] and Mohamed on(coriandrum sativum L.) and Carum carvi L. The obtained results can be attributed to the role of these elements in enhancing the metabolic processes, which reflected as enhancing the development of lateral buds to growing branches. Plant Fresh and Dry Weight: In both seasons the application of Fe, Zn and Mn either individually or in compination increased the fresh and dry weight of plants compared with those of untreated plants (control). Such increments were statistically significant in both seasons. The highest value of plant fresh weight in the first season (313.30gm) and in the second season (323.30gm) resulted from spraying the plants with the three elements( Fe 1+ Zn 1+ Mn 1) together at 50 ppm for each. These are in agreement with those obtained [5] [6] by El-Khateeb on Ruta graveolens L., El-Ghadban [8] on Mentha viridis L., Swaefy on Mentha peperita L. Fruit Yield: Number of Umbels per Plant: The data on fruit yield expressed as number of umbles/plant and fruit yield plant (gm) are illustrated intable (6). It is clear that, the number of umbels/plant was significantly increased with zinc or manganese at 50 or 100 ppm with no significant differences between the two applied concentrations from each one. Also application of iron + zinc or iron + manganese or zinc + manganese each at 50 ppm 719

4 Table 5: Effect of trace elements on plant height, number of branches, fresh and dry weights per plant of Trachyspermum ammi L. (ajowan) plants Under Sinai Conditions Plant height (cm.) Number of branches Fresh weight (gm.) Dry weight (gm.) Treatments First season Second season First season Second season First season Second season First season Second season Control Fe Fe Zn Zn Mn Mn Fe 1+ Zn Zn 1 + Mn Fe 1 + Mn Fe 1+ Zn 1+ Mn L.S.D. at 5% Table 6: Effect of Trace Elements on number umbels, fruits yield (gm) per plant, oil percentage and oil yield /plant ml of Trachyspermum ammi L (ajowan ) plants Under Sinai Conditions Number of umbels per plant Fruits yield per plant (gm) Oil percentage Oil yield /plant (ml) Treatments First season Second season First season Second season First season Second season First season Second season Control Fe Fe Zn Zn Mn Mn Fe 1+ Zn Zn 1 + Mn Fe 1 + Mn Fe 1+ Zn 1+ Mn L.S.D. at 5% Fruits yield per plant (gm): resulted in similar increase, but the striking increase was that of iron + zinc + manganese each at 50 ppm. The fruit yield of the plants showed similar trend. However it appear that solo iron application at 50 or 100 ppm was not effective in this regard. This may assure the need of the three elements to enhance the metabolic processes, which reflect on growth and flowering. The obtained results are in line with those [9] obtained by Abdel-El-Latif on Pimpinella anisum L. and Swaefy of Trachyspermum ammi L. In both seasons, the spraying of the trace elements significantly increased in the weight of fruits / plant 720

5 Table 7: Effect of trace elements on essential oil components of Trachyspermum ammi (ajowan ) plants under Sinai conditions Treatments á - pinene 8 - Terpinene P - Cymene Thymol Others components Control Fe Fe Zn Zn Mn Mn Fe 1+ Zn Zn 1 + Mn Fe 1 + Mn Fe 1+ Zn 1+ Mn Table 8: Effect of trace elements on nitrogen, phosphorus, potassium and total carbohydrates percentage in herb of Trachyspermum ammi L (ajowan ) plants under Sinai conditions. Nitrogen %in herb.) Phosphorus % in herb Potassium % in herb Tl Total carbohydrates % in herb Treatments First season Second season First season Second season First season Second season First season Second season Control Fe Fe Zn Zn Mn Mn Fe 1+ Zn Zn 1 + Mn Fe 1 + Mn Fe 1+ Zn 1+ Mn compared to control plants, which had produced and 69.60gm/ plant in the first and second season, respectively except Fe1 and Fe2 treatments. In the first season, the highest weight of fruits (91.28gm/ plant) was produced on plants received Fe + Zn + Mn. In the second season, similar trend was observed with the second season. These results are in agreement with [9] those obtained by Abd El-latif on Pimpinella anisum L., Swaefy on Trachyspermum ammi L. Oil Percentage: Data in Table (6) revealed that Trachyspermum ammi L plant responded well to spraying of the trace elements. All treatments increased oil percentage as compared with untreated plants. The values ranged, in the first season, from 5.36 % to 5.68 % while control plants had %. The highest oil percentage (5.68 %) was determined in the plants received Fe + Zn + Mn. Moreover, there was a significant difference between control plants and 721

6 Table 9: Effect of trace elements on iron, zinc and manganese (ppm) in herb of Trachyspermum ammi L (ajowan) plants under Sinai conditions. Iron in herb (ppm) Zinc in herb (ppm) Manganese in herb (ppm) Treatments First season Second season First season Second season First season Second season Control Fe Fe Zn Zn Mn Mn Fe 1+ Zn Zn 1 + Mn Fe 1 + Mn Fe 1+ Zn 1+ Mn micro nutrients treatments except Fe1 treatments in the first season. In the second season, spraying the plants with Fe, Zn and Mn either alone or in combinations showed significant increment in oil percentage compared to control. The highest oil percentage (5.78 %) was found in the plants supplied with Fe + Zn + Mn while control plants had 5.35%. These results are in [6] agreement with those obtained byel-ghadban on [10] Mentha viridis L., Abd El-Salam on Foeniculum [11] [12] vulgare Mill., Mohamed on geranium, Mohamed on(coriandrum sativum L.) andcarum carvi L. Oil Yield per Plant: The data in Table (6) clear that in both seasons oil yield of fruits per plant of Trachyspermum ammi was increased significantly as a result of the the trace elements application each alone or except iron alone. The differences between control plants and those received micro nutrients treatments were significant. In the first season, there was a gradual increase in oil yield with spraying the plants with Zn and Mn either alone or in combinations, resulting in the highest oil yield (5.19 ml/ plant) with Fe + Zn + Mn treatment. In the second season, the trend was, generally similar to that observed in the first one. the highest oil yield / Fed. There is no column showing oil yield / fed. was obtained from of application Fe + Zn + Mn treatment. Similar findings [8] were reported by Swaefy on Mentha peperita L., [9] [11] Abd El-latif on Pimpinella anisum L., Mohamed [12] on geranium, Mohamed on Coriandrum sativum L. and Carum carvi L., Swaefy on Trachyspermum ammi L. Essential Oil Components: Date in Table (7) show that the main component of essential oil was found to be thymol, and ranged from to % followed by ñ-cymene, and ranged from from %, ã- terpinene from 20.11to21.76% then, á-pinene from 1.87to2.11%. The above mentioned constituents were increased in response to application of Zink or manganese. However, it show that the application of the three elements together at 50 ppm was effective in enhancing Thymol and 8 - Terpinene. Elements Content: The data in Table (8) showed the effect of application of the micro nutrients on the herb content of N,P,K, Fe, Zn and Mn Nitrogen Content in Herb: Most of the micro nutrients application enhanced the nitrogen percentages in both seasons, the highest nitrogen percentage (2.36%) was detected in the plants received Fe 1+ Zn 1, while the control plants contained 1.12%. In the second season, the trend was, generally similar to that observed in the first One. The highest N percentage (2.42%) was determined in the plants from spraying the plants with the Fe 1+ Zn 1.The results are in [10] agreement with that obtained by Abd El-Salam on [12] Foeniculum vulgare Mill., Mohamed on Coriandrum sativum L. andcarum carvi. Phosphorus Content: In both seasons, the application of the micro nutrients increased phosphorus percentage. The highest phosphorus percentage ( %) was detected in the plants recviced Fe 1+ Zn 1+ Mn 1 as compared to the control plants which showed the 722

7 lowest P percentage (0.19 and0.22%) for the first season and second season, respectively. The results are in agreement with the data obtained by Swaefy (8) on Mentha peperita L., Abd El- [10] [12] Salam on Foeniculum vulgare Mill., Mohamed oncoriandrum sativum L. andcarum carvi L. Potassium Content: Data recorded in Table (8) showed in both seasons, that all combinations of the micro nutrients treatments increased the potassium content in ajowan plants compared to control or each micro element alone. These result agreed with those [11] stated by Mohamed on geranium Total Carbohydrates Content: Data in Table (8) showed that in the two seasons, all treatments of micro elements increased total carbohydrates content comparing with control. The highest carbohydrates percentage (31.18 and 32.07%) resulted from Fe 1+ Zn 1+ Mn 1 while the lowest carbohydrates content was found in control plants ( and 20.31%) for the first season and second season respectively. These results are in agreement with those reported [11] [12] by Mohamed on geranium, Mohamed on Coriandrum sativum L. and Carum carvi L., Kassem [13] on Rosemary. Iron, zinc and manganese content: Data in Table (9) showed that the content of three elements was generally increased, especially when they were applied together. These results were in agreement with those [5] obtained by El-Khateeb on Ruta graveolens L., [8] [11] Swaefy on Mentha peperita L., Mohamed on geranium. Swaefy on Trachyspermum ammi L., [13] Kassem on Rosemary. REFERENCES 1. Lawless, J The Encyclopedia of Essential Oils Element Book, Ltd. Long Mead, Shoftesbury. Dorest. Great Britain nd 2. Hewitt, G.J., Plant physiology 2 Ed.Stewart Acad. press. New York., 3: Hopkins, W.G., Introduction to plant physiology. New York: John Wiley, pp: Marschner, H., Mineral nutrition of higher plants. 2.ed. New York: Academic Press, pp: El-Khateeb, M.A., M. Farahat and N.AE. Boselah, The effect of trace elements on growth, yield and chemical constituents of rue plants(ruta graveolensl.). Egypt. J. Appl. Sci., 9(7): El-Ghadban, E.A.E., The effect of some trace elements on growth and oil yield of spearmint (Menth viridis, L.).M. Sc. Thesis, Fac. Agric., Cairo Univ., Egypt. 7. Nagiub, N.Y., E.N. Abu Zeid and L.K Balbaa, Response of yield and essential oils of dill to foliar application spraying with some micronutrients. Egypt. J. Appl. Sci., 13(1): Swaefy, H.M.F.A., Effect of chemical fertilization and some trace elements on growth and productivity of Mentha piperita L. plant. M. Sc. Thesis, Fac. Agric. Cairo Univ., Egypt. 9. Abd El-latif, M.Z.M., Response of Pimpinella anisum L. to planting density and phosphorus,zinc and manganese fertilization treatments. M. Sc. 10. Abd El-Salam, Inas. Z., Physiological studies on (Foeniculum vulgare Mill.).. plant. Ph. D. Thesis, Fac. Agric., Cairo Univ., Egypt. Thesis, Fac. Agric., Minia Univ., Egypt. 11. Mohamed, Abd El-Kawy, Mohamed, A comparison of three geranium species and their response to NPK fertilization and micronutrients. M. Sc. Thesis, Fac. Agric. Cairo Univ., Egypt. 12. Mohamed, I.M., Physiological studies on Coriandrum sativum L. and (Carum carvi L). Ph. D. Thesis, Fac. Agric., Cairo Univ., Egypt 13. Kassem, Aabeir, H.M., Effect of planting distances and some trace elements on rosemary plant. Ph. D. Thesis, Fac. Agric., Cairo Univ., Egypt. 14. Swaefy, H.M.F.A Physiological studies ontrachyspermum ammi L. (carum copticum. BENTH) Plant Ph. D. Thesis, Fac. Agric., Cairo Univ., Egypt. 15. Chapman, H.D. and P.F. Pratt, Methods of Soil, Plants and Water Analysis. University of California, Division of Agricultural Sciences. 16 Costat, Software program for the design and analysis of agronomic research experiments. Chort. Software, Costat 3-30, Berkeley, CA., U.S.A.C.F. 17. A.O.A.C., Official Methods of Analysis of Association of Official Agriculural Chemists. th Washington, D. C., 10 ed. 18. British Pharmacopoeia, Determination of Volatile Oil in Drugs. The Pharmaceutical Press, London. 19. Robert Adams, Identification of Essential Oils by Gas Chromatography Mass Spectrometry. Allurd Pub., USA. 20. Hach, C.C., S.V. Brayton and A.B. Kapelove, Powerful kejeldahl nitrogen method using proxy mono sulfuric acid. J. Agric., Food Chem., 33: Murphy, J. and J.H. Riley, A modified single solution for the determination of phosphate in natural waters. Annal. Chem. Acta, 27:

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