Effect of NPK, Bio and Organic Fertilizers on Growth, Herb ...journalrepository.org/media/journals/ARRB_32/2018/... · Keywords: Anatomical structure, biofertilizers, essential oil

_____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected];

Annual Research & Review in Biology 26(2): 1-15, 2018; Article no.ARRB.41038 ISSN: 2347-565X, NLM ID: 101632869

Effect of NPK, Bio and Organic Fertilizers on Growth, Herb Yield, Oil Production and Anatomical

Structure of (Cymbopogon citratus, Stapf) Plant

A. A. El-Sayed1, A. S. El-Leithy1, H. M. Swaefy1* and Z. F. M. Senossi2

1Department of Ornamental Horticulture, Faculty of Agriculture, Cairo University, Egypt. 2Department of Horticulture, Faculty of Agriculture, Omar Al-Mukhtar University, Libya.

Authors’ contributions

This work was carried out in collaboration between all authors. Author AAES designed the study.

Author ASEL wrote the protocol and managed literature searches. Author HMS managed the analyses of the study and wrote the first draft of the manuscript. Author ZFMS performed the field

research. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/ARRB/2018/41038 Editor(s):

(1) Olatunde Samuel Dahunsi, Environmental Biotechnologist, Landmark University, Nigeria. (2) George Perry, Dean and Professor of Biology, University of Texas at San Antonio, USA.

Reviewers: (1) Alexis Valerio, Valery Ramirez, Universidad Nacional Experimental del Táchira (UNET), Venezuela.

(2) Arindam Sarkar, Bidhan Chandra Krishi Viswavidyalaya, India. Complete Peer review History: http://www.sciencedomain.org/review-history/24522

Received 26th

February 2018 Accepted 4th May 2018

Published 8th

May 2018

ABSTRACT

This study was carried out at the Department of Ornamental Horticulture, Faculty of Agriculture, Cairo University, in Egypt during the two successive seasons of 2015 and 2016. The experiment was designed using a complete randomized blocks design. Eco-friendly agriculture has its priority for safe products, so the objective of this study was to investigate the effect of bio fertilizers (Nitrobien and Phosphorien) at 1, 2 and 4 g/plant, compost and poultry manure at the rates of 5, 10 and 15 ton/ feddan (feddan= 4200 m

2) as alternative to NPK (recommended dose), on growth and

anatomical characters of Cymbopogon citratus plants. The results showed that the highest number of tillers per plant (49.07) and leaf area (83.99 cm

2) obtained by the high rate of poultry treatment

(15 ton/feddan) at the second cut in the second season. Thus the result obtained showed that the organic fertilizers had a better effect on total herb fresh and dry weights per plant than bio fertilizer treatments. The poultry manure at the rate of 10 ton/feddan resulted in the maximum total herb fresh yield with value 56.53 ton per feddan and total herb dry yield with value 16.94 ton per feddan, in the

Original Research Article

El-Sayed et al.; ARRB, 26(2): 1-15, 2018; Article no.ARRB.41038

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second season. The essential oil production per feddan at the first cut (in August) was more than at the second cut (in October) in both seasons. The highest oil yield per feddan was recorded in the second season by the poultry manure at the rate of 10 ton/feddan with values 82.26 and 51.85 l/feddan at the first and second cut, respectively. The anatomical study showed that the best thickness of the leaf sheath at the midrib region was due to treating the lemongrass plants by poultry at the rate of 10 ton/feddan. The increasing ratio was 16.7% compared to control. This treatment showed the best results in the total number of vascular bundles, in addition to the No. of large bundles and their dimensions. From these results, it could be recommended that poultry manure was the best treatment for lemongrass growth and getting highly clean yield.

Keywords: Anatomical structure, biofertilizers, essential oil yield, lemongrass, organic fertilizers.

1. INTRODUCTION Cymbopogon citratus, belongs to the Poaceae family, plants are extensively adapted to various agro-climatic zones in India and mostly grow wild as natural vegetation, and fluctuations of weather conditions mainly influence their performance. Essential oil of lemongrass is used in the flavouring of food, in cosmetic as well as in pharmaceutical industries [1]. Lemongrass is a fast growing, herbaceous, perennial grass; which has a feeble branched rhizome from which erect stem grows. The stem has many light green leaves, each consisting of a leathery, cylindrical sheath and a long narrow leaf blade. Leaf blades are sharp to touch because they contain silica cells in the outer cell layer. The young erect stem with the leaf bases is used in cooking since the desired aroma is found mainly in the leaf bases [2]. The lemongrass stalks (pale green or yellow section) are used as a grocery and in cooking after removing the lower bulb, dried leaves used as a tea. Lemongrass plant is outstanding ornamental grass which can grow in the garden borders, along walkways, and in containers. It forms a dense clump of fragrant foliage. Lemongrass may take 7-9 months from planting time before the first cut can be harvested. The most economical way to extract the essential oil is to transport the harvested biomass directly to the distillery. The essential oil contains 70-90% citral, which is a mixture of the two isomers geranial and neral, the ratio between them usually being in the 40:60 to 60:40 ranges [3]. Study on an elite strain of Cymbopogon citratus coded RRL (J) CCA [4] found that the composition percentage of the essential oil of C. citratus which myrcene was 1.08%, Linalool 0.12%, neral ( citral b) 30.19%, geranial ( citral a) 49.01% and geranyl acetate 2.38%.

Using lemongrass essential oil as alternatives to synthetic fungicides to control Botrytis cinerea on

apples [5] indicated that the antifungal activity of lemongrass oil could be attributed to disruption of membrane integrity and membrane permeability. Also, Lemongrass has excellent antioxidant properties and has been reported to have sedative, spasmolytic, carminative, antibacterial, antimutagenic, antimalarial, antifungal, larvicidal, antinociceptive, anti- inflammatory, and antioxidant properties [6]. Using biofertilizers is effective agriculture practice for sustainable farming.In North Western desert of Egypt in Siwa Oasis region [7] on lemongrass found that using a mixture of Azotobacter chroococcum, Bacillus megaterium and Saccharomyces cerevisiae caused an increment in herb biomass and oil yield per plant. Using nitrogen-fixing bacteria leads to the synthesis of vitamins, auxins, and gibberellins which stimulate the plant growth. Azospirillum spp. is a nitrogen-fixing, plant growth promoting bacteria (PGPB) that can associate with several Poaceae plants such as Rice, Wheat, and Pennisetum. Analyses of field experiments have shown a success rate of inoculation with Azospirillum ranging from 60 to 70%, with statistically significant increases in yield [8]. Phosphate solubilizing bacteria is effective in the production of growth promoting substance and releasing phosphorus by converting insoluble forms of phosphorus to an accessible form which led to increasing the P uptake by the plant [9]. [10] Studied the response of Foeniculum plants to bio-organic manure as partial or full substitute for chemical fertilizers. They found that bio-organic was the best treatment for quality of the essential fennel oil. Soil fertility decline happens as a result of exploiting the soil by the production of plants without restoring the consumed nutrients, so soil fertility needs to be maintained by using organic fertilizers such as compost and poultry manure that cause nutrient exchanges between organic


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fertilizers, soil, and water. Poultry manure is applied in Egyptian village as a natural source of nutrients to plants; also it improves biological and physical properties of the soil. Compost had pronounced effect on Dracocephalum moldavica growth characters and essential oil yield [11]. Growth, yield and nutritional quality of Capsicum annuum fruits were evaluated in response to application of three poultry manure rates (0, 5, and10 t/ha). Plant height, the number of leaves and number of branches increased with increasing the poultry rates. Also, 10t/ha Poultry manure resulted in the highest percentage of volatile oil, alkaloid and flavonoid compared to other treatments [12].

Eco-friendly agriculture has priority for safe products, so this work concerns with improving the productivity of lemongrass by using organic and biofertilizers as the alternative to the chemical fertilizers.

2. MATERIALS AND METHODS

This study was carried out at the Department of Ornamental Horticulture, Faculty of Agriculture, Cairo University, in Egypt during the two successive seasons of 2015 and 2016.

The objective of this study was to investigate the effect of NPK (recommended dose), biofertilizers (Nitrobien and Phosphorien) compost and poultry manure on growth, herb yield, essential oil production and anatomical structure of Cymbopogon citratus plants.

2.1 Experimental Procedures 2.1.1 Plant material Uniform tillers of lemongrass (Cymbopogon citratus) plants were obtained from Medicinal and Aromatic Plants Department, Agriculture Research Center, El-Dokki, Cairo, Egypt. The rooted tillers were pruned and inserted in prepared soil plots on 4

th April 2015 and 2016

seasons. The physical and chemical characteristics of the soil experiment field were determined according to [13] and are shown in Tables 1 and 2. 2.1.2 Soil preparation

The soil was prepared on 2nd March 2015 and 2016 for the first and second seasons, respectively. Compost and poultry manure fertilizers were added at 5, 10 and 15 ton/fed (feddan= 4200 m

2) during soil preparation. The

chemical analysis of the used compost and poultry manure are shown in Table 3.

2.1.3 Cultivation procedures

The experiment included 14 treatments in a complete randomized blocks design, each experimental unit (plot) was 2×1.5 m (3.0 m2) and divided into 3 rows with 60 cm apart and 50 cm between the plants, i.e. the plots contained 12 plants. Then, the treatments replicated three times (42 plots).

Table 1. Mechanical analysis of the experimental soil

Sand Silt Clay Texture 56.4 19.75 23.85 SCL

Table 2. Chemical analysis of the experimental soil

pH E.C. O.M. Soluble anions (meq/l) Soluble cations (meq/l) mg/kg

(dSm-1

) % CO3-- HCO3

- Cl

-- SO4

-- Ca

++ Mg

++ Na

+ K

+ N P K

7.39 5.57 0.6 - 4.0 28.0 29 25 11 23.7 1.3 149.6 6.2 420 Table 3. The chemical analysis of the compost and poultry manure used during 2015 and 2016

Chemical characters Compost Poultry 2015 2016 2015 2016

pH 7.57 6.99 7.50 7.80 Total nitrogen (%) 0.92 1.28 3.21 2.80 Total phosphorus (%) 0.12 0.31 0.71 0.80 Total potassium (%) 1.42 0.56 1.15 1.17 C/N ratio 17 :1 18:1 18:1 19:1 Organic matter (% ) 26.5 39.4 63.6 60.6


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2.1.4 Fertilization treatments NPK fertilizers were used as the recommended dose according to the Ministry of Agriculture and Land Reclamation, Egypt [14] as 400 kg ammonium sulphate(20.5%N): 200 kg calcium superphosphate (15.5% P2O5):100 kg potassium sulphate (48% K2O)/ feddan (feddan=4200 m2) were added in two doses, the first dose was added 45 days after transplanting, while the second dose was added after half month from the first cut. Biofertilizers were obtained from General Organization for Agricultural Equalization, Agricultural Research Center, Giza, Egypt. Nitrobein contains nitrogen-fixing bacteria (Azotobacter spp. and Azospirillum spp.). Phosphorein, contains Bacillus megatherium. Biofertilizers (both Nitrobien and Phosphorien) were added at the rate of 1, 2 and 4 g/plant, and divided into two portions, the first was added after 45 days from transplanting and the second was added after half month from the first cut, in the first and second seasons. Compost and Poultry manure were added 15 days before planting in both seasons.

The experiment contained 14 treatments as the following:

1 - Control (without fertilizer) 2 - NPK (recommended dose) at

(400:200:100 kg/ feddan, respectively) 3 - Nitrobien at 1 g/plant. 4 - Nitrobien at 2 g/plant 5 - Nitrobien at 4 g/plant 6 - Phosphorien at 1 g/plant 7 - Phosphorien at 2 g/plant 8 - Phosphorien at 4 g/plant 9 - Compost at 5 ton/ feddan. 10 - Compost at 10 ton/ feddan. 11 - Compost at 15 ton/ feddan. 12 - Poultry manure at 5 ton/ feddan. 13 - Poultry manure at 10 ton/ feddan. 14 - Poultry manure at 15 ton/ feddan.

2.2 Recorded Data

The plants were harvested two times (on 10th August and 5

th October, for the first and the

second cuts, respectively, in the first season 2015 and on 8th August and 6th October for the first and the second cuts, respectively, in the second season 2016). The plants were harvested by cutting vegetative parts at 20 cm above the soil surface.

Data were recorded in the two seasons as the following:

2.2.1 Vegetative growth Data on plant height (cm), number of tillers/plant, leaf area (cm

2), herb fresh and dry weights (g /

plant and ton/ feddan). 2.2.2 Essential oil yield Essential oil yield (L / feddan) was recorded in the two seasons and was estimated as follows: - Essential oil yield/ feddan (L) Oil yield per feddan = oil yield/ plant (ml) × number of plants/feddan

1000 - Essential oil yield/ plant (ml) Essential oil yield per plant = oil percentage × fresh herb weight/plant. Essential oil % = Essential oil vol. (Measuring pipette reading) × 100 Weight of sample 2.2.3 Anatomical study For anatomical study, samples 1 cm long were taken from the middle portion of the Lemongrass leaf sheath of the following treatments: (1) Control: Without fertilization, (2) NPK: Recommended dose, (3) Nitrobein: (2 g/ plant), (4) Phosphorein: (2 g/ plant), (5) Compost: (10 ton/ feddan), (6) Poultry: (10 ton/ feddan) at the age of 3 months. The anatomical specimens were killed and fixed in F.A.A. (10 ml Formalin - 5 ml Acetic acid - 85 ml Alcohol 70%), washed in 50% ethyl alcohol, dehydrated in normal butyl alcohol series and embedded in paraffin wax (55ºC m.p.) [15]. Cross sections 20 µm thick were cut using rotary microtome, stained by light green/safranin combination and mounted in Canada balsam. Sections were examined and counts and measurements of different tissues were calculated. Photomicrographs were also taken.

2.3 Statistical Analysis of Data Data recorded on plant height (cm), number of tillers/plant, leaf area (cm

2), herb fresh & dry

weight (g/plant and ton/feddan) were statistically analyzed, and separation of means were performed using the Least Significant Difference


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(L.S.D.) test at the 5% level, as described by [16].

3. RESULTS AND DISCUSSION

3.1 Plant Height (cm)

Data presented in Fig. 1 revealed that the tallest plants were obtained from the treatment of phosphorein at the rate of 2 g/plant with the mean values 134.8 and 144.1 cm at the first and second cuts of the first season. These findings are in agreement with [17] who studied the effect of chemical fertilizer and bio fertilizer (Azotobacter, Azospirillum, Phosphate Solubilizing Bacteria and mixture of Aza + Azo + PSB) on Plumbago zeylanica. The highest effect on plant height was obtained with Phosphate solubilizing Bacteria. While in the second season, the tallest plants were obtained by using poultry at the highest rate (15 ton/feddan) with the mean values 150.5 and 155 cm in the first and second cuts, respectively compared to the other treatments. On the other hand, the shortest plants recorded from control plants compared to all treatments at both cuts in the two seasons. Similar result recorded by [18] who applied chicken manure at four rates (0, 5, 10 and 15 t/ha) to Zea mays saccharata plants. They found

that chicken at the rate of 15t/ha increased plant height by 81%. The effect of poultry manure on increasing the plant height may be due to N supply as a precursor of protein synthesis and as a vacuolar osmoticum. The osmotic compounds in the cell sap are important in order to allow cell elongation.

3.2 Number of Tillers per Plant The data presented in Fig. 2 showed that the number of tillers per plant of lemongrass was significantly affected by fertilization treatments compared to untreated plants at both cuts, in both seasons. Poultry manure treatments had a superior effect on the number of tillers per plant especially at medium and high doses (10 and 15 ton/feddan), respectively. The maximum numbers were 28.97 and 35.33 tillers per plant recorded by poultry at medium rate treatment (2g/plant) at both cuts in the first season, respectively. Moreover, in the second season, the maximum numbers of tillers per plant were 48.10, 49.07 obtained by the high rate of poultry treatment (4g/plant) at the first and second cuts, respectively. Control plants had the minimum number of tillers per plant at both cuts of the two seasons.

Fig. 1. Effect of NPK, bio and organic fertilizers on plant height (cm) of Cymbopogon citratus during 2015 and 2016 seasons

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1 , 2 , 4 g/ plant ), respectively, Phosph:Phosphorein1, 2 and3 (1, 2, 4 g/ plant), respectively, Compost: 1, 2 and 3 (5 ,10 , 15 ton/

feddan), respectively, Poultry: 1, 2 and 3 (5, 10, 15 ton/ feddan), respectively


6

In general, the highest values of tillers/plant were recorded at both cuts in both seasons by poultry and compost at different rates than the other treatments and unfertilized plants. Similar results recorded by [19] who applied poultry manure to at the rate of 0, 2, 4 6 t ha-1 and 250 kg ha-1 NPK 15-15-15 fertilizer. Growth and yield parameters of Telfaria occidentalis plants were increased by poultry manure treatments. Poultry manure at the rate of 6 t ha-1 and NPK fertilizer increased number of branches by 7.72 and 6.89%, respectively. Also, [20] studied the effect of compost rates (2, 4 and 8 ton /feddan) on Rosmarinus officinalis, L. during two successive seasons. The results showed that compost at 8 ton/ fed gave the highest number of branches/ plant, at the second and third cuts of the first and second seasons.

3.3 Leaf Area (cm2) Data presented in Fig. 3 showed that all fertilizer treatments led to significant increase in leaf area compared with control plants at the first cut of the first season. While in the second season did not have the same trend except with poultry at low and high rates. Concerning the second cut, the most of treatments increased leaf area compared with control plants without significant difference

between them except with compost at (low and medium rates) and poultry (medium and high rates) which had large leaf area with a clear significant difference in the first season. While in the second season, most treatments significantly increased leaf area. The largest leaf area (83.99 cm

2) was obtained from poultry3 (15 ton/feddan)

treatment in the second cut of the second season. The increment in leaf area may be due to cell expansion and cell division which in turn increase the leaf size. This result was in agreement with [21] who studied the effect of chicken manure at rates of 10 and 20 m3/feddan, humic acid at rates of 125 and 250 ppm and their interactions on Ocimum spp. The results in both seasons pointed out that the treated plants with chicken manure at 10 m

3 (100 g/pot) then

sprayed with humic acid at 125 ppm at the second cut of Ocimum species significantly increased plant height, number of branches, leaf area and fresh weight compared to control plants (untreated).

3.4 Herb Fresh Weight (g/ plant) Data in Table 4 show that in both seasons, the first cut produces fresh herb per plant more than the second cut in most cases except with compost treatments in the first season and

Fig. 2. Effect of NPK, bio and organic fertilizers on number of tillers/ plant of Cymbopogon citratus during 2015 and 2016 seasons

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1 , 2 , 4 g/ plant), respectively, Phosph: Phosphorein1, 2 and3 (1, 2, 4 g/ plant),respectively, Compost: 1, 2 and 3 (5 ,10 , 15 ton/



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control plants in both seasons. Results revealed that organic fertilizers had the better effect on total fresh herb per plant than bio fertilizer treatments. The highest total fresh herb per plant, in both seasons, was obtained by poultry at the medium rate with the mean values (3067, 4240 g/plant) in the first and second seasons, respectively. On the other hand, the control treatment recorded the lowest total herb fresh weight per plant in both seasons. Generally increasing herb yield per plant in the first cut compared to the second one can be explained by the effect of temperature during the growth period, besides the length of the growth period of the first cut (5 months) compared to (two months) for the second one. The growth period of the first cut was from March to July while it was from August to the first week of October in the second cut. [22] stated that plant growth responses to environment and temperature affect a range of enzymatically catalyzed. Also, the activation energy required for different reactions may differ widely. These findings were in harmony with those mentioned by [23] who studied the effect of organic manure (farmyard manure and poultry manure at a rate of 75 m3 /ha as fertilizer of each) and biofertilizers (Nitraboein and Halex-2 at a rate of

988 g/ha of each) as well as their interactions on growth and herb yield of Majorana hortensis L. The results indicated that the application of a poultry manure as an organic fertilizer to marjoram plants recorded the maximum values of herb fresh and dry yields in the early cut.

3.5 Herb Fresh Weight (ton/ feddan)

Data presented in Table 5 show that organic fertilizers produce more total herb fresh yield per feddan than NPK treatment except with compost at the low and medium rate in the first season and compost1 in the second season which decreased fresh yield/feddan with a non-significant difference. Adding NPK fertilizers increased total herb fresh yield per feddan compared with bio fertilizers in both seasons. The lowest herb fresh yield per feddan was obtained with control plants in both cuts of both seasons. Similar results were observed by [24] who studied the effect of organic fertilization, foliar spray of yeast and their interaction on three-lobed sage plants. The interaction effect proved that the highest compost manure level at 20 m3/feddan combined with a foliar spray of the highest yeast concentration at 10 g/liter gave maximum increments in vegetative growth parameters and yields of herb over control treatment, which recorded the lowest

Fig. 3. Effect of NPK, bio and organic fertilizers on leaf area (cm2) of Cymbopogon citratus during 2015 and 2016 seasons

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1 , 2 , 4 g/ plant), respectively, Phosph:Phosphorein1, 2 and 3 (1, 2, 4 g/ plant),respectively, Compost: 1, 2 and 3 ( 5 ,10 , 15 ton/

feddan), respectively, Poultry: 1, 2 and3 (5, 10, 15 ton/ feddan), respectively

40

45

50

55

60

65

70

75

80

85

First Season- FirstCut First Seson -Second Cut

Second Season- First Cut Second Season-Second Cut


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Table 4. Effect of NPK, bio and organic fertilizers on fresh herb (g/ plant) of Cymbopogon citratus during 2015 and 2016 seasons

Second season, 2016 First season, 2015 Treatments

Total 2nd

cut 1st

cut Total 2nd

cut 1st

cut 1566.5 915.8 650.7 1604.4 811.0 793.4 Control 2557.0 1189.0 1368.0 2566.1 960.1 1606.0 NPK 2483.0 1209.0 1274.0 2003.3 831.3 1172.0 Nitro1 2025.3 736.3 1289.0 2415.0 909.0 1506.0 Nitro2 2066.0 932.0 1134.0 2340.9 957.9 1383.0 Nitro3 1638.0 777.7 860.3 2215.1 931.1 1284.0 Phosph1 2245.3 823.3 1422.0 2031.3 953.3 1078.0 Phosph2 1980.5 958.5 1022.0 2048.1 920.1 1128.0 Phosph3 2205.0 1013.0 1192.0 2409.0 1331.0 1078.0 Compost1 2848.0 975.0 1873.0 2483.0 1378.0 1105.0 Compost2 3046.0 1225.0 1821.0 2672.0 1444.0 1228.0 Compost3 3143.0 1215.0 1928.0 2912.0 1267.0 1645.0 Poultry1 4240.0 1669.0 2571.0 3067.0 1439.0 1628.0 Poultry2 3764.0 1460.0 2304.0 3022.0 1466.0 1556.0 Poultry3 993.2 100.8 211.8 731.8 84.95 212.2 LSD at 0.05

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1 , 2 , 4 g/ plant), respectively, Phosph:Phosphorein1, 2 and3 (1, 2, 4 g/ plant ),respectively, Compost: 1, 2 and3 ( 5 ,10 , 15 ton/

feddan), respectively, Poultry: 1, 2 and3 (5, 10, 15 ton/ feddan), respectively Table 5. Effect of NPK, bio and organic fertilizers on fresh herb (ton/ feddan) of Cymbopogon

citratus during 2015 and 2016 seasons

Second season,2016 First season,2015 Treatments Total 2nd cut 1st cut Total 2nd cut 1st cut 20.88 12.20 8.675 21.39 10. 81 10.58 Control 34.07 15.84 18.24 34.21 12.80 21.41 NPK 33.11 16.12 16.99 26.71 11.08 15.63 Nitro1 27.00 9.817 17.18 32.19 12.12 20.07 Nitro2 27.55 12.43 15.12 31.22 12.77 18.44 Nitro3 21.84 10.37 11.47 29.54 12.41 17.12 Phosph1 29.94 10.98 18.96 27.08 12.71 14.37 Phosph2 26.40 12.78 13.62 27.30 12.27 15.03 Phosph3 29.40 13.50 15.90 32.11 17.75 14.37 Compost1 37.53 13.00 24.53 33.11 18.37 14.74 Compost2 39.28 15.00 24.28 35.17 19.25 15.92 Compost3 41.90 16.19 25.70 38.81 16.89 21.93 Poultry1 56.53 22.25 34.28 40.89 19.19 21.71 Poultry2 50.18 19.47 30.71 40.28 19.54 20.74 Poultry3 13.24 5.00 11.16 9.57 3.33 7.23 LSD at 0.05

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1 , 2 , 4 g/ plant), respectively, Phosph:Phosphorein1, 2 and 3 (1, 2, 4 g/ plant),respectively, Compost: 1, 2 and3 (5 ,10 , 15 ton/

feddan), respectively, Poultry: 1, 2 and 3 (5, 10, 15 ton/ feddan), respectively parameters. [25] found significant increase of Potato yield parameters with application of poultry manure compared to control. Adding poultry manure at the rate of 4t/ha to the Basil and Mint plants caused a significant increase in fresh leaf yield per plot (the size of a plot was 2.0mx1.0m).Unfertilized Basil plants produced 272.62 gram per plot while the fresh leaf yield reached 417.5 g/plot

with poultry manure treatment. Also, the fresh leaf yield of Mint plants increased from 249.74 to 338.9 g/plot by using 4t/ha poultry manure [26]. From the previous results, compost or poultry could be used instead of chemical fertilizers to produce approximately the same amount of herb fresh weight per feddan but more safety products for a healthy environment.


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3.6 Herb Dry Weight (g/ plant) Data presented in Table 6 showed that organic fertilizer treatments had a superior effect on total dry herb per plant without significant difference between them in the first season. While, in the second season, poultry at medium rate gave the highest total dry herb per plant (1270.4 g) followed by the other organic treatments with a clear significant difference. The effect of organic manure on increasing herb dry yield may be due to increasing water holding capacity, porosity, and total nitrogen of soil. Also, organic fertilizers caused enhancement of microbial activity in the soil which causes the availability of micronutrients for plant growth [27]. These results agreed with those stated by [28] on black cumin (Nigella sativa L.), studied the interactive effects of biofertilizer application on quality and quantity yields. Four levels of animal manure (0, 10, 20 and 30 ton/ha) and two levels of Azotobacter (non-application and application) were studied. The results indicated that in the 20 ton/ha animal manure and Azotobacter application, yield components were significantly higher.

3.7 Herb Dry Yield (ton /feddan)

The data presented in Table 7 indicated that herb dry yield per feddan followed the same trend of

herb dry weight per plant. There was a gradual increase in total dry yield with increasing compost rate. Among different bio fertilizers treatments, nitrobein had a better effect on total dry yield per feddan than phosphorein in most cases. Poultry manure at medium rate had the heaviest total herb dry yield per feddan; 11.687 and 16.94 ton in the first and second season, respectively. The control plants had the lowest total dry yield per feddan 6.458 ton in the first season, while in the second season control plants produced 6.585 ton per feddan followed by phospho1 with the mean value 6.239 ton without significant difference between them. These results are in agreement with the results reported by [29] who found that the addition of the compost at the rate of 5, 10, 20, 40 and 60 t ha-1, positively influencing the production of tillers and biomass production of lemongrass plant. [30] who studied the effect of utilizing chemical nitrogen with or without bio-fertilizers (biogein and /or nitrobein) on guar plant. Results revealed that different sources of nitrogen or bio-fertilizers increased the dry weight of aerial part and leaves per plant. Also [31] found significant effect of bio-fertilizers on biomass yield of Dracocephalum moldavica plant. [32] evaluated four organic amendments: Leaf compost, vegetable compost, poultry manure and sewage sludge for their effect on three varieties of Java citronella, Cymbopogon winterianus Jowitt (Manjusha, Mandakini, and Bio-13). Poultry manure applied

Table 6. Effect of NPK, bio and organic fertilizers on dry yield (g /plant) of Cymbopogon


Second season, 2016 First season,2015 Treatment Total 2

nd cut 1

st cut Total 2

nd cut 1

st cut

494.1 288.0 206.1 484.4 256.4 228.0 Control 754.2 367.3 386.9 771.6 313.1 458.5 NPK 721.1 359.1 362.0 557.4 239.7 317.7 Nitro1 609.0 208.6 400.4 707.2 279.1 428.1 Nitro2 654.6 313.0 341.6 677.3 287.5 389.8 Nitro3 467.9 219.3 248.6 606.2 289.1 317.1 Phosph1 612.7 253.6 359.1 591.6 281.1 310.5 Phosph2 575.0 286.4 288.6 648.1 303.3 344.8 Phosph3 639.2 259.2 380.0 708.2 400.7 307.5 Compost1 813.5 286.1 527.4 739.1 427.0 312.1 Compost2 821.3 290.6 530.7 778.3 412.4 365.9 Compost3 857.4 311.5 545.9 860.1 383.8 476.3 Poultry1 1270.4 503.9 766.5 876.5 431.4 445.1 Poultry2 1173.9 472.2 701.7 858.0 426.5 431.5 Poultry3 43.7 25.2 29.4 186.0 26.4 31.6 LSD at 0.05

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1 , 2 , 4 g/ plant), respectively, Phosph: Phosphorein1, 2 and3 (1, 2, 4 g/ plant),respectively, Compost: 1, 2 and 3 (5 ,10 , 15 ton/



10

Table 7. Effect of NPK, bio and organic fertilizers on dry yield (ton /feddan) of Cymbopogon citratus during 2015 and 2016 seasons

Second season, 2016 First season, 2015 Treatments

Total Cut 2nd 1ST

Cut Total Cut 2nd 1st

Cut 6.585 3.840 2.745 6.458 3.418 3.040 Control 10.06 4.897 5.158 10.29 4.174 6.113 NPK 9.581 4.826 4.754 7.431 3.195 4.236 Nitro1 8.120 2.782 5.338 9.43 3.722 5.708 Nitro2 8.727 4.173 4.554 9.03 3.833 5.197 Nitro3 6.239 2.924 3.315 8.081 3.854 4.227 Phosph1 8.169 3.381 4.788 7.888 3.748 4.140 Phosph2 7.666 3.818 3.848 8.641 4.044 4.597 Phosph3 8.523 3.456 5.067 9.443 5.343 4.100 Compost1 10.85 3.815 7.032 9.854 5.693 4.161 Compost2 10.95 3.874 7.075 10.38 5.499 4.879 Compost3 11.432 4.154 7.278 11.468 5.118 6.350 Poultry1 16.94 6.718 10.22 11.687 5.752 5.935 Poultry2 15.65 6.296 9.355 11.44 5.686 5.753 Poultry3 0.475 0.281 0.336 0.310 0.237 0.255 LSD at 0.05

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1, 2, 4 g/ plant), respectively, Phosph:Phosphorein1, 2 and 3 (1, 2, 4 g/ plant),respectively, Compost: 1, 2 and 3 (5, 10,

15 ton/ feddan), respectively, Poultry: 1, 2 and 3 (5, 10, 15 ton/ feddan), respectively at 100 t ha−1 followed by sewage sludge increased the herbage and dry matter yield significantly. On Kalmegh (Andrographis paniculata) plants [33] found that chicken manure at the rate of 12.5 ton/ha resulted in significant increase in dry weight yield.

3.8 Essential Oil Yield (l/ feddan) The data presented in Table (8) revealed that the essential oil yield per feddan was significantly affected by different application treatments compared to untreated plants at both cuts in both seasons. In most cases, the essential oil production per feddan at the first cut was more than the production at the second one in both seasons. In the first season, the highest oil yield per feddan was obtained from the high rate of poultry manure with values 62.24 and 42.41 l/feddan at the first and second cut, respectively. While in the second season, the highest oil yield per feddan was recorded by the medium rate of poultry manure with values 82.26 and 51.85 l/feddan at the first and second cuts, respectively. Concerning to the effect of different applications on total essential oil yield per feddan, the maximum oil yield per feddan (134.11 l/feddan) was obtained from poultry manure at the medium rate in the second season. The increase in essential oil yield per feddan was due to the increase of fresh herb biomass. Similar results recorded by [12] who found that using10t/ha poultry manure with

Capsicum annuum plants resulted in the highest percentage of volatile oil. [21]who found that using poultry manure at 10m3 (100 g/pot) plus humic acid at 125 ppm as a foliar application with Ocimum species plants significantly increased essential oil yield per feddan compared to control plants. Also, [34] found that applying 6 kg m

-

2 poultry manure to Achillea millefolium plants produced a greater accumulation of shoot biomass and greater yield of essential oil.

3.9 Anatomical Study Lemongrass leaf bases (leaf sheaths) have preferential interest to consumers, where their aroma and taste are acceptable so, it is important to study the anatomical structure of leaf sheath. 3.9.1 Anatomical structure of lemongrass leaf

sheath As shown in Figs. (4-9), the leaf sheath is composed of two epidermal layers on adaxial and abaxial surfaces. Each is uniseriate, composed of a row of compactly- rectangular cells. The lower epidermal cells are smaller in size than the upper ones with terraces and channels. Patches of mechanical tissue (sclerenchyma) are located under the lower epidermal cells. The ground tissue is composed of different sizes of parenchyma cells. The vascular bundles are collateral and closed and


11

arranged in parallel series. They are located in the ground tissue. The bundle is composed of xylem (on the upper side) and phloem (on the lower side). The xylem is made of tracheary elements and the phloem of sieve tubes and companion cells. The bundle remains surrounded by a row of colorless parenchyma cells (bundle sheath). Majority of the bundles are small, but also large bundles are found between them. 3.9.2 Structural differences among leaf

sheaths due to treatments Results in Table 9 and Figs. 4-9 clarify the structural differences among Lemongrass leaf sheaths due to the different treatments compared to control plants. The anatomical study showed that the best thickness of the leaf sheath at the midrib region was due to treating the Lemongrass plants by using Poultry treatment. The increasing ratio was 16.7% compared to control. This increase is mainly attributed to the increase in thickness of the upper epidermis and the ground tissue being; 132.1% and 15.2%, respectively compared to control. The same treatment also showed the best results in both numbers of vascular bundles and large bundles, as well as large bundle dimensions, being; 32.3, 60.0, 82.1 x 43.0%, respectively compared to the control. Although the compost treatment recorded the lowest increase in

thickness of the leaf sheath in the midrib region compared to the control (2.1%), which is mainly attributed to the slight increase in the thickness of the ground tissue and the decrease in number of rows of ground parenchyma than the control being; 2.2 and -31.3%, respectively compared to the control, there was a significant increase in No. of the vascular bundles, No. of large bundles, large bundle dimensions and the average thickness of the parenchyma cell of the ground tissue being; 25.8, 50.0, 35.5 x 63.7 and 48.6%, respectively compared to the control. On the other hand, the Nitrobien treatment recorded the lowest increment in most characters relative to the control being; 12.9%, 5.0%, 10.4% and 5.3% for No. of the vascular bundles, No. of large bundles, mean thickness of the parenchymal cell in the ground tissue and thickness of the upper epidermis, respectively. While, the increasing ratios in the other treatments ranged between 2.1-14.6% for the leaf sheath thickness at the midrib region, 21.0-25.8% for No. of the vascular bundles,10-50% for No. of the large bundles, 14.3-26.2% for No. of the small bundles, 14.5-35.5% for length of the large bundle, 9.0-63.7% for width of the large bundle, 25% for thickness of the mechanical tissue (sclerenchyma), 2.2-15.2% for thickness of the ground tissue, 15.9-48.6% for mean thickness of the parenchymal cell of the ground tissue and 18.3-30.5% for upper epidermis thickness compared to the control.

Table 8. Effect of NPK, bio and organic fertilizers on oil yield (L /feddan) of Cymbopogon


Second season, 2016 First season, 2015 Treatment Total 2

nd cut 1

st cut Total 2

nd cut 1

st cut

30.71 16.23 14.48 35.55 14.39 21.16 Control 70.78 31.23 39.56 80.60 23.43 57.17 NPK 59.99 31.11 28.88 50.53 16.63 33.91 Netro1 51.84 27.79 24.05 84.48 24.24 60.24 Netro2 56.76 31.07 25.69 70.56 21.33 49.23 Netro3 42.83 17.93 24.89 56.96 22.72 34.24 Phosph1 62.37 20.09 42.28 49.54 23.25 26.29 Phosph2 63.31 25.56 37.75 43.51 18.40 25.11 Phosph3 65.00 22.56 42.44 66.70 23.60 43.10 Compost1 77.79 23.79 54.00 54.53 27.56 26.97 Compost2 63.41 22.87 40.55 61.63 28.88 32.75 Compost3 89.53 29.64 59.89 71.04 30.91 40.13 Poultry1 134.11 51.85 82.26 82.21 35.11 47.11 Poultry2 113.81 42.24 71.57 104.65 42.41 62.24 Poultry3 10.80 2.50 10.03 4.54 2.08 3.57 LSD at 0.05

Control: Without fertilization, NPK: Recommended dose, Nitro: Nitrobein 1, 2 and 3 (1, 2, 4 g/ plant), respectively, Phosph: Phosphorein 1, 2 and 3 (1, 2, 4 g/ plant), respectively, Compost: 1, 2 and 3 (5,

10, 15 ton/ feddan), respectively, Poultry: 1, 2 and 3 (5, 10, 15 ton/ feddan), respectively


12

Table 9. Average transverse sections through the middle portion of leaf sheaths of lemongrass plant as affected by NPK, bio and organic fertilizers (Averages of 10 readings)

Characters Control

(1) NPK (2)

% to control

Nitro. (3)

% to control

Phosph (4)

% to control

Compost (5)

% to control

Poultry (6)

% to control

Midrib region thickness 829.4 950.4 14.6 915.8 10.4 898.6 8.3 846.7 2.1 967.7 16.7 No. of vascular bundles: 62.0 75.0 21.0 70.0 12.9 75.0 21.0 78.0 25.8 82.0 32.3 No. of large bundles 20.0 22.0 10.0 21.0 5.0 25.0 25.0 30.0 50.0 32.0 60.0 No. of small bundles 42.0 53.0 26.2 49.0 16.7 50.0 19.1 48.0 14.3 50.0 19.1 Large bundle dimensions 24.2

x19 22.5 x20.7

-7.0 x9.0

22.5 x22.5

-7.0 x18.4

27.7 x24.2

14.5 x27.4

32.8 x31.1

35.5 x63.7

34.6 x34.6

43.0 x82.1

Mechanical tissue thickness 69.1 86.4 25.0 86.4 25.0 86.4 25.0 86.4 25.0 69.1 0.0 Rows No. of ground parenchyma 16 14 -12.5 16 0.0 15 -6.3 11 -31.3 13 -18.8 Ground tissue thickness 794.9 915.8 15.2 881.3 10.9 864 8.7 812.2 2.2 915.8 15.2 Mean thickness of parenchymal cell 49.8 65.3 31.1 55.0 10.4 57.7 15.9 74.0 48.6 70.5 41.6 Upper epidermis thickness 13.1 17.3 30.5 13.8 5.3 15.5 18.3 17.3 30.5 30.4 132.1 Lower epidermis thickness 13.1 10.4 -20.6 10.4 -20.6 10.4 -20.6 11.4 -13.0 11.4 -13.0

(1) Control: Without fertilization (2) NPK: Recommended dose (3) Nitro: (Nitrobein, 2 g/ plant) (4) Phospho: (Phosphorein, 2 g/ plant) (5) Compost: (10 ton/ feddan) (6) Poultry: (10 ton/ feddan)


13

Fig. (4-9). Transverse sections of Lemongrass leaf sheath (3 months old) through the midrib region as affected by NPK, bio and organic fertilizers (X40)

(1) Control: Without fertilization (2) NPK: Recommended dose (3) Nitrobein: (2 g/ plant) (4) Phosphorein: (2 g/ plant) (5) Compost: (10 ton/ feddan) (6) Poultry: (10 ton/ feddan). Details: gt, ground tissue; lep, lower epidermis; lvb, large vascular bundle; ph, phloem;

sc, sclerenchyma; svb, small vascular bundle; uep, upper epidermis; x, xylem

From the above-mentioned results, it is clear that the chosen treatments under study showed a relative superiority over the control in most anatomical characteristics of the leaf sheath, and the Poultry treatment was the best of all. These results were consistent with the morphological results under study, where it was the best treatment for all the morphological characteristics studied, while the control was the lowest one for most characters.

4. CONCLUSION From previous results, it could be concluded that poultry manure was the superior treatment for Cymbopogon citratus (DC) Stapf, plants, which had a significantly positive effect upon most of

the studied characters. Poultry manure at the medium rate (10 ton/feddan) resulted in the maximum values of total fresh and dry yields, and essential oil yield per feddan as well as it has superiority over the control in most anatomical characteristics of the leaf sheath.

ACKNOWLEDGEMENTS

The authors wish to thank Dr. Atef Zakaria Sabh, Professor of Agricultural Botany, Faculty of Agriculture, Cairo University, for his assistance in the anatomical study.

COMPETING INTERESTS

Authors have declared that no competing interests exist.


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Effect of NPK, Bio and Organic Fertilizers on Growth, Herb ...journalrepository.org/media/journals/ARRB_32/2018/... · Keywords: Anatomical structure, biofertilizers, essential oil