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International Journal of Agriculture: Research and Review. Vol., 3 (4), 844-853, 2013 Available online at ISSN 2228-7973 ©2013 ECISI Journals

THE STUDY OF DIFFERENT LEVELS OF ZEOLITE APPLICATION ON QUANTITATIVE AND QUALITATIVE PARAMETERS IN BASIL (Ocimum basilicum L) UNDER DROUGHT CONDITIONS MOHAMMAD GHANBARI1, SIAVASH ARIAFAR2 1-Department of Plant Breeding, Faculty of Agriculture Research, University of Zabol, Zabol, Iran. 2-M.Sc Student of Agroecology, Agriculture faculty, University of Zabol, Zabol, Iran. *Corresponding Author: MOHAMMAD GHANBARI ABSTRACT: It has been proven that in aromatic plants, growth parametersare markedly influenced by drought stress. In order to investigate the effects of drought stress on growth parameters and mucilage yield in Basil (Ocimum basilicum) an experiment was conducted during 2011-2012 in Institute of Agriculture Research, Zabol, Iran. Plantswere treated with different levels of drought stress(100%fc,75%fc,50%fc) and four levels of zeolite (0,1.5,2,2.5 g/kgsoil), According to the result, the drought stress significantly influenced the growth traits such as plant height, stem diameter, fresh shoot weight, dry shoot weight, dry root weight, fresh root weight, leaf number, leaf area index, seed number, flower number, water use efficiency seed yield ,1000 kernel weight and mucilage yield. Also result indicated that zeolite reduced drought intensity in 75% fc and 50%fc, but these effects were not achieved in 100% fc. Mean comparison for mucilage yield revealed that the highest amount was belong to drought stress with75%fc and 2.5gr zeolite. To sum up, zeolite application in dry lands exposed to drought stress would improve growth parameters and mucilage yield in medicinal and aromatic plants. Keywords: Drought stress, Mucilage yield, Ocimum basilicum, Zeolite to different environmental stresses which limit their growth, productivity and essential oil INTRODUCTION yield. Changes in essential oils and mucilage Ocimum basilicum is one of the most extracted from aromatic plants and their important aromatic plant belongs to the genus composition were observed with water stress Ocimum L., which has an estimated containing (Sabihet al., 1999).Drought stress caused 150 different Species (Javanmardi et al., 2002). significant decrease in fresh and dry biomass, SomeOcimumspecies are used in traditional nutrient content and essential oil medicine for different application such as production(Simon et al., 1992). Razmjoo et al., Common cold, Digestive diseases and etc., (2008) reportedthat drought stress in chamomile especially in many Asian and African countries reduced some growth parameters and essential (Yusuf et al., 1994). It representedO.basilicum oil yield. is an important source of mucilage that used in However in other studies, it was shown that the food and costumesindustries. Basil seeds some growth parameters in O.bacilium were have a high mucilagethat are hydrophilic and increased in drought stress (Grieve, 1999). after hydration formgelatinous material. Basil Essential oil and prolincontents of Sweet basil seeds absorb excesswater in large intestine and increased in response to water stress. The caused a normal excretion. Also some of essential oil yield of Bacil was increased by researches showed its roles in reduce blood subjecting plants toward water stress just before cholesterol (Omidbeygi, 1995). The main harvesting (Baeck et al., 2001). Hasani et component of Basil oil are Linalool, Methyl, al.,(2003) reported water deficit reduced plant Chavicol ,Camphor and Methyl eugenol (Gill height, stem diameter, leaf area, shoot weight, &Randhawa., 1996; Marotti et al .,1966; Pino et mucilage yield, in O.basilicum. Effect of al., 1994).Aromatic plants are usually exposed different irrigation regimes in O.basilicum

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presented with increase irrigation intervals plant growth was reduced also irrigation intervalsaffected mucilage and oil content (Raffat&Saleh1997). Researchers and farmers use some methods to reduce injurious effects of drought stress.Zeolite application is one of the possible approaches to reduce effects of drought stressin plant production. Zeolite consists, alkali and alkaline materials and crystalline alumino silicate ' that have internal surface area for keeping water in drought durations (Silberbush et al., 1993). Zeolite application has may result include decrease leaching of nutrient in soil, prevent of the loss of soil moisture, increasecation exchange capacity and improve soil conditioner. The aim of this study was the evaluation different traits of basil subjected to different level of drought stress using under zeolite application in green house condition. MATERIAL AND METHODS This experiment was conducted in the Institutes of Agriculture Researchin Zabol, Iran. Physical and chemical properties of the soil used in this studyare seen in Table 1.Afactorial experiment was arranged in completely randomized block with four replications. Seeds of Ocimum bacilicum ware provided by Institute of Forest and Grass, Ministry of Agriculture Karaj, Iran.Theinvestigated factors were drought stress with three levels include irrigation at 100% field capacity, 75% field capacity and 50% field capacity and the second factor was zeolite in four levels including 0, 1, 2, and 2.5 g per one kg soil. Seeds were cultured in pots with 30 cm diameter and 60 cm height, for zeolite application referred to Tso (1999) and Urotadze et al., (2002). Normal irrigation was identified for all treatments from the beginning of planting time until complete establishment of plants(appearance of sixth leaves.In order to determine soil moisture used TDR (devise measures volumetric moisture) according to soil moisture curve, volumetric moisture, root depth and pots surface, water wasgiven to target pots in three levels of drought stress (100%fc, 75%fc and 50%fc) also in all stages water amount was measured by counter, all agricultural practices were done according to the recommendation of the Institute of Agriculture Research. At full maturity, the plants were harvested by cutting 5 cm above the soil surfaceand several parameters including plant height, stem diameter, fresh shoot weight, dry shoot weight, fresh root weight, dry wet weight, leaf number, leaf area index, seed number, flower number, WUE,


1000 kernel, seed yield were measured. Water Use Efficiency (WUE) was computed for each treatment as the division of seed yield per amount of water.Harvested seed of each treatment were transferred to the laboratory to measure mucilage, in order to determination the mucilage content, 1 g dry seed was heated with 10 ml Hcl until the solution of mucilage was separated, than seeds were washed with 5ml hot water and extracted solution were added to mucilage solution. Mucilage was obtained by adding 60 ml ethanol to pervious solution and kept in the refrigerator for five hours. The mucilage content was calculated as percentage. The data were transformed in m2 (square meter) and were analyzed by variances analysis (ANOVA), using the SAS GLM procedure (SAS institute 2002). Means were compared by Duncan test at 5% probability level. Figures were drawn by Excel Software.

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year 2012

pH 8

Table1- Soil characteristics of the experimental site EC(dS/m) N(%) P(ppm) K(ppm) Sand(%) 7.8 0.053 7.8 190 63

RESULT Plant height and stem diameter The results at Table 2 demonstrated that drought stress and zeolite application significantly affect plant height. The highest means of height was observed in 100% of field capacity and for zeolite factor the highest means of height was obtained in 2.5 g zeolite. Interaction effect had a significant different at 5% probability for plant height. Mean comparison demonstrated thatin drought levels (75%fc and 50%fc) with increasing amount of zeolite the mean height and stem diameter was increased, but at 100%fc there are no significant differences.The highest plant height in interaction treatments belonged to 100%fc in 2.5g zeolite(73cm) and the lowest mean of plant height was observed in absent of zeolite and 50% fc.(18cm). Also variance analysis for stem diameter showed there no significant difference at 5%probability in all treatment. Leaf number and leaf Area Index (LAI) The result atTable 1 indicated that drought stress, zeolite and their interaction had significant differences for leaf number and leaf area. With increasing drought intensity from 100%fc to 50% fc the leaf number decreased from 703.6 to 466.3 leaves in m2 and the LAI had significant reduction from 0.45mm2 to 0.34 mm2. Mean comparison ofzeolite indicated that the highest means of leaf number and leaf area belonged to 2.5 g zeolite. Analysis of interaction effects for leaf number and LAIindicated in 100% fc showed that there were no significant differences among zeolites, but in 75%fc and 50%fc, with increase zeolite from 0 to 2.5 g average of leaf number and LAI have increased.Interaction effect of 2.5g zeolite and 100% fc had the most effective rather than treatments. However for Leaf Area Index interaction effect of 100% fc and 1.5 g zeolite had the most effective rather than treatments. Fresh and Dry root weight Drought stress had a significant effect on the fresh and dry root weight (Table1). Mean comparison presented that the maximum dry

Silt(%) 20

Clay(%) 17

root weight (2.84g) and fresh root weight (5.25g) was obtained in 50%fc. It seems with intensity drought stress from 100%fc to 50% fc plant tries with increasing root length gets to enough moisture so root weight has increased. There were no significant difference for zeolite application in root wet weight but zeolite application had a significant effect on dry root weight. Using 2.5 g zeolite as compared with others level, caused the lowest average for dry root weight (2.4g). there was no interaction effect for fresh root weight in contrast root dry weight was impressed by interaction effect results of table 4 showed treatment of %50 fc and zeolite control caused the highest mean in root dry weight(4.3g). Fresh and dry shoot weight The variance analysis in Table 2illustrated that the effect of drought stress was significant on both fresh and dry leaf weight, by increasing the discharge of soil humidity from 100%fc to 50%fc fresh and dry leaf weight had significant decrease. The effect of zeolite application was significant for fresh and dry leaf weight, and thehighest mean on both fresh and dry leaf weightwas observed in 2g zeolite. The mean offresh and dry leaf weight for zeolite application in different drought stresslevels has been shown in Table 4 on basis results drought intensity was decreased by increasing the zeolite application, in 50%fc with increase zeolite the mean of dry shoot weight reached from 43.7g to 64.2g and fresh shoot weight reached from 91.3 to 127.5 The same result was obtained in 75% fc but in 100%fc there were nosignificant difference among different amount of zeolite. It seems that the zeolite application significantly reduced the effects of drought on shoot growth. Seed number and Flower number The seed number and flower number were affected by drought stress levels. Plant in 100% and 75 % fc had the highest mean in both of traits and use of 2.5g zeolite in 1kg soil had more effective than others level of zeolite. According to results in Table 4, the maximum flower number and seed numberwere available in 2.5g zeolite and 100%fc and followed in 100%fc and 2g zeolite .The minimum content of


Intl. J. Agric: Res & Rev. Vol., 3 (4), 844-853, 2013

this important trait pertained to 50%fc in zeolite control. Water Use Efficiency (WUE) The information presented in the Table 2 indicated that the effect of zeolite had been significant on WUE, the highest WUE obtained in 75% fc. The effect of Zeolite had significant effect and the highest WUE obtained in 1.5 g and 2 g zeolite. Also interaction effect had significant different on WUE and the highest WUE obtained in 75% fc in 2.5g zeolite. Table 4 also demonstrates that in 75%fc and 50%fc by increasing the zeolite, amount of WUE increased which was significant. Although the lowest WUE was observed in the absent of zeolite and 50%fc but in 100% fc with increasing zeolite from 0 g to 2.5 g.there was no significant change in the WUE . Seed yield and 1000 kernel weight According to Table 2 drought stress, zeolite application and their interaction hadsignificant effect in seed yield. Mean comparisonfor droughtstress showed irrigation at 100% Fc and 75% fc had the highest seed yield and mean comparison for zeolite application showedexception control treatment no significant difference was observed among other treatments. Mean comparison for interaction effect showed the highest seed yield was obtained in100%fc and1.5 g zeolite application.Result of variance analysis showed only zeolite application had significant different on 1000 kernel weight. Mean comparison of zeolite levels showed that the highest mean was observed in 1.5g.Result of interaction effects in figure 2 presented with increase zeolite amount in pots the stress intensity is reduced, so that seed yield in 2.5g zeolite and 75%fc is almost equal with irrigation at 100%fc. Also in 50%fc with increase zeolite,seed yield was reached from34.7 g to 56.1 g that is almost equal 75%fc,this result showedpositive effects zeolite in decrease drought intensity. Mucilage percentage In the present study, drought stress had significant effect on mucilage content. The mucilage percentage was increased by drought stress. With increasing drought intensity from 100% fc to 50% fc mucilage content was reached from 10.1% to 10.85%. Zeolite levels had significant effect in mucilage percentage. With increasing the amount of zeolite from 0 to 2.5 g, mucilage content was reduced from 10.5% to 10.2%. Mean comparison of interaction effect indicated that with increasing


zeolite level from 0g to 2.5 g in each of drought stress points (50%, 75% and 100%fc) amount of mucilage contents was significantly reduced. The highest mucilage percentage was observed by 50%fc in absent of zeolite(11.15%). Figure 1 presented interaction effect on mucilage percentage, lines showed in all drought treatments the highest mucilage percentage belongs to zeolite control and with increase zeolite from 0 to 2.5 g mucilage content had significant decline. So that the lowest mucilage content was observed in 100% fc and 2.5g zeolite application. DISCUSSION It seems almost in all traits there are no difference between 100%fc and 75%fc. In other word,treatment of 75%fc is a mild stress and Basil could tolerate without significant reduction in growth and yield. But irrigation in %50fc is an intense stress and caused significant decrease in growth parameters and yield. Farahani(2009) reportedthat depending upon the plant species and plant genotypes, drought stress can increase, decrease or have not effect on the levels of metabolites. In another study Khalid(2006) mentionedthat drought reduced fresh and dry weight in two genotypes of Ocimumbasilicum. Similarly, Azizi et al., (2008) pointed that increasing drought stresses caused reduction both in fresh and dry weight Moldavian Balm. The results of effect of drought in other medicinal plant indicated that water deficit during the vegetative stages can result in reduction in water use efficiency, shorter plants, smaller leaf areas and least leaves of Mint (Abbaszadeh., 2008), Yarrow (SharifiAshoorabadi et al.,2005), Chicory(Taheri et al., 2008 and Rahmani et al,. 2008). With increasing drought levels, seed yield was reduced;Reduction in seed yield may be due to disturbance in photosynthesis carbohydrate production under stress condition and suppuration of the plant growth (Flexas and Medrano,2002). In this study with increasing the drought level from 100%fc to 500%fcalmost all traits and seed yield were reduced butmucilage percentage was increased. The results of an experiment showed that shoot yield was reduced under drought stress but mucilage percentage was increased under stress(Aliabadiet al., 2009).In Table3 the highest value for mucilage percentage was observed in 50%fc and highest seed yield belong to 100%fc but the maximummucilage yield was observed in 75%fc,therefore irrigation in 75%fc is suitable point for having balance

Intl. J. Agric: Res & Rev. Vol., 3 (4), 844-853, 2013

between mucilage percentage and seed yield that lead to the highest mucilage yield. Also results showed that the effect of zeolite levels was significant for all traits and zeolite has had different effects on growth parameters.Positive effects of zeolite were reported by other researchers and it has been mentioned thatthese effects could be aresult of high potentially of zeolite to absorb water and conserve water in the soil and improve itsphysical properties. Recently, Ghanbari and Ariafar(2013) reported thatzeolite application can improve shoot yield and thereforeoil yield under drought stress conditions, with less damaging of drought stress in medicinal and aromatic plant. Interaction effect between zeolite and drought stress showed that increasing of zeolite caused significant reduction in drought intensity on growth parameter. Result of interaction effects of zeoliteand drought stress were similar withother literature (Goksey et al., (2004), Muire et al., (2007), andAfsharmanesh, (2009). High content of polymer with water supply caused opening stomata for a long time, sub squinty good fixation of CO2 resulted an increase of dry matter in medicinal plant (Khadem et al., 2010). Similar to our results, incorporation of zeolite into soil increased wheat biomass (Howard et al., 1999).Zeolite application decreased the adverse effect of stress and caused increase in WUE,effect of

water stress on WUE depends on plant species, phonological stage of plant in drought period and stress intensity. (Kumari., 1988). CONCLUSION In this studyzeolite application and water stress hada significant effect on seed yield and mucilage yield which are the most important parameter in medicinal plants.It can be mentioned that 2.5gof zeolite with 75% of water losses are lead to improving higher mucilage yield and generally the productivity of this plants. Zeolite application can improve seed yield and follow musilage yield under drought stress conditions and it can persist less damaging of drought stress in medicinal and aromatic plant farming. In addition this may be recommended as a treatment for soils in arid and semi-arid regions to enhance thedrought tolerance in medicinal and aromatic plants. ACKNOWLEDGMENT This study has been financed by University of Zabol.The author likes to appreciate Dr. Mahdi Ghanbari and Dr. IssaKhamari for their suggestion that highly improved this paper. I also thank Mrs. FarzaneNikjouand Sara Ghanbari for their help and encouragement.


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Source Variation


Rep Zeolite Drought stress Zeolite×Drought stress Error


Plant height

3 3 2

45.96 2294.1** 2881.6**

Table1- Variance analysis of main and interaction effects indifferent traits of Ocimumbasilicum Means of Squares Fresh Dry Dry Leaf Diameter Flower Seed Seed root root shoot LAI number stem number number yield weight weight weight 6321.9 0.043 1407.6 378452* 0.612 0.21 194.1 0.008 648.5 267212.9** 0.182 140074.3** 1288157** 0.856 0.32* 2718.6** 0.015** 1828.2* 228905.7** 0.325 175052** 2084702** 2.85** 1.37* 1954.3** 0.084* 8000.6**















5122.7 60226.3 0.734 0.11 100.5 0.004* *, **: significant at 5 % and 1%level of probability. Respectively.

0.04 0.09* 0.38**

Fresh shoot weight 630** 3706* 19645**

1000 Kernel weight 120.5* 401.4** 40.4












Mucilage percentage 0.024 0.023* 0.07*

Table2- Meancomparison ofdrought stress and zeoliteindifferent traits of Ocimumbasilicum Main Effects Drought Stress(%fc) 100 75 50 Zeolite(g/kg soil) 0 1 2 2.5

Plant height (cm)

Leaf number

Demeter Stem

Flower number

Seed number

Fresh root weight(g/m2)

Dry root weight(g/m2)

Seed Yield(g/m2)


Dry shoot weight(g/m2)


Fresh shoot weight

1000 Kernel weight(g)

Mucilage percentage

68.87a 56.31b 35.68b

703.6a 610.6b 466.3c

2.45a 2.19a 2.21a

708a 661a 508b

2761a 2606a 2073b

4.86ab 5.06a 5.25a

2.84b 3a 3.32a

a70.86a a65.1a 54.56b

0.45a 0.47a 0.34b

95.19a 85.7a 52.6b

0.66a 0.7a 0.37b

177.9a 162.4b 111c

26.9a 29a 25a

10.1c 10.3b 10.85a

41.8d 50.25c 62.3b

441.7c 507.7b 648.4ab

2.10a 2.30a 2.37a

499c 617b 625b

2058d 2415c 2620b

4.46a 4.55a 4.85a

2.7a 2.6ab 2.41b

b48.8 a63.2 a70.8

0.37b 0.42a 0.43a

62.9c 85.4ab 90.1a

0.44b 0.6a 0.63a

125.9c 157.2ab 1671.4a

23b 34.6a 24b

10.5a 10.5a 10.3ab















Means followed by the same letter(s) within a column are not statistically different at the p= 5% level


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Table 4- Mean comparisonof Interaction effectsin different traits of Ocimumbasilicum. Interaction effects Drought stress (%fc) 100 100 100 100 75 75 75 75 50 50 50 50

Plant height (cm/m2)

Leaf number

Diameter Stem(mm/m2)

Root wet weight(g/m2)

Root dry weight ( g/m2)

Flower number

Seed number

Seed weight ( g/m2)

777.25a 785a 762.7a 769.75a 470.25de 535.5cd 680.9ab 756a 357.7f 402.7ef 501.7cde 603bc

2.43a 2.31a 2.4a 2.65a 1.85a 2.29a 2.35a 2.28a 1.03a 1.32a 1.35a 1.9a

3.89a 4.02a 4.16a 4.23a 4.93a 4.45a 4.5a 4.07a 5.56a 4.65a 4.38a 4.35a

2.64d 2.75cd 2.73d 3.04cd 3.71b 3.21bc 3.3bc 3c 4.3a 3.3bc 3.14c 3.15c

622.5bc 687.5bc 770.5ab 855a 537d 610cd 655c 705bc 437.5e 480e 487.5e 530de

2700ab 2858ab 2875a 2812ab 2105def 2505cd 2625bc 2790bc 1770f 1882ef 2180cde 2460bcd

71.4a 71.5a 71.3a 71a 50.4b 62.2ab 64.3ab 65.4ab 34.7d 54.7c 56.1b 54.1c

LAI (mm2)

Shoot dry weight(g/m2)

WUE (g/m3)

Shoot wet weight ( g/m2)

1000 Kernel weight(g)

Mucilage percentage (%)

157.8ab 174a 175.1a 174.7a 128.6cd 146.4bc 142.4d 152.2b 91.3e 101.2e 123.8d 127.5cd

26.8a 31.6a 31.8a 31.3a 23.9a 24.1a 26.9a 29.1a 18.4a 22.1a 25.2a 24.3a

10.13e 10.04e 10.06e 10.06e 10.56c 10.51c 10.28d 10.17de 11.15a 10.85b 10.6c 10.4c

Zeolite(g/kg) soil) 0 1.5 2 2.5 0 1.5 2 2.5 0 1.5 2 2.5

64.5a 67a 68a 73.5a 52.25c 55.25bc 66a 65.75a 18.75f 26ef 34de 38d

0.5a 0.51a 0.52a 0.52a 0.36bcd 0.40bcd 0.42abc 0.45ab 0.30d 0.36bcd 0.37bcd 0.32cd

85.02abc 104.4a 101.8ab 109.4a 60.1cd 72.5c 75.7c 74.8c 43.7de 49.4de 63cd 64.2cd

0.6a 0.62a 0.64a 0.6a 0.43b 0.62a 0.65a 0.68a 0.30b 0.36b 0.44b 0.36b

Means followed by the same letter(s) within a column are not statistically different at the p= 5% level

Figure1- Interaction effects of drought stress and zeolite on mucilage percentage

Figure2-Interaction effects of drought stress and zeolite on seed yield


Intl. J. Agric: Res & Rev. Vol., 3 (4), 844-853, 2013

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