The Variations in the Phytochemical Attributes of Nerium Oleander, Ricinus Communis, Calotropis Procera and Withania Somnifera From Three Ecological Zones of Pakistan

The Variations in the Phytochemical Attributes of Nerium Oleander, Ricinus Communis, Calotropis Procera and Withania Somnifera From Three Ecological Zones of Pakistan | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The Variations in the Phytochemical Attributes of Nerium Oleander , Ricinus Communis , Calotropis Procera and Withania Somnifera From Three Ecological Zones of Pakistan Khola Tazeen, Zahoor Ahmad Sajid, Khalid Farooq Akbar This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3308843/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract The phyto-climatic diversity in a region presents the life form of vegetation. Plants produce vast and diverse variety of phytochemicals to communicate and minimize the climatic threats. In the current study, the variations in the phytochemical attributes of four plant species ( Nerium oleander, Ricinus communis, Calotropis procera and Withania somnifera) from three ecological zones of Pakistan (sandy desert, irrigated & rain-fed plains) were investigated. The phenolic acids, flavonoids and tannins were quantitatively analyzed by methanolic extraction and standardized methods. The results revealed that zone rainfed lands and zone sandy desert showed the maximum amount of phytochemicals in all species and also exhibited the extreme environmental conditions. The irrigated plains had less stressful environmental condition and maximum amount of soil organic matter that resulted in minimum amount of phytochemicals in plant species. Previous research also supports the evidence that environmental stresses like temperature stresses, water stresses, high light intensities increase, and organic matter hinders the accumulation of phytochemicals. This study proved that same species in different ecological zones show different amounts of phytochemicals due to environmental variations, although environmental stresses exert the effects on plants to produce more phytochemicals. Ecological zonation flavonoids phenolic acids phytoclimatic diversity tannins Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction The life form of vegetation in a region can be measured by the plant-environment analogy (Khan et al., 2013). Different ecological factors influence differently the vegetation, plant community, structure, and morphology of plants to adapt to their environment for their better survival of life. These influences could lead to long term adaptive strategies (Salick et al., 2009; Khan et al., 2013). Long term existence of species needs to be differentiated as they have back-and-forth in their abilities to respond constrains of their environment (Tilman, 1988). Ecological attributes that can affect biota of a region are, slope, altitude, topography, soil, habitat, vegetation type, plant community (Ahmad et al., 2008), solar radiation, temperature (Javaid et al., 2010), moisture stress, salinity, water, and precipitation (Shukat & Burhan, 2000). The phyto-climatic gradient of a region influences the plants to grow healthy i.e., to adapt to their environment (Khan et al., 2013). The effect of climatic diversity on the synthesis of phytochemicals has widely perceived a considerable amount of attention (Krasteva et al., 2013). Plants produce a vast and diverse variety of organic compounds that help them to communicate and overcome the environmental threats (Hussain & Reigosa, 2014; Pavarinia et al., 2012). The main function of plant organic compounds is thought to produce phyto-climatic adaptive mechanisms (Kliebenstein, 2004; Hussain et al., 2012). Different biotic and abiotic stresses influence the accumulation of plant organic metabolites (Naghdibadi et al., 2004; Pavarinia et al., 2012). The diversity in accumulation of phytochemicals is strongly susceptible to temperature, light intensity, altitude, and water stresses (Nikolic and Zlatkovic 2010; Radusiene et al., 2012). For the current investigation four plant species were selected from different zones. The Nerium oleander belongs to Apocynaceae family. It is a poisonous evergreen shrub that grows to about 6 to 7 meters in height. It loves warmth and tolerates all types of soils. (Huxley et al., 1992). Ricinus communis belongs to family Euphorbiaceae , it is an evergreen glabrous, soft-woody shrub. It grows throughout the warm temperate and tropical regions. (Salihu et al., 2012). Caloropis procera belongs to Asclepiadaceae family, it is erect, tall, large, branched and with milky latex shrub (Begum et al., 1987). Withania somnifera belongs to family Solanaceae, an erect, evergreen, tormentor shrub, 30-150 cm in height and with stout and fleshy taproots. (Welman, 2011). Materials and methods Study area: Pakistan has a great climatic diversity due to variation in topography, altitude, and season throughout the region. PARC (1980) have subjected the country into ten agro-ecological zones, based on physiographic characteristics (Muhammad, 1986; PARC, 1980) (Fig. 1). Only three zones were selected for the present studies. The site of sampling from investigated zones is presented in table 1 . Table 1 Ecological conditions of selected sites with their respective zones No. Name of zones Sites Altitude Ecological conditions of sites 1 Sandy Desert Hasilpur (cholistan) 140 m or 459 feet Arid climate, annual rain fall 100-200 mm, temperature max. 50 o C and mini. 13 o C and sandy soil (Noureen et al., 2008) 2 Northern Irrigated plain Sahiwal 175 m or 574 feet Semi-arid climate, annual rain fall 200 mm, temperature max. 45 o C and mini. 7 o C and sandy, loam-clay, loam soil (Arshad et al., 2007). 3 Barani lands (rainfall) Kalar kahar 644 m or 2114 feet Hot semi-arid climate, annual rain fall 485 mm, temperature max. 38 o C and mini. 4 o C and sandy, loam-clay, loam soil (Ahmed & Zaman, 2012). Soil Sampling and Determination of various parameters Collection of soil sample: All the soil samples were collected from the barren soil as well as under the canopies of selected plants. Soil samples were taken from the depth of 10-15 of the topsoil using a soil digger. Organic matter determination: Organic matter of soil was determined by the standard method, referred to as the Walkley-Black method explained in Estefan et al. (2013), 10 ml of 1 N potassium dichromate solution and 20 ml of concentrated H 2 SO 4 were added in 1gm of air-dried soil sample and swirl the beaker to mix the suspension. After 30 minutes, about 200 ml DI water and 10 ml concentrated H 3 PO 4 were added using a dispenser and allowed the mixture to cool. Then 10-15 drops of diphenylamine indicator added, and titrated with 0.5 M ferrous ammonium sulfate solution, until the color changed from violet-blue to green. Whole the process was repeated thrice, a blank solution containing all reagents but no soil, also treated in the same way as the soil suspensions. Quantitative Analysis Preparation of extract: Powder of each plant leaf sample (10 g) was soaked separately in 75% methanol for 24 hours. The soaked mixture was filtered and obtained residue was re-extracted with the same volume of solvent and filtered again and volume was made up to 100 ml in measuring flask. Determination of phenolic acids: Total phenolic acid content was estimated by the method described by Taga et al. (1984). Each extract (1ml) was mixed with 10% Folin-ciocalteu reagent (1ml) followed by the addition of saturated solution of Na 2 CO 3 (2 ml). The mixture was kept for 30 minutes, and absorbance was measured λ=760 nm by spectrophotometer. The total phenolic acid contents were calculated as gram gallic acid equivalent 1g/100g dry weight by using the regression equation from the standard curve of Gallic acid. Determination of flavonoids: The total flavonoid contents were estimated according to the method of Jia and Tang in 1999 with a slight modification. An aliquot (1ml) of each extract (0.1 mg/ml) was added to 4ml ethanol and 0.3 ml NaNO 2 . After ten minutes 10% AlCl 3 (0.3 ml) was added at 25 o C. Then 4% NaOH (4 ml) and 30% ethanol (0.4 ml) were thoroughly mixed in it. It was kept standing for 12 minutes and absorbance was measured at λ=510 nm against blank. The contents of total flavonoids were calculated as 1g/100 g dry weight by the calibration curve of catechin anhydrate. Determination of tannins: The total tannins content in ground samples were determined by the following method of Makkar as modified by Fagbemi et al., in 2005. To the methanolic extract of each sample (1ml), Folin –Ciocalteu’s reagent (0.5ml), saturated Na 2 CO 3 solution (1ml) and distilled water (8 ml) were added, mixed well, and allowed to stand for 30 minutes at room temperature. The contents were centrifuged, and absorbance was recorded at λ=725 nm by using UV-Visible spectrophotometer. The tannins contents in each sample were calculated as g/100 g dry weight by the calibration curve of tannic acid. Results The results show that in zone 1 N. oleander contains the highest amount of phenolic acids (3.251 g) and in zone 2 it contains the minimum (0.45 g) amount, while in zone 3 it has 1.966 g phenolic acids, as arranged in the order zone 1>zone 3>zone 2. It contains the maximum flavonoids (0.132 g) in zone 1 and the minimum (0.048 g) in zone 2, while in zone 3 it has 0.091 g, as arranged in the order zone 1>zone 3>zone 2. It contains maximum tannins (1.369 g) in zone 1 and minimum (0.984 g) in zone 2, while in zone 3 it has 1.021 g, as arranged in the order zone 1>zone 3>zone 2. These phytochemicals are significantly varied in each zone at the level of 0.05 (Fig. 2). The results show that R. communis has phenolic acids in the decreasing order, zone 3>zone 2>zone 1 as maximum (2.166 g) in zone 3 and minimum (1.897 g) in zone 1. It is found to contain the maximum (0.083 g) and the minimum amounts (0.063 g) of flavonoids in zone 1 and zone 2 respectively while in one 3 it has 0.081 g, as arranged in the decreasing order zone 1>zone 3>zone 2. It has maximum (1.168 g) and minimum amounts (0.854 g) of tannins in zone 3 and zone 2 respectively while in zone 1 it has tannins 0.944 gm, as arranged in the decreasing order zone 3>zone 1>zone 2. All these phytochemicals are significantly varied at the significance level of 0.05 in all samples (Fig. 3) The results show that C. procera contains phenolic acids in the order zone 2>zone 1>zone 3, as the lowest amount (1.04 g) is present in zone 3 and the highest amount (2.102 g) in zone 1. It contains flavonoids in the order zone 2>zone 3>zone 1, as the lowest amount (0.061 g) is found in zone 1 and the maximum amount (0.074 g) in zone 2. It contains tannins in the order zone 2>zone 1>zone 3, as the least amount (0.836 g) is found in zone 3 and maximum (1.225 g) in zone 2. These phytochemicals are significantly varied at the significance level of 0.05 in all zones (Fig. 4). The results show that W. somnifera contains the highest amount (3.39 g) of phenolic acids in zone 3 and the lowest amount (1.136 g) in zone 1, as in the decreasing order zone 3>zone 2>zone 1. It contains the highest amount (0.011 g) in zone 2 and the minimum (0.088 g) in zone 3 as in the decreasing order zone 2>zone 1>zone 3. It has the highest amount (1.351 g) in zone 3 and minimum amount (0.912 g) in zone 1, as in the decreasing order zone 3>zone 2>zone 1. The two phytochemicals phenolic acids and tannins are significantly varied at the significance level of 0.05 and flavonoids show significance at the level of 0.91 so; it is not significantly varied in all samples of W. somnifera (Fig. 5). Discussion The phytochemical analysis of all plant samples suggests that the quantities of phytochemicals of each species are significantly varied in studied zones. Previous research revealed that environmental stresses cause the major effect on synthesis of phytochemicals. In the current study plant species selected from three different ecological zones, which are divided on the base of different ecological conditions by PARC, these different ecological conditions can be responsible for varied phytochemical contents of same species in different zones. The results revealed that maximum phytochemical contents were found in zone 1 and zone 3 and minimum amounts in zone 2. Other researchers reported that both these zones exhibit extreme climatic conditions, as zone 3 is present at the 644 m altitude, zone 2 is at 172 m and zone 1 is at the 140 m. So, zone 3 because of highest altitude faces low temperature, high UVB light, high precipitation, and low atmospheric pressure (Ahmed & Zaman, 2012), while zone 1 has the lowest altitude and faces droughts, low precipitation, and high temperature (Noureen et al., 2008). Environmental stresses i.e., temperature, light intensity, altitude, and water stresses strongly influence the accumulation of phytochemicals (Nikolic and Zlatkovic 2010; Radusiene et al., 2012). UV-B-treated plants produce significant amounts of flavonoids and phenolics (Gould, 2004). UVB radiation increase phenylalanine ammonia lyase (PAL) activities which can facilitate phenolic accumulation and induce stresses tolerance (Tekelmariam & Black, 2004). It is also reported that flavonoids increase with light intensity forms (Erhard & Gross, 2005). Phonoilc and flavonoid concentrations are also influenced by irrigation or precipitation in berries (Cuadra and Harborne, 1996). High temperature also induces increased phytochemical contents in several plant species (Sayre et al., 1953). Bray (2002) reported that chalocane synthase (CHS) an enzyme that is involved in the biosynthesis of flavonoids and phenylalanine that (PAL) is the key enzyme in the phenolic biosynthesis that were up regulated by drought stress in Arabdopsis (Bray, 2002). High temperatures, water stresses, extreme light intensities and poor soil quality increase the tannin content of plants (Rhoades, 1979). High temperatures and UV light have been shown to influence flavonoid gene expression (Shamir and Levi-Nissim, 1997). Results indicate that zone 1 has lowest and zone 2 has maximum amount of soil organic matter from open soils as well as under plant canopies (Fig. 6 & 7). Other research also support the evidence of less soil organic matter in zone 1 than in zone 2 (Ahmed & Zaman, 2012; Noureen et al., 2008). The zone 1 is a deserted area and consists of less vegetation, as vegetation plays very important role in enrichment of nutrient poor sandy soil (Noureen et al., 2008). Soils generally show a rapid decrease in organic matter from the surface downwards and away from the vegetation (karim et al., 2009). On the other hand, zone 2 is cultivated area, as plant residues help to increase the soil organic matter of a region (karim et al., 2009). In zone 3 soil organic matter is present as more than in zone 1 and less than in zone 2, other research also supports this evidence (Ahmed & Zaman, 2012). It shows that soil organic matter in zone 2 is more than other zones; it can be due to its highly cultivated soil. The results indicate that phenolic acids of R. communis, C. procera and W. somnifera are weak positively and flavonoid and tannin of C. procera are strong positively correlated with the soil organic matter. But all other phytochemicals of four plants are negatively correlated with the soil organic matter (Table 2). Zone 2 has different environmental conditions than zone 1 and zone 3, for example it doesn’t faces environmental stresses and has high soil organic matter that is less in other zones. This zone has less stress able conditions of temperature, precipitation and light intensity that less influence the accumulation of phytochemicals by plants. The result indicates that plants produce lowest amounts of phytochemicals in zone 2 than in other zones. The results also reveal that soil organic matter is negatively correlated with the phytochemicals. So, it predicts that high organic matter has negative impact on the phytochemicals in zone 2. It is reported that higher soil fertility actually hinders the accumulation of phytochemicals (Oh, 2008). Fernandez-Esobar et al., (2006) suggested that application of organic matter to soil increases microbial activity that leads to the fixation of plant nutrients including N, while excessive nitrogen produce negative impacts on phenolic contents (Fernandez-Esobar et al., 2006). So, it reveals that in zone 2 no environmental stresses and high soil organic matter lead to the less accumulation of phytochemicals by plants. Conclusion All these results lead to the conclusion that different plant species and even same specie produces different amounts of phytochemicals in response of different ecological conditions. Plants produce high amounts of phytochemicals in response of environmental stresses like temperature stresses, water stresses, high light intensities and low soil organic matter. But the most favorable conditions for the phytochemicals accumulation are low temperature, moderate organic matter high UVB light and droughts, in other words at high altitudes plants contain more phytochemicals. Declarations Contributing Author Details; S.A.K. and K.T. perform the experiments and wrote the main manuscript. Z.A.S. supervise the work. KFA. Proofread the Manuscript ZAS & KFA reviewed/finalize the manuscript. Competing Interest: It is declared that there is no competing interest. Ethical Approval: NA Approval for Reproducing Image: NA Funding: Authors declare that funding was not received from any source for this research work. Availability of data and materials: All the data to support this study are included in the manuscript. References Ahmad, I., M. Hussain, M. Sajid, A. Ahmad and M. Hameed. 2008. Spatio-Temporal Effects on association of plant species in Soone valley of Pakistan. Pak. J. Bot., 40(5): 1865-1876. Ahmed, S. and N. Zaman. 2012. Evaluation of groundnut germplasm against root rot disease in agro-ecological conditions of district Chakwal, Punjab-Pakistan. Afri J Microbiol., 6: 5862-5865. Arshad, M., M.Y. Ashraf, M. Ahamad and F. Zaman. 2007. Morpho-genetic variability potential of Cenchrus ciliaris L. from Cholistan desert. Pakistan. Pak. J. Bot., 39: 1481-1488. Begum, VH., and J. Sadique. 1987. Effect of Withania somnifera on glycosaminoglycan synthesis in carrageenin-induced air pouch granuloma. Biochem Med. Metab. Biol. 38:272-277. Bray, E. A. (2002). Classification of differentially expressed water deficit stress in Arabidopsis thaliana an analysis using microarray and differential expression data. Ann Bot., 89:603-811. Cuadra, P. and J. B. Harborne. 1996. Changes in epicuticular flavonoids and photosynthetic pigments as a plant response to UV-B radiation. Z. Naturforsch. C, 51, 671. Erhard, D. and E. M. Gross. 2005. Do environmental factors influence composition of potential allelochemicals in the submersed freshwater macrophyte Elodea nuttallii (Hydrocharitaceae)? First publ. in: Verhandlungen der Internationalen Vereinigung für Limnologie 29 pp. 287-291. Estefan, G., R. Sommer, and J. Ryan. 2013. Third Edition- Methods of Soil, Plant, and Water Analysis: A manual for the West Asia and North Africa region. Fagbemi T.N., A. A. Oshodi and K. O. Ipinmoroti. 2005. Processing effects on some antinutritional factors and in vivo multienzymes protein digestibility (IVPD) of three tropical seeds. Pak. J. Nutrition., 4(4):250-256. Frenandz-Escobar, R. G., B. Novelo, M. P. Aguilera and M. Uceda, 2006. Olive oil quantity decreases with N over fertilization. Hortscience 41:215-219. Gould, K.S. 2004. Nature’s Swiss army knife: the diverse protective roles of anthocyanins in leaves. J. Biomed. Biotechnol., 5: 314. Hussain, I. H. and M. J. Reigosa. 2014. Evaluation of herbicide potential of sesquiterpene lactone and flavonoid: impact on germination, seedling growth indices and root length in Arabidopsis thaliana . Pak. J. Bot., 46(3): 995-1000. Hussain, Md. S., S. Fareed, S. Ansari, M. A. Rahman, I. Z. Ahmad, and M. Saeed. 2012. Current approaches toward production of secondary plant metabolites. J Pharm Bioallied Sci. 4(1): 10–20. Huxley, A., M. Griffiths, and M. Levy. 1992. The new RHS Dictionary of Gardening. Macmillan. ISBN 0-333-47494-5. Javaid A., S, Shafique and S. Shafique. 2010. Seasonal pattern of seed dormancy in Parthenium hysterophorus L. Pak. J. Bot., 42(1): 497-503. Jia Z.S. and Tang. 1999. The flavonoids content in Mulberry and their Scavenging effects on superoxide Radicals. J. Food Chem., 64: 555-559. Karim, B., A. Mukhtar, H. Mukhtar and M. Athar, 2009. Effect of the Canopy Cover on the Organic and Inorganic Content of Soil in Cholistan Desert. Pak. J. Bot., 41(5): 2387-2395. Khan, S.M., S. Page, H. Ahmad, Zahidullah, H. Shaheen, M. Ahamd and D. Harper. 2013. Phyto-climatic gradient of vegetation and habitat specificity in the high elevation Western Himalayas. Pak. J. Bot., 45(SI): 223-230. Kliebenstein, D. J. 2004. Secondary metabolites and plant /environment interactions: a view through Arabidopsis thaliana -tinged glasses. Plant Cell Environ, 27: 675–684. Krasteva, I., A. Nedelcheva, D. Pavlova, P. Zdraveva, S. Nikolov and K. Mitov, 2013. Influence of serpentine soils on the flavonoid content of Hypericum populations growing in Bulgaria. 7(25): 1762-1765. Muhammad, A. 1986. Cropping pattern in different ecological zones of Pakistan, PARC In: Khan, G. 2004. Technical report on the characterization of the agro ecological context in which FANGR (farm animal genetic resource) are found development and application of decision support tools to conserve and sustainably use genetic diversity in indigenous livestock and wild relatives, Islamabad, Pakistan. Naghdibadi. H., D. Yazdani, S. M. Ali., and F. Nazari 2004. Effects of spacing and harvesting time on herbage yield and quality/quantity of oil in thyme, Thymus vulgaris L. Ind. Crop. Pro, 2004; 19: 231-236. Nikolic, G. S. and Zlatkovic, Z. S. 2010. Assaying the variation in secondary metabolites of St. John’s wort for its better use as an antibiotic. Journal of Medicinal Plants Research Vol. 4(3) pp. 211-224. Noureen S., M. Arshad, K. Mahmood and M. Y. Ashraf. 2008. Improvement in fertility of nutritionally poor sandy soil of cholistan desert, Pakistan by Calligonum polygonoides Linn. Pak. J. Bot., 40(1): 265-274. Oh M., 2008. Kansas university Munhattan Kansas Uml number: 3310843 PARC. 1980. Agro-ecological regions of Pakistan In: Khan, A. G. 2004. Technical report on the characterization of the agro ecological context in which FANGR (farm animal genetic resource) are found development and application of decision support tools to conserve and sustainably use genetic diversity in indigenous livestock and wild relatives, Islamabad, Pakistan. Pavarinia, D. P., S. P. Pavarinib, M. Niehuesa and N. P. Lopesa. 2012. Exogenous influences on plant secondary metabolite levels. http://dx.doi.org/10.1016/j.anifeedsci.2012.07.002 Radušienė, J., B. Karpavičienė and Ž. Stanius. 2012. Effect of external and internal factors on secondary metabolites accumulation. In: St. John’s Worth 2012, 18(2): 101–108 Rhoades, D.F. 1979. Evolution of plant chemical defence against herbivores. In: Herbivores: their interactions with secondary plant metabolites (Rosenthal G.A. and Janzen D.H., eds.). Acad. Press, NY, USA, pp. 3-54. Sayre, C. B., W. B. Roinson and T. Wishnetsky, 1953. Effect of temperature on the color of lycopene and carotene content of detached and wine ripened tomatoes. Proc Am. Soc. Hortic. Sci. 61:381-387. Salick, J., Z. Fangb, and A. Byg. 2009. Eastern Himalayan alpine plant ecology, Tibetan ethnobotany, andclimate change. Global. Environ. Chang, 19: 147-155. Salihu, B. Z., Gana, K. Andrew and O. B. Apuyor. 2012. Castor oil plant ( Ricinus communis L.). botany, ecology and uses. Int. J. Sci. Res 3:358-362 Shamir, O., M. and A. Levi-Nissim. 1997. Temperature effects on the leaf pigmentation of Cotinus coggygria ‘‘Royal Purple’’, J. Hortic. Sci., 72: 425. Shaukat, S. S. and N. Burhan. 2000. Fecundity, seed characteristics and factors regulating germinination of Rhynchosia minima (L.) D.C., Pak. J. Bot.,32(1):211-226,2000. Taga, M.S., E.E. Millar and D.E. Pratt. 1984. Chia seed as a source of natural lipid antioxidants. J. Chemi soci., 61, 982-931. Tilman, D. 1988. Princeton, N.J.: Princeton University Press, Series:Monographs in population biology, 26. Teklemariam, T. A. and T. J. Black. 2004. Phenylalanine ammonia-lyase induced freezing tolerance in Jack Pine ( Pinus banksiana ) seedlings treating with low ambient levels of ultraviolet-B radiation. Physical plant 122:244-253. Welman, M. 2011. National Herbarium, Withania somnifera Pretoria December 2011. Plant Strategies and the Dynamics and Structure of Plant Communities. (MPB-26), Volume 26 Princeton University Press. Table 2 Variables Soil organic matter N. oleander R. communis C. procera W. somnifera N. oleander Soil organic matter 1 Phenolic acids -.989 * Flavonoids -.983 Tannins -.805 R. communis Soil organic matter .999 * 1 Phenolic acids .154 .113 Flavonoids -.983 -.990 * Tannins -.462 -.498 C. procera Soil organic matter 1.000 ** .997 * 1 Phenolic acid .499 .534 .473 Flavonoids .971 .960 .977 Tannins .846 .867 .830 W. somnifera Soil organic matter -.047 -.088 -.018 1 Phenolic acids .471 .435 .498 .859 Flavonoids -.998 * -1.000 ** -.995 * .118 Tannins -.177 -.217 -.148 .991 * Table. 2 Correlation between phytochemicals of different plants and soil organic matter Soil samples were collected from the depth of 10-15 of the topsoil using a soil digger. 3 soils samples were collected and mixed from each data. Additional Declarations The authors declare no competing interests. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3308843","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":231845124,"identity":"2344ac07-a11f-449c-ae8c-5f823f6a8d15","order_by":0,"name":"Khola Tazeen","email":"","orcid":"","institution":"University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Khola","middleName":"","lastName":"Tazeen","suffix":""},{"id":231845125,"identity":"5c52ef1a-fcb7-41b5-880d-59367387d2f5","order_by":1,"name":"Zahoor Ahmad Sajid","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIiWNgGAWjYFACHjCZAOFUgEkDhABeLWwg6gzJWhjbiNAi33724IOPexjy+OWbnz3mnXcnsYG9eZsE4440nFoYe/KSDWc8YyiWbGMzN+bd9iyxgedYmQTjmRycWpgZcsykeQ4wJG44xmAmzbvtcGKDRI6ZBGNbBU4tbPxvYFrYv0nzzgFqkX+DXwuPBNwWHqAtDSBbeEBacDtMQuKNseGMAxJAv+SUSc459sy4jSet2CLxDG7vy/fnGD74cMAmj5/5+DaJNzV3ZPvZD2+88XFHMk4tMMvAJBMPwwFIlCY2ENIBBYw/gFogLGK1jIJRMApGwUgAAGsLUEutMfkCAAAAAElFTkSuQmCC","orcid":"","institution":"University of the Punjab Lahore","correspondingAuthor":true,"prefix":"","firstName":"Zahoor","middleName":"Ahmad","lastName":"Sajid","suffix":""},{"id":231845126,"identity":"5e2f9bd7-ea2d-4a9a-9fa2-1c288a9c1268","order_by":2,"name":"Khalid Farooq Akbar","email":"","orcid":"","institution":"University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Khalid","middleName":"Farooq","lastName":"Akbar","suffix":""}],"badges":[],"createdAt":"2023-08-30 05:44:24","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-3308843/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-3308843/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":53422953,"identity":"e7a3c4bd-5416-4fd6-9726-7393cbb8813a","added_by":"auto","created_at":"2024-03-25 19:19:15","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":89539,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMap of Pakistan with agro-ecological zones (PARC1980)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/8df7d63df997ad738a44c8ba.png"},{"id":53422951,"identity":"ddb943c8-b4ba-4a31-89fc-29bf3467899b","added_by":"auto","created_at":"2024-03-25 19:19:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":10733,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ephytochemical variation of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eN. oleander\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e in 3 ecological zones\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ezone 1-sandy desert; zone 2-northern irrigated plane; and zone 3-barani lands\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/0dd92aaf2ae61b6e24ffcfeb.png"},{"id":53423363,"identity":"a84aa369-203d-4f80-98f7-612942ea00c3","added_by":"auto","created_at":"2024-03-25 19:27:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":13101,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePhytochemicals variation of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. procera\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e in 3 ecological zones\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/8532b01d602024edb0ed77be.png"},{"id":53422949,"identity":"47373079-48a7-4e1c-9228-8732f7cbbca7","added_by":"auto","created_at":"2024-03-25 19:19:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":10691,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePhytochemical variation of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eR. communis\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e in 3 ecological zones\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/38592b574a6deb9468dffbf7.png"},{"id":53422954,"identity":"559f958b-1ac2-4d79-a3c1-23c93a2d56b3","added_by":"auto","created_at":"2024-03-25 19:19:15","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":11545,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePhytochemical variation of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eW. somnifera\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e in 3 ecological zones\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/8250dece55ed60f0e8868eac.png"},{"id":53423364,"identity":"b8170591-6a6f-4839-b114-7d23e6b43acd","added_by":"auto","created_at":"2024-03-25 19:27:15","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":11752,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSoil organic matter % in three ecological zones.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/2a553d7cc6f4216180c08d55.png"},{"id":53423365,"identity":"5b283f84-17da-407a-88dc-84c84caf9ec2","added_by":"auto","created_at":"2024-03-25 19:27:15","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":19623,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMeans and standard deviation of soil organic matter under plant canopies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eN. oleander\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e, \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eR. communis\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e, \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. procera\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e and \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eW. somnifera\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/08332def6209aa1cafd2f189.png"},{"id":53423742,"identity":"2c35fd60-caab-479e-a846-f40cb672a3f3","added_by":"auto","created_at":"2024-03-25 19:35:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":570308,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3308843/v2/2d52df85-5377-4a21-9aaa-346b2169763f.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eThe Variations in the Phytochemical Attributes of \u003cem\u003eNerium Oleander\u003c/em\u003e, \u003cem\u003eRicinus Communis\u003c/em\u003e, \u003cem\u003eCalotropis Procera\u003c/em\u003e and \u003cem\u003eWithania Somnifera\u003c/em\u003e From Three Ecological Zones of Pakistan\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe life form of vegetation in a region can be measured\u0026nbsp;by the plant-environment analogy (Khan et al., 2013). Different ecological factors influence differently the vegetation, plant community, structure, and morphology of plants to adapt to their environment for their better survival of life. These influences could lead to long term adaptive strategies (Salick et al., 2009; Khan et al., 2013). Long term existence of species needs to be differentiated as they have back-and-forth in their abilities to respond constrains of their environment (Tilman, 1988). Ecological attributes that can affect biota of a region are, slope, altitude, topography, soil, habitat, vegetation type, plant community (Ahmad et al., 2008), solar radiation, temperature (Javaid et al., 2010), moisture stress, salinity, water, and precipitation (Shukat \u0026amp; Burhan, 2000).\u003c/p\u003e\n\u003cp\u003eThe phyto-climatic gradient of a region influences the plants to grow healthy i.e., to adapt to their environment (Khan et al., 2013). The effect of climatic diversity on the synthesis of phytochemicals has widely perceived a considerable amount of attention (Krasteva et al., 2013). Plants produce a vast and diverse variety of organic compounds that help them to communicate and overcome the environmental threats (Hussain \u0026amp; Reigosa, 2014; Pavarinia et al., 2012). The main function of plant organic compounds is thought to produce phyto-climatic adaptive mechanisms (Kliebenstein, 2004; Hussain et al., 2012). Different biotic and abiotic stresses influence the accumulation of plant organic metabolites (Naghdibadi et al., 2004; Pavarinia et al., 2012). The diversity in accumulation of phytochemicals is strongly susceptible to temperature, light intensity, altitude, and water stresses (Nikolic and Zlatkovic 2010; Radusiene et al., 2012). For the current investigation four plant species were selected from different zones. The \u003cem\u003eNerium oleander\u003c/em\u003e belongs to \u003cem\u003eApocynaceae\u003c/em\u003e family. It\u0026nbsp;is a\u0026nbsp;poisonous evergreen shrub\u0026nbsp;that grows to about 6\u0026thinsp;to 7\u0026thinsp;meters in height. It loves warmth and tolerates all types of soils.\u0026nbsp;(Huxley et al., 1992). \u0026nbsp;\u003cem\u003eRicinus communis\u0026nbsp;\u003c/em\u003ebelongs to family\u0026nbsp;\u003ca href=\"http://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN\u0026search_value=28031\"\u003e\u003cem\u003eEuphorbiaceae\u003c/em\u003e\u003c/a\u003e, it is an evergreen glabrous, soft-woody shrub.\u0026nbsp;It grows throughout the warm temperate and tropical regions. (Salihu et al., 2012). \u003cem\u003eCaloropis procera\u0026nbsp;\u003c/em\u003ebelongs to\u0026nbsp;\u003cem\u003eAsclepiadaceae\u003c/em\u003e family,\u0026nbsp;it is erect, tall, large, branched and with milky latex shrub (Begum et al.,\u0026nbsp;1987).\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cem\u003eWithania somnifera\u003c/em\u003e belongs to family Solanaceae, an erect, evergreen, tormentor shrub, 30-150 cm in height and with stout and fleshy taproots. (Welman, 2011).\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003e\u003cstrong\u003eStudy area:\u0026nbsp;\u003c/strong\u003ePakistan has a great climatic diversity due to variation in topography, altitude, and season throughout the region. PARC (1980) have subjected the country into ten agro-ecological zones, based on physiographic characteristics (Muhammad, 1986; PARC, 1980) (Fig. 1). Only three zones were selected for the present studies. The site of sampling from investigated zones is presented in table 1\u003cstrong\u003e.\u003c/strong\u003e\u0026nbsp; \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp style='margin-top:6.0pt;margin-right:0in;margin-bottom:6.0pt;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eTable 1 Ecological conditions of selected sites with their respective zones\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"width: 4.9e+2pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 35.2pt;border: 1pt solid black;padding: 0in 5.4pt;height: 6.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eNo.\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59pt;border-width: 1pt 1pt 1pt medium;border-style: solid solid solid none;border-color: black black black currentcolor;border-image: none;padding: 0in 5.4pt;height: 6.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eName of \u0026nbsp;zones\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 55.15pt;border-width: 1pt 1pt 1pt medium;border-style: solid solid solid none;border-color: black black black currentcolor;border-image: none;padding: 0in 5.4pt;height: 6.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSites\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.05pt;border-width: 1pt 1pt 1pt medium;border-style: solid solid solid none;border-color: black black black currentcolor;border-image: none;padding: 0in 5.4pt;height: 6.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eAltitude\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 288.1pt;border-width: 1pt 1pt 1pt medium;border-style: solid solid solid none;border-color: black black black currentcolor;border-image: none;padding: 0in 5.4pt;height: 6.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eEcological conditions of sites\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 35.2pt;border-width: medium 1pt 1pt;border-style: none solid solid;border-color: currentcolor black black;border-image: none;padding: 0in 5.4pt;height: 19.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 19.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSandy Desert \u0026nbsp;\u0026nbsp;\u003c/span\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 55.15pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 19.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eHasilpur (cholistan)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.05pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 19.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e140 m or\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e459 feet\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 288.1pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 19.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eArid climate, annual rain fall 100-200 mm, temperature max. 50 \u003csup\u003eo\u003c/sup\u003eC and mini. 13 \u003csup\u003eo\u003c/sup\u003eC and sandy soil (Noureen et al., 2008)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 35.2pt;border-width: medium 1pt 1pt;border-style: none solid solid;border-color: currentcolor black black;border-image: none;padding: 0in 5.4pt;height: 27.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eNorthern Irrigated plain\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 55.15pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSahiwal\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.05pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e175 m or\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;574 feet\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 288.1pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSemi-arid climate, annual rain fall 200 mm, temperature max. 45 \u003csup\u003eo\u003c/sup\u003eC and mini. 7 \u003csup\u003eo\u003c/sup\u003eC and sandy, loam-clay, loam soil (Arshad et al., 2007).\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 35.2pt;border-width: medium 1pt 1pt;border-style: none solid solid;border-color: currentcolor black black;border-image: none;padding: 0in 5.4pt;height: 27.45pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e3\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.45pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eBarani lands (rainfall)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 55.15pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.45pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eKalar kahar\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.05pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.45pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e644 m or\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e2114 feet\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 288.1pt;border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor black black currentcolor;padding: 0in 5.4pt;height: 27.45pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eHot semi-arid climate, annual rain fall 485 mm, temperature max. 38 \u003csup\u003eo\u003c/sup\u003eC and mini. 4 \u003csup\u003eo\u003c/sup\u003eC and sandy, loam-clay, loam soil (Ahmed \u0026amp; Zaman, 2012).\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:200%;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;line-height:200%;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSoil Sampling and Determination of various parameters\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCollection of soil sample:\u0026nbsp;\u003c/strong\u003eAll the soil samples were collected from the barren soil as well as under the canopies of selected plants. Soil samples were taken from the depth of 10-15 of the topsoil using a soil digger.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOrganic matter determination:\u0026nbsp;\u003c/strong\u003eOrganic matter of soil was determined by the standard method, referred to as the Walkley-Black method explained in\u0026nbsp;Estefan et al. (2013), 10 ml of 1 \u003cem\u003eN\u0026nbsp;\u003c/em\u003epotassium dichromate\u0026nbsp;solution and 20 ml of concentrated\u0026nbsp;H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e were added in 1gm of air-dried soil sample and swirl the beaker to mix the suspension. \u0026nbsp;After 30 minutes, about 200 ml DI water and 10 ml concentrated H\u003csub\u003e3\u003c/sub\u003ePO\u003csub\u003e4\u0026nbsp;\u003c/sub\u003ewere added\u003csub\u003e\u0026nbsp;\u003c/sub\u003eusing a dispenser and allowed the mixture to cool. Then 10-15 drops of diphenylamine\u0026nbsp;indicator added, and titrated with 0.5 \u003cem\u003eM\u0026nbsp;\u003c/em\u003eferrous ammonium sulfate\u0026nbsp;solution, until the color changed from violet-blue to green. Whole the process was repeated thrice, a blank solution containing all reagents but no soil, also treated in the same way as the soil suspensions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cbr\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eQuantitative Analysis\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of extract:\u0026nbsp;\u003c/strong\u003ePowder of each plant leaf sample (10 g) was soaked separately in 75% methanol for 24 hours. The soaked mixture was filtered and obtained residue was re-extracted with the same volume of solvent and filtered again and volume was made up to 100 ml in measuring flask.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetermination of phenolic acids:\u0026nbsp;\u003c/strong\u003eTotal phenolic acid content was estimated by the method described by Taga et al. (1984).\u003cem\u003e\u0026nbsp;\u003c/em\u003eEach extract (1ml) was mixed with 10% Folin-ciocalteu reagent (1ml) followed by the addition of saturated solution of Na\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e (2 ml). The mixture was kept for 30 minutes, and absorbance was measured \u0026lambda;=760 nm by spectrophotometer. The total phenolic acid contents were calculated as gram gallic acid equivalent 1g/100g dry weight by using the regression equation from the standard curve of Gallic acid.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetermination of flavonoids:\u0026nbsp;\u003c/strong\u003eThe total flavonoid contents were estimated according to the method of Jia and Tang in 1999 with a slight modification. An aliquot (1ml) of each extract (0.1 mg/ml) was added to 4ml ethanol and 0.3 ml NaNO\u003csub\u003e2\u003c/sub\u003e. After ten minutes 10% AlCl\u003csub\u003e3\u003c/sub\u003e (0.3 ml) was added at 25\u003csup\u003eo\u003c/sup\u003eC. Then 4% NaOH (4 ml) and 30% ethanol (0.4 ml) were thoroughly mixed in it. It was kept standing for 12 minutes and absorbance was measured at \u0026lambda;=510 nm against blank. The contents of total flavonoids were calculated as 1g/100 g dry weight by the calibration curve of catechin anhydrate.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetermination of tannins:\u0026nbsp;\u003c/strong\u003eThe total tannins content in ground samples were determined by the following method of Makkar as modified by Fagbemi et al., in 2005.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eTo the methanolic extract of each sample (1ml), Folin \u0026ndash;Ciocalteu\u0026rsquo;s reagent (0.5ml), saturated Na\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e solution (1ml) and distilled water (8 ml) were added, mixed well, and allowed to stand for 30 minutes at room temperature. The contents were centrifuged, and absorbance was recorded at \u0026lambda;=725 nm by using UV-Visible spectrophotometer. The tannins contents in each sample were calculated as g/100 g dry weight by the calibration curve of tannic acid.\u0026nbsp;\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe results show that in zone 1 \u003cem\u003eN. oleander\u003c/em\u003e contains the highest amount of phenolic acids (3.251\u0026nbsp;g)\u0026nbsp;and in zone 2 it contains the minimum (0.45\u0026nbsp;g) amount,\u0026nbsp;while in zone 3 it\u003cem\u003e\u0026nbsp;\u003c/em\u003ehas\u0026nbsp;1.966\u0026nbsp;g\u0026nbsp;phenolic acids, as arranged in the order zone 1\u0026gt;zone 3\u0026gt;zone 2. It contains the maximum flavonoids (0.132\u0026nbsp;g)\u0026nbsp;in zone 1 and the minimum (0.048\u0026nbsp;g)\u0026nbsp;in zone 2, while in zone 3 it has\u0026nbsp;0.091\u0026nbsp;g, as arranged in the order zone 1\u0026gt;zone 3\u0026gt;zone 2. It contains maximum tannins (1.369\u0026nbsp;g)\u0026nbsp;in zone 1 and minimum (0.984\u0026nbsp;g) in zone 2,\u0026nbsp;while in zone 3 it has\u0026nbsp;1.021\u0026nbsp;g, as arranged in the order zone 1\u0026gt;zone 3\u0026gt;zone 2. These\u0026nbsp;phytochemicals are significantly varied in each zone at the level of 0.05 (Fig. 2).\u003c/p\u003e\n\u003cp\u003eThe results show that \u003cem\u003eR.\u003c/em\u003e\u003cem\u003e\u0026nbsp;communis\u003c/em\u003e has phenolic acids in the decreasing order, zone 3\u0026gt;zone 2\u0026gt;zone 1 as maximum (2.166 g) in zone 3 and minimum (1.897 g) in zone 1.\u003cem\u003e\u0026nbsp;\u003c/em\u003eIt is found to contain the maximum (0.083\u0026nbsp;g)\u0026nbsp;and the minimum amounts (0.063\u0026nbsp;g) of flavonoids\u0026nbsp;in zone 1 and zone 2 respectively while in one 3 it has\u0026nbsp;0.081\u0026nbsp;g, as arranged in the decreasing order zone 1\u0026gt;zone 3\u0026gt;zone 2.\u0026nbsp;It has\u0026nbsp;maximum (1.168\u0026nbsp;g)\u0026nbsp;and minimum amounts (0.854\u0026nbsp;g) of tannins\u0026nbsp;in zone 3 and zone 2 respectively while in zone 1 it has tannins\u0026nbsp;0.944\u0026nbsp;gm, as arranged in the decreasing order zone 3\u0026gt;zone 1\u0026gt;zone 2.\u0026nbsp;All these phytochemicals are significantly varied at the significance level of 0.05 in all samples\u0026nbsp;(Fig. 3)\u003c/p\u003e\n\u003cp\u003eThe results show that \u003cem\u003eC. procera\u003c/em\u003e contains phenolic acids in the order zone 2\u0026gt;zone 1\u0026gt;zone 3, as the lowest amount (1.04\u0026nbsp;g)\u0026nbsp;is present in zone 3 and the highest amount (2.102\u0026nbsp;g)\u0026nbsp;in zone 1. It contains flavonoids in the order zone 2\u0026gt;zone 3\u0026gt;zone 1, as the lowest amount (0.061\u0026nbsp;g)\u0026nbsp;is found in zone 1 and the maximum amount (0.074\u0026nbsp;g)\u0026nbsp;in zone 2.\u003cem\u003e\u0026nbsp;\u003c/em\u003eIt contains tannins in the order zone 2\u0026gt;zone 1\u0026gt;zone 3, as the least amount (0.836\u0026nbsp;g)\u0026nbsp;is found in zone 3 and maximum (1.225\u0026nbsp;g)\u0026nbsp;in zone 2.\u0026nbsp;These phytochemicals are significantly varied at the significance level of 0.05 in all zones (Fig. 4).\u003c/p\u003e\n\u003cp\u003eThe results show that \u003cem\u003eW. somnifera\u003c/em\u003e contains the highest amount (3.39\u0026nbsp;g) of phenolic acids\u0026nbsp;in zone 3 and the lowest amount (1.136\u0026nbsp;g)\u0026nbsp;in zone 1, as in the decreasing order zone 3\u0026gt;zone 2\u0026gt;zone 1. It contains the highest amount (0.011\u0026nbsp;g)\u0026nbsp;in zone 2 and the minimum (0.088\u0026nbsp;g)\u0026nbsp;in zone 3 as in the decreasing order zone 2\u0026gt;zone 1\u0026gt;zone 3. It has the highest amount (1.351\u0026nbsp;g)\u0026nbsp;in zone 3 and minimum amount (0.912\u0026nbsp;g)\u0026nbsp;in zone 1, as in the decreasing order zone 3\u0026gt;zone 2\u0026gt;zone 1.\u0026nbsp;The two phytochemicals phenolic acids and tannins are significantly varied at the significance level of 0.05 and flavonoids show significance at the level of 0.91 so; it is not significantly varied in all samples of \u003cem\u003eW. somnifera\u003c/em\u003e (Fig. 5).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe phytochemical analysis of all plant samples suggests that the quantities of phytochemicals of each species are significantly varied in studied zones. Previous research revealed that environmental stresses cause the major effect on synthesis of phytochemicals. In the current study plant species selected from three different ecological zones, which are divided on the base of different ecological conditions by PARC, these different ecological conditions can be responsible for varied phytochemical contents of same species in different zones.\u003c/p\u003e\n\u003cp\u003eThe results revealed that maximum phytochemical contents were found in zone 1 and zone 3 and minimum amounts in zone 2. Other researchers reported that both these zones exhibit extreme climatic conditions, as zone 3 is present at the 644 m altitude, zone 2 is at 172 m and zone 1 is at the 140 m. So, zone 3 because of highest altitude faces low temperature, high UVB light, high precipitation, and low atmospheric pressure (Ahmed \u0026amp; Zaman, 2012), while zone 1 has the lowest altitude and faces droughts, low precipitation, and high temperature\u0026nbsp;(Noureen et al., 2008). Environmental stresses i.e., temperature, light intensity, altitude, and water stresses strongly influence the accumulation of phytochemicals (Nikolic and Zlatkovic 2010; Radusiene et al., 2012). UV-B-treated plants produce significant amounts of flavonoids and phenolics (Gould, 2004). UVB radiation increase phenylalanine ammonia lyase (PAL) activities which can facilitate phenolic accumulation and induce stresses tolerance (Tekelmariam \u0026amp; Black, 2004). It is also reported that flavonoids increase with light intensity forms (Erhard \u0026amp; Gross, 2005). Phonoilc and flavonoid concentrations are also influenced by irrigation or precipitation in berries (Cuadra and Harborne, 1996). High temperature also induces increased phytochemical contents in several plant species (Sayre et al., 1953). Bray (2002) reported that chalocane synthase (CHS) an enzyme that is involved in the biosynthesis of flavonoids and phenylalanine that (PAL) is the key enzyme in the phenolic biosynthesis that were up regulated by drought stress in Arabdopsis (Bray, 2002). High temperatures, water stresses, extreme light intensities and poor soil quality increase the tannin content of plants (Rhoades, 1979). High temperatures and UV light have been shown to influence flavonoid gene expression (Shamir and Levi-Nissim, 1997).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults indicate that zone 1 has lowest and zone 2 has maximum amount of soil organic matter from open soils as well as under plant canopies (Fig. 6 \u0026amp; 7). Other research also support the evidence of less soil organic matter in zone 1 than in zone 2 (Ahmed \u0026amp; Zaman, 2012; Noureen et al., 2008). The zone 1 is a deserted area and consists of less vegetation, as vegetation plays very important role in enrichment of nutrient poor sandy soil (Noureen et al., 2008). Soils generally show a rapid decrease in organic matter from the surface downwards and away from the vegetation (karim et al., 2009).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOn the other hand, zone 2 is cultivated area, as plant residues help to increase the soil organic matter of a region (karim et al., 2009). In zone 3 soil organic matter is present as more than in zone 1 and less than in zone 2, other research also supports this evidence (Ahmed \u0026amp; Zaman, 2012). It shows that soil organic matter in zone 2 is more than other zones; it can be due to its highly cultivated soil.\u003c/p\u003e\n\u003cp\u003eThe results indicate that phenolic acids of \u003cem\u003eR.\u003c/em\u003e\u003cem\u003e\u0026nbsp;communis,\u003c/em\u003e \u003cem\u003eC. procera\u0026nbsp;\u003c/em\u003eand \u003cem\u003eW. somnifera\u0026nbsp;\u003c/em\u003eare weak positively and flavonoid and tannin of\u0026nbsp;\u003cem\u003eC. procera\u0026nbsp;\u003c/em\u003eare strong positively correlated with the soil organic matter. But all other phytochemicals of four plants are negatively correlated with the soil organic matter (Table 2).\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eZone 2 has different environmental conditions than zone 1 and zone 3, for example it doesn\u0026rsquo;t faces environmental stresses and has high soil organic matter that is less in other zones. This zone has less stress able conditions of temperature, precipitation and light intensity that less influence the accumulation of phytochemicals by plants. The result indicates that plants produce lowest amounts of phytochemicals in zone 2 than in other zones. The results also reveal that soil organic matter is negatively correlated with the phytochemicals. So, it predicts that high organic matter has negative impact on the phytochemicals in zone 2.\u0026nbsp;It is reported that higher soil fertility actually hinders the accumulation of phytochemicals (Oh, 2008).\u0026nbsp;Fernandez-Esobar et al., (2006) suggested that application of organic matter to soil increases microbial activity that leads to the fixation of plant nutrients including N, while excessive nitrogen produce negative impacts on phenolic contents (Fernandez-Esobar et al., 2006). So, it reveals that in zone 2 no environmental stresses and high soil organic matter lead to the less accumulation of phytochemicals by plants.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eAll these results lead to the conclusion that different plant species and even same specie produces different amounts of phytochemicals in response of different ecological conditions. Plants produce high amounts of phytochemicals in response of environmental stresses like temperature stresses, water stresses, high light intensities and low soil organic matter. But the most favorable conditions for the phytochemicals accumulation are low temperature, moderate organic matter high UVB light and droughts, in other words at high altitudes plants contain more phytochemicals.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eContributing Author Details;\u0026nbsp;\u003c/strong\u003eS.A.K. and K.T. perform the experiments and wrote the main manuscript. Z.A.S. supervise the work. KFA. Proofread the Manuscript ZAS \u0026amp; KFA reviewed/finalize the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interest:\u0026nbsp;\u003c/strong\u003eIt is declared that there is no competing interest.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval:\u0026nbsp;\u003c/strong\u003eNA\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eApproval for Reproducing Image:\u003c/strong\u003e NA\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eAuthors declare that funding was not received from any source for this research work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials:\u0026nbsp;\u003c/strong\u003eAll the data to support this study are included in the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003eAhmad, I., M. Hussain, M. Sajid, A. Ahmad and M. Hameed. 2008. Spatio-Temporal Effects on association of plant species in Soone valley of Pakistan. Pak. J. Bot., 40(5): 1865-1876.\u003c/p\u003e\n\u003cp\u003eAhmed, S. and N. Zaman. 2012. Evaluation of groundnut germplasm against root rot disease in agro-ecological conditions of district Chakwal, Punjab-Pakistan.\u0026nbsp;Afri J Microbiol., 6: 5862-5865.\u003c/p\u003e\n\u003cp\u003eArshad, M., M.Y. Ashraf, M. Ahamad and F. Zaman. \u0026nbsp;2007. Morpho-genetic variability potential of \u003cem\u003eCenchrus ciliaris\u003c/em\u003e L. from Cholistan desert. Pakistan. Pak. J. Bot., 39: 1481-1488.\u003c/p\u003e\n\u003cp\u003eBegum, VH., and J. Sadique. 1987. Effect of \u003cem\u003eWithania somnifera\u003c/em\u003e on glycosaminoglycan synthesis in carrageenin-induced air pouch granuloma. Biochem Med. Metab. Biol. 38:272-277.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBray, E. A. (2002). Classification of differentially expressed water deficit stress in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e an analysis using microarray and differential expression data. Ann Bot., 89:603-811.\u003c/p\u003e\n\u003cp\u003eCuadra, P. and J. B. Harborne. 1996. Changes in epicuticular flavonoids and photosynthetic pigments as a plant response to UV-B radiation. Z. Naturforsch. C, 51, 671.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eErhard, D. and E. M. Gross. 2005. Do environmental factors influence composition of potential allelochemicals in the submersed freshwater macrophyte \u003cem\u003eElodea nuttallii\u0026nbsp;\u003c/em\u003e(Hydrocharitaceae)? First publ. in: Verhandlungen der Internationalen Vereinigung f\u0026uuml;r Limnologie 29 pp. 287-291.\u003c/p\u003e\n\u003cp\u003eEstefan, G., R. Sommer, and J. Ryan. 2013. Third Edition- Methods of Soil, Plant, and Water Analysis: A manual for the West Asia and North Africa region.\u003c/p\u003e\n\u003cp\u003eFagbemi T.N., A. A. Oshodi and K. O. Ipinmoroti. 2005. Processing effects on some antinutritional factors and in vivo multienzymes protein digestibility (IVPD) of three tropical seeds. Pak. J. Nutrition., 4(4):250-256.\u003c/p\u003e\n\u003cp\u003eFrenandz-Escobar, R. G., B. Novelo, M. P. Aguilera and M. Uceda, 2006. Olive oil quantity decreases with N over fertilization. Hortscience 41:215-219.\u003c/p\u003e\n\u003cp\u003eGould, K.S. 2004. Nature\u0026rsquo;s Swiss army knife: the diverse protective roles of anthocyanins in leaves. J. Biomed. Biotechnol., 5: 314.\u003c/p\u003e\n\u003cp\u003eHussain, I. H. and M. J. Reigosa. 2014. Evaluation of herbicide potential of sesquiterpene lactone and flavonoid: impact on germination, seedling growth indices and root length in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e. Pak. J. Bot.,\u0026nbsp;46(3): 995-1000.\u003c/p\u003e\n\u003cp\u003eHussain, Md. S., S. Fareed, S. Ansari, M. A. Rahman, I. Z. Ahmad, and M. Saeed. 2012. Current approaches toward production of secondary plant metabolites. J Pharm Bioallied Sci. 4(1): 10\u0026ndash;20.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHuxley, A., M. Griffiths, and M. Levy. 1992. The new RHS Dictionary of Gardening. Macmillan. ISBN 0-333-47494-5.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eJavaid A., S, Shafique and S. Shafique. 2010. Seasonal pattern of seed dormancy in \u003cem\u003eParthenium hysterophorus\u003c/em\u003e L. Pak. J. Bot., 42(1): 497-503.\u003c/p\u003e\n\u003cp\u003eJia Z.S. and Tang. 1999. The flavonoids content in Mulberry and their Scavenging effects on superoxide Radicals. J. Food Chem., 64: 555-559.\u003c/p\u003e\n\u003cp\u003eKarim, B., A. Mukhtar, H. Mukhtar and M. Athar, 2009. Effect of the Canopy Cover on the Organic and Inorganic Content of Soil in Cholistan Desert. Pak. J. Bot., 41(5): 2387-2395.\u003c/p\u003e\n\u003cp\u003eKhan, S.M., S. Page, H. Ahmad, Zahidullah, H. Shaheen, M. Ahamd and D. Harper. 2013. Phyto-climatic gradient of vegetation and habitat specificity in the high elevation Western Himalayas. Pak. J. Bot., 45(SI): 223-230.\u003c/p\u003e\n\u003cp\u003eKliebenstein, D. J. 2004. Secondary metabolites and plant /environment interactions: a view through \u003cem\u003eArabidopsis thaliana\u003c/em\u003e-tinged glasses. Plant Cell Environ, 27: 675\u0026ndash;684.\u003c/p\u003e\n\u003cp\u003eKrasteva, I., A. Nedelcheva, D. Pavlova, P. Zdraveva, S. Nikolov and K. Mitov, 2013.\u0026nbsp;Influence of serpentine soils on the flavonoid content of \u003cem\u003eHypericum\u0026nbsp;\u003c/em\u003epopulations growing in Bulgaria. 7(25): 1762-1765.\u003c/p\u003e\n\u003cp\u003eMuhammad, A. 1986. Cropping pattern in different ecological zones of Pakistan, PARC In: Khan, G. 2004. Technical report on the characterization of the agro ecological context in which FANGR (farm animal genetic resource) are found development and application of decision support tools to conserve and sustainably use genetic diversity in indigenous livestock and wild relatives, Islamabad, Pakistan.\u003c/p\u003e\n\u003cp\u003eNaghdibadi. H., D. \u0026nbsp;Yazdani, S. M. Ali., and F. Nazari 2004. Effects of spacing and harvesting time on herbage yield and quality/quantity of oil in thyme, \u003cem\u003eThymus vulgaris\u003c/em\u003e L. Ind. Crop. Pro, 2004; 19: 231-236.\u003c/p\u003e\n\u003cp\u003eNikolic, G. S. and Zlatkovic, Z. S. 2010. Assaying the variation in secondary metabolites of St. John\u0026rsquo;s wort for its better use as an antibiotic. Journal of Medicinal Plants Research Vol. 4(3) pp. 211-224.\u003c/p\u003e\n\u003cp\u003eNoureen S., M. Arshad, K. Mahmood and M. Y. Ashraf. 2008. Improvement in fertility of nutritionally poor sandy soil of cholistan desert, Pakistan by \u003cem\u003eCalligonum polygonoides\u003c/em\u003e Linn.\u0026nbsp;Pak. J. Bot., 40(1): 265-274.\u003c/p\u003e\n\u003cp\u003eOh M., 2008. Kansas university Munhattan Kansas Uml number: 3310843 PARC. 1980. Agro-ecological regions of Pakistan In: Khan, A. G. 2004. Technical report on the characterization of the agro ecological context in which FANGR (farm animal genetic resource) are found development and application of decision support tools to conserve and sustainably use genetic diversity in indigenous livestock and wild relatives, Islamabad, Pakistan.\u003c/p\u003e\n\u003cp\u003ePavarinia, D. P., S. P. Pavarinib, M. Niehuesa and N. P. Lopesa. 2012.\u0026nbsp;Exogenous influences on plant secondary metabolite levels. http://dx.doi.org/10.1016/j.anifeedsci.2012.07.002\u003c/p\u003e\n\u003cp\u003eRadu\u0026scaron;ienė, J., B. Karpavičienė and Ž. Stanius. 2012. Effect of external and internal factors on secondary metabolites accumulation. In: St. John\u0026rsquo;s Worth 2012, 18(2): 101\u0026ndash;108\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRhoades, D.F. 1979. Evolution of plant chemical defence against herbivores. In: Herbivores: their interactions with secondary plant metabolites (Rosenthal G.A. and Janzen D.H., eds.). Acad. Press, NY, USA, pp. 3-54.\u003c/p\u003e\n\u003cp\u003eSayre, C. B., W. B. Roinson and T. Wishnetsky, 1953. Effect of temperature on the color of lycopene and carotene content of detached and wine ripened tomatoes. Proc Am. Soc. Hortic. Sci. 61:381-387.\u003c/p\u003e\n\u003cp\u003eSalick, J., Z. Fangb, and A. Byg. 2009. Eastern Himalayan alpine plant ecology, Tibetan ethnobotany, andclimate change. Global. Environ. Chang, 19: 147-155.\u003c/p\u003e\n\u003cp\u003eSalihu, B. Z., Gana, K. Andrew and O. B. Apuyor. 2012.\u0026nbsp;Castor oil plant (\u003cem\u003eRicinus communis\u0026nbsp;\u003c/em\u003eL.). botany, ecology and uses. \u0026nbsp;Int. J. Sci. Res 3:358-362\u003c/p\u003e\n\u003cp\u003eShamir, O., M. and A. Levi-Nissim. 1997. Temperature effects on the leaf pigmentation of \u003cem\u003eCotinus coggygria\u003c/em\u003e \u0026lsquo;\u0026lsquo;Royal Purple\u0026rsquo;\u0026rsquo;, J. Hortic. Sci., 72: 425.\u003c/p\u003e\n\u003cp\u003eShaukat, S. S. and N. Burhan. 2000. Fecundity, seed characteristics and factors regulating germinination of \u003cem\u003eRhynchosia minima\u003c/em\u003e (L.) D.C., Pak. J. Bot.,32(1):211-226,2000.\u003c/p\u003e\n\u003cp\u003eTaga, M.S., E.E. Millar and D.E. Pratt. 1984. Chia seed as a source of natural lipid antioxidants. J. Chemi soci., 61, 982-931.\u003c/p\u003e\n\u003cp\u003eTilman, D. 1988. Princeton, N.J.: Princeton University Press, Series:Monographs in population biology, 26.\u003c/p\u003e\n\u003cp\u003eTeklemariam, T. A. and T. J. Black. 2004. Phenylalanine ammonia-lyase induced freezing tolerance in Jack Pine (\u003cem\u003ePinus banksiana\u003c/em\u003e) seedlings treating with low ambient levels of ultraviolet-B radiation. Physical plant 122:244-253.\u003c/p\u003e\n\u003cp\u003eWelman, M. 2011. National Herbarium, \u003cem\u003eWithania somnifera\u003c/em\u003e Pretoria December 2011. Plant Strategies and the Dynamics and Structure of Plant Communities. (MPB-26), Volume 26 Princeton University Press.\u003c/p\u003e"},{"header":"Table 2","content":"\u003ctable style=\"width:514.5pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" style=\"border: 1pt solid windowtext;padding: 0in 5.4pt;height: 12.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; Variables\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" style=\"border-width: 1pt 1pt 1pt medium;border-style: solid solid solid none;border-color: windowtext windowtext windowtext currentcolor;border-image: none;padding: 0in 5.4pt;height: 12.9pt;vertical-align: bottom;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; Soil organic matter\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 23.6pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eN. oleander\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 23.6pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eR.\u003c/span\u003e\u003c/em\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;communis\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 23.6pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eC. procera\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 23.6pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eW. somnifera\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"border:solid windowtext 1.0pt;border-top:none;padding:0in 5.4pt 0in 5.4pt;height:12.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eN. oleander\u0026nbsp;\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003eSoil organic matter\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt medium medium;border-style: none solid none none;border-color: currentcolor windowtext currentcolor currentcolor;border-image: none;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt medium medium;border-style: none solid none none;border-color: currentcolor windowtext currentcolor currentcolor;border-image: none;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt medium medium;border-style: none solid none none;border-color: currentcolor windowtext currentcolor currentcolor;border-image: none;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003ePhenolic acids\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.989\u003csup\u003e*\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt medium medium;border-style: none solid none none;border-color: currentcolor windowtext currentcolor currentcolor;border-image: none;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt medium medium;border-style: none solid none none;border-color: currentcolor windowtext currentcolor currentcolor;border-image: none;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt medium medium;border-style: none solid none none;border-color: currentcolor windowtext currentcolor currentcolor;border-image: none;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003eFlavonoids\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New 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style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cem\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eW. somnifera\u003c/span\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003eSoil organic matter\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: 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style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.018\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 11.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003ePhenolic acids\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp 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style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 10.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003eFlavonoids\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 10.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.998\u003csup\u003e*\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 10.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-1.000\u003csup\u003e**\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 10.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.995\u003csup\u003e*\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 10.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e.118\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cspan style='font-size:16px;font-family: \"Times New Roman\",serif;color:black;'\u003eTannins\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.177\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.217\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e-.148\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border-width: medium 1pt 1pt medium;border-style: none solid solid none;border-color: currentcolor windowtext windowtext currentcolor;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:center;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;color:black;'\u003e.991\u003csup\u003e*\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:10.0pt;margin-left:0in;line-height:200%;font-size:15px;font-family:\"Calibri\",sans-serif;text-align:justify;'\u003e\u003cstrong\u003e\u003cspan style='font-size:16px;line-height:200%;font-family:\"Times New Roman\",serif;color:black;'\u003eTable. 2 Correlation between phytochemicals of different plants and soil organic matter\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003eSoil samples were collected from the depth of 10-15 of the topsoil using a soil digger.\u003c/span\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:15px;font-family:\"Calibri\",sans-serif;'\u003e\u003cspan style='font-size:16px;font-family:\"Times New Roman\",serif;'\u003e3 soils samples were collected and mixed from each data.\u003c/span\u003e\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Ecological zonation, flavonoids, phenolic acids, phytoclimatic diversity, tannins","lastPublishedDoi":"10.21203/rs.3.rs-3308843/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3308843/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe phyto-climatic diversity in a region presents the life form of vegetation. Plants produce vast and diverse variety of phytochemicals to communicate and minimize the climatic threats. In the current study, the variations in the phytochemical attributes of four plant species (\u003cem\u003eNerium oleander, Ricinus communis, Calotropis procera \u003c/em\u003eand \u003cem\u003eWithania somnifera)\u003c/em\u003efrom three ecological zones of Pakistan (sandy desert, irrigated \u0026amp; rain-fed plains) were investigated. The phenolic acids, flavonoids and tannins were quantitatively analyzed by methanolic extraction and standardized methods. The results revealed that zone rainfed lands and zone sandy desert showed the maximum amount of phytochemicals in all species and also exhibited the extreme environmental conditions. The irrigated plains had less stressful environmental condition and maximum amount of soil organic matter that resulted in minimum amount of phytochemicals in plant species. Previous research also supports the evidence that environmental stresses like temperature stresses, water stresses, high light intensities increase, and organic matter hinders the accumulation of phytochemicals. This study proved that same species in different ecological zones show different amounts of phytochemicals due to environmental variations, although environmental stresses exert the effects on plants to produce more phytochemicals.\u003c/p\u003e","manuscriptTitle":"The Variations in the Phytochemical Attributes of Nerium Oleander, Ricinus Communis, Calotropis Procera and Withania Somnifera From Three Ecological Zones of Pakistan","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2024-03-25 19:19:10","doi":"10.21203/rs.3.rs-3308843/v2","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":1,"date":"2023-09-08 18:24:30","doi":"10.21203/rs.3.rs-3308843/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f35eaf9c-ebc6-4051-82d6-d581906237c1","owner":[],"postedDate":"March 25th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-09-08T18:24:32+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-25 19:19:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-3308843","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3308843","identity":"rs-3308843","version":["v2"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}