Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species

Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species

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CHNAES-00666; No of Pages 9 Acta Ecologica Sinica xxx (2019) xxx

Contents lists available at ScienceDirect

Acta Ecologica Sinica journal homepage: www.elsevier.com/locate/chnaes

Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species Abbas Tahan a, Mohammad Jafari b,⁎, Damoun Razmjoue c, Seyed akbar Javadi a a b c

Department of Range Management, Science and Research Branch, Islamic Azad University, Tehran, Iran Faculty of Natural Resources, University of Tehran, Iran Medicinal Plants Research Center, Yasuj University of Medical Sciences, Yasuj, Iran

a r t i c l e

i n f o

Article history: Received 9 December 2018 Received in revised form 3 June 2019 Accepted 5 August 2019 Available online xxxx Keywords: Ajuga chamaecistus Ging Phytochemistry Essential oil Ecological conditions PCA

a b s t r a c t Ajuga chamaecistus belongs to Lamiaceae plant family. Plant samples were collected from four habitats during vegetative and flowering periods for comparing the essential oil yield and composition. After drying, the essential oil was extracted by distillation method. Essential oil yields of plants in four habitats were calculated which the analysis and identification of essential oil composition was performed using GC and GC–MS. Ultimately, the PCA was used to determine the relationship between plant major components and environmental factors. The results showed that β-Pinene and Linalool compounds had the highest percentage of essential oil composition. Environmental factors including elevation, annual precipitation, mean annual temperature, annual evaporation, sand and clay content were identified as the most effective factors in the isolation of A. chamaecistus habitats. It appears that the difference between ecological and soil factors of four habitats can have a significant effect on the type and percentage of essential oil components. © 2019 Published by Elsevier B.V. on behalf of Ecological Society of China.

Contents 1. 2.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1. Specifications of the studied areas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2. Examining environmental factors of A. chamaecistus plant habitats . . . . . . . . . . . . . . . . . . . . . . . . 2.3. Study of phytochemical characteristics of A. chamaecistus . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.4. Process of calculating the quatz inhibition index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.5. Data analysis and examining the relationship between environmental factors and plant compounds . . . . . . . . . 3. Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1. Results of examining A. chamaecistus environmental factors in the studied habitats . . . . . . . . . . . . . . . . . 3.2. The results of examining the phytochemical characteristics of A. chamaecistus in studied habitats . . . . . . . . . . 3.3. Results of PCA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.4. The results of correlation coefficient between soil and climate variables with essential oil yield and major components 4. Discussion and conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1. Introduction Medicinal plants are the most important sub-products of rangelands. In general, medicinal plant is a plant that all or some of its components

⁎ Corresponding author. E-mail address: [email protected] (M. Jafari).

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are freshly dried or processed to detect, treat, prevent, assist physiological action and maintain human or animal health [31]. The diversity of climates in Iran has led to a great diversity of medicinal and essential herbs during so many years by adapting their biological conditions to ecological stresses showing large-scale adaptation. Thus, the vast array of natural resources of Iran, given the diversity of native medicinal plants, has brought about a suitable base for the production and

https://doi.org/10.1016/j.chnaes.2019.08.001 1872-2032/© 2019 Published by Elsevier B.V. on behalf of Ecological Society of China.

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

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A. Tahan et al. / Acta Ecologica Sinica xxx (2019) xxx

By examining the effect of soil pH on the distribution of plant species in Brazil, Viani et al. [35] concluded that in soils with high PH, this factor is important in the distribution of plant species. Pinke et al. [28] examined the effects of environmental factors on the composition of weed species in western Hungary and found that all variables studied in this study had a significant effect on the composition of species of weed plants. Geographical condition had the greatest effect on vegetation, and then soil pH, average annual precipitation, soil texture, average annual temperature and factor height were significant. The results of Angiolini et al. [2] in Italy showed that soil moisture in F.C., pH, CaCO3, NaCl,organic matter and soil moisture content were the most important parameters affecting the vegetation pattern. Moreover, Mirazadi et al. [23] studied the effect of ecological factors on the percentage of shrub essential oil yield in forest habitats of Lorestan reporting that the quantitative and qualitative differences between the essential oil of this region and other natural habitats of this shrub can be attributed to the difference in ecological characteristics such as moisture, elevation or other terrestrial and geographic factors. Pourfathi et al. [29] conducted a study in the north of Iran and found that soil pH, potassium, and elevation played a significant role in the distribution of Artemisia fragrans species. Modiri et al. [24] extracted and identified the chemical compounds in the essential oils of different species of Ziziphora Clinopodioide in different habitats in Iran, and found a significant difference between the essential oil yields of Fars and Garmabedred specimens. According to the studies done on the internal and external references, carried out by the authors so far, no studies have been conducted on the effect of environmental conditions on the phytochemical properties of Ajuga chamaecistus Ging. Hence, the purpose of the study was to examine the effect of environmental factors affecting the phytochemical properties of A. chamaecistus in Fars to identify and introduce the superior habitats of this plant besides introducing the major components of essential oil in its separate habitats.

Table 1 Specifications of studied areas. Sites studied variable

Eqlid

Abade

Bavanat

Pasargad

Longitude

E 52° 41′ 9.27″ 30° 54′ 21.79″ N 12.9 315 45% 1500

52° 39′ 2.16″ E 31° 9′ 38.88″ N 14.3 124 30% 1700

53° 39′ 1.73″ E 30° 27′ 14.9″ N 14.8 256 40% 2330

53° 0′ 0″ E 30° 0′ 0″ N 15.6 352 71% 1800

45

12

28

35

2300

1800

2530

2100

latitude Average temperature Average precipitation Average relative humidity Anuual average evaporation The average number of frosty days Elevation above sea level

processing of plants. Vegetative and yield of plants in ecosystems are affected by various factors like species, climate, soil environment, elevation and geographical location. Each of these factors has ablilty to affect a significant change on the quantity and quality of products [33]. Some of the researchers believe that genotypes have a drastic effect on the variety of chemical compounds of medicinal plants [36]. As environmental factors can affect the essential oils of medicinal plants, this study examined the effect of some environmental factors (climate, soil and elevation) on the quantity and quality of essential oils of Ajuga chamaecistus. The Ajuga chamaecistus Ging. is a species of the Lamiaceae. the family of Lamiaceae has about 300 species distributed in Asia, Africa, Australia, Europe and North America [4,12]. This genus has six species in Iran. Some Ajuga species have uses in traditional medicine for treating pain, diabetes, inflammation, hypertension, toothache and fever. Moreover, it is an antifungal, antimicrobial, anesthetic and good for urinary tract enhancement [7,9]. Several studies have been conducted on the mentioned species in their habitats inside and outside Iran regarding phytochemical properties and to understand the effect of environmental factors on the percentage of essential oils and plant compounds. Majority of current studies show the presence of various chemical compounds such as βPinene, Linalool, Bicyclogermacrene, pHytol, α-Pinene, α-Thujen, Caryophyllene oxide, Germacrene-D, 1-Octen-3-ol, α-Bergamotene, δCadinene and α-Copaene and various amounts of other compounds in the mentioned plant [3,8,25].

2. Materials and methods 2.1. Specifications of the studied areas The studied areas were Eghlid, Abadeh, Bavanat and Pasargad, all of which are located in Fars province, Iran. The ecological characteristics of the studied areas are presented in Table 1.

Table 2 Physicochemical properties of soil of studied areas. Soil properties

Depth of soil

EC

PH

OC

N

P

K

Clay

Silt

Sand

Eqlid

0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50 0–10 10–50

0.95 ± 0.01 0.45 ± 0.01 1.28 ± 0.02 0.69 ± 0.01 1.05 ± 0.03 0.49 ± 0.02 0.61 ± 0.01 0.89 ± 0.04 0.65 ± 0.00 0.6 ± 0.01 0.67 ± 0.01 0.75 ± 0.00 1.6 ± 0.01 0.5 ± 0.04 0.93 ± 0.00 0.45 ± 0.02 0.99 ± 0.02 0.53 ± 0.01 1 ± 0.05 0.5 ± 0.01 0.9 ± 0.02 0.4 ± 0.01 0.97 ± 0.02 0.6 ± 0.00

7.52 ± 0.04 7.84 ± 0.07 7.72 ± 0.01 8.08 ± 0.02 7.61 ± 0.01 7.91 ± 0.04 7.8 ± 0.03 8.15 ± 0.08 8.03 ± 0.05 8.02 ± 0.07 8 ± 0.03 7.9 ± 0.04 7.5 ± 0.06 7.2 ± 0.07 7 ± 0.04 7.13 ± 0.02 7.2 ± 0.07 7.16 ± 0.03 7.6 ± 0.02 8.2 ± 0.04 8.5 ± 0.03 8.2 ± 0.06 7.8 ± 0.09 8.1 ± 0.05

3.23 ± 0.08 1.14 ± 0.04 0.85 ± 0.01 0.66 ± 0.01 2.11 ± 0.05 0.91 ± 0.04 1.2 ± 0.08 0.39 ± 0.00 0.9 ± 0.01 0.62 ± 0.00 1.7 ± 0.07 0.52 ± 0.00 1.12 ± 0.08 0.59 ± 0.02 3.3 ± 0.01 1.17 ± 0.04 2.9 ± 0.05 0.68 ± 0.01 0.85 ± 0.01 0.65 ± 0.02 1.5 ± 0.00 0.66 ± 0.01 0.9 ± 0.01 0.52 ± 0.00

0.31 ± 0.001 0.1 ± 0.001 0.07 ± 0.003 0.05 ± 0.000 0.25 ± 0.000 0.19 ± 0.004 0.13 ± 0.000 0.03 ± 0.001 0.08 ± 0.001 0.05 ± 0.000 0.17 ± 0.001 0.06 ± 0.000 0.1 ± 0.002 0.04 ± 0.000 0.31 ± 0.002 0.1 ± 0.000 0.22 ± 0.004 0.07 ± 0.002 0.04 ± 0.000 0.32 ± 0.002 0.06 ± 0.000 0.1 ± 0.000 0.06 ± 0.000 0.32 ± 0.001

12 ± 0.20 3.5 ± 0.05 4 ± 0.08 2.5 ± 0.04 9 ± 0.09 2.1 ± 0.02 7.5 ± 0.09 2 ± 0.03 2.5 ± 0.05 1.5 ± 0.02 6 ± 0.06 5 ± 0.04 14.5 ± 0.60 12.5 ± 0.43 12 ± 0.49 4 ± 0.01 11 ± 0.34 7 ± 0.60 12.5 ± 0.87 10.5 ± 0.56 11 ± 0.78 2.5 ± 0.05 11.9 ± 0.79 6.5 ± 0.24

208 ± 12.01 208 ± 11.43 96 ± 5.97 42 ± 3.68 101 ± 9.69 150 ± 8.37 83 ± 6.59 110 ± 5.56 40 ± 2.45 117 ± 12.92 70 ± 3.78 100 ± 4.62 210 ± 12.78 180 ± 6.67 220 ± 14.96 205 ± 16.84 201 ± 14.08 197 ± 9.91 200 ± 14.34 160 ± 7.54 205 ± 9.23 208 ± 12.56 207 ± 13.05 175 ± 8.21

24 ± 1.24 32 ± 1.43 30 ± 2.05 26 ± 1.49 27 ± 1.78 29 ± 1.98 19 ± 1.19 14 ± 1.05 30 ± 1.48 25 ± 2.07 22 ± 1.76 17 ± 1.48 19 ± 1.86 25 ± 1.35 24 ± 1.05 32 ± 2.09 18 ± 1.04 28 ± 1.63 28 ± 1.95 32 ± 1.76 18 ± 1.01 16 ± 1.06 20 ± 1.57 27 ± 1.79

32 ± 2.47 40 ± 2.56 42 ± 2.78 42 ± 1.39 39 ± 1.87 40 ± 2.71 40 ± 2.24 38 ± 1.97 45 ± 2.76 35 ± 1.78 41 ± 2.58 34 ± 1.98 32 ± 2.08 34 ± 1.12 33 ± 2.56 41 ± 2.54 33 ± 1.65 37 ± 2.92 48 ± 2.45 37 ± 2.24 43 ± 2.54 36 ± 1.52 45 ± 2.54 36 ± 2.58

44 ± 2.57 28 ± 1.75 28 ± 1.32 32 ± 2.45 39 ± 2.09 30 ± 3.12 50 ± 3.76 45 ± 2.97 55 ± 3.78 30 ± 2.43 51 ± 2.96 37 ± 2.70 72 ± 3.46 43 ± 2.28 45 ± 2.85 26 ± 1.47 55 ± 2.76 32 ± 2.07 29 ± 1.83 46 ± 2.06 45 ± 2.41 52 ± 2.94 30 ± 2.30 49 ± 2.61

Abade

Bavanat

Pasargad

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

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Table 3 Essential oil content in the essential oil of A. chamaecistus in the studied regions at two vegetative and flowering stages. Sites studied

Growth stage

Essential oil percentage

Flowering stage

Abade

Eqlid

Pasargad

Bavanat

Abade

Eqlid

Pasargad

Bavanat

0.94

1.18

0.86

0.8

1.74

1.65

1.24

1.17

2.2. Examining environmental factors of A. chamaecistus plant habitats In the studied habitats, the elevation was measured using the VISTA CX global Pposiotion System (GPS). Climate data was collected from the nearest, the most aligned and the nearst weather stations. Characteristics of these habitats are seen in Table 1. The soil samples of two depths (0−10) and (10–50) were collected and some soil parametres were measured. Some physical and chemical properties of soil were obtained in the following methods. Soil texture was measured using hydrometric method [5], soil acidity by potentiometric method, electrical conductivity (EC) using sonar and via saturation extraction method [22], soil nitrogen was by Kjeldahl method in three stages of digestion, distillation and tetrazation, P [27] and potassium (K) by ammonium acetate extraction method was measured at pH 7.

of 250 μm and a static phase layer thickness of 0.25 μm. The oven temperature increased from 60 °C to 230 °C at 3 °C/min rate. 2.4. Process of calculating the quatz inhibition index The inhibition index of each compound can be measured from the chromatogram of a mixture of the object with at least two normal alkanes, the time of their inhibition on both sides of the desired inhibition time. Normal alkanes are the standards for which the quatz index is calibrated and, according to the definition of the quatz index, a normal alkan is 100 times the number of carbon in it. The quatz index is independent of the temperature and dimensions of the GC column. The quatz Inhibition Indicator is measured from the following equation:  KI ¼ 100 n þ 100

t ðxÞ −t ðnÞ t ðnþ1Þ −t ðnÞ



2.3. Study of phytochemical characteristics of A. chamaecistus One hundred gram of crushed plant with 250 ml of distilled water was poured into celvenger balloon. After boiling, the temperature was kept constant at 80 °C and distillation was done for one and half hour. Essential oil was extracted by Clevenger apparatus (extracting the essential oil starts from the fall of the first drop). The essential oil was washed with sodium sulfate and the pure essential oil was weighed and kept in a glass container at 4 °C in the refrigerator until it was injected into GC and GC–MS devices [1]. GC and GC–MS devices were used to detect essential oils. Identification of the spectra was done by calculating the retention index (RI) and the mass spectra. The relative percentages of each of the composites were obtained according to the surface of the curve under its peak in the chromatogram spectrum [6,30]. We used 6890 N Agilent technologies gas chromatograph, equipped with a DB0–5 column with a length of 30 m, a 250 μm internal diameter, and a static phase layer thickness of 25.0 μm. We also used GC–MS 7890A gas chromatograph attached to a mass spectrometer equipped with a HB-5MS column of 30 m in length, an internal diameter

Table 4 The percentage of essential oil composition in the four studied habitats in the vegetative stage.

We have the following: t(x): Inactive time of sampling. t(n): The previous normal alkane inhibition time. t(n+1): The next normal alkane inhibition time. KI: Quatz Inhibition Index. n: The number of previous normal carbon alkanes. 2.5. Data analysis and examining the relationship between environmental factors and plant compounds Following the necessary measurements in each area, all variables were entered into Excel software version 2010 in a worksheet and the results obtained from the average percentage of essential oil of A. chamaecistus were drawn in a chart. As the studied areas and measurement criteria were different, the results were analyzed using SPSS software in a completely randomized block design. Ultimately, Pearson correlation coefficient was used to determine the relationship between the traits and ecological factors. Finally, various habitats were compared in terms of plant composition and the most appropriate ecological Table 5 The composition of the oils in the four habitats studied in the flowering stage.

No

Compounds

Retention index

Abade

Eqlid

Bavanat

Pasargad

No

Compounds

Retention index

Abade

Eqlid

Bavanat

Pasargad

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

α-Thujen α-Pinene Sabinene β-Pinene 1-Octen-3-ol Myrcene α-Phellandrene Limonene Linalool α-Terpineol Eugenol α-Copaene α-Bergamotene (Z)- β-Farnesene Germacrene-D Bicyclogermacrene δ-Cadinene Caryophyllene oxide Spathulenol Phytol

932 936 965 972 975 980 995 1020 1090 1170 1335 1374 1430 1442 1480 1492 1512 1517 1585 1935

2.45 3.72 – 11.17 2.73 0.24 – 1.15 9.65 – 0.98 5.73 5.84 2.01 6.47 4.96 2.74 3.14 1.85 3.72

3.15 2.94 0.29 12.17 3 0.76 0.08 1.19 10.17 1.06 0.36 4.16 6.92 2.92 2.89 5.07 3.84 2.98 2.03 3.58

3.73 4.16 – 9.96 3.92 1.09 – 0.89 11.03 – – 4.90 6.36 1.99 3.76 5.92 3 3.66 1.83 4.01

3.13 4.01 – 10.21 3.26 – – 0.89 11.06 0.45 – 4.71 5.12 1.12 3.15 5.06 3.85 4.01 1.31 5.45

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

α-Thujen α-Pinene Sabinene β-Pinene 1-Octen-3-ol Myrcene α-Phellandrene Limonene Linalool α-Terpineol Eugenol α-Copaene α-Bergamotene (Z)- β-Farnesene Germacrene-D Bicyclogermacrene δ-Cadinene Caryophyllene oxide Spathulenol Phytol

932 936 965 972 975 980 995 1020 1090 1170 1335 1374 1430 1442 1480 1492 1512 1517 1585 1935

4.73 5.06 1.12 16.23 3.51 2.04 0.45 2.07 13.14 1.43 1.16 3.02 3.21 2.11 3.96 6.76 3.51 4.28 2.92 5.29

4.16 4.99 1.32 16.14 3.02 2.11 0.32 2.84 14.26 1.73 1 3.69 6.02 3.03 4.02 6.19 4.28 3.46 2.16 4.98

5.64 4.27 1.07 13.84 4.51 2.74 – 2.42 15.17 1.09 – 3.85 4.05 2.02 4.12 5.43 3.07 4.09 2.16 6.18

3.39 4.83 – 17.37 3.62 2.18 – 2 14.02 1.08 – 3.13 3.05 2.06 3.81 5.94 4.2 4.51 2.15 6.73

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

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Table 6 Table of coefficients of main components in all variables (major components and percentage yield of essential oil of vegetative stage, soil and climate). Variable

PC1

PC2

PC3

PC4

PC5

PC6

PC7

ESANS Phytol Caryophyllene oxide δ-Cadinene Bicyclogermacrene Germacrene-D α-Bergamotene α-Copaene Linalool 1-Octen-3-ol β-Pinene α-Pinene α-Thujen EC PH OC N1 P K Clay Silt Sand TEMP PRE EVA DAYS H1

−0.135 0.049 0.168 0.05 0.254 −0.189 0.038 −0.132 0.215 0.285 −0.178 0.133 0.288 0.2 −0.217 0.208 0.248 0.278 0.273 0.105 −0.195 0.061 −0.094 −0.166 −0.223 −0.126 0.266

−0.303 −0.174 0.174 −0.238 0.076 0.217 −0.235 0.291 0.055 0.065 −0.262 0.303 −0.058 −0.174 −0.003 −0.189 −0.162 0.063 0.016 −0.317 −0.038 0.336 −0.198 −0.211 0.084 −0.104 −0.12

0.046 −0.322 −0.245 −0.268 0.168 0.163 0.275 0.109 −0.25 0.042 0.088 −0.059 0.025 0.201 −0.255 0.168 −0.095 −0.093 −0.136 0.062 −0.28 0.034 0.287 −0.21 0.228 −0.324 0.085

0.005 −0.143 −0.11 −0.256 −0.081 0.004 0.071 0.032 −0.069 −0.048 −0.081 0.029 −0.076 0.277 −0.07 −0.084 0.054 −0.165 0.382 −0.096 −0.071 0.078 0.033 −0.052 0.035 0.732 0.208

−0.293 −0.142 0.105 0.204 0.056 −0.062 −0.065 −0.108 0.097 0.045 0.203 0.04 −0.035 0.02 −0.124 0.151 0.131 0.058 0.023 −0.123 −0.056 0.33 0.222 0.57 0.446 0.055 −0.017

0.181 0.293 0.452 −0.219 −0.056 −0.115 0.02 −0.099 −0.014 0.154 0.233 0.068 0.098 −0.031 0.139 0.243 −0.273 0.073 0.104 −0.225 0.045 −0.166 0.153 −0.264 0.403 0.066 0.013

0.061 0.052 0.185 0.305 −0.015 0.022 0.364 0.561 0.023 0.107 −0.192 −0.126 0.014 −0.018 0.055 0.232 0.117 −0.178 0.15 0.25 0.021 0.101 −0.121 −0.07 0.192 0.089 −0.301

Bold indicates variables having the highest coefficients for each component.

conditions were determined in terms of the variables examined. Indirect gradient method of PCA method was used to determine the relationship between species and environmental factors, and the Euclidean index was used to determine the similarity between the regions according to different factors and using the Wards method. The cluster analysis (dendrogram) obtained was interpreted [15]. 3. Results 3.1. Results of examining A. chamaecistus environmental factors in the studied habitats According to Table 1, it is shown that A. chamaecistus species extends at an elevation of 1800–2500 m in the studied regions. The lowest mean temperature is in Eghlid (12.9 °C) and the highest is related to Pasargad (15.6 °C). The average annual precipitation in the habitats of the plant is about 125–135 mm and the relative humidity of the studied areas is about 30–71%. The average annual evaporation potential in the studied regions is between 1500 and 2330 mm per year. Moreover, the minimum and maximum number of ice days is, respectively, in Abadeh (12) and Eghlid (45). Table 2 shows the physicochemical properties of the soil of the studied areas. In each of the parameters, the minimum and maximum values are specified as bold. 3.2. The results of examining the phytochemical characteristics of A. chamaecistus in studied habitats Table 3 shows the yield of A. chamaecistus essential oil in the studied habitats in two stages of vegetative and flowering. In the vegetative stage, the cities of Eghlid, Abadeh, Pasargadae and Bavanat had the highest percentage of essential oil yield. In the flowering stage, Abadeh, Eghlid, Pasargad and Bavanat, respectively, had the highest percentage of essential oil yield. This plant has had the greatest yield at flowering stage and in Abadeh (1.74), and the highest percentage of essential oil yield. Moreover, the lowest percentage of return essential oil yield in vegetative stage was in Bavanat (0.8).

According to Tables 4 and 5, we see that the major components of A. chamaecistus essential oil in Abadeh, Eghlid, Pasargad and Bavanat in vegetative and flowering stage are β-Pinene, Linalool, Bicyclogermacrene, Phytol, α-Pinene, α-Thujen, Caryophyllene oxide, Germacrene-D, 1-Octen-3-ol, α-Bergamotene, δ-Cadinene and α-Copaene.

3.3. Results of PCA Taking into account all the variables (major components and percentage of yield of essential oil of the vegetative stage, soil and climate), we see that the first three components cover almost all the changes. The first component manages 43.3% and the second 30.3% of the changes. In the first component, 1-Octen-3-ol and α-Thujen have the highest coefficients in the second component of clay and sand, and in the third component Phytol, the most coefficients (Table 6). Cluster analysis also shows two distinct groups, one including Eghlid, Bavanat and Pasargad and the second group Abadeh. In the first group, Eghlid and Bavanat have 87.5% similarity and Pasargad is similar by 6.6% to these two areas. Abadeh is not similar to these three areas (Fig. 1). Moreover, by examining all variables (major components and percentage of yield of essential oil of flowering stage, soil and climate), the main components of the first component are analyzed with 50.6% and the second component is 28.3% of the variation. The pH and Linalool had the highest coefficients in the first component; Phytol had the highest coefficient in the second component and evaporation of the highest coefficient in the third component. Cluster analysis also shows two distinct groups, one of which includes Eghlid, Bavanat and Pasargad. The highest similarity was for Eghlid and Bavanat with 87.5% similarity. Pasargad is 76.6% similar to these two areas and Abadeh is not similar to any of the cities in terms of the variables mentioned (Table 7). By all the classifications and analyses of the main components, we find that β-Pinene, Linalool and Phytol in the variable of the major components of the essential oil and the percentage of gravel and evaporation in the soil and elevation are the most important variables in the classification of areas (Fig. 2).

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

A. Tahan et al. / Acta Ecologica Sinica xxx (2019) xxx

5

Score Plot 3 A BA DH

2 BA VA NT PA SA R

Second Component

1 0 -1 -2 -3 -4

EGHLID

-5 -4

-3

-2

-1

0

1

2

3

4

5

First Component

A

Loading Plot 0

0.4

Sand α-Pinene

α-Copaene

0.3

Second Component

Germacrene-D

0.2

Caryophyllene oxide Bicyclogermacrene

0.1 0.0

EVA PH

α-Thujen

Silt

DA YS

-0.1 -0.2 -0.3

P 1-Octen-3-ol K

Linalool

PRE

0

H1 TEMP

Phytol α-Bergamotene

β-Pinene ESANS

N1

EC OC

δ-Cadinene Clay

-0.4 -0.2

B

-0.1

0.0 0.1 First Component

0.2

0.3

Dendrogram with Ward Linkage (Sim=60%)

Similarity

-10.56

26.29

63.15

100.00 ABADH

C

EGHLID BAVANT Observations

PASAR

Fig. 1. Ranking of sites in all variables (A), Loaded chart in all variables (B), Dendrograms for determining the similarity between all variables (C), major components and yield percent of vegetative stage, soil and climate).

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

6

A. Tahan et al. / Acta Ecologica Sinica xxx (2019) xxx

Table 7 Table of coefficients of main components in all variables (major components and percentage yield of essential oil of flowering stage, soil and climate). Variable

PC1

PC2

PC3

PC4

PC5

PC6

PC7

ESANS Phytol Caryophyllene oxide δ-Cadinene Bicyclogermacrene Germacrene-D α-Bergamotene α-Copaene Linalool 1-Octen-3-ol β-Pinene α-Pinene α-Thujen EC PH OC N1 P K Clay Silt Sand TEMP PRE EVA DAYS H1

−0.176 0.043 −0.066 −0.157 −0.237 0.206 0.073 0.238 0.257 0.202 −0.24 −0.252 0.188 0.231 −0.26 0.228 0.18 0.202 0.185 0.111 −0.249 0.056 0.013 −0.2 −0.116 −0.203 0.25

0.21 −0.351 −0.35 0.147 0.077 0.141 0.344 0.168 0.019 −0.229 −0.015 0.13 −0.006 0.187 −0.033 0.188 0.06 −0.161 −0.129 0.298 −0.014 −0.323 0.312 0.169 0.054 0.017 0.079

−0.206 0.073 0.006 0.296 −0.182 −0.218 0.061 0.037 0.129 −0.086 0.194 −0.013 −0.302 0.019 0.111 0.06 0.306 0.208 0.267 0.164 0.164 −0.167 −0.212 0.204 −0.373 0.276 0.133

−0.125 0.081 −0.035 0.012 −0.253 0.067 0.021 −0.134 0.1 −0.211 0.086 0.048 −0.103 −0.157 −0.175 −0.023 0.072 −0.048 −0.026 0.127 0.14 −0.099 −0.181 0.237 0.086 −0.735 −0.263

−0.001 0.199 −0.307 −0.174 0.14 −0.217 −0.114 0.149 −0.057 0.063 −0.054 −0.152 0.152 −0.075 −0.063 0.043 −0.07 0.117 0.009 0.018 −0.13 −0.018 −0.229 0.636 0.349 0.096 0.207

0.022 −0.158 −0.12 0.122 0.238 0.062 0.148 −0.145 0.013 0.228 −0.12 0.086 0.151 −0.029 −0.096 0.04 −0.008 −0.049 −0.083 −0.008 −0.098 0.453 −0.071 0.359 −0.574 −0.06 −0.207

−0.2 −0.097 0.272 −0.338 0.16 0.036 0.379 0.066 −0.017 −0.148 −0.284 0.126 −0.17 −0.337 0.084 0.066 −0.036 −0.061 0.162 −0.205 0.212 0.042 0.11 0.134 −0.054 −0.095 0.391

Bold indicates variables having the highest coefficients for each component.

3.4. The results of correlation coefficient between soil and climate variables with essential oil yield and major components Table 8 shows correlation between soil and climate variables with essential oil percentage and major components (vegetative stage) indicates that Caryophyllene oxide parameters have a negative significant relationship with temperature at 5% level. Germacrene-D has a significant negative relationship with total nitrogen at 5% level and αThujen has a significant positive relationship at the 5% level with sea level elevation. According to Table 9 (correlation between soil and climate variables with essential oil percentage and major components of flowering stage), we see that the percentage of essential oil with phosphorus has a negative significant relationship at 5% level. δ-Cadinene has a significant positive correlation with percipitation at 5% level. Bicyclogermacrene has a significant negative correlation at level of 5% with potassium and phosphorus. α-Bergamotene has a significant positive correlation of 5% with clay. α-Copaene has a significant positive correlation with OC at the level of 1%. β-Pinene has a positive relationship with pH at the level of 5% and frost days with silt percentage at the level of 1%. Linalool has a significant positive relationship with elevation at the level of 5%. α-Thujen has a negative significant relationship with frosty days at the level of 1%.

4. Discussion and conclusions The study of climatic characteristics indicated that the process of producing effective materials in the studied plant does not have the same trend in the four habitats. Essential oil yields in this study showed that the percentage of essential oil yields differed from 0.8% (Bavanat) to 1.74% (Abadeh). Given the results of phytochemistry studies in the four study areas, Abadeh and Eghlid with 20 chemical compositions the most compoition, whereas Bavanat with 18 compounds and Pasargad with 17 compounds ranked the next in terms of the number of plant compounds. Some studies have been conducted on the percentage of the essential oil yield of A. chamaecistus and other species of this genus, of which the following can be mentioned. Ardekani et al. (2013) studied Ajuga chamaecistus Ging. subsp. tomentella rech. f collected

from Khojir National Park and stated that its essential oil percentage was 0.3% and the number of compounds was 43 compounds. Javidnia et al. [13] stated the essential oil yield of Ajuga austro-iranica Rech as 0.01%. The results of the table of the chemical composition of Ajuga chamaecistus resulted in the identification of 12 major combinations in two stages of vegetative and flowering including β-Pinene, Linalool, Bicyclogermacrene, Phytol, α-Pinene, α-Thujen, Caryophyllene oxide, Germacrene-D, 1-Octen-3-ol, α-Bergamotene, δ-Cadinene and αCopaene. β-Pinene and Linalool were also identified as the two major compounds with the highest levels. Here, a comparison was required between the chemical compounds identified in the felted lobster plant in the recent study with some of the phytochemical studies carried out on the plant that is present in other areas. Mazloomifar et al. [31] examined the major components of Ajuga chamaecistus Ging. Α-Thujen, αPinene, β-Pinene, Linalool, α-Copaene, α-Bergamotene, Germacrene-D, Bicyclogermacrene, δ-Cadinene and Phytol. The main components of this plant were β-Pinene and Linalool. Venditti et al. [34] introduced the major components of the Ajuga chamaepitys (L.) plant β-Pinene, 1Octen-3-ol, Linalool, and Germacrene-D. Karami [16] studied the composition of essential oil of Ajuga comata Stapf collected from southern Zagros, Iran and found that the major components of this plant were Caryophyllene oxide, Germacrene-D and δ-Cadinene. The results of the effects of soil chemical and physical factors on the studied areas showed that when soil variables were analyzed and evaluated, the chemical factors such as OC, EC, PH and potassium content were significantly different from the factors of climate, major components and essential oil yield. Then, it was found that the effects of physical factors on soil, the effect of factors such as sand and clay was higher than the rest. With these interpretations, one can state that the role of soil on secondary metabolites of medicinal plants is unavoidable. In this regard, to prove this, the following studies can be made Yan et al. [37] conducted a study and indicated that low pH and high exchangeable nitrogen were the most important soil factors affecting the essential oil quality in Changhui Mountains of China. The results of a study on Achillea ptarmica show a significant relationship between the amount of essential oil and soil and biochemical factors [10]. In some studies on Stachys, it was found that different climatic and soil factors

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

A. Tahan et al. / Acta Ecologica Sinica xxx (2019) xxx

7

Score Plot EGHLID

4

Second Component

3 2 1 0 ABADH BAVA NT

-1 -2

PASAR

-3 -3

-2

-1

0

A

1 2 First Component

3

4

5

6

Loading Plot 0.4

α-Bergamotene TEMP Clay

0.3 ESANS PRE δ-Cadinene

Second Component

0.2 α-Pinene

0.1

Bicyclogermacrene

ECOC α-Copaene Germacrene-D

α-Thujen

Silt PH β-Pinene

0.0

H1

N1

EVA

DAYS

-0.1

Linalool

K P

-0.2

1-Octen-3-ol

-0.3

Caryophyllene oxide

Sand Phytol

-0.4 -0.3

-0.2

-0.1

0.0 First Component

0.1

0.2

0.3

B Dendrogram with Ward Linkage (Sim=60%)

Similarity

-10.56

26.30

63.15

100.00 ABADH

C

EGHLID BAVANT Observations

PASAR

Fig. 2. Ranking of sites in all variables (A), Loading graph in all variables (B), Dendrograms for determining the similarity between all variables (C), (major components and percentage yield of essential oil of flowering stage, soil and climate).

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

8

A. Tahan et al. / Acta Ecologica Sinica xxx (2019) xxx

Table 8 Correlation table between soil and climate variables with percentage of essential oil yield and major components (vegetative stage).

ESANS Phytol Caryophyllene oxide δ-Cadinene Bicyclogermacrene Germacrene-D α-Bergamotene α-Copaene Linalool 1-Octen-3-ol β-Pinene α-Pinene α-Thujen

EC

PH

OC

N1

P

K

Clay

Silt

Sand

TEMP

PRE

HUM

EVA

DAYS

H1

0.19 −0.08 −0.20 0.10 0.72 −0.54 0.81 −0.59 0.08 0.63 0.09 −0.21 0.79

0.27 0.45 0.00 0.35 −0.94 0.19 −0.57 0.14 −0.11 −0.80 0.30 −0.24 −0.77

0.21 0.02 −0.15 0.19 0.69 −0.62 0.78 −0.66 0.15 0.64 0.08 −0.23 0.82

−0.01 0.59 0.41 0.64 0.52 −0.95⁎ 0.25 −0.85 0.71 0.71 −0.22 0.01 0.90

−0.63 0.28 0.80 0.21 0.76 −0.61 −0.18 −0.35 0.89 0.94 −0.77 0.63 0.89

−0.52 0.44 0.79 0.38 0.66 −0.73 −0.17 −0.49 0.93 0.88 −0.69 0.52 0.89

0.66 0.39 −0.36 0.58 0.18 −0.74 0.79 −0.89 0.01 0.19 0.52 −0.67 0.52

0.32 0.57 0.03 0.48 −0.93 0.05 −0.54 0.00 −0.03 −0.75 0.32 −0.29 −0.69

−0.94 −0.53 0.55 −0.70 0.44 0.52 −0.57 0.75 0.25 0.40 −0.85 0.94 0.05

0.74 −0.40 −0.96⁎ −0.19 −0.07 0.17 0.89 −0.12 −0.82 −0.34 0.81 −0.77 −0.17

0.73 0.68 −0.28 0.71 −0.87 −0.25 −0.05 −0.42 −0.15 −0.73 0.68 −0.71 −0.49

0.95 0.55 −0.56 0.70 −0.57 −0.44 0.47 −0.68 −0.28 −0.52 0.87 −0.94 −0.18

0.21 −0.76 −0.70 −0.72 −0.34 0.90 0.17 0.72 −0.92 −0.63 0.42 −0.22 −0.75

0.36 0.80 0.15 0.73 −0.82 −0.28 −0.46 −0.30 0.19 −0.57 0.30 −0.33 −0.43

−0.09 0.14 0.20 0.24 0.81 −0.71 0.52 −0.64 0.47 0.85 −0.24 0.07 0.96⁎

⁎ Significance at a level of 5%.

play an important role in the quality and quantity of essential oils [19,20]. Except for the soil that showed a significant effect on the quantity and quality of the chemical compounds of plants, some climate and physiographic variables also showed their effect on the effects of elevation factors such as elevation, percipitation, temperature and evaporation. The results of the following studies also confirm this: In studying, Chaharmahal and Bakhtiari and south of Isfahan, Farhang et al. [38] concluded that environmental factors such as annual temperature, annual freezing days and annual evaporation were the most effective factors in the isolation of field grassland habitats. The results of the present study are similar to Soleimani (2011) [32] that investigated the relationship between phytochemicals of mountain tea and environmental factors in western region of Isfahan regarding the effect of environmental factors on the production of A. chamaecistus material. This is because of its many effects on the environmental factors evaluated in this study on the production of active substances in the current study having also been proven. Among these similar environmental factors, one could mention the elevation and the average annual precipitation. In the study of Jaymand and Rezaei [14], it was also stated that the elevation of the vegetative area could cause changes in the physical condition of the plant and the amount of effective substances. The results of the correlation coefficient between the studied variables showed a direct correlation between the chemical composition of the plant and elevation, as well as the inverse relationship between the compounds with temperature and freezing days. There was a significant relationship between the percentages of essential oil yield in different habitats with phosphorus. There was a significant positive correlation between some components of essential oil with precipitation, clay content, OC content, pH, and elevation, as well as some compounds with negative potassium, phosphorus and ice days. The results

are in line with the results of Mirazadi et al. [23], Najafpour Navaei and Mirza [26] which show a significant relationship between elevation as one of the physiographic components and essential oil content. In a study of three different elevations, Mahzooni Kachpi et al. [18] found a significant correlation between some components of Salvia verticillata and potassium, phosphorus, acidity and organic carbon content of soil at 5% level. Hossein zadegan and Bakhshi [11] examined the impact of some ecological factors on essential oil of Teucrium and reported a significant correlation between the height of the sea surface, temperature, rainfall and electrical conductivity with some compounds of Teucrium polium L. plant. The results of this study and the researchers mentioned confirmed that the yield of the plants in ecosystems is affected by various factors such as type of species, climate of the region, soil type, elevation, and geographic location, each of which could have a significant effect on quantity and quality of the plants. 5. Conclusion By determining the effect of environmental factors on the distribution of vegetation, the production capacity can be obtained in similar ecological conditions. Knowledge of soil properties of any plant has an effective role in suggesting species compatible with soil conditions in similar areas, because soil is actually an important factor of determining the type of vegetation within a climate. Therefore, the results of this research can be used to improve the vegetation cover of areas with similar conditions. According to the above mentioned, it can be stated that A. chamaecistus species is a valuable medicinal plant, which is necessary for cultivation and improvement. Also, the presence of this plant in other similar areas can indicate the suitable places for its cultivation. Meteorological and climatic data can also help identify areas susceptible

Table 9 Correlation table between soil and climate variables with percentage of essential oil yield and major components (flowering stage).

ESANS Phytol Caryophyllene oxide δ-Cadinene Bicyclogermacrene Germacrene-D α-Bergamotene α-Copaene Linalool 1-Octen-3-ol β-Pinene α-Pinene α-Thujen

EC

PH

OC

N1

P

K

Clay

Silt

Sand

TEMP

PRE

HUM

EVA

DAYS

H1

−0.28 −0.36 −0.71 −0.25 −0.66 0.83 0.73 1.00 0.85 0.30 −0.76 −0.61 0.55

0.44 −0.02 0.33 0.71 0.70 −0.90 −0.31 −0.87 −0.84 −0.71 0.97⁎ 0.85 −0.85

−0.32 −0.34 −0.71 −0.17 −0.69 0.77 0.74 0.99⁎⁎

−0.69 0.07 −0.31 0.20 −0.86 0.19 0.44 0.73 0.86 0.24 −0.26 −0.58 −0.07

−0.98⁎ 0.64 0.25 −0.26 −0.96⁎ 0.14 −0.15 0.49 0.84 0.74 −0.41 −0.87 0.17

−0.96⁎ 0.57 0.19 −0.09 −0.95⁎ 0.05 −0.06 0.49 0.83 0.61 −0.30 −0.79 0.02

0.02 −0.67 −0.89 0.37 −0.35 0.43 0.95⁎

0.38 −0.04 0.27 0.79 0.63 −0.92 −0.23 −0.79 −0.76 −0.74 0.99⁎⁎ 0.83 −0.92

−0.46 0.83 0.81 −0.77 −0.20 0.02 −0.85 −0.27 0.03 0.80 −0.33 −0.50 0.45

0.72 −0.92 −0.85 −0.03 0.36 0.64 0.76 0.40 −0.06 −0.40 −0.31 0.28 0.38

0.51 −0.49 −0.26 0.96⁎ 0.53 −0.63 0.32 −0.39 −0.53 −0.95 0.86 0.84 −0.87

0.53 −0.81 −0.73 0.83 0.32 −0.14 0.77 0.12 −0.17 −0.88 0.46 0.62 −0.54

0.80 −0.37 −0.05 −0.32 0.80 0.19 −0.10 −0.39 −0.67 −0.23 −0.04 0.48 0.34

0.19 −0.05 0.15 0.92 0.38 −0.89 −0.06 −0.58 −0.51 −0.73 0.96⁎ 0.70 −0.99⁎⁎

−0.63 −0.01 −0.43 −0.22 −0.90 0.62 0.50 0.94 0.98⁎

0.87 0.27 −0.70 −0.60 0.47

0.78 0.55 −0.29 −0.22 −0.10 −0.01

0.49 −0.68 −0.79 0.41

⁎ Significance at a level of 5%. ⁎⁎ Significant at 1% level.

Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001

A. Tahan et al. / Acta Ecologica Sinica xxx (2019) xxx

to planting. Differences in the type and percentage of essential oil components can be due to genetic or non-genetic changes in response to environmental differences in the ecosystem of habitats, such as soil chemical composition and physiographic factors. The results of this study showed that ecological factors, like genetic factors, can affect the production and quantities of chemical compounds present in medicinal plants.

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Please cite this article as: A. Tahan, M. Jafari, D. Razmjoue, et al., Relationship among some ecological factors and chemical composition of Ajuga chamaecistus Ging. plant species..., Acta Ecologica Sinica, https://doi.org/10.1016/j.chnaes.2019.08.001