craniotome furcata (link) o. kuntze and its...
TRANSCRIPT
151
Chapter- 5
Effect of soil properties and microclimatic
conditions on essential oil composition of
Craniotome furcata (Link) O. Kuntze and its
chemosystematics
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5.1. Introduction
Craniotome furcata (Link.) O. Kuntze (Syn. C. versicolar, Anisomeles (Link)
furcata, A. nepalensis, Nepeta versicolor) belonging to family the Lamiaceae is an
erect, perennial, branched and soft hairy herb. It has 1-2 m tall stem with subwoody
base. Petiole is 2.5-7 cm long and leaves are stalked, cordate, broadly ovatewith dentate
margin. The flowers are numerous, white pink or yellow, crowed in small stalked cyme
forming narrow terminal panicles (14-18 cm). The lower floral leaves are leaf like while
spatulated upper leaves. The calyx (1.5 mm) is ovoid, glandular and 5-toothed. The
bracteoles are shorter than calyx tube. The corolla (3-4 mm) is reddish or purple-red in
colour. It is longer than the calyx, limb 2-lipped, upper lip very short, erect, hood like,
longer, spreading, 3-lobed and mid-lobe largest. The stamens are four, in unequal pairs,
ascending under the upper lip, outer or interior pair longer than the inner1,2
. Only one
species of C. furcata has been reported in India3,4
. It is distributed in China, Bhutan,
India, Korea, Laos, Myanmar, Nepal, Sikkim, Taiwan and Vietnam at a height of 1500-
2300 m 1,5
.
C. furcata has been used as folk medicine. The leaf juice is applied for
treatment of wounds 6. Joshi (2010)
7and Joshi et al. (2010)
8 reported the antimicrobial
and antioxidant activity in the essential oil and extract of C. furcata respectively.
Joshi and Pande (2009)9
reported the essential oil composition of C. furcata for
the first time from India. They reported germacren D (30.9%) as the major component
along with germacrene D-4-ol (12.1%), α-cadinol (6.4%), 3-octanone (6.1%),
germacrene A (5.8%) and epi-α-cadinol (4.0%). Three saponins, craniosaponin A and
buddlejasaponins Ia and I were isolated from the n-butanol soluble fraction of C.
furcata for the first time. Among them, craniosaponin A was identified as a new
compound10
. Cranioside A and B, mussaeniside and ningpogenin were isolated from the
ethyl acetate fraction of C. furcata and among them, cranioside A and B were identified
as new compounds11
.
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The essential oil composition of medicinal and aromatic plants is not constant
but varies quantitatively and qualitatively. Essential oil quality depends upon different
environmental factors like nature of soil, climatic conditions such as light, altitude,
moisture, growing and harvesting time etc. 12
. Important factors, which affects the
essential oil composition of aromatic and medicinal plants play a very important role in
the biogenetic pathways of different secondary metabolites of oil. Therefore, it is
necessary to investigate the relationship among the metal content in soil, plant and
environmental factors with the active constituents of aromatic and medicinal plants to
examine which factor is responsible for any variation of active constituents of aromatic
and medicinal plant.
Literature search revealed that no work has been done on the chemosystematics
of the plant Craniotome furcata (Link.) O. Kuntze with respect to soil and
microclimatic conditions in Uttarakhand region. Therefore, the objective of the present
work is to examine the effect of macro and micronutrient in soil and plant and
microclimatic conditions on essential oil composition of C. furcata,
5.2. Collection of plant material and soil samples
Fresh plant material of C. furcata (Link.) O. Kuntze along with its soil samples
(0-20 cm) were collected in September to November, 2010 from ten locations viz.
Bhowali (29º23'N: 79º31’E), Ramgarh (29º23'N: 79º30'E), Rushi village (29º23'N:
79º30'E), Nainital (29º23'N: 79º30'E), Jeolikot (29º23'N: 79º30'E), Mussoorie (30º 27'
N: 78º 06' E), Mukteshwar (29°28'N: 79°39'E), Kilbury (29º23'N: 79º30'E), Binsar
(29°37'N: 79º40'E) and Munsiyari (30°04'37"N: 80°23'04"E) in Kumaun Himalaya
(Uttarakhand, India). The plants were in full blooming stage. The botanical
identification of the specimen was done at Botany Department, Kumaun University,
Nainital and deposited in Botanical Survey of India, Dehradun (Voucher no. 34806).
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5.3. Fractionation of the oil and identification of major
compounds
The essential oils of C. furcata (5.0 mL) were fractionated by column
chromatography (CC) on a column packed with 100 g silica gel (230-400 mesh) in n-
hexane (Scheme 5.1 and 5.2). The fractions (CF # 1 and CF# 2) obtained by column
chromatography were analyzed by spectroscopy (1H and
13C NMR) and MS to
determine their identity.
5.3.1. Flow sheet (1) for CC of essential oil C. furcata
Essential oil
(5.0 mL)
n-hexane 5% Et2O 10% Et2O 15% Et2O 20 % Et2O
in n-hexane in n-hexane in n-hexane in n-hexane
Fr (1-12) Fr (13-20) Fr (21-26) Fr (28-33) Fr (34-43)
A B C D E
CF # 01
Scheme 5.1 Isolation of compound from C. furcata (Link.) from Kilbury.
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5.3.2. Flow sheet (2) for CC of essential oil C. furcata (Link.).
Essential oil
(5.0 mL)
n-hexane 5% Et2O 10% Et2O 15% Et2O 20 % Et2O
in n-hexane in n-hexane in n-hexane in n-hexane
Fr (1-12) Fr (13-20) Fr (21-26) Fr (27-33) Fr (34-43)
A B C D E
Recolumn 10% Et2O
CF # 02
Scheme 5.2 Isolation of compound from C. furcata (Link.) from Rushi.
5.4. Results and Discussion
5.4.1. Characterization of the constituents
1) Characterization of CF#01:
Physico-chemical data
IR vmax cm-1
: 2927, 2871, 1689,1463, 1383, 980, 734 (Figure 5.1).
EIMS (70eV): 240(M+),189, 161 (100%), 133, 119, 105, 91, 79, 77, 67, 65, 43.
1H NMR (300MHz, CDCl3-TMS) (Figure 5.2):
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δ 0.89 (6H, d), 1.52 (3H, s), 4.80 (2H, dd), 4.80 (2H, dd), 5.24 (2H, m), 5.81 (1H, s).
13CNMR (75MHz, CDCl3-TMS) (Figure 5.3):
δ 129.8 (d, C-1), 29.7 (t, C-2), 34.5 (t, C-3), 148.8 (s, C-4), 135.6 (d, C-5), 133.2 (d, C-
6), 52.9 (d, C-7), 26.5 (t, C-8), 40.7 (t, C-9), 133.7 (s, C-10), 32.7 (d, C-11), 19.7 (q, C-
12), 20.9 (q, C-13), 15.8 (q, C-14), 109.0 (t, C-15).
The compound CF#01 was obtained as viscous liquid. The EIMS of compounds
shows molecular ion peak at m/z 204, corresponding to molecular formula C15H24. The
1H- NMR spectrum showed a doublet at δ 0.89 (6H, d) which revealed the presence of
isopropyl group in the molecule. The signal at δ 1.52 (3H, s) was attributed to a methyl
group attached to an olefinic carbon. The 13
C spectra of compounds represent fifteen
carbons. Based on these spectral data, the compound CF#01 was identified as
germacrene D. Finally, its identity was confirmed by comparison of its spectral data
with those reported in literature13, 14
.
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Figure 5.1 IR Spectrum of CF #1
Figure 5.2 1H NMR Spectrum of CF #1
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Figure 5.3 13
C NMR Spectrum of CF#1
2) Characterization of CF#02:
Physico-chemical data
IR vmax cm-1
: 3533, 2932, 1385, 1366, 1199, 980, 787, 764 (Figure 5.4).
EIMS (70eV): 222(M+), 204, 189, 161 (100%), 147, 133 123, 119, 105, 91, 81, 79, 77,
67, 43.
1H NMR (300MHz, CDCl3-TMS) (Figure 5.5):
δ 0.799 (d, 3H, Me-13), 0.819 (d, 3H, Me-12), 1.195 (s, 3H, Me-15), 1.542 (s, 3H, Me-
14), 4.933 (brd, 1H, H-1), 5.183 (dd, 1H, H-6), 5.233 (d, 1H, H-5).
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13CNMR (75MHz, CDCl3-TMS) (Figure 5.6):
δ 128.8 (d, C-1), 23.6 (t, C-2), 41.2 (t, C-3), 73.1 (s,C-4), 140.0 (d, C-5), 125.7 (d, C-6),
52.8 (d, C-7), 39.6 (t, C-8), 25.9 (t, C-9), 132.5 (s, C-10), 33.0 (d, C-11), 20.6 (q, C-12),
18.9 (q, C-13), 16.7 (q, C-14), 30.7 (q, C-15).
The compound CF#02 was obtained as light green liquid. The IR spectrum of
compound has a broad absorption band at 3533 cm-1
showing the presence of OH
group. The EIMS of the compound displayed (M+) at m/z 222 corresponding to the
molecular formula C15H26O. The 1H –NMR showed doublets for the signals at δ 0.819
(3H, d) and 0.799 (3H,d) for two methyl of isopropyl group. A signal at δ 1.195 (3H, s)
was attributed to a methyl attached with quaternary carbon bearing alcoholic group.
Another broad singlet at δ 1.542 (3H, s) appears for methyl attached to the carbon
having endocyclic double bond, which is confirmed by the broad doublet at δ 4.933
(1H, brd, H-1). One doublet appear at δ 5.233 (1H, d) and one doublet signal at δ 5.183
(1H, dd), clearly indicate the presence of one other endocyclic double bond. 13
C NMR
showed the presence of total fifteen carbons. On the basis of the above spectral data
CF#02 was characterized as germacrene D-4-ol. Finally, its identity was confirmed by
comparison of its spectral data with those reported in literature 15, 16.
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Figure 5.4 IR Spectrum of CF #2
Figure 5.5 1H NMR Spectrum of CF #2
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Figure 5.6 13
C NMR Spectrum of CF#2
5.4.2. Chemosystematics of Craniotome furcata.
The essential oils of C. furcata (Link.) O. Kuntze collected from ten sites were
analyzed by GC and GC/MS. The structures of major components are shown in
Figure 5.7. Ward’s hierarchical clustering analysis of major constituents of essential
oils was conducted in order to classify chemotypes (Figure 5.8). The result of cluster
analysis showed four groups on the basis of difference in their main chemical
constituents and allowing them to be characterized into four distinct chemotypes
(Table 5.1). Group one consisted oils of C. furcata collected from Binsar (Figure
5.9) and Munsiyari (Figure 5.10) (Chemotype I) was significantly rich in δ-elemene
(9.9-11.1%) and germacrene D (52.8-59.8%) while the second group consisted
Jeolikot (Figure 5.11), Mussoorie (Figure 5.12), Mukteshwar (Figure 5.13) and
Kilbury (Figure 5.14) (Chemotype II) which was further divided in to two subgroups
on the basis of their similarity in dendrogram. The main components of the first
subgroup (Jeolikot and Mussoorie) were δ-elemene (3.4-7.9%), germacrene D (36.7-
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36.8%), α-zinziberene (3.5-12.8%) and α-cadinol (1.1-9.3%) while the components of
second subgroup (Mukteshwar and Kilbury) was germacrene D (42.9-46.2%), α-
zinziberene (5.5-5.8%), germacrene B (1.6-11.9%) and α-muurolol (8.5-10.5%), so
from above discussion it is possible to integrate these in to two subgroups. The oil
from Nainital (Figure 5.15) and Rushi village (Figure 5.16) (Chemotype III) showed
the presence of γ-cadinene (6.6-9.6%), germacrene D-4-ol (10.0-24.8%), α-muurolol
(2.4-5.2%), α-cadinol (9.2-11.9%), oplopanon (5.2-6.2%) and α-bisabolol oxide A
(6.1-10.6%) were placed into group III. The plants having high content of δ-elemene
(3.0-6.8), germacrene D (13.3-17.5%), α-zinziberene (5.1-14.0%), germacrene B (3.5-
15.6%), α-muurolol (8.1-15.2%) and α-cadinol (2.3-8.6 %) from Bhowali (Figure
5.17) and Ramgarh (Figure 5.18) (Chemotype IV) belongs to group fourth.
The cluster analysis classified the essential oils into four chemotypes on the
basis of presence or absence of chemical markers.
Chemotype-I: δ-elemene and germacrene D
Chemotype-II: Subgroup-I: δ-elemene, germacrene D, α-zinziberene and α-cadinol
Subgroup-II: germacrene D, α-zinziberene, germacrene B and α-muurolol
Chemotype-III: γ-cadinene, germacrene D-4-ol, α-muurolol, α-cadinol, oplopanon and
α-bisabolol oxide A
Chemotype-IV: δ-elemene, germacrene D, α-zinziberene, germacrene B, α-muurolol
and α-cadinol
These four chemotypes showed the chemical variability in essential oil
composition of C. furcata collected from ten regions. Earlier reports showed the
presence of germacrene D, germacrene D-4-ol, epi-α-cadinol, and α-cadinol as major
constituents from Nainital9,17
.
It has been well documented that germacrene D plays an important role as a
precursor of various sesquiterpenes such as cadinenes and selinenes18,19
. Plant
terpenes have been found to show anti-herbivore defenses20
. Germacrene D has also
been reported to have deterrent effects against herbivores and insecticidal activity
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against mosquitoes21
, as well as repellent activity against aphids22
and ticks23
.
Therefore, the common presence of germacrene D in Craniotome furcata may
be useful as a source of other terpenes that could make the plants inedible for
herbivores24
.
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H
H
δ-elemene (5) α-cubebene (6) germacrene D (16)
MF- C15H24 MF- C15H24 MF- C15H24
FW- 204 g/mol FW- 204 g/mol FW- 204 g/mol
(Z)
H
H
α-Zinziberene (17) (Z)-α-Bisabolene (23) ץ-cadinene (26)
MF- C15H24 MF- C15H24 MF- C15H24
FW- 204 g/mol FW- 204 g/mol FW- 204 g/mol
OH HO
Germacrene B (30) Germacrene D-4ol (31) α-Muurolol (34)
MF- C15H24 MF- C15H26O MF- C15H26O
FW- 204 g/mol FW- 222 g/mol FW- 222 g/mol
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H
H
OH
O
HO
H
α-Cadinol (35) α-Bisabolol oxide A (41)
MF- C15H26O MF- C15H26O2
FW- 222 g/mol FW- 238 g/mol
Figure 5.7 Structures of major constituents
Figure 5.8 Agglomerative hierarchical clustering analysis by SPSS 16.0 for the
chemical abundances of 12 essential oil components in the 10
populations of C. furcata.
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S.No
.
Compoundsa
RIb
RIc
Chemotype I Chemotype II Chemotype III Chemotype IV
Binsar
(9)
Munsiy
ari (10)
Jeoliko
t (1)
Musso
orie (5)
Muktesh
war (4)
Kilbury
(8)
Nainita
l (6)
Rushi
Village
(7)
Bhow
ali
(2)
Ramgar
h (3)
1 1-octene-3-ol AA 979 974 0.4 0.2 0.8 3.2 1.7 3.1 0.7 0.2 1.4 0.3
2 sabinene
MH 975 946 - - - - - 0.4 - - - -
3 (Z)- β-
ocimene
MH 1037 1032 - - 1.4 1.3 1.7 - - - - 0.3
4 (E)- β-
ocimene
MH 1050 1044 0.9 0.9 - - - - 1.6 0.4 - -
5 δ-elemene SH 1338 1335 11.1 9.9 7.9 3.4 2.7 3.3 2.2 1.7 3.0 6.8
6 α-cubebene SH 1348 1345 1.9 0.9 1.1 1.3 3.2 1.5 3.3 0.5 3.3 5.4
7 α-copaene SH 1376 1374 2.0 1.7 - - - - - - 1.7 -
8 β-bourbonene SH 1388 1387 3.5 4.5 - - - - - - - -
9 β–elemene SH 1389 1388 4.2 3.2 1.4 1.6 0.5 - 3.8 2.5 0.5 1.1
10 (E)-
caryophyllene
SH 1419 1417 3.3 2.7 1.03 0.7 1.2 1.0 4.2 1.6 1.9 0.5
11 γ-elemene SH 1436 1434 1.9 2.1 - 0.3 - - - - - -
12 α-guainene SH 1439 1437 - 0.8 - 0.7 0.7 - - - - 1.3
13 (Z)-β-
farnesene
SH 1442 1440 - - 0.9 - - - - - - -
14 9-epi-(E)-
caryophyllene
SH 1466 1464 - - - 1.2 - - - - - -
15 ar-curcumene SH 1480 1479 - - 1.7 3.5 - - - - - -
Table 5.1 Chemotypes of C. furcata collected from different sites
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16 germacrene D SH 1485 1484 52.8 59.8 36.7 36.8 46.2 42.9 4.0 4.5 13.3 17.5
17 α-zinziberene SH 1493 1493 - - 12.8 3.5 5.5 5.8 3.7 1.8 5.1 14.0
18 epi-cubibol OS 1494 1493 1.7 - - - - - - - - -
19 bicyclogerma
crene
SH 1500 1500 - - - - - 0.9 - - - -
20 α-muurolene SH 1500 1500 - - - 0.8 - 3.3 0.7 0.7 - -
21 (E)-β-guaiene SH 1502 1502 - - - - - 1.4 - - - -
22 α-fernesene SH 1505 1505 - - 1.3 0.6 - 1.1 2.8 1.0 - -
23 (Z)-α-
bisabolene
SH 1507 1506 - - - - - 10.3 - - - -
24 germacrene A SH 1509 1508 2.8 0.3 - - - - - - 2.78 -
25 δ-amorphene SH 1512 1511 - - 2.1 3.8 - - - - - -
26 γ-cadinene SH 1513 1513 - - 0.9 0.9 - 6.8 6.6 9.6 8.8 -
27 β-
sesquiphellan
drene
SH 1522 1521 - - 3.0 2.3 1.0 1.1 2.6 0.9 4.5 3.1
28 δ-cadinene SH 1523 1522 1.4 0.6 1.0 1.3 - - - - 2.5 -
29 hedycaryol OS 1548 1546 - - - 0.3 - - - - - -
30 germacrene B SH 1561 1559 1.5 1.7 1.0 4.1 11.9 1.6 4.8 6.9 3.5 15.6
31 germacrene
D-4-ol
OS 1575 1574 - - - 4.4 1.5 10.0 24.8 2.9 2.2
32 1,10-di-epi-
cubinol
OS 1619 1618 - - - - - 0.8 - - - -
33 α-muurolol OS 1646 1644 - - 2.7 3.8 8.5 10.5 5.2 2.4 15.2 8.1
34 β-eudesmol OS 1650 1649 - - 2.6 - - - - - - -
35 α-cadinol OS 1654 1652 0.7 - 1.1 9.3 1.5 0.9 9.2 12.0 8.6 2.3
36 ar-tumerone OS 1669 1668 - - - 0.6 3.2 - 4.6 1.3 3.3 1.9
37 epi-β-
bisabolol
OS 1671 1670 - - - 1.6 1.2 - 3.7 1.9 3.6 2.2
38 khusinol OS 1680 1679 - - 1.2 - - - - - -
39 (Z)-(E)-α- OS 1690 1690 1.1 0.6 - - - - - - - -
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aMode of identification: Retention Index, coinjection with standards/Peak enrichment with known oil constituents,
bRetention indices determined on the Equity-5
column using an n-alkane homologous series (C9–C24); cretention indices from the literature (Adams, 2007), Bold type indicates major components, %), AA=
aliphatic alcohol, MH= monoterpene hydrocarbon, OM= oxygenated monoterpene, SH= sesquiterpene hydrocarbon, OS= oxygenated sesquiterpene.
bergamotol
40 oplopanone OS 1740 1739 - - - 0.7 2.7 - 5.2 6.2 1.7 3.3
41 α-bisabolol
oxide A
OS 1749 1748 - - - - - - 6.1 10.6 - -
Total 91.2 89.9 81.4 93.2 94.9 96.7 81.4 93.2 94.9 96.7
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Figure 5.9 GC of the essential oil of C. furcata collected from Binsar.
Figure 5.10 GC of the essential oil of C. furcata collected from Munsiyari.
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Figure 5.11 GC of the essential oil of C. furcata collected from Jeolikot.
Figure 5.12 GC of the essential oil of C. furcata collected from Mussoorie.
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Figure 5.13 GC of the essential oil of C. furcata collected from Mukteshwar.
Figure 5.14 GC of the essential oil of C. furcata collected from Kilbury.
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Figure 5.15 GC of the essential oil of C. furcata collected from Nainital.
Figure 5.16 GC of the essential oil of C. furcata collected from Rushi
village.
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Figure 5.17 GC of the essential oil of C. furcata collected from Bhowali.
Figure 5.18 GC of the essential oil of C. furcata collected from Ramgarh.
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5.4.3. Physicochemical properties of soil
Physicochemical properties of the soil are given in Table 5.2. Soils were
classified as loamy sand and sandy loam. Soils were acidic to neutral (pH 5.42 to 7.85).
Most of the soil EC, OC %, CEC and WHC values are within the limits. The content of
macro and micronutrients in soil falls within the permissible limits.
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Table 5.2. Physicochemical properties of soil used in the study
Sites
Binsar Munsiyari Jeolikot Mussoorie Mukteshwar Kilbury Nainital
Rushi
Bhowali
Ramgarh
General soil properties
Sand (%) 66 70 78 68 82 84 78 76 80 70
Silt (%) 20 26 15 26 16 14 12 18 15 22
Clay (%) 16 8 4 6 4 2 8 6 5 8
Texture Sandy loam Sandy loam Loamy sand Sandy loam Loamy sand Loamy sand Loamy sand Loamy sand Loamy sand Sandy loam
Other soil properties
pH (1:2) 5.83±0.27 5.42±0.010 7.85±0.02 7.62±0.33 6.1±0.80 6.83±0.06 6.84±0.69 7.44±0.04 6.32±0.03 6.41±0.07
O.C. % 1.71±0.10 4.17±0.04 1.68±0.03 3.04±0.14 3.85±0.04 1.2±0.00 3.23±0.20 2.65±0.35 2.69±0.06 3.15±0.04
EC 0.111±0.000 0.11±0.001 0.42±0.07 0.156±0.011 0.054±0.022 0.23±0.040 0.074±0.001 0.34±0.02 0.78±0.01 0.195±0.01
CEC 10.26±0.030 27.28±0.070 10.78±0.010 18.13±0.060 16.3±0.100 31.97±0.040 25.76±0.080 38.11±0.010 13.64±0.100 15.06±0.130
W HC 37.92±0.070 40.36±0.010 43.52±0.030 58.51±0.100 38.24±0.080 49.69±0.010 39.57±0.050 46.08±0.010 42.88±0.070 35.11±0.040
Total content (mg kg-1)
Zn 57.663±0.028 38.67±0.002 25.05±0.16 54.281±0.05 26.875±0.10 62.657±0.03 47.377±0.26 91.677±0.01 42.007±0.11 41.718±0.05
Fe 561.849±0.228 519.33±0.230
521.664±0.01 522.604±0.01 516.859±0.26 566.618±0.04 542.206±0.03
559.248±0.05 556.288±0.03 528.632±0.06
Mn 15.5±0.130 24.63±0.090 10.70±0.10 19.825±0.29 10.7±0.010 25.783±0.01 14.67±0.06 15.55±0.08 11.484±0.05 10.825±0.01
Cu 158.336±0.02 165±0.73 192.105±0.08 220.897±0.09 182.15±0.02 275.888±0.07 303.703±0.09 348.023±0.03 255.251±0.12 221.418±0.05
Available content (mg kg-
1)
Zn 2.710±0.05 0.784±0.03 3.954±0.03 0.984±0.05 1.706±0.01 9.452±0.01 7.3680.03 12.262±0.05 1.486±0.03 1.104±0.01
Fe 22.65±0.04 36.68±0.07 32.53±0.04 91.70±1.03 29.00±0.08 29.94±0.11 28.77±0.06 57.98±0.30 35.38±0.40 33.28±0.08
Mn 7.08±0.06 14.28±0.11 6.68±0.05 17.11±0.88 3.00±0.73 14.56±0.21 15.43±0.42 17.39±0.20 9.58±0.07 10.00±0.23
Cu 0.150±0.03 1.119±0.01 0.96±0.02 1.820±0.03 0.320±0.04 2.256±0.11 1.830±0.02 7.328±0.08 0.610±0.07 0.400±0.00
Macronutrient content (%)
N (av) 0.005±0.10 0.008±0.03
0.009±0.05 0.012±0.34 0.012±0.71 0.011±0.69 0.010±0.63 0.014±0.01 0.009±0.25 0.012±0.09
N(tot) 0.20±0.01 0.28±0.01 0.20±0.04 0.25±0.03 0.24±0.01 0.21±0.04 0.18±0.05 0.35±0.03 0.13±0.02 0.30±0.06
P (av) 0.0026±0.00 0.0011±0.00 0.0014±0.00 0.0009±0.00 0.0007±0.00 0.0037±0.00 0.0019±0.001 0.0024±0.01 0.0006±0.00 0.0033±0.00
K (av) 0.0163±0.00 0.016±0.001 0.0076±0.00 0.0176±0.004 0.0046±0.001 0.0255±0.00 0.0132±0.00 0.009±0.002 0.0242±0.001 0.0193±0.001
*(av)=Available, (tot)= Total, EC= Electrical conductivity (dS cm-1), WHC= Water holding capacity, CEC= Cation exchange capacity (c mol kg-1), O.C.%= Organic carbon %
Estelar
177
5.4.4 Microclimatic conditions and oil properties
Microclimatic conditions and oil properties are shown in Table 5.3.
Table 5.3 Microclimatic conditions and oil properties
Microclim
atic and
other
properties
Binsar Muns
iyari
Jeoli
kot
Musso
Orie
Mukte
shwar
Kilb
ury
Nai
nital
Rushi Bho
wali
Ramg
arh
Altitude
(m)
2300 2386 1490 2000 2265 2200 2100 1600 1706 1789
Temperatu
re (0C)
23 18 30 25 20 22 23 28 28 23
Plant
height
(inch)
54.67
±1.53
37.67
±2.52
46
±2.65
35.33
±3.51
34.33
±2.52
26.67
±1.53
27.33
±1.53
39.33
±3.51
26
±2.66
34
±2.00
Month of
collection
in 2010
Octo
ber
Octo
ber
Septe
mber
Octo
Ber
Novem
ber
Novem
ber
Novem
ber
Septem
ber
Novem
ber
Novem
ber
Oil colour yellow yellow yellow yellow yellow light
green
light
green
yellow yellow yellow
Oil % 0.43 0.45 0.30 0.52 0.38 0.36 0.34 0.35 0.32 0.32
5.4.5. Correlation among major constituents
Simple correlation matrix (r) among major constituents of different chemotypes
given in (Table 5.4). δ-elemene is positively correlated with germacrene D (r=0.641,
P≤0.05) and negatively correlated with γ-cadinene (r=-0.678, P≤0.05), oplopanon (r=-
0.648, P≤0.05) and α-cadinol (r=-0.724, P≤0.05). Germacrene D showed negative
correlation with γ-cadinene (r=-0.695, P≤0.05), germacrene D-4-ol (r=-0.682, P≤0.05),
α-cadinol (r=-0.784, P≤0.01), oplopanon (r=-0.733, P≤0.05) and α-Bisabolol oxide A
(r=-0.678, P≤0.05). γ-cadinene is positively correlated with Germacrene D-4-ol
(r=0.635, P≤0.05), α-cadinol (r=0.675, P≤0.05) and α-Bisabolol oxide A (r=0.640,
P≤0.05). Germacrene D-4-ol is positively correlated with α-cadinol (r=0.780, P≤0.01),
oplopanon (r=0.866, P≤0.01) and α-Bisabolol oxide A (r=0.966, P≤0.01) while α-
cadinol is positively correlated with oplopanon (r=0.694, P≤0.05) and α-Bisabolol oxide
A (r=0.702, P≤0.05). Oplopanon showed positive correlation with α-Bisabolol oxide A
(r=0.906, P≤0.01).
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178
5.4.6. Effect of macronutrient on essential oil composition
Correlation matrix (r) between macronutrients and major constituents of essential oil
are given in Table 5.5 Available N is negatively correlated with δ-elemene (r=-0.734,
P=<0.05).
5.4.7. Effect of micronutrient on essential oil composition
5.4.7.1. Effect of zinc (Zn)
Total Zn in soil is positively correlated with Germacrene D-4-ol (r=0.735,
P≤0.05) and α-Bisabolol oxide A (r=0.695, P≤0.05) and available Zn is positively
correlated with γ-cadinene (r=0.722, P≤0.05), Germacrene D-4-ol (r=0.703, P≤0.05)
and α-Bisabolol oxide A (r=-0.778, P≤0.01) while total Zn in plants is positively
correlated with γ-cadinene (r=0.642, P≤0.05) (Table 5.6). Carbon dioxide and glucose
are the main precursors of monoterpene biosynthesis. Saccharides are also a source of
energy and reducing power for terpenoid synthesis. As zinc is involved in
photosynthesis and saccharide metabolism, and as CO2 and glucose is the most likely
sources of carbon utilized in terpene biosynthesis, the role of zinc becomes very
important in the terpenoid biosynthesis25
. As zinc is an essential micronutrient for plants
by acting either as a metal component of various enzymes or as a functional, structural,
or regulatory cofactor associated with saccharide metabolism, photosynthesis, and
protein synthesis26
.
5.4.7.2. Effect of iron (Fe)
Total iron in soil showed positive correlation with γ-cadinene (r=0.699, P≤0.05)
(Table 5.7).
5.4.7.3. Effect of copper (Cu)
. Available Cu and showed positive correlation with γ-cadinene (r=0.639,
P≤0.05), germacrene D-4-ol (r=0.912, P≤0.01), oplopanon (r=0.871, P≤0.01) and α-
bisabolol oxide A(r=0.705, P≤0.05) while Cu in plant is positively correlated with
Estelar
179
germacrene D-4-ol ( r=0.705, P≤0.01) and α-bisabolol oxide A (r=0.712, P≤0.05)
(Table 5.8).
5.4.7.4. Effect of manganese (Mn)
Total Mn in soil is negatively correlated with δ-elemene (r=-0.766, P=<0.01)
and germacrene D (r=-0.825, P=<0.01) while positively correlated with γ-cadinene
(r=0.886, P≤0.01), germacrene D-4-ol (r=-0.791, P=<0.01), α-cadinol (r=-0.752,
P=<0.05), oplopanon (r=0.748, P≤0.05) and α-bisabolol oxide A (r=-0.800, P=<0.01)
while Mn in plants is positively correlated with γ-cadinene (r=0.716, P≤0.05) (Table
5.9). Duarte et al. (2010)27
suggested that γ-cadinene, limonene, and caryophyllene
oxide have a strong relationship with micronutrient balance in soils (Zn, Cu, Fe, Mn) in
Eugenia dysenterica.
5.4.8. Effect of plant properties and microclimatic conditions on essential oil
composition
Altitude is positively correlated with germacrene D (r=0.644, P≤0.05)and
negatively with temperature (r=-0.909, P≤0.01) while plant height is positively
correlated with δ-elemene (r=0.723, P≤0.05) and negatively correlated with α-muurolol
(r=-0.759, P≤0.05) (Table 5.10).
Comparison of volatile constituents of C. furcata from ten locations shows that
there is some quantitative difference between the essential oil components which may
be due to the environmental factors. The only distinct feature is altitudinal variation
with Jeolikot being located at 1490 m, Rushi, 1600 m, Mussoorie, 2000 m, Nainital,
2100 m, Kilbury, 2220 m, Mukteshwar, 2265, Binsar, 2300 m and Munsiyari, 2386 m,
having germacrene D 13.3%, 17.5%, 36.7%, 36.8 %, 46.2%, 42.9%, 52.8% and 59.8 %
respectively. This clearly indicates a continuous increasein the percentage of
germacrene D with increasing altitude, Vakou et al. (1993)28
reported that altitude
influenced the oil content of O.vulgare ssp. hirtum from Grees..
5.4.9. Effect of soil physical properties on essential oil composition
Estelar
180
Soil CEC is positively correlated with germacrene D-4-ol (r=0.651, P≤0.05) and
α-bisabolol oxide A (r=0.683, P≤0.05). Sand is positively correlated with α-muurolol
(r=0.637, P≤0.05) (Table 5.11).
5.4.10. Micro, macro nutrients and microclimatic conditions
Total zinc in soil is positively correlated with available Zn in soil (r=0.728,
P≤0.05), Zn concentration in plant (r=0.937, P≤0.01), total iron in soil (r=0.695,
P≤0.05), available Cu in soil (r=0.831, P≤0.01), Cu in plant (r=0.781, P≤0.01) and total
Mn in soil (r=0.676, P≤0.05) (Table 5.12). Available Zn in soil is positively correlated
with Zn concentration in plant (r=0.767, P≤0.01), available Cu in soil (r=0.815,
P=<0.01), Cu in plant (r=0.738, P≤0.01), and total Mn in soil(r=0.810, P≤0.01). Zinc
concentration in plant is positively correlated with total iron in soil (r=0.741, P≤0.05),
available Cu in soil (r=0.755, P≤0.05), Cu in plant (r=0.727, P≤0.05) and total Mn in
soil(r=0.723, P≤0.05). Available Fe in soil is positively correlated with Fe in plant
(r=0.776, P≤0.01). Iron in plant is negatively correlated with altitude (r=-0.682,
P≤0.05). Available Cu in soil is positively correlated with Cu in plant (r=0.862,
P≤0.01), total Mn in soil (r=0.775, P≤0.01) and available Mn in soil (r=0.646, P≤0.05).
Copper in plant is positively correlated with total Mn in soil (r=0.694, P≤0.05),
available Mn in soil (r=0.808, P≤0.01) and Mn in plant (r=0.730, P≤0.05). Total Mn in
soil is positively correlated with available Mn in soil (r=0.633, P≤0.05) and Mn in plant
(r=0.662, P≤0.05). Available Mn in soil is positively correlated with Mn in plant
(r=0.750, P≤0.05). Altitude is negatively correlated with temperature (r=-0.909,
P≤0.01).
5.4.11. Physical properties and micronutrients
Percent organic carbon is negatively correlated with total iron in soil (r=-0.643,
P≤0.05) (Table 5.13). Cation exchange capacity is positively correlated with total Zn in
soil (r=0.688, P=<0.05), available Zn in soil (r=0.764, P≤0.05), Zn concentration in
plant (r=0.680, P≤0.05), available Cu in soil (r=0.805, P≤0.01), Cu in plant (r=0.977,
P≤0.05), total Mn in soil (r=0.696, P≤0.05) and Mn in plant (r=0.685, P≤0.05). Water
Estelar
181
holding capacity is positively correlated with available iron (r=0.812, P≤0.01). Sand is
negatively correlated with silt and clay percentage (r=-0.812, P≤0.01 and r=-0.770,
P≤0.01 respectively).
5.4.12. Physical properties, macronutrients and microclimatic conditions
Soil pH is negatively correlated with altitude (r=-0.697, P≤0.05) and
positively correlated with temperature (r=-0.742, P≤0.05) (Table 5.14). Silt is
positively correlated with oil percentage (r=-0.741, P≤0.05). Clay is positively
correlated with plant height (r=0.638, P≤0.05) while negatively correlated with
available nitrogen (r=-0.628, P≤0.05).
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182
Table-5.4 Simple correlation matrix (r) among major constituents.
1 2 3 4 5 6 7 8 9 10 11 12
S.N δ-elemene α-
cubeben
e
germacrene
D
α-zinziberene Z-α-
bisabolene
γ-cadinene germacrene
B
germacrene
D-4-ol
α-muurolol α-cadinol oplopanon α-bisabolol
oxide A
1 1.00 0.137 0.641* -0.018 -0.195 -0.678
* -0.275 -0.549 -0.564 -0.702
* -0.648
* -0.488
2 1.00 -0.184 0.335 -0.206 -0.240 0.581 -0.308 0.362 -0.177 -0.059 -0.289
3 1.00 -0.235 0.199 -0.695* -0.335 -0.682* -0.364 -0.789** -0.733
* -0.678*
4 1.00 0.043 -0.198 0.396 -0.264 0.360 -0.234 -0.223 -0.287
5 1.00 0.298 -0.255 -0.208 0.354 -0.288 -0.249 -0.160
6 1.00 -0.183 0.635* 0.441 0.675
* 0.627 0.640
*
7 1.00 0.143 0.276 0.041 0.292 0.046
8 1.00 -0.172 0.780**
0.866**
0.966**
9 1.00 0.163 0.032 -0.226
10 1.00 0.694* 0.702
*
11 1.00 0.906**
12 1.00 * Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
Table-5.5 Correlation matrix (r) between macronutrients and major constituents of essential oil
1
N
(av)
2
N(total)
%
3
P2O5
%(av)
4
K2O %
(av)
5
δ-
eleme
ne
6
α-
cubebe
ne
7
germacre
ne D
8
α-
zinzibere
ne
9
Z-α-
bisabolen
e
10
γ-
cadinene
11
germacre
ne B
12
germacre
ne D-4-ol
13
α-
muurolol
14
α-
cadinol
15
oplopan
on
16
α-
bisabolol
oxide A
1 1.00 0.588 0.093 -0.141 -0.732* -0.159 -0.479 0.277 0.112 0.307 0.602 0.552 0.308 0.477 0.456 0.416
2 1.00 0.278 -0.326 0.009 -0.247 -0.031 -0.026 -0.131 -0.149 0.416 0.518 -0.457 0.085 0.238 0.421
3 1.00 0.349 0.110 0.221 -0.111 0.208 0.586 0.139 0.041 0.095 -0.020 -0.234 -0.031 0.164
4 1.00 0.032 0.205 -0.014 -0.032 0.521 0.269 -0.314 -0.341 0.493 -0.44 -0.427 -0.353
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
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183
Table-5.6 Correlation matrix (r) among zinc (Zn) in soil and plant with major constituents in soil
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level
Table-5.7 Correlation matrix (r) among iron (Fe) in soil and plant with major constituents in soil
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
1
Zn
Total
2
Zn
DTPA
3
Zn
Plant
4
δ-
elemene
5
α-
cubebene
6
germacrene
D
7
α-
zinziberene
8
Z-α-
bisabole
ne
9
γ-
cadine
ne
10
germacr
ene B
11
germacrene D-
4-ol
12
α-
muurolol
13
α-
cadinol
13
oplopano
n
14
α-
bisabolol
oxide A
1 1.00 0.728* 0.937** -0.306 -0.311 -0.355 -0.463 0.251 0.594 -0.175 0.735* -0.167 0.525 0.541 0.695*
2 1.00 0.767** -0.477 -0.411 -0.452 -0.157 0.454 0.722* -0.185 0.703* -0.044 0.380 0.387 0.778**
3 1.00 -0.458 -0.254 -0.349 -0.288 0.513 0.642* -0.096 0.611 0.091 0.449 0.461 0.561
1
Fe
total
2
Fe
DTPA
3
Fe
Plant
4
δ-
elemene
5
α-
cubebene
6
germacrene
D
7
α-
zinziberene
8
Z-α-
bisabolene
9
γ-
cadinene
10
germacrene
B
11
germacrene
D-4-ol
12
α-
muurolol
13
α-
cadinol
14
oplopanon
15
α-
bisabolol
oxide A
1 1.00 -0.209 -0.055 -0.175 0.036 -0.294 -0.324 0.479 0.699* -0.363 0.293 0.248 0.234 0.230 0.343
2 1.00 0.776** -0.339 -0.462 -0.135 -0.169 -0.169 0.048 -0.053 0.363 -0.163 0.587 0.033 0.185 3 1.00 -0.340 -0.233 -0.442 0.186 -0.269 0.242 0.134 0.443 0.135 0.613 0.113 0.232
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184
Table-5.8 Correlation matrix (r) among copper (Cu) in soil and plant with major constituents in soil
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
Table-5.9 Correlation matrix (r) among manganese (Mn) in soil and plant with major constituents in soil
* Correlation is significant at the 0.05 level. ** Correlation is significant at the 0.01 level.
1
Cu
total
2
Cu
DTPA
3
Cu
Plant
4
δ-
elemene
5
α-
cubebene
6
germacrene
D
7
α-
zinziberene
8
Z-α-
bisabolene
9
γ-
cadinene
10
germacrene
B
11
germacrene
D-4-ol
12
α-
muurolol
13
α-
cadinol
14
oplopanon
15
α-
bisabolol
oxide A
1 1.00 0.202 0.588 0.105 -0.537 0.449 -0.516 0.607 0.065 -0.517 -0.100 -0.228 -0.175 -0.292 -0.066
2 1.00 0.862** -0.483 -0.581 -0.480 -0.280 0.096 0.639* -0.073 0.912** -0.201 0.622 0.664* 0.871** 3 1.00 -0.383 -0.578 -0.277 -0.444 0.328 0.591 -0.164 0.705* -0.170 0.423 0.505 0.712*
1
Mn
total
2
Mn
DTPA
3
Mn
Plant
4
δ-
elemene
5
α-
cubebene
6
germacrene
D
7
α-
zinziberene
8
Z-α-
bisabolene
9
γ-
cadinene
10
germacrene
B
11
germacrene
D-4-ol
12
α-
muurolol
13
α-
cadinol
14
oplopanon
15
α-
bisabolol
oxide A
1 1.00 0.633* 0.662* -0.766** -0.203 -0.825** -0.042 0.243 0.886** 0.019 0.791** 0.296 0.754* 0.748* 0.800**
2 1.00 0.750* -0.336 -0.480 -0.348 -0.363 0.217 0.481 -0.334 0.533 -0.165 0.560 0.285 0.528
3 1.00 -0.280 -0.353 -0.505 -0.351 0.069 0.716* -0.374 0.521 0.042 0.513 0.468 0.592
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185
Table-5.10 Correlation matrix (r) between microclimatic conditions and major constituents of essential oil
1
Alti
tude
2
Oil
%
3
Temp
4
Plant
height
5
δ-
eleme
ne
6
α-
cubebene
7
germacre
ne D
8
α-
Zinziberene
9
Z-α-
bisabolene
10
γ-
cadinene
11
germacrene
B
12
germacrene
D-4-ol
13
α-
muurolol
14
α-
cadinol
15
oplopano
n
16
α-Bisabolol
oxide A
1 1.00 0.588 -0.909** 0.001 0.262 0.115 0.644* -0.574 0.239 -0.372 -0.039 -0.395 -0.203 -0.437 -0.243 -0.319
2 1.00 -0.437 0.233 0.212 -0.274 0.553 -0.615 -0.086 -0.418 -0.207 -0.137 -0.461 -0.004 -0.335 -0.223
3 1.00 0.131 -0.234 -0.197 -0.548 0.348 -0.186 0.434 -0.244 0.350 0.143 0.486 0.188 0.291 4 1.00 0.723* -0.068 0.440 -0.128 -0.369 -0.369 -0.211 -0.070 -0.759* -0.354 -0.262 -0.066
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
Table 5.11 Correlation matrix (r) between physical properties of soil and major constituents of essential oil
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
pH EC OC% CEC Moist
ure
conten
t
sand silt clay δ-
elem
ene
α-
cube
bene
germ
acren
e D
α-
Zinzi
beren
e
Z-α-
bisabo
lene
γ-
cadinen
e
germacr
ene B
germacre
ne D-4-
ol
α-
muurolol
α-
cadinol
oplopano
n
α-
Bisabolo
l oxide A
1 1.00 0.240 -0.411 0.131 0.621 0.200 -0.228 -0.457 -0.450 -0.432 -0.442 0.379 0.072 0.294 -0.147 0.406 -0.017 0.482 0.204 0.351
2 1.00 -0.312 -0.161 0.129 0.326 -0.298 -0.330 -0.200 -0.085 -0.402 0.237 -0.027 0.546 -0.254 0.056 0.557 0.307 0.006 -0.010 3 1.00 0.080 -0.254 -0.187 0.414 0.013 -0.102 0.104 -0.014 -0.193 -0.560 -0.260 0.500 0.109 -0.089 0.159 0.258 0.064
4 1.00 0.287 0.271 -0.064 -0.311 -0.452 -0.549 -0.254 -0.393 0.415 0.584 -0.096 0.651* -0.097 0.355 0.531 0.683*
5 1.00 0.026 0.205 -0.415 -0.389 -0.647 0.026 -0.184 0.333 0.237 -0.410 0.154 0.018 0.404 -0.142 0.038
6 1.00 -0.812** -0.770** -0.630 -0.160 -0.246 0.232 0.493 0.543 0.081 0.081 0.637* 0.070 0.322 0.123
7 1.00 0.341 0.461 -0.114 0.428 -0.220 -0.312 -0.565 0.038 -0.133 -0.474 -0.128 -0.438 -0.264
8 1.00 0.634 0.421 0.172 -0.400 -0.431 -0.344 -0.122 -0.054 -0.544 -0.137 -0.085 0.004
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
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186
Table 5.12 Simple correlation matrix of micro, macronutrients and microclimatic conditions
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
N
(av)
N (t)
%
P2O5
%
K2O % Zn Zn
DTPA
Zn
Plant
Fe Fe
DTPA
Fe
Plant
Cu Cu
DTPA
Cu
Plant
Mn Mn
DTPA
Mn
Plant
Altitude Oil
%
Temper
ature
Plant
height 1 1.00 0.588 0.093 -0.141 0.307 0.341 0.425 -0.151 0.477 0.568 -0.055 0.556 0.473 0.584 0.355 0.141 -0.342 -0.133 0.092 -0.516
2 1.00 0.278 -0.326 0.440 0.231 0.336 -0.218 0.375 0.381 0.163 0.575 0.552 0.161 0.342 0.083 -0.072 0.199 -0.173 0.206
3 1.00 0.349 0.480 0.511 0.584 0.540 -0.280 -0.173 0.247 0.224 0.314 0.305 0.215 0.056 0.025 -0.260 -0.088 0.051
4 1.00 0.181 -0.092 0.304 0.490 0.011 0.081 0.446 -0.201 0.078 0.113 0.332 0.307 0.130 0.078 -0.078 -0.423
5 1.00 0.728* 0.937**
0.695* 0.366 0.349 0.360 0.831
** 0.781
** 0.676
* 0.670 0.475 -0.108 0.142 0.194 0.033
6 1.00 0.767**
0.615 -0.013 0.006 0.217 0.815**
0.738* 0.810
** 0.481 0.475 -0.232 -0.341 0.275 -0.139
7 1.00 0.741* 0.311 0.313 0.377 0.755
* 0.727
* 0.723
* 0.602 0.363 -0.108 0.036 0.182 -0.161
8 1.00 -0.209 -0.055 0.195 0.376 0.373 0.525 0.252 0.369 -0.050 -0.228 0.245 -0.064
9 1.00 0.776**
0.229 0.420 0.311 0.222 0.589 0.114 -0.225 0.587 0.258 -0.066
10 1.00 -0.169 0.456 0.208 0.349 0.370 0.172 -0.682* 0.075 0.598 -0.090
11 1.00 0.202 0.588 0.034 0.620 0.366 0.548 0.561 -0.472 -0.146
12 1.00 0.862**
0.775**
0.646* 0.556 -0.367 -0.064 0.341 -0.053
13 1.00 0.694* 0.808
* 0.730
* 0.012 0.098 -0.073 -0.267
14 1.00 0.633* 0.662
* -0.427 -0.381 0.391 -0.524
15 1.00 0.750* -0.037 0.309 0.035 -0.340
16 1.00 -0.157 -0.143 0.112 -0.436
17 1.00 0.588 -0.909**
0.001
18 1.00 -0.437 -.233
19 1.00 0.131
20 1.00
* Correlation is significant at the 0.05 level.,
** Correlation is significant at the 0.01 level.
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Table 5.13 Correlation matrix among soil physical properties and micronutrients
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
pH EC OC% CEC (WHC) Sand Silt Clay Zn Zn
DTPA
Zn
Plant
Fe Fe
DTPA
Fe
Plant
Cu Cu
DTPA
Cu
Plant
Mn Mn
DTPA
Mn
Plant 1 1.00 0.240 -0.411 0.131 0.621 0.200 -0.228 -0.457 0.245 0.441 0.312 -0.006 0.537 0.542 -0.148 0.477 0.157 0.487 0.324 0.034
2 1.00 -0.312 -0.161 0.129 0.326 -0.298 -0.330 0.014 0.028 0.054 0.328 0.015 0.486 -0.308 0.094 -0.087 0.268 0.062 0.301
3 1.00 0.080 -0.254 -0.187 0.414 0.013 -0.296 -0.431 -0.427 -0.643* 0.171 0.051 -0.078 -0.095 0.053 -0.154 0.048 0.132
4 1.00 0.287 0.271 -0.064 -0.311 0.688* 0.764* 0.680* 0.329 0.207 0.074 0.586 0.805** 0.977** 0.696* 0.743 0.685*
5 1.00 0.026 0.205 -0.415 0.371 0.207 0.447 0.076 0.812** 0.518 0.489 0.372 0.339 0.273 0.581 0.125
6 1.00 -0.812** -0.770** -0.116 0.441 0.119 0.193 -0.332 -0.198 -0.098 0.137 0.120 0.421 -0.159 0.109
7 1.00 0.341 0.047 -0.520 -0.112 -0.420 0.539 0.362 0.344 -0.071 0.056 -0.452 0.244 -0.087
8 1.00 0.141 -0.243 -0.091 0.178 -0.188 -0.260 -0.071 -0.217 -0.209 -0.355 -0.103 -0.104
9 1.00 0.728* 0.937** 0.695* 0.366 0.349 0.360 0.831** 0.781** 0.676* 0.670* 0.475
10 1.00 0.767** 0.615 -0.013 0.006 0.217 0.815** 0.738* 0.810** 0.481 0.475
11 1.00 0.741* 0.311* 0.313 0.377 0.755* 0.727* 0.723* 0.602 0.363
12 1.00 -0.209 -0.055 0.195 0.376 0.373 0.525 0.252 0.369
13 1.00 0.776** 0.229 0.420 0.311 0.222 0.589 0.114
14 1.00 -0.169 0.456 0.208 0.349 0.370 0.172
15 1.00 0.202 0.588 0.034 0.620 0.366
16 1.00 0.862** 0.775** 0.646* 0.556
17 1.00 0.694* 0.808** 0.730*
18 1.00 0.633 0.662*
19 1.00 0.750*
20 1.00
* Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
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Table 5.14 Correlation matrix of soil physical properties, soil macronutrients and microclimatic conditions
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 pH EC OC% CEC WHC Sand Silt Clay
N (av)
N(total)
%
P2O5
%
K2O % Altitude Oil % Temperat
ure
Plant
height
1 1.00 0.240 -0.411 0.131 0.621 0.200 -0.228 -0.457 0.424 0.083 0.040 -0.208 -0.697* -0.176 0.742
* -0.031
2 1.00 -0.312 -0.161 0.129 0.326 -0.298 -0.330 -0.002 -0.385 -0.228 0.325 -0.692* -0.488 0.726
* -0.218
3 1.00 0.080 -0.254 -0.187 0.414 0.013 0.282 0.325 -0.539 -0.302 0.285 0.279 -0.503 -0.258
4 1.00 0.287 0.271 -0.064 -0.311 0.508 0.501 0.323 0.036 0.087 0.31 -0.168 -0.367
5 1.00 0.026 0.205 -0.415 0.303 0.032 -0.128 0.234 -0.131 0.487 0.293 -0.183
6 1.00 -0.812**
-0.770**
0.319 -0.336 -0.031 -0.059 -0.167 -0.598 0.141 -0.590
7 1.00 0.341 0.013 0.533 -0.135 0.103 0.240 0.741* -0.322 0.322
8 1.00 -0.628* 0.015 0.157 0.009 0.333 0.318 -0.200 0.638
*
9 1.00 0.588 0.093 -0.141 -0.342 -0.133 0.092 -0.516
10 1.00 0.278 -0.326 -0.072 0.199 -0.173 0.206
11 1.00 0.349 0.025 -0.260 -0.088 0.051
12 1.00 0.130 0.078 -0.078 -0.423
13 1.00 0.588 -0.909**
0.001
14 1.00 -0.437 0.233
15 1.00 0.131
16 1.00 * Correlation is significant at the 0.05 level.
** Correlation is significant at the 0.01 level.
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5.5. Conclusions
Aerial parts of ten samples of Craniotome furcata family Lamiaceae, collected
from different locations in Central Himalayas, India was analyzed by GC and GC/MS
for their essential oil composition. Cluster analysis was done to differentiate plants
collected from different locations on the basis of their main constituents. Macro and
micronutrients (N, P, K, Zn, Cu, Fe and Mn) in soil and plant samples were also
determined. Statistical analysis of correlation coefficient was done to correlate different
environmental and soil factors with major constituents. The results of the present
investigation are summarized in this section.
Chemosystematics
Cluster analysis classified wild C. furcata in to four groups on the basis of major
constituents. The genus is classified into four chemotypes as follows:
Chemotype I: Binsar and Munsiyari (δ-Elemene and germacrene D)
Chemotype II: Jeolikot, Mussoorie, Mukteshwar and Kilbury (Germacrene D)
Chemotype III: Rushi and Nainital (α-Bisabolol oxide A, α-cadinol and
germacrene D-4-ol)
Chemotype IV: Bhowali and Ramgarh (α-Zinziberene, α-muurolol and
germacrene D)
Correlation among major constituents
δ-Elemene was positively correlated with germacrene D. Germacrene D-4-ol
was positively correlated with α-cadinol and α-bisabolol oxide A while α-cadinol
correlated with α-bisabolol oxide A
Effect of macronutrient and micronutrients on essential oil composition
Correlation analysis revealed that micronutrients in soil and plant affected
essential oil composition. Available nitrogen was negatively correlated with δ-elemene.
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Total and available zinc, available copper and total manganese in soil was positively
correlated with α-bisabolol oxide A, suggesting the role of nitrogen, zinc, copper and
iron in their biosynthesis in C. furcata.
Effect of plant characteristics and microclimatic conditions on essential oil
composition
Altitude was positively correlated with plant height with δ-elemene and
negatively correlated with α-muurolol.
Thus, it can be suggested that essential oil composition of Craniotome furcata
was affected by variation in soil properties and microclimatic conditions. The four
chemotypes were detected on the basis of germacrene D content. At higher altitude,
more germacrene D was synthesized in C. furcata. Nitrogen and Iron in soil
negatively affect synthesis of δ-elemene while zinc, copper and manganese in soil
positively affect the synthesis of α-bisabolol oxide A in C. furcata. The percentage of
δ-elemene was found to be more in taller plants.
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