2. Medicinal plants are powerful source of new drugs, now
contributing about 90% of the newly discovered pharmaceutical
products.
And these products provides health coverage of over 80% of the
world population.(Lambert,2012)
Production of medicinal plants is affected by both biotic and
abiotic stresses.
Drought stress has both positive and negative effect on
production of medicinal plants.
Positive effects- increases the secondary metabolites
Negative effect- reduction in growth and total yield.
11/11/2016 2
3. IMPACT OF DROUGHT STRESS ON SECONDARY
METABOLITE PRODUCTION OF MEDICINAL CROPS
CHANDRAKANT BALLOLLI
UHS15PGM564
Department of PSMA
K.R.C.COLLEGE OF HORTICULTURE,ARABHAVI11/11/2016 3
UNIVERSITY OF HORTICULTURAL
SCIENCES, BAGALKOT
SEMINAR-I ON
5. • Drought is a period or condition of unusuall dry weather
within a geographic area where there is a lack of precipitation.
• Drought is governed by various factors, the most prominent
being extremes in temperature.
11/11/2016 5
6. • Plants experience drought stress either when the water supply
to their roots becomes limiting or when the transpiration rate
becomes intense
• High Evaporation, low air humidity, high temperature, high
irradiance, strong wind
11/11/2016 6
8. TERMINOLOGY
Water potential:
• Free energy content of water which satisfies all requirements
in plants.
• Expressed in terms of pressure units such as MPa. 1 bar =
105 Pascal,(0.1 Mpa).
Relative water content (%):
• It is the appropriate measure of plant water status in terms of
the physiological consequences of cellular water deficit.
• RWC(%)= [(W-DW)/(TW-DW)]x100.
Field water capacity:
• The amount of moisture held in the soil after excess water has
drained away.
11/11/2016 8
9. CONSEQUENCESOFDROUGHT
o Diminished crop and livestock production
o Famine due to lack of water for irrigation
o Habitat damage
o Mass migration
o Shortages of water for industrial users
11/11/2016 9
10. • Production of stress proteins
• Accumulation of osmotically active compounds
• ROS production and development of anti-oxidative systems
• Changes in enzyme activities
• Changes in biosynthesis and catabolism of phytohormones
• Changes in absorption and transport of ions
11/11/2016 10
11. o Changes the leaf anatomy and ultra structure
o Inhibition of growth, cell division, changes in cell wall
synthesis
o Increase in root to shoot ratio
o Acceleration of ageing
o Inhibition of photosynthesis
o Decrease in plant biomass yield
11/11/2016 11
12. Water stress and protein synthesis
1) Protein in plants decreased due to suppressed synthesis
2) Synthesis of other proteins and mRNAs
3) Synthesis of specific stress proteins
• A) Proteins taking part in signal transduction and gene
expression. Eg: protein kinases
• B) Proteins participating in stress tolerance. Eg: LEA and
HSP
11/11/2016 12
14. ABA accumulation
• Increased ABA is a common plant response to stress.
• Increased ABA closure of stomata
• ABA - maintaining plant growth by suppressing ethylene
evolution
• In water-stressed plants increased ABA is an early response to
decreased leaf .
• Generally ABA is produced in the root tips.
11/11/2016 14
15. • Proline is the most widely distributed osmolyte
• Proline accumulation increases under drought condition
.
• It increases the osmotic potential.
• Roles: Osmotic adjustment, membranes protection, a reservoir
of nitrogen and carbon source for post stress growth.
11/11/2016 15
16. • Decreases the Ca 2+
• Decrease in K+ concentration-due to membrane
damage and disruption in ion homeostasis.
• Disruption in N-metabolism.
11/11/2016 16
18. Drought stress
PLANT RESPONSES TO DROUGHT
STRESS
11/11/2016
18
Shao et al., 2008
Physiological Responses Biochemical Responses Molecular Responses
Recognition of root signals Decreased efficiency of Rubisco Stress responsive gene expression
Loss of turgor and osmotic
adjustment
Accumulation of stress
metabolites like glutathione,
glybet, polyamines
Increased expression in ABA
biosynthetic genes
Reduced leaf water potential Increase in antioxidant enzymes
like SOD,CAT,APX,POD
Drought stress tolerance
Decrease in stomatal
conductance
Reduced ROS accumulation Synthesis of specific proteins like
LEA,HSP
Reduced internal co2
concentration
Decline in net photosynthesis
19. 19
Causes of growth reduction under drought stress
CAUSES
Drought stress
(water scarcity)
Reduced cell
elongation and
enlargement
Diminished
growth
Loss of turgor
11/11/2016
20. How secondary metabolites are increased as a response
to drought?
During drought stress due to the incipient water deficiency the
stomata gets closure and the uptake of CO2 markedly
decreases.
As result, the consumption of reduction equivalents
(NADPH+H+) for the CO2-fixation via Calvin cycle declines
considerably, generating a massive oversupply of
NADPH+H+.
As a consequence, metabolic processes are pushed towards the
synthesis of highly reduced compounds like isoprenoids,
phenols and alkaloids.
11/11/2016 20
Selmar et al.,2013
21. Phytochemicals
• Classification of Phytochemicals
1) Primary metabolites
2) Secondary metabolites, depending on their role in plant metabolism
• Primary metabolites :
Common sugars
Amino acids
Proteins
Purines and pyrimidines of nucleic acids,
Chlorophyll’s etc.
11/11/2016 21
24. Secondary metabolites Use Plant species
Ajmalicine Anti hypertensive Catharanthus roseus.
Artemisinin Antimalarial Artemisia annua.
Ajmaline Anti hypertensive Rauvolfia serpentina.
Codeine Sedative Papaver somniferum.
Digoxin Heart stimulant Digitalis lanata.
Diosgenin Steroidal precursor Dioscorea deltoidea..
Forskolin Bronchial asthma Coleus forskolii.
11/11/2016 24Raja et al.,2015
25. Secondary metabolites Use Plant species
Morphine Sedative Papaver somniferum.
Quinine Antimalarial Cinchona ledgeriana.
Vincristine Antileukemic Catharanthus roseus.
Vinblastine Antileukemic C.roseus.
Taxol Anticancer Taxus brevifolia.
Emetine Emetic Cephalis ipecaccuanha.
Capsaicin Counterirritant Capsicum frutescens.
11/11/2016 25
Raja et al.,2015
26. Drought stress increase in the contents of
secondary metabolites in medicinal plants
Papaver somniferum Morphine alkaloids Strong increase Szabo et al. (2003)
Catharanthus
roseus
Ajmalicine Higher content in
stressed plants
Abdul et al., 2008
Salvia officinalis Monoterpenes Higher content in
stressed plants
Nowak et al., 2010
Artemisia annua Artemisinin Significant increase Jose et al., 2010
Withania somnifera Withanolide Strong increase Sonal et al., 2010
Andrographis
panniculata
Andrographolide Higher content in
stressed plants
Priyanka et al.,
2009
Thymus capitatus Phenolics Higher content in
stressed plants
Delitala et al.,1986
2611/11/2016
28. • Major bitter constituent.
• Diterpene lactone used in
hypertension, diarrhoea,
ulcers, malaria, HIV,
hepatitis, diabetes, cancer
and kidney disorders.
11/11/2016 28
29. Studies of soil moisture stress on biochemical,
active ingredient content and productivity of
Kalmegh (Andrographis panniculata )
Dwivedi et al., 2009
29
Objectives:
To study the effect of soil moisture
stress on biochemical, active
ingredient content and productivity of
Kalmegh
11/11/2016
30. Fig.1 Effect of different soil moisture stress on proline content in Kalmegh
30
Dwivedi et al., 2009
STANDARD
50% WATERING
11/11/2016
31. Fig. 2 Effect of different soil moisture stress on nitrogen % in
Kalmegh
0
0.5
1
1.5
2
2.5
3
3.5
4
control 75% MS 50% MS 25% MS 0% MS
Dwivedi et al., 2009
Nitrogen%
11/11/2016
32. Fig. 3 Effect of different soil moisture stress on Protein
% in Andrographis panniculata
0
5
10
15
20
25
control 75% MS 50% MS 25% MS 0% MS
32
Protein%
Dwivedi et al., 2009
11/11/2016
33. Fig.4 Effect of different soil moisture stress on
androgapholide content in Kalmegh
33
0
0.05
0.1
0.15
0.2
0.25
control 75% MS 50%MS 25%MS 0%MS
Andrographolide%
Dwivedi et al., 2009
11/11/2016
34. 17
18
19
20
21
22
23
control 75% MS 50% MS 25% MS 0% MS
Dwivedi et al., 2009
Fibre%
Fig. 5 Effect of different soil moisture stress on fibre
% in Andrographis panniculata
11/11/2016
35. Fig. 6 Effect of different soil moisture stress on dry
herbage yield in Andrographis panniculata
0
5
10
15
20
25
30
35
control 75% MS 50%MS 25% MS 0% MS
35
Dryherbageyield(q/ha)
Dwivedi et al., 2009
11/11/2016
36. Fig. 7 Effect of different soil moisture stress on seed
yield in Andrographis panniculata
0
5
10
15
20
25
30
35
40
control 75% MS 50% MS 25% MS 0% MS
36
Seedyield(q/ha)
Dwivedi et al., 2009
11/11/2016
37. Methyl chavicol, Linalool and Eugenol-
• These aroma compounds possesses a wide range of biological
properties such as antibacterial, antifungal and Nematicidal and
antioxidant properties.
• Basils are used for head-aches, coughs, stomach-ache and kidney
malfunctions.
11/11/2016 37
38. Influence of water stress on growth, essential oil, and
chemical composition of herbs (Ocimum sp.)
Kh.A. Khalid
Department of Cultivation and Production of Medicinal and Aromatic Plants, National
Research Centre, Dokki, Giza, Egypt
Received March 31, 2006; accepted June 19, 2006
INTERNATIONAL
Agrophysics
www.ipan.lublin.pl/int-agrophysics
Int. Agrophysics, 2006, 20, 289-296
11/11/2016 38
Objectives:
Toinvestigatethepossibleeffectofwaterstressonthevegetative
growth,essentialoilcontent,andchemicalcontentofOcimum
basilicumandOcimumamericanum.
39. Treatment details.
4 levels of water stress Two species.
125% field water capacity. Ocimum basilicum
100% field water capacity. O.americanum
75% field water capacity.
50% field water capacity.
11/11/2016 39
40. 40
Ocimum sp. Water stress
treatments (%)
Total herb fresh weight (g /plant) Total herb dry weight (g /plant)
1st season 2nd season 1st season 2nd season
Ocimum
basilicum L.
125
100
75
50
872
2162
2773
1393
794
2172
2703
1381
176
432
554
279
179
438 541
276
Average of Ocimum basilicum L. 1800 1764 360 359
Ocimum
americanum L.
125
100
75
50
1162
2650
3428
2109
1201
2512
3572
2379
233
534
686
418
240 551
730
427
Average of Ocimum americanum 2337 2416 468 487
Average of
Ocimum
basilicum &
Ocimum
americanum
125
100
75
50
1017
2406
3101
1751
998
2342
3138
18808
205
483
620
349
210 495
635
352
LSD at 0.05
Ocimum sp. 37.89 41.23 22.32 18.56
Water stress treatments 50.36 56.2 25.1 19.25
Ocimum sp. x Water stress 40.23 45.23 30.12 29.45
Table.1 Effect of water stress treatments, Ocimum sp. and their interactions
on the fresh and dry weights of Ocimum sp. during both seasons Khalid et al., 2006
11/11/2016
41. 41
Ocimum sp. Water stress
treatments
(%)
Essential oil percentage Total yield of essential oil
(g /plant)
1st season 2nd season 1st season 2nd season
Ocimum
basilicum L.
125 100
75 50
0.36
0.24
0.33
0.38
0.32
0.29
0.31
0.35
0.63
1.04
1.83
1.10
0.6
1.2
1.68
0.97
Average of Ocimum basilicum L. 0.33 0.32 1.15 1.11
Ocimum
americanum L.
125
100
75 50
0.29
0.23
0.25
0.30
0.27
0.21
0.22
0.25
0.68
1.23
1.72
1.25
0.65
1.16
1.61
1.07
Average of Ocimum americanum 0.27 0.24 1.22 1.12
Average of O.
basilicum & O.
americanum
125 100
75
50
0.33
0.24
0.29
0.34
0.03
0.25
0.27
0.30
0.66
1.14
1.78
1.18
0.63
1.18
1.65
1.02
LSD at 0.05
Ocimum sp. 0.0001 0.001 0.01 0.01
Water stress treatments 0.005 0.004 0.03 0.02
Ocimum sp. x Water stress 0.008 0.007 0.04 0.03
Table.2 Effect of water stress treatments, Ocimum sp. and their interactions on the
essential oil content of Ocimum sp. during both seasons Khalid et al., 2006
11/11/2016
42. 42
Table.3 Effect of water stress treatments on the chemical constituents of
essential oil extracted from Ocimum basilicum L. herb during both seasons
Compounds Water stress treatments
Ocimum americanum L.
125% 100% 75% 50%
ᾳ-Pinene 0.5 0.4 0.3 0.2
Camphene 0.2 0.1 0.2 0.3
Sabenene 0.2 0.3 0.2 0.4
ᵦ-Pinene 0.5 1.6 1.8 1.0
Myrcene 1.4 1.3 1.2 1.2
Limonene 1.2 1.3 1.2 1.1
1,8-Cineol 9.4 8.7 8.8 9.9
Ocimene 1.3 1.2 1.4 1.1
Terpinolene 1.1 1.2 1.3 1.1
Linalool 33.4 32.5 32.8 35.6
Camphore 1.6 1.5 1.8 1.9
Methyl-chavicol 35.0 34.0 34.6 34.5
Geraniol 5.8 5.1 5.6 6.0
Eugenol 1.5 2.7 2.6 1.0
Methyl-eugenol 1.5 3.2 3.9 1.0
ᵦ-Caryophyelene 1.8 1.6 1.8 0.7
Germacrene D 1.3 1.0 1.2 1.1
Geranyl iso-butyrate 0.9 1.0 1.1 1.0
Cadinol 1.4 13.0 1.2 0.9
Khalid et al., 2006
11/11/2016
43. 43
Compounds
Water stress treatments
Ocimum americanum L.
125% 100% 75% 50%
ᾳ-Pinene 1.09 1.27 1.07 0.10
ᵦ-Pinene 1.21 1.23 0.34 0.25
Myrcene 0.20 1.30 0.10 0.15
ᾳ-Terpinene 1.12 2.30 0.16 0.22
Limonene 1.00 1.55 0.12 0.12
1,8-Cineol 2.90 2.81 4.10 3.60
Camphore 2.00 1.96 2.30 2.20
Linalool 1.94 1.97 1.90 2.10
Linalyl acetate 1.78 1.85 0.75 0.66
Farnesene 10.60 9.74 13.20 12.40
ᵦ-Bisabolene 4.90 3.75 5.90 5.90
Methyl-chavicol 22.90 20.70 24.90 25.60
Terpneol 15.70 13.90 14.60 14.00
Methyl-eugenol 0.63 3.71 0.90 1.00
Eugenol 29.40 26.10 27.65 29.90
Iso-eugenol 1.74 3.11 1.14 1.00
farnesol 0.89 2.75 0.87 0.80
Table.4 Effect of water stress treatments on the chemical constituents of
essential oil extracted from Ocimum americanum L. herb during both seasons
Khalid et al., 2006
11/11/2016
44. 44
Water stress
treatments
Proline(/mg) N (%) Protein(%)
1st
season
2nd
season
1st season 2nd
season
1st
season
2nd season
Ocimum basilicum 125
100
75
50
6.97 3.80
4.67 6.13
7.27
4.10 5.00
6.43
2.27
2.81
2.51
2.39
2.50
2.93
2.47
2.35
14.19
17.57
15.70
14 .94
15.63
18.31
15.43
14.69
Over all Ocimum basilicum L. 5.40 5.70 2.49 2.60 15.60 16.00
Ocimum americanum L. 125
100
75
50
8.67 4.10
4.20 6.20
8.97
4.40 4.53
6.50
2.09
2.72
3.49
2.31
2.44
2.72
3.57
2.41
13.10
17.00
21.81 14.43
15.25
17.00
22.31
15.07
Over all Ocimum amer.L. 5.80 6.10 2.65 2.79 16.59 17.41
Avg. of O. americanum and
O. basilicum
125
100
75
50
7.80 4.00
4.40 6.20
8.10
4.30 4.80
6.50
2.18
2.77
3.00
2.35
2.47
2.83
3.02
2.38
13.65
17.29
18.76
14.69
15.44
17.66
18.87
14.88
LSD at 0.05
Ocimum sp. 0.14 0.14 0.05 0.03 0.52 0.20
Water stress treatment 0.17 0.17 0.06 0.04 0.63 0.24
Ocimum sp. x water stress 0.18 0.28 0.09 0.07 1.03 0.40
Table.5 Effect of water stress treatments, Ocimum sp. and their interactions
on the chemical composition of Ocimum sp. during both seasons
Khalid et al., 2006
11/11/2016
45. Objectives :
To investigate the effect of different levels of
irrigation water on yields and quality
characteristics of purple basil.
11/11/2016 45
46. Table.6 Effect of different irrigation treatments on
plant height (cm) of Ocimum basilicum L.
Treatment 2007 2008
Average Average
I50 31.8 27.2
I75 33.4 30.8
I100 38.1 32.5
I125 39.2 38.0
LSD 0.05 4.507 6.9
Ekren et al., 2012
11/11/2016 46
47. Table.7 Effect of different irrigation
levels on yields of Ocimum basilicum L.
Treatments
2007 I50 I75 I75 I125 LSD 0.05
The green herb yield (kg/da) 908 1288 1998.3 2399.3 572.81
The drug herb yield(kg/da) 203.13 282.63 401.43 467.70 109.491
The drug leaves yield(kg/da) 131.03 171.03 253.77 308.00 66.723
2008
The green herb yield (kg/da) 841.3 1231.9 1539.0 2139.5 572.81
The drug herb yield(kg/da) 141.50 222.10 276.90 366.67 109.491
The drug leaves yield(kg/da) 112.77 177.67 212.50 293.13 66.723
11/11/2016 47
Ekren et al., 2012
48. Table.8 Effect of different irrigation treatments on
essential oil (%) of Ocimum basilicum L.
Treatments 2007 2008
Average Average
I50 1.00 1.21
I75 0.83 1.14
I100 0.81 1.00
I125 0.94 1.04
LSD (0.05) N.S 0.107
Ekren et al., 2012
11/11/2016 48
49. Table.9 Average plant height, yield, and oil ratio for the study
years.
I50 I75 I75 I125 LSD( 0.05)
Plant height (cm) 29.5 32.1 35.27 38.6 2.294
Green herb yield (kg da−1) 874.7 1260.1 1768.6 2269.4 405.039
Drug herb yield (kg da−1) 172.3 252.4 339.2 417.2 77.422
Drug leaves yield (kg da−1) 121.9 174.4 233.1 300.6 47.18
Essential oil ratio (%) 1.10 0.99 0.91 0.99 0.078
Ekren et al., 2012
11/11/2016 49
50. 11/11/2016 50
Also called as Raubasine
Used in the treatment of
hypertension and circulatory
disorders.
51. 51
Alterations in osmoregulation , antioxidant
enzymes and indole alkaloid levels in
Catharanthus roseus exposed to water deficit
Jaleel et al., 2007
Objectives:
To study effect of water deficit condition
on osmoregulation, antioxidant enzymes
and indole alkaloid levels in catharanthus
roseus.
11/11/2016
52. Jaleel et al., 2007
Fig. 8 Effect of
drought on
(a) Aminoacid
(b) Proline
(c) Glycine betaine
contents in leaves and
roots of Catharanthus
roseus plants.
Values are given as mean ±
S.D. of six samples in each
group. Values are not sharing
a common superscript (a and
b) differ significantly at P≤0.05
(DMRT).
11/11/2016
53. Jaleel et al., 2007
Fig. 9 Effect of drought
on
(a) Glutamyl kinase
and
(b) Proline oxidase
activities in leaves and
roots of C. roseus plants
Values are given as
mean±S.D. of six samples in
each group. Values are not
sharing a common
superscript (a and b) differ
significantly at P≤0.05
(DMRT).
11/11/2016
54. Jaleel et al., 2007
Fig. 10 Effect of drought on total indole alkaloid accumulation
in leaves and roots of C. roseus plants.
Values are given as mean ± S.D. of six samples in each group. Values are not sharing a
common superscript (a and b) differ significantly at P≤0.05 (DMRT).11/11/2016
55. Objectives: To evaluate the drought stress effects on reactive oxygen metabolism
in terms of antioxidant molecules and antioxidant enzymes and total root alkaloid
almalicine content in drought stressed C.roseus
11/11/2016 55
56. Treatment details.
o Control –frequently irrigated till harvest.
o 3 levels of water stress- 10, 15, 20 days by
withholding the irrigation prior to harvest.
11/11/2016 56
57. Fig.11 Effect of drought stress on ajmalicine content of C.
roseus plants.
Values are given as mean±S.D. of six experiments in each group. Values not sharing a
common superscript (a, b) differ significantly at P ≤0.05 (DMRT).
Jaleel et al., 200811/11/2016 57
58. 58
Fig . 12 Effect of drought on (a) H2O2 and (b) LPO contents of
Catharanthus roseus plants.
Jaleel et al., 2008
Values are given as mean ± S.D. of six experiments in each group. Values not
sharing a common superscript (a, b) differ significantly at P≤0.05 (DMRT).11/11/2016
59. Fig.13 Effect of
drought on
Ascorbic Acid(a),
Reduced
Glutathione (b)
and α-
tocopherol (c)
contents of C.
roseus plants.
Values are given as
mean±S.D. of six
experiments in each
group. Values
not sharing a common
superscript (a, b) differ
significantly at P≤0.05
(DMRT).
Jaleel et al., 2008
11/11/2016 59
60. Fig. 14 Effect of
drought on APX
(a), SOD (b) and
CAT (c) activities
of C. roseus
plants.
Values are given as
mean±S.D. of six experiments
in each group. Values
not sharing a common
superscript (a, b) differ
significantly at P≤0.05
(DMRT).
Jaleel et al., 2008
11/11/2016 60
61. ARTEMISININ
o It is having antimalarial activity.
o Used against cancer.
o Used in anti-parasitic drugs.
11/11/2016 61
62. Objectives: To evaluate the effect of water deficit on biomass
and artemisinin accumulation in A. annua.
11/11/2016 62
64. Fig.16 Effect of
different levels of
water deficit on the
dry leaves artemisinin
content (B)and plant
artemisinin content (D)
in A. annua
• The irrigation was stopped
84 days after sowing to the
treatments with water
deficit.
• Means ± SE, n = 6, values
significantly different at
Student t test. * (p = 0.084)
and ** (p = 0.023).
Marchese et al., 201011/11/2016
5.2
64
15
65. Fig.17 Effect of different levels of water deficit on
artemisinin concentration.
Marchese et al., 201011/11/2016 65
29
13.3
66. Table 10. Effect of different levels of water deficit on the
dry stem biomass accumulation and leaf-to-stem ratio.
a- Irrigation was suspended 84 days after sowing; b -Average + SD of 6
replicates; c -Irrigated control; ** Significant (p ≤ 0.022) by the Student t test
Marchese et al., 201011/11/2016 66
1.83±0.55**
67. CONCLUSION
Drought stress affect the growth , development and yield of
many crops in general and medicinal plants in particular .
Even though it reduces the growth and total yield of medicinal
plants but it increases the production of secondary metabolites.
Drought stress increases the Ajmalicine content in Periwinkle ,
Glycirrhizin content in Liquorice, Andrographolide content in
Kalmegh , Artemisinin content in Annual wormwood.
So there is a need of studying relationship between drought and
secondary metabolite biosynthesis.
We can utilize the drought prone areas for the production of
medicinal plants.
11/11/2016 67
