Influence of soil water management on plant growth, essential oil yield and oil

composition of rose-scented geranium (Pelargonium spp.)

by

Bahlebi Kibreab Eiasu

Submitted in partial fulfilment of the requirements for the degree

Doctor of Philosophy: Horticultural Science

Department of Plant Production and Soil Science

In the Faculty of Natural and Agricultural Sciences
University of Pretoria
Pretoria

Supervisor: Dr J.M. Steyn

Co-supervisor: Prof. P. Soundy

March 2009

University of Pretoria
 TABLE OF CONTENTS

Page

LIST OF TABLES. vii

LIST OF FIGURES... ix

LIST OF SYMBOLS AND ABBREVATIONS....... xiii

ACKNOWLEDGEMENTS....... xv

ABSTRACT.... xvi

CHAPTER 1
GENERAL INTRODUCTION. 1

CHPTER 2
LITERATURE REVIEW..
5
2.1 ESSENTIAL OILS.. 5

2.1.1 Nature and composition of essential oils. 5

2.1.2 Importance of essential oils to the host plant. 6

2.1.3 Role of essential oils in history and at present... 7

2.2 ROSE-SCENTED GERANIUM (PELARGONIUM SPECIES)... 9

2.2.1 Botany and origin... 9

2.2.2 Uses of geranium oil... 10

2.2.3 History of Pelargonium species cultivation for oil production... 11

2.2.4 Geranium oil yield and composition. 11

2.3 IRRIGATION.... 18

2.3.1 History of irrigation... 18

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 2.3.2 Contribution of irrigated agriculture... 19

2.3.3 Freshwater scarcity.... 20

2.3.4 Coping with freshwater scarcity in agriculture.. 21

CHAPTER 3
GROWTH, ESSENTIAL OIL YIELD AND OIL COMPOSITION OF
ROSE-SCENTED GERANIUM GROWN AT DIFFERENT MAXIMUM
ALLOWABLE SOIL WATER DEPLETION LEVELS.......... 25

3.1 ABSTRACT..... 25

3.2 INTRODUCTION... 26

3.3 MATERIALS AND METHODS.... 28

3.3.1 Study area and duration..... 28

3.3.2 Plant culture.. 28

3.3.3 Field layout and treatments........ 29

3.3.4 Irrigation monitoring.. 30

3.3.5 Agronomic practices.... 33

3.3.6 Data recorded...... 33

3.4 RESULTS AND DISCUSSION...... 35

3.4.1 Soil water depletion patterns.. 35

3.4.2 Plant growth parameters.... 41

3.4.3 Essential oil yield and quality parameters............. 47

3.4.4 Water use and water-use efficiency.... 50

3.5 CONCLUSIONS AND RECOMMENDATIONS. 53

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CHAPTER 4
RESPONSE OF ROSE-SCENTED GERANIM GROWTH, ESSENTIAL
OIL YIELD AND OIL COMPOSITION TO A ONE-MONTHIRRIGATION-
WITHOLDING PERIOD... 54

4.1 ABSTRACT... 54

4.2 INTRODUCTION................. 55

4.3 MATERIALS AND METHODS. 56

4.3.1 Site description... 56

4.3.2 Plant culture... 57

4.3.3 Field layout and treatments.......... 57

4.3.4 Irrigation monitoring......... 58

4.3.5 Agronomic practices...... 59

4.3.6 Data recorded. 60

4.4 RESULTS AND DISCUSSION... 60

4.4.1 Soil water content during the irrigation-withholding periods... 60

4.4.2 Herbage growth parameters..... 62

4.4.3 Essential oil yield and quality parameters... 68

4.4.4 Water use and water use-efficiency (WUE). 73

4.5 CONCLUSIONS AND RECOMMENDATIONS.......... 76

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CHAPTER 5
RESPONSE OF ROSE-SCENTED GERANIUM GROWTH, ESSENTIAL
OIL YIELD AND OIL COMPOSITION TO IRRIGATION FREQUENCY
AND A TERMINAL ONE-WEEK WATER-WITHHOLDING PERIOD..
77

5.1 ABSTRACT.... 77

5.2 INTRODUCTION. 78

5.3 MATERIALS AND METHODS. 79

5.3.1 Growth system description 79

5.3.2 Plant culture. . 79

5.3.3 Treatments and experimental design... 80

5.3.4 Irrigation monitoring. 81

5.3.5 Fertiliser application.. 82

5.3.6 Data recorded. 82

5.4 RESULTS AND DISCUSSION.... 83

5.4.1 Effects of irrigation frequency on herbage yield in the tunnel trials. 83

5.4.2 Effects of irrigation frequency on essential oil yield components in the

tunnel trials. 84
5.4.3 Effects of irrigation frequency and withholding irrigation on herbage
yield parameters in the tunnel trials.... 86
5.4.4 Response of essential oil yield parameters to irrigation frequency,
growing medium and withholding irrigation in the tunnel trials.. 88
5.4.5 Response of herbage yield parameters to irrigation levels in the
glasshouse trials. 91
5.4.6 Response of essential oil yield parameters to irrigation levels in the
glasshouse trials.. 92
5.4.7 Water use and water-use efficiency (WUE). 95

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5.5 CONCLUSIONS.... 97

CHAPTER 6
RESPONSE OF ROSE-SCENTED GERANUIM LEAF PHYSIOLOGYAND
MORPHOLOGY TO IRRIGATION FREQUENCY..................... 98

6.1 ABSTRACT.... 98

6.2 INTRODUCTION. 99

6.3 MATERIALS AND METHODS.. 100

6.3.1 Data recorded. 101

6.4 RESULTS AND DISCUSSION................ 103

6.4.1 Leaf physiological response to water stress. 103

6.4.2 Leaf morphological response to water stress....... 108

6.5 CONCLUSIONS.... 117

CHAPTER 7
GENERAL DISCUSSION. 118

REFERENCES 124

APPENDIX A
SOIL AND WEATHER DATA FOR THE EXPERIMENTAL SITES 146

APPENDIX B
NEUTRON PROBE CALIBRATION PROCESSES. 151

APPENDIX C
SUMMARISED ANALYSIS OF VARIANCE (ANOVA) TABLES..... 155

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 LIST OF TABLES

Page

Table 3.1: Regression equations used to determine soil water content. 31

Table 3.2: Days of regrowth cycle and amount of irrigation applied to the
different maximum allowable soil water depletion treatments in the
open field... 36
Table 3.3: Days of regrowth cycle and amount of irrigation applied to the
different maximum allowable soil water depletion treatments in the
rain shelter.... 37
Table 3.4: Percentage available soil water depleted and amount of irrigation
applied (mm) per soil layer for the different maximum allowable soil
water depletion levels....... 40
Table 3.5: Leaf area index of rose-scented geranium grown under different
maximum allowable depletion levels (MAD) of plant available soil
water for the final harvests...... 42
Table 3.6: Fresh leaf mass ratio (as a percentage of total fresh herbage mass) for
rose-scented geranium grown at different maximum allowable
depletion levels (MAD) of plant available soil water recorded at final
harvests.. 43
Table 3.7: Leaf and stem dry matter contents (%) of rose-scented geranium
grown at different maximum allowable depletion levels (MAD) of
plant available soil water recorded at final harvests. 45
Table 3.8: Average water use and water-use efficiency (expressed on essential oil
yield and herbage dry mass basis) of rose-scented geranium grown at
different maximum allowable depletion levels of plant available soil
water.............. 51
Table 4.1: Maximum LAI of rose-scented geranium that was water-stressed for
one month at different regrowth stages.. 63

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Table 4.2: Dry matter content (percentage) of rose-scented geranium grown
under one-month at different regrowth stages.. 65

Table 4.3: Fresh leaf mass to total fresh biomass ratio (%) of rose-scented
geranium that was water-stressed for one-month at different regrowth
stages.. 68

Table 4.4: Total irrigation applied and amount of water used by rose-scented
geranium that was water-stressed at different regrowth stages.. 74

Table 5.1: Fresh herbage yield of rose-scented geranium grown under different
irrigation frequencies in the tunnel. 83

Table 5.2: Essential oil yield of rose-scented geranium as affected by irrigation

frequency in the tunnel. 85

Table 5.3: Water use and water-use efficiency (on essential oil yield basis) of
rose-scented geranium grown under different irrigation frequencies
and a one-week irrigation-withholding period.. 96
Table 6.1: Non-glandular trichome and stomatal density from leaves of rose-
scented geranium grown under different irrigation frequencies. 113

Table 6.2: Glandular trichome density on the abaxial and adaxial surfaces of
rose-scented geranium leaves.. 115

Table 6.3: Response of petiole and leaf length to irrigation frequency. 116

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 LIST OF FIGURES

Page

Figure 3.1: Temporal variations in plant available soil water (ASW) content in
the layers of the root zone of rose-scented geranium in the open-
field trial.. 38
Figure 3.2: Temporal variations in plant available soil water (ASW) content in
the layers of the root zone of rose-scented geranium in the rain-
shelter trial.. 39
Figure 3.3: Leaf area index of rose-scented geranium as affected by different
maximum allowable depletion (MAD) levels of plant available soil
water 41
Figure 3.4: Herbage dry mass of rose-scented geranium grown at different
maximum allowable depletion (MAD) levels of plant available soil
water 44
Figure 3.5: Fresh herbage yield of rose-scented geranium grown at different
maximum allowable depletion (MAD) levels of plant available soil
water 46
Figure 3.6: Essential oil content of rose-scented geranium grown at different
maximum allowable depletion (MAD) levels of plant available soil
water 47
Figure 3.7: Essential oil yield of rose-scented geranium grown at different
maximum allowable depletion (MAD) levels of plant available soil
water 48
Figure 3.8: Major essential oil components (percentage of essential oil yield) of
rose-scented geranium grown under different maximum allowable
depletion levels of plant available soil water 50

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Figure 4.1: Available soil water content per soil layer in the root zone of rose-
scented geranium during the one-month irrigation-withholding
periods.. 61
Figure 4.2: Leaf area index growth trends of rose-scented geranium that was
water-stressed for one month at different regrowth stages............ 62
Figure 4.3: Dry matter accumulation of rose-scented geranium that was water-
stressed for one month at different regrowth stages... 64
Figure 4.4: Fresh herbage yield of rose-scented geranium that was water-
stressed for one month at different regrowth stages... 66
Figure 4.5: Essential oil content (% oil on fresh herbage mass basis) of rose-
scented geranium that was water-stressed for one month at
different regrowth stages... 69
Figure 4.6: Essential oil yield (kg/ha) of rose-scented geranium that was water-
stressed for one month at different regrowth stages... 70
Figure 4.7: Essential oil composition of rose-scented geranium that was water-
stressed for one month at different regrowth stages... 72
Figure 4.8: Water-use efficiency (WUE) (kg/ha/mm) of rose-scented geranium
that was water-stressed for one month at different regrowth stages. 75
Figure 5.1: Plant arrangements in the tunnel trial. 81
Figure 5.2: Drainage-collecting container in the glasshouse pot trials.. 82
Figure 5.3: Essential oil content of rose-scented geranium as affected by
irrigation frequency in the tunnel. 85
Figure 5.4: Rose-scented geranium fresh (A) and dry (B) herbage mass as
affected by irrigation frequency and a one-week stress period
before harvest in the tunnel... 87
Figure 5.5: Rose-scented geranium oil content (percentage oil on fresh herbage
mass basis) as affected by irrigation frequency and a one-week
stress period before harvest in two growing media in the tunnel.. 88

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Figure 5.6: Essential oil yield of rose-scented geranium as affected by
irrigation frequency, a one-week stress period before harvest and
growing media in the tunnel.. 89
Figure 5.7: Chemical composition (%) of essential oil of rose-scented geranium
as affected by irrigation frequencies and a one-week stress period
in the tunnel. 90
Figure 5.8: Fresh (A) and dry (B) herbage mass of rose-scented geranium as
affected by irrigation frequency and a one-week irrigation-
withholding period in the glasshouse 91
Figure 5.9: Effect of irrigation frequency on essential oil yield of rose-scented
geranium in the glasshouse 92
Figure 5.10: Essential oil content (percentage oil on fresh herbage mass basis) of
rose-scented geranium as affected by irrigation frequency and one-
week irrigation-withholding treatments... 93
Figure 5.11: Essential oil yield of rose-scented geranium grown in different
irrigation frequencies and a one-week water stress period 94
Figure 5.12: Composition (percentage of essential oil) of rose-scented geranium
as affected by irrigation frequency and irrigation withholding for
the week prior to harvesting treatments in the glasshouse. 95
Figure 6.1: Effect of irrigation frequency on stomatal conductance of rose-
scented geranium recorded during a one-week irrigation-
withholding period.. 103
Figure 6.2: Effect of irrigation frequency on transpiration rate of rose-scented
geranium leaves recorded during a one-week irrigation-
withholding period.. 105
Figure 6.3: Rose-scented geranium canopy size as affected by irrigation
frequency. 105
Figure 6.4: Effect of irrigation frequency on relative water content of rose-
scented geranium leaves observed during a one-week irrigation-
withholding period.. 106

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Figure 6.5: After-effect of irrigation frequency on leaf water potential of rose-
scented geranium leaves recorded during a one-week irrigation-
withholding period.. 107
Figure 6.6: Large (L) and small (S) glandular trichomes on a leaf surface
viewed under a scanning electron microscope. 109
Figure 6.7: Leaf trichomes observed under a light microscope. 109
Figure 6.8: Morphology of glandular trichomes observed under a scanning
electron microscope 111
Figure 6.9: Glandular trichomes and stomata in immature leaves (A, B and C)
and in mature leaves (D and E). 112
Figure 6.10: Effect of irrigation frequency on stomatal opening observed under
a scanning electron microscope. 114
Figure 6.11: Leaf size of rose-scented geranium as affected by irrigation
frequency. 116

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LIST OF SYMBOLS AND ABBREVIATIONS

A Plot area
ASW Available soil water
cv. Cultivar
D Water deep percolation
DM Dry mass
ET Evapotranspiration
FAO Food and Agricultural Organization of the United Nations (Rome, Italy)
FM Fresh mass
GC Gas chromatography
G:C ratio Geraniol to citronellol ratio
Gs Stomatal conductance
H LA Harvested land area
I Irritation water applied
IW:CPE ratio Irrigation water to cumulative pan evaporation ratio
Litre
LAI Leaf area index
LA (LA) Leaf area
MAD Maximum allowable depletion
n Number of soil layers
P Rainfall (precipitation)
PRD Partial root zone drying
R Water runoff
RA Neutron probe reading in air
RS Neutron probe reading in soil
Rt Transpiration rate
RWC Relative water content
Rz Plant root zone
SANDA South African National Department of Agriculture
SFE Supper fluid extraction
S1 Initial soil water content of a cropping season (regrowth period)
S2 Final soil water content of a cropping season (regrowth period)

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THRIP Technology and Human Resources for Industry Programme (South Africa)
UIDEA Ugandas Investment in Developing Export Agriculture (Uganda)
VI Volume of irrigation water
WUE Water use efficiency

d Depleted water
FCi Volumetric soil water content at field capacity
i Measured volumetric soil water content
PWPi Volumetric soil water content at plant permanent wilting point
W Water potential

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 ACKNOWLEDGEMENTS

I would like to thank the following people and institutions:

My supervisor, Dr J.M. Steyn, for his valuable advice, readiness for consultation and
much-needed technical support.
My co-supervisor Prof. P. Soundy for his guidance, inspiration and encouragement. I
enjoyed his generous grooming for seven years.
The Technology and Human Resources for Industry Programme (THRIP), Biosys
Plant Extracts (Pty) Ltd, and Clive Teubes CC for funding the research.
Biosys Plant Extracts (Pty) Ltd staff members, particularly Mr R.A. Mojela, for
conducting and/or co-ordinating the oil distillation and GC analysis.
All my friends and colleagues in Pretoria, including Michael Gebre-Meskel, Melake
Kesete, Musie Bokurezion and Yamane Manna (Wedi-Mann), for their unreserved
support, sharing of knowledge and advice: their wonderful company made me feel at
home.
The managers, technicians and daily labourers at the Hatfield Experimental Farm of
the University of Pretoria for their technical assistance.
My wife, Rosa Beraki, for her encouragement, support and understanding: she was
always there for me.
My beloved parents, who offered me the opportunity of going to school, which they
themselves never had.
My aunts, Milashu Eyasu and Ghiday Eyasu, who brought me up as if I was their own
child: without their being there for me, my attending school would have been hard.

I am also grateful to the Almighty God, who is the ultimate guide of this work and my life as a
whole. I feel guilty remembering those countless days and nights I spent worrying about the
staggered path of worldly life and apparently dark futures ahead of me, forgetting that the
perfect guide has always been with me.

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 INFLUENCE OF SOIL WATER MANAGEMENT ON PLANT GROWTH,
ESSENTIAL OIL YIELD AND OIL COMPOSITION OF ROSE-SCENTED
GERANIUM (PELARGONIUM SPP.)
by

Bahlebi Kibreab Eiasu

SUPERVISOR: Dr J.M. Steyn

CO-SUPERVISOR: Prof. P. Soundy

DEGREE: PhD
ABSTRACT

Introducing effective irrigation management in arid and semi-arid regions, like most areas of
South Africa, is an indispensable way of maximising crop yield and enhancing productivity of
scarce freshwater resources. Holistic improvements in agricultural water management could be
realised through integrating the knowledge of crop-specific water requirements. In order to
develop effective irrigation schedules for rose-scented geranium (Pelargonium capitatum x P.
radens), greenhouse and field experiments were conducted at the Hatfield Experimental Farm of
the University of Pretoria, Pretoria, South Africa, from 28 October 2004 to 2006.

Results from 20, 40, 60 and 80% maximum allowable depletion (MAD) levels of the plant
available soil water (ASW) indicated that plant roots extracted most of the soil water from the top
40 cm soil layer, independent of the treatment. Both essential oil yield and fresh herbage mass
responded positively to high soil water content. Increasing the MAD level to 60% and higher
resulted in a significant reduction in herbage mass and essential oil yields. An increase in the
degree of water stress apparently increased the essential oil concentration (percentage oil on fresh
herbage mass basis), but its contribution to total essential oil yield (kg/ha oil) was limited. There
was no significant relationship between MAD level and essential oil composition. For water
saving without a significant reduction in essential oil yield of rose-scented geranium, a MAD of
40% of ASW is proposed.

xvi
Response of rose-scented geranium to a one-month irrigation withholding period in the second or
third month of regrowth cycles showed that herbage mass and oil yield were positively related.
Herbage yield was significantly reduced when the water stress period was imposed during the
third or fourth month of regrowth. A remarkable essential oil yield loss was observed only when
the plants were stressed during the fourth month of regrowth. Essential oil content (% oil on fresh
herbage mass basis) was higher in stressed plants, especially when stressed late, but oil yield
dropped due to lower herbage mass. The relationship between essential oil composition and
irrigation treatments was not consistent. Water-use efficiency was not significantly affected by
withholding irrigation in the second or in the third month of regrowth. With a marginal oil yield
loss, about 330 to 460 m3 of water per hectare per regrowth cycle could be saved by withholding
irrigation during the third month of regrowth. The overall results highlighted that in water-scarce
regions withholding irrigation during either the second or the third month of regrowth in rose-
scented geranium could save water that could be used by other sectors of society.

In greenhouse pot experiments, rose-scented geranium was grown under different irrigation
frequencies, in two growth media. Irrigation was withheld on 50% of the plants (in each plot) for
the week prior to harvesting. Herbage and essential oil yields were better in the sandy clay soil
than in silica sand. Essential oil content (% oil on fresh herbage mass basis) apparently increased
with a decrease in irrigation frequency. Both herbage and total essential oil yields positively
responded to frequent irrigation. A one-week stress period prior to harvesting significantly
increased essential oil content and total essential oil yield. Hence, the highest essential oil yield
was obtained from a combination of high irrigation frequency and a one-week irrigation-
withholding period. In the irrigation frequency treatments, citronellol and citronellyl formate
contents tended to increase with an increase in the stress level, but the reverse was true for
geraniol and geranyl formate.

Leaf physiological data were recorded during the terminal one-week water stress in the
glasshouse pot trial. Upon rewatering, stomatal conductance (Gs) and transpiration rate (Rt) were
significantly lower in the less often irrigated than in the more often irrigated treatments, while
leaf water potential (w) and relative water content (RWC) were the same for all plants,
indicating that water stress had an after-effect on Gs and Rt. At the end of the stress period, Gs, Rt,

xvii
w and RWC were lower in the plants from the more often irrigated than from the less often
irrigated treatments. Irrespective of irrigation treatment, one type of non-glandular and two types
(different in shape and size) of glandular trichomes were observed. In water stressed-conditions,
stomata and trichome densities increased, while the total number of stomata and trichomes per
leaf appeared to remain more or less the same. Water stress conditions resulted in stomatal
closure.

Keywords: Citronellol; citronellyl formate; essential oil content; geraniol; geranyl formate;
herbage yield; irrigation-withholding period; maximum allowable depletion level; relative water
content; stomatal conductance; transpiration rate; trichomes; water potential; water stress; water
use; water-use efficiency

xviii