Monringa

“EXPLORING THE POTENTIAL OF MORINGA OLEIFERA L.
AS AN OIL SEED TREE”
MUHAMMAD IDREES FAISAL

M.Sc. (Hons.)Agronomy
2006-ag-1875
A thesis submitted in partial fulfillment of requirements for the degree of
DOCTOR OF PHILOSPHY
IN
CROP PHYSIOLOGY
DEPARTMENT OF AGRONOMY
FACULTY OF AGRICULTRE,
UNIVERSITY OF AGRICULTURE, FAISALABAD, PAKISTAN
2017
i
To,
The Controller of Examinations,
University of Agriculture,
Faisalabad.
We, the Supervisory Committee, certify that the contents and form of thesis submitted by
Mr. Muhammad Idrees Faisal, Regd. No. 2006-ag-1875 have been found satisfactory and
recommend that it may be processed for evaluation of External Examiner (s) for the award of
degree.
SUPERVISORY COMMITTEE
Chairman _________________________
Dr. Shahzad Maqsood Ahmed Basra
Member _________________________
Dr. Irfan Afzal
Member _________________________
Dr. Abdul Wahid
ii
Declaration
I hereby declare that the contents of the thesis, “Exploring the potential of Moringa oleifera L.
as an oil seed tree’’ are product of my own research and no part has been copied from any
published source (except the references, standard mathematical and genetic models/ equations/
formulae/ protocols etc.). I further declare that this work has not been submitted for award of any
other diploma/degree. The University may take action if information provided is found inaccurate
at any stage. (In case of any default the scholar will be proceeded against per HEC plagiarism
policy).
Muhammad Idrees Faisal

2006-ag-1875
iii
DEDICATED
TO
LOVE OF MY LIFE
"AMA G”
Today whatever I
achieved in my life is
because of Allah
blessing and your
prayers.
iv
ACKNOWLDGEMENTS
All worships and praises are only due to the Lord of creation, the most beneficent,
merciful and compassionate, Whose blessings and exaltation flourished my thoughts
and thrived my ambitions to have the cherish fruit of my modest effort in the form
of this manuscript.
I offer my humblest thanks and countless salutations to the Holy Prophet
Muhammad (PBUH), who is forever, a torch of guidance for the entire humanity.
I owe my deepest gratitude to my great supervisor Dr. Shahzad M.A Basra,
Professor, Department of Agronomy, University of Agriculture Faisalabad, who in
spite of his busiest tiring routine work provided his dexterous and valuable
suggestions throughout research efforts.
Thanks are extended to the members of my supervisory committee Dr. Irfan Afzal,
Associate Professor Department of Agronomy and Dr. Abdul Wahid, Professor,
Department of Botany, University of Agriculture Faisalabad for their sincere
cooperation and invigorating encouragement during the course of present
investigation. I am very grateful to Dr. Selina Wang, Research Director, UC Davis
Olive Center for providing all necessary resources during my stay in USA. I am also
very thankful to my dear fellows Muhamad Sohail Saddiq and Shahid Iqbal
Department of Agronomy, University of Agriculture Faisalabad for their valuable
suggestions and guidance during my research activities and thesis write up. I am also
grateful to Uncle Muhammad Iqbal and his family for providing all necessary
resources during my experiment at Khanewal.
I can’t forget prayers of my beloved mother and great support of my brothers
(Abdul Wahab and Muhammad Ismail) and sisters for the strenuous efforts done
by them in enabling me to join the higher ideals of life and also their financial and
moral support, patience and prayers they had made for my success.
Muhammad Idrees Faisal
v
TABLE OF CONSENTS
1. Introduction 1
2. Review of Literature 5
2.1 Origin and distribution 5
2.2 Morphology and physical characters 5
2.2.1 Stem 5
2.2.2 Branch 6
2.2.3 Leaves and young shoots 6
2.2.4 Flowers, fruits and seeds 6
2.2.5 Wood and bark 6
2.2.6 Root type 6
2.3 Genetic diversity and taxonomy 7
2.4 Growth and development 7
2.5 Cultivation and production 7
2.6 Genetic and breeding 8
2.7 Ecology 9
2.8 Insect, pests and pathogens 10
2.9 Nutritional value of moringa 10
2.10 Economic importance 10
2.10.1 Wood 11
2.10.2 Human consumption 11
2.10.3 Water purification 11
2.10.4 Medicinal uses 12
2.10.5 Moringa as fodder for livestock 12
2.10.6 Crop growth enhancer 13
2.11 Moringa seeds oil (Ben oil) 13
vi
2.12 Effect of climate on seed production 14
2.13 Climatic factors and seed composition 15
2.14 Seed composition 15
2.15 Physio-chemical characteristics of moringa oil 16
2.16 Oxidative stability of oil 16
2.17 Fatty acid composition 17
2.18 Sterol composition 17
2.19 Moringa oil benefits 18
2.19.1 Health benefits 18
2.19.2 Medicinal and nutraceutical action 18
2.19.3 Biodiesel production 18
2.19.4 Cosmetic industry 19
3 Materials and Methods 20
3.1 Experiment duration 20
3.2 Wild tree selection 20
3.3 Pod harvest and seed yield data collection 20
3.4 Oil extraction 20
3.5 Degumming of oils 21
3.6 Experiment 1: Exploring the potential of wild grown Moringa oleifera as oil 21
seed tree at different locations of Punjab
3.6.1 Pod length (cm) 21
3.6.2 Pod weight (g) 21
3.6.3 Number of seeds per pod 21
3.6.4 Seed weight per pod (g) 22
3.6.5 Number of pods per tree 22
3.6.6 Seed weight per tree (kg) 22
vii
3.6.7 1000 seeds weight (g) 22
3.6.8 Refractive Index 22
3.6.9 Saponification value 23
3.6.10 Iodine Value 24
3.6.11 Free fatty acid 25
3.6.12 Peroxide value 27
3.6.13 p-anisidine value 28
3.6.14 Specific extension of oil 29
3.6.15 Fatty Acid Profile (FAP) 30
3.6.16 Sterol Composition 31
3.7 Experiment II: Exploring the potential of vegetatively propagated Moringa 35

oleifera as an oil seed tree
3.7.1 Experimental Detail 35
3.7.2 Stem cutting propagation: 35
3.7.3 Soil analysis: 36
3.7.4 Pod harvest and seed yield data collection: 36
3.7.5 Oil extraction: 36
3.7.6 Degumming of oils: 37
3.7.7 Average pod weight (g) 37
3.7.8 Average pod length (cm) 37
3.7.9 Average number of seeds per pod 37
3.7.10 Average seed weight per pod (g) 37
3.7.12 Average seed weight per tree (kg) 37
3.7.13 1000 seeds weight (g) 37
3.7.14 Refractive index 37
viii
3.7.15 Saponification Value 37
3.7.16 Iodine Value 38
3.7.17 Free Fatty Acids 38
3.7.18 Peroxide Value 38
3.7.19 p-Anisidine Value 38
3.7.20 Specific extinctions 38
3.7.21 Fatty Acid Profile (FAP) 38
3.7.22 Sterol Composition 38
3.8 Statistical Analysis 38
4 Results and Discussion 39
4.1 Experiment 1 39
4.1.1 Pod length 39
4.1.2 Pod weight 39
4.1.4 Seed weight per pod 39
4.1.6 Seeds weight per tree 40
4.1.7 1000 seeds weight 40
4.1.8 Oil yield per tree 40
4.1.9 Seed oil content 40
4.1.12 Iodine value 41
ix
4.1.17 Fatty acids composition 42
4.1.18 Sterols Composition 42
Discussion 42
4.2 Experiment 2 61
4.2.1 Pod length 61
4.2.2 Pod weight 61
4.2.4 Seed weight per pod 61
4.2.6 Seeds weight per tree 62
4.2.7 1000 seeds weight 62
4.2.8 Oil yield per tree 62
4.2.9 Oil yield per hectare 62
4.2.10 Seed oil content 62
4.2.13 Iodine value 63
4.2.18 Fatty acids composition 64
4.2.19 Sterols Composition 64
Discussion 64
Summary 83
x
Literature Cited 85
xi
LIST OF TABLES
Table Page
2.1 Chemical composition of Moringa oleifera (g/100 g of dry weight) 16
3.1 Physio-chemical characteristics of soil 36

4.1 Mean sum of squares of data of pod length, pod weight, number of seeds pod-1 45
and Seed weight pod-1 of wild moringa landraces during year 2014
4.2 Mean sum of squares of data of pod length, pod weight, number of seeds pod -1 45
4.3 Mean sum of squares of data of number of pods tree-1, seed weight tree-1, 1000 46
seeds weight, oil yield tree -1 and oil yield hectare-1 of wild moringa landraces
during year 2014
4.4 Mean sum of squares of data of number of pods tree -1, seed weight tree-1, 1000 46
seeds weight, oil yield tree-1 and oil yield hectare-1 of wild moringa landraces
during year 2015
4.5 Mean sum of squares of data of oil content, refractive index, saponification 47
value, iodine value and free fatty acids of wild moringa landraces during year
2014
2015
4.7 Mean sum of squares of data of oil peroxide value, p-anisdine value, K232, and 49
K268 of wild moringa landraces during year 2014
and seed weight pod-1 of cultivated moringa landraces during year 2014
and seed weight pod-1 of cultivated moringa landraces during year 2015
during year 2014
during year 2015
xii
2014
2015
4.16 Mean sum of squares of data of oil peroxide value, p-anisdine value, K232, 71
and K268 of wild moringa landraces during year 2015
4.1.1 Effect of different landraces on pod length (cm) of wild grown moringa trees 50
4.1.2 Effect of different landraces on pod weight (g) of wild grown moringa trees 50
4.1.3 Effect of different landraces on number of seeds per pod of wild grown moringa 51
trees
4.1.4 Effect of different landraces on seeds weight per pod (g) of wild grown moringa 51
trees
4.1.5 Effect of different landraces on number of pods per tree of wild grown moringa 52
trees
4.1.6 Effect of different landraces on seeds weight per tree (kg) of wild grown 52
moringa trees
4.1.7 Effect of different landraces on 1000 seeds weight (g) of wild grown moringa 53
trees
4.1.8 Effect of different landraces on oil yield per tree (kg) of wild grown moringa 53
trees
4.1.9 Effect of different landraces on seed oil content (%) of wild grown moringa 54
trees
4.1.10 Effect of different landraces on refractive index (at 40°C) of wild grown 54
moringa trees
4.1.11 Effect of different landraces on saponification value (mg of KOH/g of oil) of 55
wild grown moringa trees
4.1.12 Effect of different landraces on iodine value (g of I/100 g of oil) of wild grown 55
moringa trees
4.1.13 Effect of different landraces on free fatty acid (% oleic acid) of wild grown 56
moringa trees
4.1.14 Effect of different landraces on peroxide value (meq of O 2 kg-1 of oil) of wild 56
grown moringa trees
xiii
4.1.15 Effect of different landraces on p-anisidine value of wild grown moringa trees 57
4.1.16 Effect of different landraces on K232(K1%

1cm ) of wild grown moringa trees 57
4.1.17 Effect of different landraces on K232(K1%

1cm ) of wild grown moringa trees 58
4.1.18 Fatty acids composition (grams per 100 g of Fatty Acids) of wild grown moringa 59
tree landraces in selected locations of Punjab
4.1.19 Sterol composition (%) of wild grown moringa tree landraces in selected 60
locations of Punjab
4.2.1. Effect of different landraces on pod length (cm) of cultivated moringa trees 72
4.2.2. Effect of different landraces on pod weight (g) of wild grown moringa trees 72
4.2.3. Effect of different landraces on number of seeds per pod of cultivated moringa 73
trees
4.2.4. Effect of different landraces on seeds weight per pod (g) of cultivated moringa 73
trees
4.2.5. Effect of different landraces on number of pods per tree of cultivated moringa 74
trees
4.2.6. Effect of different landraces on seeds weight per tree (kg) of cultivated moringa 74
trees
4.2.7. Effect of different landraces on 1000 seeds weight (g) of cultivated moringa 75
trees
4.2.8. Effect of different landraces on oil yield per tree (kg) of cultivated moringa trees 75
4.2.9. Effect of different landraces on oil yield ha -1 (kg) of cultivated moringa trees 76
4.2.10. Effect of different landraces on seed oil content (%) of cultivated moringa trees 76
4.2.11. Effect of different landraces on refractive index (at 40°C) of cultivated moringa 77
trees
4.2.12. Effect of different landraces on saponification value (mg of KOH/g of oil) of 77
cultivated moringa trees
4.2.13. Effect of different landraces on iodine value (g of I/100 g of oil) of cultivated 78
moringa trees
4.2.14. Effect of different landraces on free fatty acid (% oleic acid) of cultivated 78
moringa trees
4.2.15. Effect of different landraces on peroxide value (meq of O2 kg-1 of oil) of 79
xiv
4.2.16. Effect of different landraces on p-anisidine value of cultivated moringa trees 79
4.2.17. Effect of different landraces on K232(K1%
1cm ) of cultivated moringa trees 80
4.2.18. Effect of different landraces on K232(K1%

1cm ) of cultivated moringa trees 80
4.2.19. Fatty acids composition (grams per 100 g of Fatty Acids) of cultivated moringa 81
tree landraces in selected locations of Punjab
4.2.20. Sterol composition (%) of cultivated moringa tree landraces in selected 82
locations of Punjab
xv
LIST OF FIGURES
Figure Page
2.1 Uses of different parts of Moringa 10
2.2 White and brown seeds of Moringa 14
xvi
ABBREVIATIONS
Abbreviations Full
FSD Faisalabad
RYK Rahim Yar Khan
BWP Bahawalpur
LAY Layyah
MUL Multan
KWL Khanewal
SFA Saturated Fatty acid
MUFA Monounsaturated Fatty acid
% Percent
mL Milliliter
cm centimeter (s)
g gram (s)
ha-1 per hectare
kg Kilogram
mM milli Molar
xvii
Abstract
This study was conducted to determine seed yield, qualitative and quantitative potential of seed
oil of wild and vegetatively cultivated moringa trees. Six locations in Punjab (Faisalabad,
Rahimyar Khan, Bahawalpur, Layyah, Mulan and Khanewal) were selected having wild grown
moringa. Data regarding seed and oil yield and quality were taken for two growing seasons (2014
and 2015). Results showed that wild trees of RYK landrace performed best among all landraces
and produced highest seed yield (4.11 and 3.77 kg/tree) during both harvests. Maximum seed oil
content (35.95 and 37.02%) and oil yield per tree (1.48 and 1.40 kg/tree) were also observed in
RYK landrace during both growing seasons. However, oil physio-chemical characteristics,
oxidative state, fatty acid and sterol composition were almost similar in all selected landraces.
However, peroxide and p-anisidine values of seed oil of selected landraces were ranged between
0.95-1.68 meq of O2 kg-1 and 3.27-5.07 respectively during both years. Oil of all landraces was
found to contain high level of monounsaturated fatty acid (oleic acid >71%) followed by palmitic,
stearic and behenic acid. β-sitosterol, campesterol, stigmasterol and ∆5 -Avenasterol collectively
contributed more than 90% in sterol composition of selected landraces. β-sitosterol was
predominant plant sterol (>55%) in oil of all landraces. All landraces propagated through stem
cuttings at a farmer’s field, Khanewal produced flowers and pods within one year of plantation
except FSD however, during second year all landraces produced seeds. Seeds and oil yield of RYK
landrace were highest among other landraces during both years of study. RYK trees produced on
average 0.49 kg oil/tree and 232 kg oil/ha during both growing seasons. However, seed oil content,
oil physio-chemical characteristics, oxidative stability, fatty acid and sterol composition had little
variability among all landraces. Monounsaturated fatty acid (oleic acid) was main fatty acid in oil
of all landraces followed by palmitic, stearic and behenic acid. Oleic acid contributes more than
71% in total fatty acids. Four sterol compounds (β-sitosterol campesterol, stigmasterol and ∆5 -
Avenasterol) contributed more than 90% to sterol composition of all landraces oil. In conclusion,
RYK landrace wild and propagated trees through stem cuttings produced highest seed and oil yield
among all selected landraces during both year of study. However, seed oil quality of all selected
landraces was similar. On basis of finding, it is concluded that RYK landrace has potential to
propagate for oil seed tree in Pakistan.
xviii
1. Chapter 1
Introduction
Agriculture is backbone of the Pakistan’s economy as it provides jobs to about half of the
populace and provides raw input to agriculture based industries. Agricultural income has
created demand for industrial products. Agriculture provides main force to economic growth
by creating additional demand of goods and services as a result of higher prices of agricultural
produce.
One of the challenges to the economy of Pakistan is the edible oil deficit. Edible oil is
a basic kitchen item, so is increasing with time. Edible oil produced locally is very less than its
demand, so Pakistan has to import edible oil to meet the local demand which is increasing by
each passing year due to increase in population. In Pakistan only 2% of the total cropped area
is covered by oilseed crops. During 2014-15 total available edible oil was 3.20 million tons
while local produce only contributed 0.573 million tons (only 18% of total availability), while
remaining 2.627 million tons edible oil was imported. During year 2014-15, Pakistan spends
US$ 2.50 billion in edible oil import (Govt. of Pakistan, 2014-15).
Despite agriculture based country there are many reasons behind this shortcoming for example
competition with major crops (e.g. wheat), less availability of seed of high yielding varieties,
lack of awareness among farmers, market uncertainty and ignorance of policy makers
regarding oilseed crops and technological deficiency in oilseed production. In current situation,
oilseed production in Pakistan meets only about 18% of the requirements (Govt. of Pakistan,
2014-15). This increasing import of edible oil contributes heavily to the ever increasing food
expenditure of household. The increasing population rate is one of major reason behind
increasing edible oil demand in Pakistan.
During year 2014-15 global edible oil consumption was around 184.08 million metric
tons (Statista, 2017); while oil seed tree crops (palm oil, palm kernel oil, coconut oil and olive
oil) contribute 42% of total world consumption. Moringa seeds contain 30-40% premium
quality, brilliant yellow, high oleic acid oil having pleasant peanut like flavor. Oil contain about
80% unsaturated fatty acid, more than 70% of which is oleic acid (Tsaknis et al., 1999; Foidl
et al., 2001; Anwar et al., 2006; Rahman et al., 2009; Ayerza, 2011). Various studies revealed
1
that seed oil contents and physio-chemical characteristics show variation depending upon
genotype and environment (Lalas and Tsaknis, 2002; Anwar et al., 2007; Ayerza, 2012).
Kleiman et al. (2006) reported that the oxidative stability of moringa oil is also higher than
other oils rich in oleic acid like sunflower, safflower, safflower, almond and apricot oils.
Moringa oil has been used as potential source of edible oil in some countries of Middle East
and Africa (Banerji and Verma, 2003). Anwar et al. (2007) reported that proper blending of
traditional edible oils with ben oil increase nutritional value and improved shelf life of edible
oils for domestic cooking and deep frying. The contents of polyunsaturated fatty acid (PUFA)
in moringa oil are less than 1% which is a huge advantage found by cosmetic industry on
moringa oil over other high oleic acid source. Due to very low content of PUFA, moringa oil
is less prone to oxidation. In cosmetics industry, ben oil is being used in manufacturing of
perfume and soap, while it is also used as lubricant in expensive watches (Banerji and Verma,
2003). Azam et al. (2005) reported that seed oil of moringa has good potential for biodiesel
production. After oil extraction, press cake can be used as animal feed, fertilizer and solid fuel.
Moringa, a multipurpose plant, belongs to family Moringaceae with 13 known species.
Moringa oleifera is most extensively diversified specie in Pakistan. It is native to sub-
Himalayan regions of Pakistan, India, Bangladesh and Afghanistan and now it is distributed
across tropics and subtropical regions in the Cambodia, African, Philippines, Central and North
America (Morton, 1991; Makkar and Becker, 1996). M. oleifera is fast growing, drought
tolerant, grows well in poor soil. It can survive under a wide range of rainfall (30-300 cm per
year) and pH (5.0-9.0). In those regions which receive annual rainfall less than 400 mm, plant
attains height of 6-7 m in a year (Odee, 1998; Palada and Chang, 2003).
Moringa (Moringa oleifera) is a valuable tree with great nutritional, medicinal,
industrial and numerous agronomic uses (Crosby, 2007). The leaves, flowers, and tender pods
of moringa are used as vegetable in my sub-tropical and tropical countries (Siddhuranj and
Backer, 2003; Anhwange et al., 2004). Moringa is known as Miracle Tree due to its diversified
uses and benefits as food, medicinal, oil, fodder, water purification and natural plant growth
enhancer (Berger et al., 1984; Olsen, 1987; Anwar et al.,, 2007; Basra et al., 2011; Yasmeen et
al., 2012; Yasmen et al., 2013; Ayerza, 2012; Basra et al., 2014; Nouman et al., 2014;
Yasmeen et al., 2014).
2
Moringa grows rapidly under favorable conditions. Tree grows up to 10 -12 m, while
sometime reaches height up to 15 m with stem diameter up to 75 cm (Parrotta, 2001; Odee,
1998). Moringa is propagated by sexual (seed) or asexual (cutting) method. Jahn et al. (1986)
reported that in semi-arid and arid climate propagation through seed is preferred to stem cutting
because some studies reveal that plant propagated through seed develop elongated root system
which goes deep in soil in search of water and nutrients. So in semi-raid climate where water
is a limiting factor for plant growth and development, propagation through seed is better than
stem cuttings. Plant propagated through stem or branch cutting, tree produce flowers and pods
within one year (Ramachandran et al., 1980). During first two years flowers and fruit yield are
generally low but after second year, one tree produced up to 1600 pods per year
(Ramachandran et al., 1980; Booth and Wickens, 1988; Morton, 1991).
M. oleifera is locally known as “Sohanjan” in Pakistan. In Pakistan only two species of family
Moringaceae: M. concanensis and M. oleifera are reported. M. concanensis is very uncommon
and only noticed in some far-off parts of Tharparkar (Sindh province). M. oleifera is very
common, wildly grown and cultivated in the many regions of the Punjab, Sindh, Balochistan
and KPK province of Pakistan (Qaiser, 1973). The tree propagated through stem cutting bears
fruit within one year and 1 kg moringa seed gives 0.33 kg seed oil. Nevertheless, high oleic
acid (>70%), low polyunsaturated fatty acid contents (<1%), high oxidative stability and
increasing use in cosmetic industry is promoting interest in its cultivation as oil seed tree crop
(Foidl et al., 2001; Lalas and Tsaknis, 2002; Kleiman et al., 2006). Ayerz (2011) and Nel
(2001) reported that environmental factors affect seed yield, seed yield related attributes and
kernel composition more than genetic factor. Anwar et al. (2006) reported that drought not
only reduces seed weight and oil yield but also affect seed composition.
Moringa oleifera is native to Pakistan, but according to best of our knowledge no study has
been conducted to explore it as oil seed tree production potential. Keeping in view the current
scenario of edible oil and potential of Moringa oleifera tree, there is dire need to explore it as
new oil seed tree. The present study has been therefore designed with following objectives:
 To explore and compare the yield potential (seed and oil) of wild grown and cultivated
Moringa oleifera.
3
 To compare the qualitative and quantitative potential of seed oil from wild grown and
cultivated Moringa oleifera.
4
2. Chapter 2
Review of Literature
2.1 Origin and distribution
Moringa oleifera Lam (Moringa) belongs to a monogeneric family of shrubs and tree. There
are many local names in Dravidian language but all drived from “Morunga”. In English it is
well-known as Horseradish tree, Never Die tree, Drumstick tree and Radish tree. In Pakistan
“Sohanjna” is most common name of this tree (Ramachandran et al., 1980; Qaiser, 1973). Out
of 13 species of Moringaceae, Moringa oleifera is the most common and widely distributed
specie native to sub-Himalayan tracts of Pakistan, India, Bangladesh and Afghanistan
(Sengupta and Gupta, 1970; Morton, 1991; Mughal et al., 1999; Paliwal and Sharma, 2011)
and now distributed to Middle East, African and Asian (Fahey, 2005; Palada et al., 2007;
Nouman et al., 2013).
Due to its diversified uses and benefits as food, medicinal, oil, fodder, water
purification and natural plant growth enhancer, Moringa is commonly known as Miracle Tree
(Berger et al., 1984; Olsen, 1987; Anwar et al., 2007; Basra et al., 2011; Yasmeen et al., 2012,
2013, 2014; Ayerza, 2011; Basra et al., 2014; Nouman et al., 2014). In world, this tree is very
famous due to nutrient rich leaves, flowers, pods, seed and roots. Many plant parts are used in
traditional medicines.
2.2 Morphology and physical characters
Moringa is a small to average sized, fast growing, evergreen to deciduous tree that can reach
up to 15 m in height with diameter up to 75 cm at chest height. It has spreading type of open
crown with fragile branches, tripinnate leaves and whitish bark. Plant grows well in tropical
and subtropical regions of world where annual precipitation and temperature ranges between
760-2500 mm and 18-28°C respectively. Plant can grow in any soil texture except heavy clay
and water logged soil with pH ranges from 4.5-8, an altitude up to 2000 m (Palada, 1996; Foidl
et al., 2001; Nouman et al., 2014).
2.2.1 Stem
Stem is usually straight and before branching, grows straight up to height of 1.5-2 m. Wood is
soften and usually not used in construction (Foidl et al., 2001).
5
2.2.2 Branch
Moringa tree has umbrella type canopy while extended branches grow in disorganized manner.
2.2.3 Leaves and young shoots
At the branch tips, 20-70 cm long mostly tripinnate leaves grow. At young stage leaves are
grayish-downy, 8-10 pairs of pinnae and pinnules are opposite and leaflets are 1-2 cm long and
0.5-1 cm wide (Morton, 1991).
2.2.4 Flowers, fruits and seeds
Pleasant fragrance flowers are bisexual, yellowish white born on 10-25 cm long dropping
axillary panicles. Single flower set in a basal with five yellowish white petals, five stamens
with five smaller sterile stamens and pistil of a one celled ovary and slender type style (Little
et al., 1964; Ramachandran et al., 1980).
The fruit are three lobed pods, usually 20-50 cm long and sometime length reached up to 1 m
or longer. Each pod contains 12-35 seeds, dark green during development and at maturity turns
brown and open into three parts. From flowering to maturity pods takes almost three months
(Palanisamy et al., 1985).
Upon maturity brown pods contains dark brown or whitish seeds about 1 cm in diameter.
Depending upon variety seed weight may differ ranging from 3-9 thousand seeds per kg (Negi,
1977). On an average each tree can produce 15-20 thousands seeds per year (Makkar and
Becker, 1996).
2.2.5 Wood and bark
Tree has soft, thick whitish-gray bark. Upon any cut, bark produce reddish brown gum which
has medicinal uses. The wood is soft and never used in furniture (World agroforestry database).
2.2.6 Root type
Lahjie and Siebert (1987) reported that seedlings usually develop a deep, tuberous taproot
system with very thin lateral roots while tree propagated through seeds have deep taproots with
wide spreading thick tuberous lateral roots and no taproots developed in trees which
propagated through stem cuttings.
2.3 Genetic diversity and taxonomy
6
Moringa, derived from Tamil language. Moringa is only genus in family Moringaceae with 13
known species (Mabberley, 1997). Moringa oleifera is diploid specie with 28 chromosomes.
Many species of moringa are well known as source of food, fiber, medicine and many other
products. These include are Moringa concanensis, longituba,drouhardii, peregrine, ovalifolia,
and Moringa stenopetala (Jahn et al., 1986; Morton, 1991). In Pakistan only M. oleifera and
M. concanensis are reported. Moringa oleifera is most commonly grown in Punjab, Sindh and
some parts of KPK province while M. concanensis is uncommon and only observed in some
far-off parts of Tharparker (In Sindh province) (Qaiser, 1973; Manzoor et al., 2007). M.
oleifera tree exhibits huge phenotypic variation within its range (Ramachandran et al., 1980;
Suthanthirapandian et al., 1989). Wild trees usually bear poor quality fruits (pods). In South
India, many cultivated varieties produce pods ranging in length from 60-120 cm. Wild trees
bear flowering only once in year while, Indian high yield varieties (PKM-1 and PKM-2)
produce flowers and pods round the year. Several varieties are cultivated in the West India.
Some genotypes produce lot of pods while others barely flower and are mainly grown for their
leaves (Ramachandran et al., 1980).
2.4 Growth and development
Moringa growth is very rapid under favorable environmental conditions. Plant increases 1-2 m
height per year during the 3-4 years. Munyanziza and Sarwatt (2003) reported that in Tunzania,
trees grown by nursery seedlings attained an average 4 m height within first year of plantation.
Generally mature moringa trees reached height of 10-12 m with stem diameter up to 75 cm at
chest height (Parrotta, 2001). When tree propagated through stem cuttings produce flowers and
pods within one year (Ramachandran et al., 1980). During first two years of plantation, tree
bears fewer flowers and fruit but when tree attain full size, a single tree can yield up to 1600
pods per year (Ramachandran et al., 1980; Booth and Wickens, 1988; Morton, 1991).
2.5 Cultivation and production
Moringa is propagated by two methods, sexual (seed) or asexual (cutting). Jahn et al. (1986)
reported that in semiarid and arid climate propagation through seeds is preferred over stem
cutting because according to Animashaun et al. (2013) in deep water table area; trees
propagated through seeds develop longer roots as compared to stem cuttings. In Sudan, mostly
moringa trees propagated through seeds while in India and some countries of West Africa,
7
vegetative propagation is more common (Palada, 1996). However, Ramachandran et al. (1980)
reported that due to genetic diversity, plants propagated through seeds produce poor quality
fruits (pods).
Depending upon variety, seeds weight differs ranging from 3-9 thousand seeds per kg. Under
ideal storage conditions (humidity 5-8%, temperature 3°C) seeds remain viable with good
germination percentage (80-90%) however, when seeds exposed to ambient temperature with
high relative humidity, seed germination percentage dropped to 8% within three months
(Morton, 1991; Roloff et al., 2009).
When seeds availability is limited and homogeneity among plants required then tree is
preferably propagation through stem cutting. Animashaun et al. (2013) reported that during
rainy season, when mature stem cuttings (1-2 m long, 6-15 cm diameter) are planted (burying
1/3 length in soil), they develops roots and attained considerable size in few months.
Moringa is a very fast growing tree. To facilitate harvesting, pruning practice is adopted to
boost horizontal growth and give the tree bush shape.
Seeds and leaves are most important parts of plant. For production of leaves, Moringa plants
are usually planted as following:
After three months of flowering, pods mature and turned brown color. Each pod contains 12 -
30 seeds covered with whitish or brownish semi permeable hull. Depending upon environment
and variety, seed production may vary. According to Ayerza (2011), a single tree can produce
15-25 thousand seeds. Average seed weight around 0.3 g. Some genotypes bear flowering
within six months of plantation while other takes one or more than one year (Paliwal and
Sharma, 2011).
2.6 Genetic and breeding
Due to cross pollination in moringa, many scientists reports heterogeneity in development and
yield. Raja et al., 2013 reported that in some genotypes flowering occur throughout the year
while others flowers only once in year. Some genotypes are deciduous while other evergreen.
Flowering phenology mostly varies depending upon variety and locality. In Northern India,
tree only flower once in a year (between month of April and July) while in Southern parts of
India, flowering recorded twice in each year. In Caribbean islands, where temperature and
rainfall almost remains constant, tree flowers throughout the year (Little et al., 1964;
Ramachandran et al., 1980). Generally flowering start within year after planting and tree
8
produce good seed crop up to 14 years (World Agroforestry Database). Usually plant
pollinated by insects, birds and honey bees. (Jyothi et al., 1990; Morton, 1991).
Many varieties of moringa are grown in India for different purposes. For example
Jafana and Chavakacheri murungai has soft and tasty fruits, Punamurungai typically grow for
leaves. (Ramachandran et al., 1980; Kumar et al., 2014). At Tamil Nadu, India two new
moringa varieties named as PKM-1 and PKM-2 have been develop to improve seeds
production. Those verities yield seeds throughout the year. After two harvests, plants pulled
out and new seedlings are planted (Saint Sauveur, 2001).
2.7 Ecology
In native region of moringa, annual temperature fluctuation is very large, mean lowest and
highest temperatures ranging from 1-3°C and 38-48°C in winter and summer season
respectively.
Moringa is very drought tolerant and mostly grown in semiarid and arid regions where
annual perception as below as 300 mm (Ramachandran et al., 1980; Booth and Wickens,
1988). In such regions soil surface may be very dry for many months but water is available to
roots due to irrigation or high water table (Champion, 1936). It grows along the rivers of its
native regions from sea level up to 1400 m; on well drained sandy to sandy loam soils often
very low in organic matter (Booth and Wickens, 1988). In its introduced regions, best growth
is recorded on sandy loam soils at elevations up to 1200 m (Ramachandran et al., 1980).
Moringa tree is very responsive to light that’s why it is usually planted at spacing of 3
× 3 m to 5 × 5 m (Ramachandran et al., 1980; Nautiyal and Venhataraman, 1987). Hedges of
moringa plants are typically established by planting it at 1 m or less distance and cut frequently
to produce more foliage (Morton, 1991). Basra et al. (2015) reported that for fodder purpose
plants should established at narrow spacing (15 × 30 cm) to harvest maximum biomass. Growth
of moringa seedlings are severely affected by drought and weeds infestation while tree is
highly susceptible to wind damage (Champion, 1936; Agrawal and Joshi, 1986).
2.8 Insect, pests and pathogens
In native region, moringa is susceptible to many insect pests especially bark eating, hairy and
green leaf caterpillars which cause severe damage to leafs while aphid, scale insects, stem
borers and fruit fly also cause damages (Ramachandran et al., 1980; Morton, 1991). Martin
9
and Ruperte (1979) reported that in many regions tree is very susceptible by termites attack.
No serious diseases reported in its native or introduced regions.
2.9 Nutritional value of moringa
Moringa pods, fresh leaves and dry leaves are highly nitrous and rich source of protein, fiber,
minerals, vitamins and amino acids. Mathur (2006) reported that moringa leaves restrain nine
times more protein then yoghurt. Fresh leaves and dry leaves of moringa contain more crude
protein then cow, buffalo, sheep and goat milk (Chandan, 2006). Moringa leaves are excellent
source of essential amino acids that required by human body.
2.10 Economic importance
All over the world because of its benefits, moringa is well known as “Miracle Tree”. Detail
uses of different parts of plant are mention in Figure 2.1.
Figure 2.1 Uses of different parts of Moringa
2.10.1 Wood
Moringa wood is soft that’s why it has little used except as a fuel and some time for light
construction work (Little et al., 1964; FAO, 1982). However in India, it is limitedly used in
textile industry (Mahajan and Sharam, 1984; Nautiyal and Venhataraman. 1987). Coarse fiber
obtains from corky bark which is used in making of mats and paper while leather industry also
used its stem gum in tanning of leather (Ramachandran et al., 1980; Nautiyal and
Venhataraman, 1987).
2.10.2 Human consumption
10
Moringa is a very important food product in tropics. Many countries, particularly India,
Pakistan, Philippines and many parts of Africa, its leaves, flowers, pods and roots are used as
highly nutritious vegetable in many countries such as Pakistan, India, Philippines (Anwar and
Bhanger, 2003; Anwar et al., 2005). Leaves are cooked as raw vegetable or mixed with flowers.
Many researchers reveals that moringa leaves are rich source of protein, calcium, potassium,
β-carotene, thiamine, riboflavin and other vitamins, particularly vitamins A and C (Rockwood
et al., 2013; Lakshmipriya et al., 2016). In Philippines it is used to enhanced mother milk
production that’s why well known as “mothers best friend” (Siddhuraju and Becker, 2003).
Tender pods (fruits) of plant are eaten as vegetable or pickled. Protein content of young pods
ranges between 5-10%. Root of plant mostly used in pickle (Martin and Ruperte, 1979;
Ramachandran et al., 1980).
2.10.3 Water purification
In many developing nations of Asia, Africa and South America availability of purified water
is very limited. Billions of peoples drink surface water to meet their water requirements.
Annually, around two million people died by diseases caught from contaminated water and
majority of those are children under five years of age (Mahmood et al., 2010). Many countries
of West Asia used moringa seed powder to purify drinking water (Berger et al., 1984; Olsen,
1987). Many rural area of Sudan also used moringa seed power for water treatment.
Polyelectrolytes are one of the active ingredients in the moringa seed. These
polyelectrolyte act as coagulant agent and bind soil particles, reducing bacterial and viral
contamination from drinking water in many rural populations of India, Myanmar, Sudan,
Malawi, and Indonesia (Jahn et al., 1986; Nyein and Aye, 1997; Mandloi et al., 2004). Water
purification through seed powder is very quick and easy method. First seed hull removed from
seed and then kernel ground to make powder. After mixing seed powder with water, allow
water to coagulate impurities. After one hour, water is filtered to obtain pure water. In other
method, seed powder packed in a cloth and suspended in water for overnight to coagulate
impurities. The cloth containing the seeds powder removed from water and the purified water
is poured leaving coagulant material in bottom. By this method up to 99% of colloids and
germs can be removed. Only two seeds required in treatment of one liter very dirty water.
2.10.4 Medicinal uses
11
In its native and introduced regions, moringa plant has many important roles in traditional
medicine and different parts of plant still uses in traditional Asian and West African medicine
(Ramachandran et al., 1980; Mossa, 1985; Booth and Wickens, 1988; Parrotta, 2001).
Different parts of plant used in traditional Indian medicine for treatment of rheumatism, ascites
and cardiac stimulants. In Ayurveda, leaves, root, stem bark, flowers and seeds are used in
treatment of different diseases (Ramachandran et al., 1980). Leaf extract of moringa has strong
antibiotic and antimalarial properties (Eilert et al., 1980; Gbeassor et al., 1990). Seed oil mostly
used externally to cure rheumatism and gout (CSIR, 1962; Parrotta, 2001). Many medicinal
important compounds have been extracted from root, root bark, stem bark and seeds (Booth
and Wickens, 1988)
2.10.5 Moringa as fodder for livestock
A number of studies reveal that, Moringa being used as livestock fodder or mix with other
fodder as a supplement (Atta-Karh, 1990; Lefroy et al., 1992; Otsyina and Dzowela, 1995).
Human and animals widely used all parts of moringa due to high nutritional value (CSIR, 1962;
Hartwell, 1971). With less input and management practices, moringa fodder can easily be
grown and increase milk and meat production. Many scientists have explored cultivation of
moringa fodder and its utilization in livestock (Richter et al., 2003; Sanchez et al., 2006;
Mendieta et al., 2011). Moringa plant is easy to use for different purpose because after root
establishment it is easy to maintain. Under abiotic stress conditions, plant roots enter deep in
soil in search of water and nutrients. Apart from its root system, moringa is very fast growing
plant with least nutrient and water requirement. This character makes moringa superior over
other crops (soybean and cotton seed cake) and different rang grasses, which relatively high
water requirement (Benavides, 1994). Depending on fertilizer, genotype, season and
environment, moringa crop produce high dry matter ranging from 4-8 t ha–1 (Palada et al.,
2007). Moringa leaves are rich source of all important nutrients and minerals important for
livestock to produce more milk and meat (Newton et al., 2010; Mendieta et al., 2011). Richter
et al. (2003) reported that low quality fodders can be enriched by adding moringa leaves which
not only increases the total dry matter and digestibility but also improve protein intake in fish.
Moringa fodder produces more biomass per unit area as compared to other forage crops. More
over moringa fodder ensure availability of fodder during dry season which extends from
December through May.
12
2.10.6 Crop growth enhancer
Moringa leaf extract (MLE) is rich source of zeatin, auxin, vitamin E, ascorbic acid, minerals,
and phenolic compounds (Foidl et al., 2001; Nouman et al., 2013; Choudhary et al., 2016).
Zeatin is the natural form of cytokine which has important role in cell elongation, cell division,
root formation, leaf senescence and apical dominance (Davies, 1995; Brault and Maldney,
1999). With increase in concentration of zeatin in plant, many physiological processed
accelerated many times (Swarup et al., 2002; Taiz and Zeiger, 2002; Nordstrom et al., 2004).
Nouman et al. (2012a) reported that seed priming with MLE increase seed emergence
percentage and early seed emergence. While different scientist reported that MLE foliar and
priming on crops increase photosynthetic and antioxidant activity which help plants to mitigate
abiotic stresses. Ultimately increase crop yield (Foidl et al., 2001; Nouman et al., 2012a;
Nouman et al., 2013; Yasmen et al., 2012; Yasmen et al.,2013)
2.11 Moringa seeds oil (Ben oil)
Fully mature dried seeds are round or triangular in shape while, kernel is covered with
semipermeable hull with tree wings and are brownish or whitish in color (Figure 2.3). Mature
dry seeds harvested from pods approximately yield 30-40% of non-drying oil (Anwar et al.,
2006; Anwar and Rashid, 2007; Uzama et al., 2011) commercially well known as “Ben oil” or
“Behen oil”. Oil is highly edible, good taste, and resembles olive oil in its fatty acid
composition (Lowell, 1999; Ramachandran et al., 1980). Ben oil has both nutritional and
industrial applications (Foidl et al., 2001; Anwar et al., 2007). Seed oil contents of moringa
seeds varies depending upon climate, genotype and extraction methods (Tsaknis et al., 1999;
Banerji et al., 2009; Ayerza, 2012). Seed also contains 38.4% crude protein. Moringa oil
contains almost 17% saturated and 83% unsaturated fatty acids. Monounsaturated fatty acid,
oleic acid contributes major part in fatty acid composition (more than 70%) while
polyunsaturated fatty acids are less than 1% which make oil less prone to oxidative damages
(Anwar and Bhanger, 2003; Anwar et al., 2006; Foidl et al., 2001).
13
Figure 2.2 White and brown seeds of Moringa
2.12 Effect of climate on seed production
Moringa seed production varies tremendously depending upon climate, genotype and soil
(Rajangam et al., 2001). Ndubuaku et al. (2014) reported that across Nigeria ranges moringa
seed yield ranges from 3-24 tons per hectare depending upon soil type, location and climate.
Traditionally moringa were cultivated only for fresh pods at low density population (Kumar et
al., 2014). Ayerza (2012) determine seed and oil potential of Indian variety Periyakulam-1
(PKM-1) in four ecosystems of Argentina and Bolivia. During different growing seasons seed
oil content percentage varies in different ecosystem; however seed yield per tree were statically
similar in all locations. However overall total oil yield per tree were significantly different in
all ecosystem. According to Ayerza (2012), seed yield affected by soil and environmental
factors.
Ayerza (2011) reported that in subtropical North West regions of Argentina, PKM-1 produce
significantly higher number of pods/tree, seed weight/pod and kernel weight than African
variety. However fatty acid composition of both varieties was similar.
2.13 Climatic factors and seed composition
Seed yield and oil yield of oil seeds crop greatly affected by genetic and environmental factors.
However environment affects significantly more on seed and kernel composition than genetic
factors. Drought directly affects seed quality and composition (Nel, 2001). Water availability
is one of most important factor in oil seed production. Anwar et al. (2006) reported that drought
14
significantly reduce seed oil content, seed weight and oil yield. Drought adversely affects
vegetative and reproduction stages. As a result decline in seed oil content and oil yield. To
tolerate drought stress, plants undergo morphological and genetic adaptions and thus affect
lipid composition (Gigon et al., 2004). Moringa seed oil content and its properties significantly
different, depending upon genotype and climate (Nel, 2001).
2.14 Seed composition
The contribution of seed oil content in total seed weight is about 30-40%. Oil extraction
methods determine the efficiency of extracting oil. The most common method of oil extraction
is the solvent extraction method using n-hexane as a solvent. The advantage of using solvent
extraction method is that it can extract larger quantity of oil from the seed; however less
quantity almost up to 69% can be extracted by cold press extraction technique (Tsaknis et al.,
1998; Lalas and Tsaknis, 2002; Ogunsina et al., 2014). The nutritional status of moringa seeds
also tell that besides oil it is also rich in protein (31.4%), carbohydrate (18.4%), fiber (7.3%)
and ash content (6.2%). Besides higher protein content, moringa seeds have ample content of
cysteine and methionine (Ogunsina et al., 2014). The higher protein digestibility (93%) of
moringa seeds is liable to its free urease activity and trypsin inhibition (Santos et al., 2005;
Oliveira et al., 1999). Table 2.1 shows the chemical composition of the M. oleifera seed.
2.15 Physio-chemical characteristics of moringa oil
At room temperature, moringa oil is liquid in state and yellow in color. The refractive index
and density of the moringa oil are similar to those of olive oil (Boskou, 2011). The oil obtained
by solvent extraction is lower in acidity and viscosity than that obtained by cold pressure
extraction. Water bounding is responsible for the higher viscosity of oil during extraction
(Tsaknis et al., 1999). The seed contact with temperature and air not only prolongs but also the
lipolytic enzymatic action enhances due to addition of the water (Sengupta and Gupta, 1970).
Moderate acidity referred toward its good resistance of cold pressed oil to hydrolysis. Due to
less unsaturation of moringa oil its iodine number is lower than olive oil (Boskou, 2011).
2.16 Oxidative stability of oil
The quality deterioration of vegetables oil is liable to oxidation, hydrolysis and polymerization
of fatty acid. Oil deteriorated because of the presence of high amount of linoleic acid in edible
oils (Che Men et al., 1999). Oils rich in monounsaturated fatty acids and oleic acid with
15
reduced contents of saturates and linoleic acid have several nutritional advantages. Oil blends
with improved fatty acid profile and stability results because of proper mixing of high-oleic
and high-linoleic oils. Anwar et al. (2007) reported that, blending of moringa oil with
conventional edible oil improves physio-chemical characteristics of oil. The fatty acid
composition of substrate oil is different from the blends. Blended also improves oxidation state
of blended sunflower and soybean oil with improved induction period. It was concluded that
oxidative stability of commercial edible oils can be enhanced using moringa oil.
2.17 Fatty acid composition
Ben oil contains up to 20% saturated fatty acid. Palmitic acid is most dominating fatty acid
follower by behenic, steric and arachidic acids. Content of behenic acid are more than other
common edible oil that’s why moringa oil commercially known as “Ben” or “Behen” oil.
Monounsaturated fatty acids are dominant in fatty acid composition. Oleic acid (18:1) is the
predominant fatty acid with contributes up to 75% of total fatty acids. While polyunsaturated
fatty acids (linoleic and linoleic acid) are less than 1% in moringa oil. Ogunsina et al. (2014)
reported that oil extraction methods have no effect on fatty acid composition. According to
Ayerza (2012) under different climatic conditions, fatty acid composition of oil almost remains
similar. Moringa oil MUFA/SFA ratio is high due to high content of oleic acid. Due to this
characteristic, oil is associated with a reduce risk of cardio vascular diseases (Schwingshackl
and Hoffmann, 2014).
2.18 Sterol composition
Sterol composition of moringa oil is different from conventional edible oils and olive oil
(Boskou, 2011). It mainly comprised of four main plant sterol compounds β-sitosterol,
compesterol, stigmasterol and delta-5-avenasterol. These four compounds account almost 92%
of total sterols. Several studies reveals that genotype and agro-climatic conditions could affect
sterol composition (Anwar and Bhanger, 2003; Rashid et al., 2008). Sterols compounds play
important role in metabolism of cholesterol and reduce chances of heart diseases while β -
sitosterol also possesses antidiabetic potential (Rashid et al., 2008; Demonty et al., 2009; Ras
et al., 2014).
2.19 Uses of moringa oil
2.19.1 Health benefits
16
The application of monounsaturated oils in varying number of foods has increased due to their
superior oxidative stability and health benefits; hence the production and need of
monounsaturated oils is rising all over the world (Corbett, 2003). Oleic acid has some
cholesterol lowering characteristics (Lokuruka, 2007). Oleic acid is rich in canola, olive,
rapeseed and peanut oil with concentration ranging from 50-80% (Corbett, 2003). To minimize
the risk of cardiovascular diseases the dietary guidelines strongly suggested that oleic acid
should be the part of daily diet (Lee et al., 2007). Through several genetic and biotechnological
methods oleic acid content has been increased in some crops like canola and sunflower
(Corbett, 2003). Partial hydrogenation is not necessary for monounsaturated oils for increasing
their shelf life and stability. Incidence of cardiovascular diseases has been positively correlated
with the consumption ratio of partially hydrogenated fats (Mensink and Katan, 1990).
Enzymatic transesterification and fraction resulted in high oleic acid fraction of ben oil
(Abdulkarim et al., 2007). High oleic acid fraction contains greater than 80% oleic acid; hence
considered as the power house of oleic acid (Rahman et al., 2009).
2.19.2 Medicinal and nutraceutical application
Moringa seed oil has been used in folk medicines since pre-historic time. It is highly rich in
oleic acid and possesses anti-inflammatory properties and also used in the treatment of breast
cancer and cardiovascular diseases (Pauwels, 2011). Oil is rich in vitamins A and E and
possesses good antibacterial properties. It also has antifungal, antiepileptic and
antihypertensive characteristics (Singha, 2010).
2.19.3 Biodiesel
Moringa seeds yields 30–40% good quality oil, rich in oleic acid. The worth of moringa oil is
far better than the sunflower oil and research studies have also demonstrated that the biodiesel
mad from moringa oil is better in quality than those of other substrates (Umer et al., 2008), as
it contains the highest number of methyl esters i.e. 67 which is the maximum for any biodiesel
fuel. As trees bears fruit after one year of plantation hence production of biodiesel can be
started after 1 year of cultivation. One hectare approximately can produced 2000 L biodiesel
annually (Brockman, 2008).The moringa oil based biodiesel has better stability as they contain
higher iodine content as compared to conventional diesel fuels. Its ignition performance is also
better in winter because of the cold filter plugging point and it also contains higher octane
number. The quality and recovery of moringa oil biodiesel is far better than other crops with
17
by-product of glycerine (Parawira, 2010). Moringa plants were milled to mesh size 5 after 30
day of cultivation, and solid mass is separated from the liquid by the process of filtration. The
liquid was then moved to a gas reactor. It was noted that 580 L of gas is produced from 1 kg
volatile solids produced poseessing 81% methane content (Fuglie, 2001).
2.19.4 Cosmetic industry
Oleic acid is the major fatty acid of moringa oil and it is extensively suggested for the
preparation of pharmaceutical ointments. It is a good cleansing agent with good cosmetic
value. The ability of mixing with other essential oils and its non-drying properties made
moringa oil excellent massage oil. The major utilization of oil is carried out in manufacturing
of different soaps and cosmetic products. Egyptian, Roman and Greek also used oil as perfume
and skin lotion (Kale and Megha, 2011).
18
3. Chapter 3
4. Materials and Methods
The study was conducted to explore comparative seed production potential and oil quality of
wild and asexually propagated (by mature stem cutting) of Moringa oleifera trees. Experiment
details regarding experiment site and materials are described here;
3.1 Experiment duration
The experiments were done during March, 2013 to July, 2015. In month of February, 2013
wild growing moringa trees were tagged in all selected locations while stem cuttings were
planted on March 30, 2013. Data regarding seed and oil yield and oil quality were taken for
two growing seasons (2014 and 2015).
3.2 Wild tree selection
On basis of morphological similarities confirmed by plant taxonomist five mature wild grown
moringa trees (at least five year old) were tagged during year 2013 at six locations of Punjab,
Pakistan (Faisalabad (FSD), Rahim Yar Khan (RYK), Bahawalpur (BWP), Layyah (LAY),
Multan (MLU) and Khanewal (KWL).
3.3 Pod harvest and seed yield data collection
Matured dry pods (when turned brown and dry) were harvested from each tree manually. Pod
length (cm), pod weight (g), number of seeds per pod, seed weight per pod (g), number of pods
per tree, seed weight per tree (kg) and 1000 seeds weight (g) were determine.
3.4 Oil extraction
Seed oil was extracted and degummed by procedure described by Tsaknis et al. (1998). After
the removing seed coat, 200 g seeds of each replication were grinded and then fed into a
Soxhelt apparatus with a 1 L round bottom flask and condenser. The extraction was performed
on a water bath up to 6 hours by using 600 mL n-hexane. After oil extracted from seed, solvent
in oil was distilled off by using rotary evaporator (EYELA, N.N. Series, Rikakikai Co. Ltd.
Tokyo, Japan).
19
3.5 Oil degumming
Oil was degummed by heating at 70°C on water bath and water was added 18% to final volume
and well mixed with a glass rod. After cooling water and oil mixture was centrifuged at 3000
rpm for 10 minutes in temperature controlled centrifuge machine (Sorval RC-3). Anhydrous
sodium sulfate was used to dried water in oil. After filtering, oil was kept in sealed bottles in
refrigerator until used for further analysis.
3.6 Experiment 1: Exploring the potential of wild grown Moringa oleifera as
oil seed tree at different locations of Punjab
Design: Randomized Complete Block Design (RCBD)
Replication: 4
Treatment: Six locations of Punjab, Pakistan where wild moringa commonly

reported.
1. Faisalabad
2. Rahim Yar Khan
3. Bahawalpur
4. Layyah
5. Multan
6. Khanewal
3.6.1 Pod length (cm)
Pod length was recorded by measuring length of twenty random pods with measuring tape and
average was calculated.
3.6.2 Pod weight (g)
Weight of twenty pods was recorded separately with the help of digital balance and average
weight was calculated.
3.6.3 Number of seeds per pod
Numbers of seeds per pod was counted manually.
20
3.6.4 Seed weight per pod (g)
After counting number of seeds per pod, seeds weight of each pod was measured by using
automatic electronic balance and average was calculated.
3.6.5 Number of pods per tree
At maturity when pods turn brown, pods from each tree were harvested manually and counted
total number of pods per tree.
3.6.6 Seed weight per tree (kg)
All pods were opened by hand and seeds were collected from each pod. Then weight of whole
seed lot was measured using automatic electronic balance.
3.6.7 1000 seeds weight (g)
1000 seeds were calculated by measuring weight of 100 seeds with digital balance and then
multiply it by ten.
Physio-chemical characterization of oils
Recommended AOCS (2009), standard methods were adopted to determine Physio-
chemical characterization of moringa seed oil.
3.6.8 Refractive Index
Refractive index was determined by standard AOCS Official Method Cc 7-25.
Apparatus
Abbe Refractometer
By using thermo stability controlled water bath, the temperature of the refractometer was
controlled to within ±0.1°C.
Calibration of the Instrument
Calibration of instrument was done by using distilled water which as refractive index of 1.3306
at 40°C.
Light Source
Tungsten lamp was used as a source of light.

Procedure
Traces of impurities and moisture were removed from oil samples through filtration.
Temperature of refractometer was adjusted at 40°C. A drop of oil sample was added in sample
21
chamber and closed it. Knob was adjusted in such a way that dark and light field crossed the
cross bar and reading was recorded.
3.6.9 Saponification value
Saponification value of the each test sample oil was estimated using official AOCS Method
(Cd 3-25).
Apparatus
1. Erlenmeyer flask-alkali resistance, 250-300 mL

2. Condensers
3. Water bath
Reagents
1. HCl, 0.5 M
2. KOH (Alcoholic) was prepared as following: 34 g of KOH was dissolved in 20 mL of
distilled water and diluted to 1 L with 95-100% pure ethanol.
3. 1% phenolphthalein indicator solution in 95% ethyl alcohol.
Procedure
1. Traces of impurities and moisture in oil samples were removed through filtration.
2. 50 mL alcoholic KOH was added in 5 g oil sample
3. Blank determination was prepare and conducted concurrently with the test portion
4. Condenser was connected and sample was boiled gently and steadily for 30 min.
5. After cool down of flask and condenser, condenser was washed with distilled water
and 1 mL of phenolphthalein indicator was added and titrate with 0.5 M HCl until pink
color just disappeared. Volume of 0.5 M HCl was recorded that used in titration.
Calculation
By using formula, saponification value (SV) was calculated;

(𝐵−𝑆)×(𝑀)
SV = × 56.1
𝑊
Where;
B = Volume of 0.5 M HCl used to titrate blank, mL
S = Volume of 0.5 M HCl used to titrate the test portion, mL
M = molarity of HCl solution
22
W = weight of test oil in grams
3.6.10 Iodine Value
Iodine value (IV) of the oil samples were estimated by following official AOCS Method (Cd 1-
25)
Apparatus
1. Glass-stoppered iodine flasks-500 mL.

2. Glass-stoppered volumetric flasks-1000 ML, for preparation of standard solutions.
3. Pipet-25 mL, for accurately dispensing 25.0 mL, of Wijs solution.
4. Volumetric dispenser-20 mL, 1 mL adjustability, for 10% potassium iodide (KI)
solution.
5. Volumetric dispenser-2 mL, 1 mL adjustability, for starch solution.
6. Volumetric dispenser-50 mL, 1 mL adjustability, for distilled water.
7. Repeater piper-with filling flask, 20 mL for cyclohexane.
8. Analytical halance-accurate to ± 0.0001 g.
9. Magnetic stirrer
10. Filter paper-Whatman no. 41H.
11. Beaker-50 mL.
12. Hot air oven.
13. Timer
Reagents Preparation:
Wij’s Solution:
Iodine trichlorides (9 g) was dissolved in mixture of acetic acid and chloroform (3:7) and
stirred for 12 h. Then Wij’s solution was standardized.
Wij’s solution standradization:
In a wide neck titration flask; 50 mL HCl and 50 mL CCl4 were taken then 25 mL Wij’s
solution was added and shacked energetically. Potassium iodide solution was titrated against
available iodine in CCl4 rosy purple layer until colorless endpoint. In another titration flask 5
mL aqueous potassium iodide (KI) solution + 150 mL distilled water was added wide in 25
mL Wij’s solution and contents in flask were titrated with Na2 S2O3.5H2O solution by using
starch as indicator until the colorless end point reached.
23
Procedure
1. Traces of impurities and moisture were removed from oil samples through filtration.
Filtration was performed in an air oven at 80°C.
2. After sample cooled down at 68-71 ± 1°C, sample was weighed into a 500 mL iodine
flask.
3. On top of sample, 15 mL CCl4 was added and spin to completely dissolved sample.
4. Wijs solution (25 mL) was added into flask containing the sample, stopper and swirl to
insured intimate mixture. Immediately set the timer for 30 min and store the flask in
the dark at a 25 ± 5°C.
5. 20 mL of KI solution and followed by 100 mL distilled water was added in flask after
removing flask from storage (30 min)
6. Then titrate sample with 9.1 N Na2 S2O3 slowly until yellow color nearly disappeared.
After adding 1 mL starch solution titration was continues until end point (blue color
disappears).
7. Blank was prepared and conducted with each group of samples simultaneously and
similar in all respects to the sample.
Calculations:
Using following equation, iodine value was calculated;

(B−S)×N×12.69
Iodine value (IV) =
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑜𝑖𝑙 (𝑔)
Where;
B = volume of titrant, mL of blank
S = volume of titrant, mL of sample
N = normality of Na2S2O3
3.6.11 Free fatty acid
Free fatty acid of the each test sample oil was estimated using official AOCS Method Ab 5 -
49.
Apparatus
1. Henry nut slicer
24
2. Cotton-clean, bleached
3. Butt-type extraction apparatus.
4. Oil sample bottle
Reagents
1. Petroleum ether
2. Isopropyl alcohol (99%)
3. 1% Phenolphthalein indicator solution.
4. Sodium hydroxide-0.25 M
Procedure
1. Oil sample (7.05 ± 0.05 g) was added into a flask.

2. Neutral alcohol (50 mL) and indicator (1 mL) was added in oil sample and titrated
against 0.25 M NaOH until faint pink color endpoint
Calculation
Free fatty acid was calculated by using following equation;
mL ×0.25 ×282×100
% FFA =
7.05 ×1000
Where-
7.05 g of extracted oil used
mL = mL of NaOH used in titration
0.25 molarity of NaOH solution
282 = equivalent weight of FFA in which results were expressed
3.6.12 Peroxide value
Peroxide value (PV) of the oil samples were determined following standard AOCS Official
Method 8b-90.
Apparatus
1. Pipet (0.5 mL)

2. Erlenmeyer flasks (250 mL)
3. Burette (25 mL)
4. Timer
25
5. Digital balance (± 0.01 gram sensitivity)
Reagents
1. 3:2 v/v acetic acid and isooctane solution

2. Saturated KI solution
3. 0.1 M sodium thiosulfate solution
4. 0.01 M sodium thiosulfate solution accurately standardized
5. Starch solution 1%
6. 10% SDS solution
Procedure
5.0 gram of oil sample was carefully dissolved in 50 mL of the 3:2 Acetic acid-
isooctane solutions. Then Swirl to dissolve the test portion. 0.5 mL of saturated KI solution
was added with pipet. Allow the solution to stand exactly 1 min, throughout shaking the
solution at least three times during 1 min, and then 30 mL of distilled water was added. Then
titrate with 0.1 M sodium thiosulfate adding it gradually and with constant and vigorous
agitation. Continued the titration until yellow color of iodine just disappeared. 0.5 mL of 10%
SDS and then 0.5 mL starch indicator solution was added. Continue the titration near the end
point to liberate all iodine from solvent layer. Added the thiosulfate solution drop wise until
the blue color just disappeared. Then blank determination was conducted.
Calculation
Peroxide value Iodine value was calculated by using following equation;

(S−B)×M × 1000
Peroxide value (mill equivalents peroxide/1000 g test portion) =
7.05 ×1000
Where-
B = volume of titrant, mL of blank
S = volume of titrant, mL of test portion
M = Molarity of sodium thiosulfate solution
3.6.13 p-anisidine value
Apparatus
1. 10 mL test tubes
2. Volumetric flask (25 mL).
26
3. Automatic pipet.
4. Spectrophotometer.
5. Glass cuvettes
Reagent
1. Isooctane (2,24 tripmethylepetane)

2. Glacial acetric acid
3. p-anisidine-analytical reagent quality
Procedure
1. Test sample (0.5 g) was diluted with isooctane into a 25 mL volumetric flask.
2. 5 mL of the fat solution was added into one test tube and exactly 5 mL of the solvent
into a second tube and shake.
3. After exactly 10 min absorbance of the solvent in the first test tube in a cuvette was
measured at 350 nm using the solution from the second test tube as blank in the
reference cuvette.
Calculation
The p-anisidine value (p-AV) of oil samples was calculated by the formula;
25 ×(1.2As−Ab
p-AV =
𝑚
Where as
As = absorbance of the fat solution after reaction with the p-anisidine reagent
Ab = absorbance of fat solution
m = mass of test portion, g
3.6.14 Specific extension of oil
Free fatty acid of the each test sample oil was estimated using AOCS official method Ch 5-
91.
Apparatus
1. Spectrophotometer
2. Quartz cuvettes
3. Graduated flasks-25ml.
4. Chromatographic column-450 mm length and approximately 10 mm i.d.
Reagents
27
1. Spectrophotometrically pure isooctane (2,2,4-trimethylpentane)
2. Basic alumina-for column chromatography
3. n-Hexane-for chromatography, analytical reagent grade
Procedure
1. Oil sample was filtered to removes impurities and traces of water at room temperature.
2. Oil sample (0.25 g) was weighted into a 25 ml graduated flask, make up to the mark
with the solvent and homogenize.
3. Cuvette was filled with the solution obtained and measures the extinctions at 232 and
268 nm.
Calculation
Specific extinctions were recorded by using following equation;
𝐸λ
Kλ =
𝐶×𝑆
Where as
Kλ = Specific extension at wavelength λ
Eλ = Extension measured at wavelength λ
C = solution concentration (g/100 mL)
S = Cuvette thickness
3.6.15 Fatty Acid Profile (FAP)
Fatty acid profile was determined by following the International Olive Council (IOC) official
method (COI/T.20/Doc. no. 33-2015).
Reagents
1. Methanol
2. N-Hexane
3. Heptane
4. Diethyl ether, stabilized for analysis
5. Acetone
6. Elution solvent
7. Potassium hydroxide
8. Silica gel cartridges
28
Apparatus
1. Screw-top test tubes (5 mL volume)

2. Automatic pipettes
Procedure
1. Oil sample (0.01 g) was weighted
2. 0.4 toluene was added and vortex
3. 3 mL methanol was 0.6 mL methanol-HCl mixture (80% Methanol and 20% HCl
(37%)) and vortex
4. Hexane (1.5 mL) and nano pure water (1 mL) was added and vortexed.
5. After stratifying, upper layer was decanted.
6. Water residues were removed by adding anhydrous sodium sulfate.
7. The clear solution was transferred into GC vials for injection in GC.
8. 0.2 μL sample was injected into Varian 450-GC.
9. Helium (flow rate of 1.5 mL/min. A 30 m × 0.25 mm × 0.1 μm) was used as carrier
gas.
10. For separation of individual fatty acid composition, DB-5 capillary colum (Agilent
Technologies, Santa Clara, CA, USA) was used.
11. The injector temperature was held at 240°C at a split ratio of 150.
12. The GC oven program was initially held isothermally at 80°C for 5 min; then ramped
at 10°C/min to 230 °C and held for 5 min and finally ramped at 20°C/min and held for
10 min.
13. FID detector temperature was 260°C. While mixture of air (flow rate: 300 mL/min),
hydrogen (flow rate: 30 mL/min) and helium (flow rate: 25 mL/min) was used as
detector gas.
14. A FAME mix was used as standards for peak identification by retention times.
3.6.16 Sterol Composition
The IOC official method (COI/T.20/Doc. No. 30/Rev.1 2013) was carried out for analysis of
sterol composition.
Apparatus
Following laboratory equipment and apparatus were used;
29
1. 250 ml flask
2. 500 ml separating funnel.
3. Complete apparatus for analysis by thin-layer chromatography using 20 x 20 cm glass
plates.
4. UV lamp
5. Micro syringes of 100 µL and 500 µL.
6. Cylindrical filter funnel
7. Vacuum conical flask with ground-glass female joint (50 mL),
8. 10 ml test tube with a tapering bottom and a sealing glass stopper.
9. Gas chromatograph
10. Fused-silica capillary column of length 20-30 m
11. Micro syringe, of 10 µL capacity
12. Calcium dichloride desiccator
Reagents
1. KOH
2. 2 M KOH ethanoic solution
3. Ethyl ether, for analysis quality.
4. Potassium hydroxide ethanolic solution, approximately 0.2 M.
5. Anhydrous sodium sulphate
6. Glass 20 x 20 plates coated with silica gel
7. Toluene
8. Acetone
9. n-Hexane
10. Ethyl ether
11. Ethanol
12. Ethyl acetate
13. 5% phytosterols, and Erythrodiol solution in Ethyl acetate as reference solutionfor
TLC (thin-layer chromatography)
14. 0.2% 2,7-dichlorofluorescein in ethanolic solution
15. Anhydrous pyridine
16. Hexamethyl disilazane
30
17. Trimethylchlorosilane
18. Sample solutions of sterol trimethylsilyl ethers.
19. 99% pure α-cholestanol
20. 0.2% α-cholestanol used as internal standard solution
21. Phenolphthalein
22. Hydrogen or helium as carrier gas
23. Hydrogen, nitrogen, helium and air as auxiliary gases
24. n-Hexane/ethyl ether in 65:35 (V/V)
Procedure
Preparation of the unsaponifiable matter
1. 0.5 mL of IS solution was added into 250 mL volumetric flask with ethanol;
2. Then IS was dried with steam of nitrogen;
3. Oil sample (5 g) was added into the flask and then 50 mL of 2 M KOH and some
boiling chips were added;
4. Flask was set on reflux system and heat to gentle boiling for 20 min until
saponification take place
5. Then 50 mL of DI water was added from top of the condenser and detach the
condenser and flask was cooled to 30°C
6. The content of flask was transferred into separation funnel. 80 mL of ethyl ether was
added into separation funnel and shake vigorously for 60 seconds; periodically
pressure was released by opening the stopcock. Allow liquid to stand until two phases
separate completely. Soap solution was p completely into a second separating funnel.
Two extractions were performed by using 60 mL ethyl ether
7. The extract of third time with 60 Ml of ethyl ether only top part was collected
8. Collected top part was pour back to separation funnel and 80 mL of DI waster was
added. After shaking, bottom part (water and KOH) was released into 400 mL beaker;
9. Again 80 mL of DI water was added into funnel, shake and release. This step was
repeated 4 times;
10. First three phases was discarded while 4 th water phase was released into a clean beaker
and test with 4-5 drops of phenolphthalein solution. If pink colored appeared then was
again with water.
31
11. Funnel was prepared with filter water on 250 mL round bottom flask. 2 spatula of
anhydrous sodium sulfate was added. Top part was pour to remove water residues;
12. Then solvent was evaporated with rotvap, water bath at 40°C, and condenser at 15°C.
13. TLC plates were prepared by immersing them into 0.2 M KOH ethanolic solution for
10 seconds and then dried under fume hood for 1 hour and then heat into oven for 1
hour;
14. After evaporation, round bottom flask was put into oven at 103°C for 15 minutes for
complete dry.
15. Hexane/ethyl ether mixture (60/40 v/v) was placed into TLC chamber to a depth of 1
cm. Chamber was closed for half an hour.
16. At that time, 1 mL of ethyl acetate was added into the cooled 250 mL round bottom
flask to dissolve the sample.
17. Line was drown on TLC plate about 2 cm from the bottom
18. 100 µL micro syringe was used to depose 0.3 mL of solution on a narrow and uniform
streak;
19. Plate was placed into TLC chamber into a cool place and chamber was closed with lid
until solvent reached approximately 1 cm from the upper edge of TLC plate;
20. TLC plate was dried with air stream under fume hood for 10 minutes;
21. 2,7-dichlorofluorescein solution was sprayed uniformly on TLC plate and leave in
fume hood for drying;
22. Plate was observed under UV light and cut bands
23. Bands are put into clean beaker and add 10 mL warm ethyl acetate to dissolve the
sterols. Then solution was transferred into 100 mL round bottom flask
24. 10 mL of ethyl ether was added into emptied beaker and dissolved for 3 times and
filtrated was collected into 100 mL flask and evaporate under rotvap until dryness
25. 0.5 mL silylation reagent was added into the flask and allowed to react for 20 minutes.
26. 300 µL clear solutions were transferred into GC injection vial.
27. Analysis was carried out on a Varian 450-GC (Varian, Palo Alto, CA, U.S.A.). GC
was equipped with flame ionization detector (FID) and a methyl phenyl polysiloxanes-
coated capillary column OV-17 (30 m 0.25 mm, 0.20-ím film thickness).The column
was operated isothermally at temperature 260°C while FID and injector temperature
32
were set at 280 and 270°C respectively. Extra pure Helium (at a flow rate of 20-35
cm/s) was used as a carrier gas and α-cholestanol was used as internal standard. Pure
sterol standard mixtures were used in identification and quantification of unknown
sterol components.
Analytical procedure
With automatic injector, 1 µL solution was injected. Recording was carried out until the TMSE of the
present triterpene dialcohols was completely eluted.
Peak identification
On basis of retention time, peaks were identified.
Concentration of each individual sterol (mg/kg) was calculated by using following formula;
𝐴𝑎 . 𝑚𝑠 . 1000
Sterol = x −
𝐴𝑏 . 𝑚
Where:
Aa = peak area for sterol a.
Ab = area of the α-cholestanol
ms = mass of α-cholestanol, in milligrams;
m = weight of oil samples, in grams.
3.7 Experiment II: Exploring the potential of vegetatively propagated
Moringa oleifera as an oil seed tree
3.7.1 Experimental Detail
Five stem cuttings from each selected location of wild grown moringa trees will be sown at the
farmer’s field Khanewal, Punjab, Pakistan. The experiment will be laid out in randomize
complete block design (RCBD) employing five replications.
Treatments
1. Faisalabad
2. Rahim Yar Khan
3. Bahawalpur
4. Layyah
5. Multan
6. Khanewal
Sowing date
33
March 30, 2013
Planting geometry
Plant to plant distance = 4.5 m

Row to row distance = 4.5 m
3.7.2 Stem cutting propagation:
Five mature stem cuttings (180-185 cm length and 12-15 cm), from each tagged tree of above
mentioned locations were obtained and propagated on March 30, 2013 at a farmer’s field at
Chack No. 7619R Khanewal. Stem cutting were planted in square 4.5 m × 4.5 m. Stem cuttings
were planted by burying 1/3 stem length in compost filled pits, after that pits were irrigated
with canal water immediately. Subsequent irrigations were done fortnightly interval with canal
and tube well water rotations until second harvest. All stem cutting sprouted within one month
and allowed to grow during 2013-2015. Pits were supplemented with nitrogen and
phosphorous @ 150:100/NP g/pit in two splits in a year followed by irrigation.
3.7.3 Soil analysis
Soil samples of the experimental site were taken from three different depths (10, 20 and 30
cm), and composite sample was prepared and sent to soil and water testing laboratory of Fauji
Fertilizer Company Multan to analyze following physio-chemical characteristics of soil.
Table 3.1 Physio-chemical characteristics of soil
Characteristic Unit Value
Textural class - Sandy clay

loam
Saturation percentage % 31.5
pH - 7.9
EC dS m-1 1.32
Available phosphorus mg kg-1 3.27
Extractable potassium mg kg-1 78
Organic matter % 0.74
34
Total nitrogen % 0.7
3.7.4 Pod harvest and seed yield data collection:
Matured dry pods (when turned brown and dry) were harvested from each tree manually in
first week of June. Pod length (cm), pod weight (g), number of seeds per pod, seed weight per
pod (g), number of pods per tree, seed weight per tree (kg) and 1000 seeds weight (g) were
determine.
3.7.5 Oil extraction
Seed oil was extracted by following procedures mentioned in section 3.4.
3.7.6 Degumming of oils:
Seed oil was degummed by following procedures mentioned in section 3.5.

Average pod weight (g)
Average pod weight was determined by procedure as mentioned in section 3.6.1.

Average pod length (cm)
Average pod length was determined by procedure as mentioned in section 3.6.2.

Average number of seeds per pod
Average number of seeds per pod was determined by procedure as mentioned in section 3.6.3.
Average seed weight per pod (g)
Average seed weight per pod was determined by procedure as mentioned in section 3.6.4.
Number of pods per tree
Number of pods per tree was determined by procedure as mentioned in section 3.6.5.
Average seed weight per tree (kg)
Average seed weight per tree was determined by procedure as mentioned in section 3.6.6.
1000 seeds weight (g)
1000 seeds weight was determined by procedure as mentioned in section 3.6.7.

Physico-chemical characteristics of oils
35
Physico-chemical characteristics of seed oil were determined by standard AOCS official
methods.
Refractive index
Refractive index was determined by standard AOCS Official Method Cc 7-25 as described in
section 3.6.8.
Saponification Value
Saponification was determined by standard AOCS Official Method Cd 3-25 as described in

section 3.6.9.
Iodine Value
Iodine value was determined by standard AOCS Official Method Cd 1-25 as described in
section 3.6.10.
Free Fatty Acids
Free fatty acids were determined by standard AOCS Official Method Ab 5-49 as described in
section 3.6.11.
Peroxide Value
Peroxide value was determined by standard AOCS Official Method Cd 8b-90 as described in
section 3.6.12.
p-Anisidine Value
p-Anisidine value was determined by standard AOCS Official Method Cd 18-90 as described
in section 3.6.13.
Specific extinctions
Specific extinctions at 232 and 270nm were determined following the standard AOCS Official
Method Ch 5-91 as described in section 3.6.14.
Fatty Acid Profile (FAP)
Fatty Acid Profile was determined by following IOC Official Method (COI/T.20/Doc. no. 33-
2015) as described in section 3.6.15.
Sterol Composition
36
Sterol Composition was determined by following IOC Official Method (COI/T.20/Doc. No.
30/Rev.1 2013) as described in section 3.6.16.
3.8 Statistical Analysis
The experiments were laid out in randomized complete block design (RCBD) with four
replications. Recorded data were analyzed using Fisher’s analysis of variance technique and
comparison of treatment means were done by Tukey’s test at 5% probability level (Steel et al.,
1997). In case of no difference (P>0.05) between the values of different locations, the latter
were excluded from the analysis of data.
37
4 Chapter 4
Results and discussion
Experiment 1
Results
Seed yield and yield attributes
Wild growing moringa trees were selected at six locations (FSD, RYK, BWP, LAY, MUL,
and KWL) to study the qualitative and quantitative potential of seed and oil. The data recorded
for seed yield and yield attributes are as under;
Pod length
Pod length was significantly (P<0.05) different among landraces during both years (Table
4.1.1). Maximum pod length was recorded in RYK trees while minimum in FSD during year
I and similar trend was seen during next season.
Pod weight
Pod weight of moringa landraces was significantly (P<0.05) different during the years 2014
and 2015 (Table 4.1.2). Maximum pod weight was observed in RYK trees during year 2014,
while minimum was noted in FSD and parallel trend was recorded during year 2015.
Number of seeds per pod
Data in Table 4.1.3 depicts that number of seeds per pod was significantly (P<0.05) different
among landraces during both the growing seasons. RYK trees produced significantly more
number of seeds per pod during 2014, while minimum was observed in FSD w,hich were
statically at par with values of BWP, LAY, MUL and KWL. Similar trend was seen during
year 2015.
Seed weight per pod
Seeds weight per pod was statically (P<0.05) different among landraces during the year 2014
and 2015 (Table 4.1.4). Maximum seed weight per pod was achieved by KWL trees during
years 2014, which was statically par with values of RYK and MUL. KWL trees also excelled
during 2015. However, minimum was noted in LAY during year I and in FSD during year II.
38
Number of pods per tree were statically (P<0.05) different among landraces in both growing
seasons (2014 and 2015) (Table 4.1.5). Maximum pods per tree were gained in RYK while
minimum in LAY during year 2014. Similar trend was seen during year 2015.
Seeds weight per tree
Data in Table 4.1.6 showed that seeds weight per tree was significantly (P<0.05) different
during both years of study (2014 and 2015). During both years maximum seeds weight per tree
was recorded in RYK trees while minimum in FSD.
1000 seeds weight
1000 seeds weight was significantly (P<0.05) different among landraces during year I and year
II (Table 4.1.7). Maximum 1000 seeds weight was observed in RYK tree, which was statically
similar with values of LAY and KWL, minimum was seen in FSD during year 2014. Maximum
1000 seeds weight was noted in RYK trees, which were statically similar with values of LAY
and BWP, MUL and KWL during year 2015.
Oil yield per tree
Seeds weight per tree was significantly (P<0.05) different during both years of study (2014
and 2015) (Table 4.1.8). Maximum oil yield per tree was recorded in RYK trees while
minimum in FSD. Similar trend was observed during second growing season.
Physio-chemical parameters
Seed oil content
Oil content was significantly (P<0.05) different among landraces during both years of study
(Table 4.1.9). During year I, maximum seed oil content was recorded in RYK tree while
minimum was observed in LAY, which was statically at par with FSD. During year 2015
maximum seed oil content was observed in RYK trees, which were statically at par with BWP,
MUL and KWL, minimum value was seen in FSD, which is statically similar with LAY.
Refractive index
The data regarding refractive index was non-significant (P>0.05) during both years of study
(Table 4.1.10). However, maximum value was noted in RYK during both seasons.
Saponification value
39
Saponification value was significantly (P<0.05) different among landraces during both years
of study (Table 4.1.11). Maximum saponification value was observed in RYK trees, which was
statically at par with LAY while minimum in BWP during year 2014. During year 2015,
highest saponification value was noticed in FSD, which was statically at par with RYK, BWP
and LAY while minimum MUL.
Iodine value
Iodine value of oil was significantly (P<0.05) different among landraces during year I while
non-significant during year II (Table 4.1.12). Maximum iodine value was noted in RYK trees
during year 2014; however, during second growing season (2015), maximum value was
recorded in FSD which was statically at par with RYK, BWP and LAY while minimum value
was seen in KWL, which was statically similar with MUL.
Free fatty acid
Free fatty acid contents of the moringa oil from the different landraces were not statically
significant (P>0.05) during both harvests.
Oxidative state of oil
Peroxide value
Peroxide value was non-significant (P>0.05) among landraces during both year of study (Table
4.1.14) while maximum value was observed in LAY during years 2014-15.
p-anisidine value
p-anisidine value was significantly (P<0.05) different among landraces during first growing
season while non-significant(P>0.05) during second growing seasons (Table 4.1.15).
Maximum p-anisidine value was recorded in RYK trees, which was statically at par with FSD,
BWP and LAY while minimum in KWL, which was statically at par with MUL during year
2014. Maximum peroxide value was observed in LAY trees during year 2015.
Specific extinction
Specific extinction of oil was non-significant (P>0.05) among landraces during both growing
seasons (2014 and 2015) (Table 4.1.16 and Table 4.1.17) however, maximum specific
extinction at K232 and K268 was recorded in LAY during the years 2014 and 2015.
Fatty acids composition
40
During both years of study (2014-15) all landraces of different locations trees produced oil
with almost identical fatty acid composition (Table 4.1.17). Monounsaturated ω-9 oleic (18:1)
acid dominate fatty acid (contribute more than 71% of total fatty acid composition) in seed oil
of all landraces, while polyunsaturated ω-6 linoleic (18:1) and linolenic fatty acids were less
than 1%. The content of saturated fatty acids, Palmitic (16:0), Stearic (18:0), Arachidic (20:0)
and Behenic (22:0) acids were ranged from 8.99-10.99, 3.98-5.91, 2.67-3.53 and 4.01-4.99%
respectively during both years of study (Table 4.1.17).
Sterol Composition
Sterol composition of all moringa landraces are given in Table 4.1.18. All moringa landraces
produced oil with nearly similar sterols composition. The β-sitosterol was the main sterols
(more than 55% of total sterol composition). The content of other major sterols compounds;
campesterol, stigmasterol and ∆5 -Avenasterol were ranged from 15.81-18.41, 15.96-18.37 and
5.04-7.63% respectively during both growing season (2014 and 2015).
Discussion
Moringa oleifera is native to Pakistan and locally known as “Sohanjna” is wildly grown in
Punjab, Sind and some part of KPK province. This study was conducted to explore seed, oil
yield and oil quality of mature wild moringa trees in six locations of Punjab (FSD, RYK, BWP,
LAY, MUL and KWL). During both years of experiment pods and oil yield of wild moringa
landraces were significantly different in all six locations. Landrace of RYK performed best
among all selected landraces during both harvests and gave maximum pod length, pod weight,
number of seeds per pod, seeds weight per pod, pods per tree and seeds weight per tree.
Rajangam et al. (2011) reported that seed production of moringa varies significantly. At
different locations of Nigeria, seed yield were ranged between 3-24 tons ha-1 depending upon
climate, soil and vegetation (Ndubuaku et al., 2014). Ayerza (2012) suggested that seed yield
of moringa may differ due to one or more environmental factors such as soil type, light,
temperature and available nutrients. Champolivier and Merrien (1996) also reported that
different ecosystem may affect seed composition, oil yield and oil quality. RYK landrace
produced maximum seeds and oil yield per tree as compared to other landraces of different
locations. Difference in yield potential of different landraces indicates genetic diversity
41
between landraces or more likely due to different climatic conditions. Ayerza (2011) reported
that seeds yield and oil yield of moringa affected by genotypes.
The seed oil content of wild trees of selected landraces was significantly different and ranged
between 32.25% - 37.02% (Table 4.1.9). RYK wild trees attained maximum seed oil content
while minimum in LAY during both years of study (2014 and 2015). Variation in seed oil
contents of wild moringa landraces may be due to genetic characteristic or more probably due
to environmental effects. Anwar et al. (2006) reported lower seed oil content in LYA as
compared to other regions of Punjab (RYK and Jhung). LAY region face drought conditions
because water deficit and less annual precipitations are characteristics of that region. This may
be the reason for low seed oil percentage in LAY landrace. Muhl et al. (2014) reported that
adequate water supply at reproductive and fruit set stage is essential for better seed yield in
moringa. Champolivier and Merrien (1996) reported that seed oil contents of moringa decline
under drought conditions. Carvalho et al. (2005) also reported that water deficiency is major
factor which causes decrease in seed oil percentage. Seed oil contents in this study were quite
comparable with earlier reported values in Pakistan (Anwar et al., 2005; Anwar et al., 2006).
However, physio-chemical characteristics of seed oil of wild landraces had less variability and
comparable with previous reported values of moringa oil (Table 4.1.14 to Table 4.1.16).
Refractive index at 40°C (1.4622-1.4631), saponification value (176.21-180.22 KOH/g of oil),
iodine value (65.46-71.96 g of iodine/100 g of oil) and free fatty acids (0.41-0.48 % oleic acid)
in current study were within range of earlier reported national and international work (Tsaknis
et al., 1998; Tsaknis et al., 1999; Lalas and Tsaknis, 2002; Anwar and Bhanger, 2003; Anwar
et al., 2006).
Peroxide value (PV), specific extinction and p-anisdine value (p-AV) are indicators of oxidative
state of oil. The data in Table 4.1.14-4.1.16 shows that seeds oil from wild moringa trees of
different locations possess identical oxidative state despite significant difference in oil yield.
Data also depicts that oil of selected landraces had good oxidative state compared with oil of
conventional crops. Peroxide value and p-AV which measure hydro peroxides and aldehydic
secondary oxidation products of oil respectively, were quite lower than palm oil thus showing
high resistance to oxidation (Mcginely, 1991). Peroxide value and p-anisdine value of different
landraces oil were higher than earlier reported values in Pakistan and other countries (Anwar
and Bhanger, 2003; Tsaknis et al., 1998; Anwar et al., 2006) however, less than Indian variety
42
PKM-1(Lalas and Tsaknis 2002). Specific extinction at 232 and 268 nm reveals the oxidative
deterioration of oil. Specific extinction values of oil from wild trees were in range of reported
values of Anwar and Bhanger (2003) and Anwar et al. (2006) and less than Indian variety
PKM-1 and African variety Mbololo (Tsakins et al., 1998; Lalas and Tsaknis 2002).
There was no significant difference in fatty acid composition of oil from wild trees of selected
landraces. Ayerza (2011) reported that climate and genotypes affects seed and oil production,
but their fatty acid composition remain identical. The content of saturated fatty acids was in
range from 21.90-22.50%. Palmitic acid is dominant saturated fatty acid followed by behenic,
stearic and arachidic acid. Behenic acid in moringa seed oil is high then conventional oil seed
that’s why moringa oil is commercially well known as “Ben oil” or “Behen oil.” All landraces
seed oil contains significantly higher monounsaturated fatty acid. Oleic acid (ω-9) is the major
monounsaturated fatty acid in this study, which accounts for more than 71% of total fatty acids
(Table 4.1.18) while polyunsaturated fatty acid (linoleic and linolenic acid) were less than 1%.
Abdulkarim et al. (2007) reported that value of monounsaturated fatty acid (oleic acid) in
moringa oil is even higher than reported values of conventional oil seed crop. Demand of high
monounsaturated fatty acid oil is increasing due to their health benefits. Oil with more oleic
acid reduces the chance of coronary heart diseases (Mensink and Katan, 1990; Schwingshackl
et al., 2014). Due to high level of monounsaturated fatty acids and less polyunsaturated fatty
acids moringa oil is more stable than other high oleic acid edible oils to oxidative damage at
ambient and high temperatures (Warner and Knowlton, 1997). Anwar et al. (2007) reported
that proper blending of moringa oil in high linoleic oils improves oil nutritional value and oil
stability to oxidation during cooking and deep frying. The content of PUFA (polyunsaturated
fatty acid) in moringa oil was less than 1% in all landraces. Cosmetic industry use moringa oil
over other high oleic acid source due to very low content of PUFA because oil is less prone to
oxidation (Kleiman et al., 2008).
There was no significant difference observed in sterol composition of oil of selected landraces
(Table 4.1.19). Sterol fractions mainly comprised of β-sitosterol, campesterol, stigmasterol and
delta 5-avenasterol. These four plant sterols comprised of 96% of total sterols. β-sitosterol is
the predominant plant sterol in all seed oils of wild trees, contribute more than 55% in total
sterols compostion. This is higher than earlier reported national and international work
(Tsaknis et al., 1999; Lalas and Tsaknis, 2002; Anwar et al., 2006; Anwar and Bhanger, 2003).
43
It is involved in metabolism of cholesterol and reduces the LDL cholesterol level in blood and
ultimately decreases risk of coronary heart diseases (Abumweis et al., 2008; Ros et al., 2014).
Anwar and Rashid (2007) reported that genotype and environment had a significant effect on
sterol composition.
Conclusion
Based on findings of this experiment, it is concluded that, there was significant difference in
seed and oil yield of wild moringa landraces grown in different regions of Punjab. RYK
landrace had maximum potential to further propagate for seed and oil. However, physio-
chemical characteristics, oxidative stability and oil quality had little variability between
landraces, and their values are much comparable with earlier reported work.
44
Table 4.1: Mean sum of squares of data of pod length, pod weight, number of seeds pod-
1
Number of Seed weight

SOV df Pod length Pod weight
seeds per pod per pod
Treatment 5 320.25* 8.74* 4.97* 1.34*
Replication 3 2.24 0.03 0.46 0.003
Error 15 37.11 0.08 0.53 0.009
*P<0.05 ns = non-significant
Table 4.2: Mean sum of squares of data of pod length, pod weight, number of seeds pod -
1

Treatment 5 30.62* 8.06* 4.55* 1.04*
Replication 3 0.40 0.29 0.32 0.02
Error 15 1.38 0.09 0.33 0.006
45
Table 4.3: Mean sum of squares of data of number of pods tree -1, seed weight tree-1, 1000
seeds weight, oil yield tree-1 and oil yield hectare-1 of wild moringa landraces during year
2014
Number of pods Seed weight 1000 seed Oil yield per

SOV df
per Tree per tree weight tree
Treatment 5 92910.0* 2.28* 60.45* 0.34*
Replication 3 779.7 0.007 49.61 0.0007
Error 15 729.6 0.003 37.57 0.0005
Table 4.4: Mean sum of squares of data of number of pods tree -1, seed weight tree-1, 1000
seeds weight, oil yield tree -1 and oil yield hectare-1 of wild moringa landraces during year
2015
Number of pods Seed weight 1000 seed Oil yield per

SOV df
per Tree per tree weight tree
Treatment 5 103452* 1.77* 6438.80* 0.30*
Replication 3 2185 0.012 35.0 0.0021
Error 15 1012 0.009 53.75 0.0026
46
Table 4.5: Mean sum of squares of data of oil content, refractive index, saponification
value, iodine value and free fatty acids of wild moringa landraces during year 2014
Oil Refractive Saponification Iodine Free fatty

SOV df
content index value value acids
Treatment 5 4.39* 7.28ns 22.44* 11.00* 0.0018ns
Replication 3 0.18 6.52 7.80 0.71 0.0077
Error 15 0.29 6.74 9.19 0.31 0.0175
value, iodine value and free fatty acids of wild moringa landraces during year 2015

SOV df
Treatment 5 14.26* 6.98ns 11.62* 6.21* 0.0030ns
Replication 3 0.61 1.04 0.46 11.35 0.0051
Error 15 1.00 2.03 1.52 1.79 0.0067
47
Table 4.7: Mean sum of squares of data of oil peroxide value, p-anisdine value, K232, and
p-anisdine
SOV df Peroxide value K232(K1%
1cm) K268(K1%
1cm)
value
Treatment 5 0.41* 0.047* 0.0043ns 0.0002ns
Replication 3 0.09 0.001 0.0015 0.0003
Error 15 0.33 0.007 0.0055 0.0015
Table 4.8: Mean sum of squares of data of oil peroxide value, p-anisdine value, K232, and
p-anisdine
1cm) K268(K1%
1cm)
value
Treatment 5 0.22 0.030 0.0018ns 0.0001ns
Replication 3 0.04 0.033 0.0011 0.0008
Error 15 0.09 0.038 0.0043 0.0010
48
Table 4.1.1 Effect of different landraces on pod length (cm) of wild grown moringa trees
Locations Year 2014 Year 2015

FSD 33.68 c 36.85 c
RYK 45.98 a 45.48 a
BWP 41.08 b 41.16 b
LAY 41.51 b 42.10 b
MUL 40.52 b 41.20 b
KWL 42.16 b 41.99 b
CVC 3.61 2.70

Means sharing
same letters are statically similar
CVC = Critical value of comparison
FSD (Faisalabad) RYK (Rahim Yar Khan)
BWP (Bahawalpur) LAY (Layyah)
MUL (Multan) KWL (Khanewal)
Table 4.1.2: Effect of different landraces on pod weight (g) of wild grown moringa trees

FSD 5.56 c 5.52 c
RYK 10.02 a 9.64 a
BWP 7.16 b 6.72 b
LAY 6.69 b 6.26 b
MUL 7.06 b 6.60 b
KWL 7.03 b 6.55 b
CVC 0.66 0.71

Means sharing
same letters are
statically similar
49
Table 4.1.3: Effect of different landraces on number of seeds per pod of wild grown
moringa trees

FSD 14.60 b 15.57 b
RYK 17.19 a 17.60 a
BWP 14.26 b 14.45 b
LAY 14.36 b 15.63 b
MUL 14.43 b 15.30 b
KWL 15.01 b 16.36 b
Means sharing same
CVC 1.68 1.32 letters are statically
similar
CVC = Critical value
of comparison
Table 4.1.4: Effect of different landraces on seeds weight per pod (g) of wild grown
moringa trees
FSD 1.83 c 2.01 c

RYK 3.39 a 3.33 a
BWP 3.15 b 3.11 b
LAY 3.10 b 3.12 b
MUL 3.26 ab 3.22 ab
KWL 3.40 a 3.37 a
Means sharing same
CVC 0.22 0.18 letters are statically
similar
50
Table 4.1.5: Effect of different landraces on number of pods per tree of wild grown
moringa trees
FSD 958.00 b 971.2 b
RYK 1167.8 a 1101.25 a
BWP 853.25 c 837.18 c
LAY 750.00 d 728.43 d
MUL 813.50 c 830.62 c
KWL 821.00 c 804.10 c
CVC 62.05 73.08

Means
sharing same
letters are statically similar
Table 4.1.6 Effect of different landraces on seeds weight per tree (kg) of wild grown
moringa trees

FSD 1.81 e 1.83 e
RYK 4.11 a 3.77 a
BWP 2.67 bc 2.25 c
LAY 2.47 d 2.20 c
MUL 2.55 cd 2.58 bd
KWL 2.77 b 2.53 b
CVC 0.13 0.22
Means sharing same letters are statically similar

51
Table 4.1.7: Effect of different landraces on 1000 seeds weight (g) of wild grown
moringa trees

FSD 130.57 c 131.10 c
RYK 234.15 a 238.64 a
BWP 217.36 b 222.01 a
LAY 227.30 ab 222.40 a
MUL 215.55 b 222.71 a
KWL 227.09 ab 233.71 a
Means sharing
CVC 14.08 0.16 same letters are
statically similar
Table 4.1.8: Effect of different landraces on oil yield per tree (kg) of wild grown
moringa trees

FSD 0.60 d 0.59 d
RYK 1.48 a 1.40 a
BWP 0.92 b 0.80 c
LAY 0.81 c 0.73 c
MUL 0.86 c 0.93 b
KWL 0.96 b 0.92 b
CVC 0.04 0.11 Means sharing
same letters are
statically similar
52
4.1.9: Effect of different landraces on seed oil content (%) of wild grown moringa trees
FSD 33.40 cd 32.25 b

RYK 35.95 a 37.02 a
BWP 34.57 bc 35.66 a
LAY 33.06 d 33.25 b
MUL 33.86 bcd 35.97 a
KWL 34.71 b 36.34 a
Means sharing
statically similar
4.1.10: Effect of different

landraces on Locations Year 2014 Year 2015 refractive
index (at 40°C) of wild grown
moringa trees FSD 1.4624 1.4626
RYK 1.4630 1.4631

BWP 1.4622 1.4626
LAY 1.4629 1.4629
MUL 1.4625 1.4630
KWL 1.4627 1.4628
CVC 0.09 0.08

53
4.1.11: Effect of different landraces on saponification value (mg of KOH/g of oil) of wild
grown moringa trees
FSD 176.96 180.22 a
RYK 177.59 179.52 a
BWP 179.08 178.68 ab
LAY 182.89 177.84 ab
MUL 176.21 176.29 b
KWL 178.71 176.01 b
CVC 6.96 2.84
Means sharing
4.1.12: Effect of different landraces on iodine value (g of I/100 g of oil) of wild grown
moringa trees

FSD 67.31 c 68.52 b
RYK 70.44 a 71.93 a
BWP 68.92 b 69.71 ab
LAY 65.46 d 68.73 b
MUL 67.92 bc 69.40 ab
KWL 68.14 bc 70.29 ab
CVC 1.28 3.08
Means sharing
same letters are
statically similar
54
4.1.13: Effect of different landraces on free fatty acid (% oleic acid) of wild grown
moringa trees
FSD 0.46 0.42

RYK 0.43 0.45
BWP 0.48 0.49
LAY 0.47 0.45
MUL 0.43 0.41
KWL 0.45 0.46
CVC 0.30 0.18

Means sharing
4.1.14: Effect of different landraces on peroxide value (meq of O 2 kg -1 of oil) of wild
grown moringa trees
FSD 1.50 1.15

RYK 1.35 1.40
BWP 1.63 1.35
LAY 1.68 1.65
MUL 1.65 1.35
KWL 1.38 0.95
CVC 0.34 0.71
Means sharing
55
4.1.15: Effect of different landraces on p-anisidine value of wild grown moringa trees

FSD 4.51 abc 3.72
RYK 5.07 a 4.07
BWP 4.97 ab 4.13
LAY 4.61 abc 4.83
MUL 4.43 bc 4.19
KWL 4.22 c 3.27
Means sharing
p < 0.05 0.20 0.45 same letters are
statically similar
4.1.16: Effect of different landraces on K232(𝐊 𝟏%

𝟏𝐜𝐦 ) of wild grown moringa trees

FSD 1.77 1.71
RYK 1.68 1.66
BWP 1.73 1.72
LAY 1.72 1.70
MUL 1.70 1.69
KWL 1.69 1.71
Means sharing
statically similar
CVC = Critical
value of comparison
56
4.1.17: Effect of different landraces on K268(𝐊 𝟏%
𝟏𝐜𝐦 ) of wild grown moringa trees

FSD 0.43 0.42
RYK 0.43 0.40
BWP 0.43 0.44
LAY 0.44 0.46
MUL 0.43 0.41
KWL 0.41 0.40 Means sharing
same letters are
CVC 0.03 0.07
statically similar
CVC = Critical
value of comparison
57
Table 4.1.18: Fatty acids composition (grams per 100 g of Fatty Acids) of wild grown moringa tree landraces in selected locations
of Punjab
Locations Year Fatty acids

16:0 16:1 18:0 18:1 18:2 18:3 20:0 20:1 22:0 24:0
2014 9.28 1.55 5.91 71.35 0.61 0.09 3.53 2.18 4.39 1.13
FSD
2015 9.64 2.02 4.54 71.76 0.63 0.12 3.03 2.31 4.77 1.18
2014 9.35 1.78 5.15 71.21 0.71 0.11 3.28 2.28 4.87 1.27
RYK
2015 9.62 1.79 4.79 72.66 0.67 0.12 3.05 2.22 4.06 1.03
2014 9.72 2.01 4.40 71.60 0.66 0.12 2.92 2.32 4.94 1.29
BWP
2015 9.75 1.86 4.82 71.61 0.65 0.12 3.13 2.29 4.61 1.15
2014 8.99 1.82 4.9 72.34 0.65 0.11 3.16 2.29 4.9 1.28
LAY
2015 9.67 1.52 5.1 72.38 0.72 0.12 3.23 2.19 4.04 1.04
2014 9.48 1.75 5.23 71.45 0.64 0.11 3.24 2.26 4.65 1.19
MUL
2015 8.99 1.57 5.35 72.34 0.75 0.12 3.38 2.27 4.19 1.05
2014 9.73 1.97 4.46 71.44 0.66 0.12 2.97 2.35 4.99 1.31
KWL
2015 10.99 1.94 3.98 72.1 0.73 0.10 2.67 2.36 4.01 1.12
2014 1.07 1.00 3.17 0.50 0.26 0.07 1.18 0.33 1.67 0.41
CVC
2015 3.08 1.04 1.53 1.58 0.21 0.08 0.99 0.27 1.43 0.39
FSD (Faisalabad), RYK (Rahim Yar Khan), BWP (Bahawalpur), LAY (Layyah), MUL (Multan), KWL (Khanewal)
58
Sterol Composition FSD RYK BWP LAY MUL KWL CVC
2014 2015 2014 2015 2014 2015 2014 2015 2014 2015 2014 2015 2014 2015
Cholesterol 0.13 0.12 0.12 0.14 0.13 0.14 0.12 0.14 0.12 0.16 0.13 0.14 0.04 0.06
24-Methylenecholesterol 0.32 0.2 0.3 0.21 0.37 0.22 0.3 0.31 0.23 0.3 0.27 0.22 0.38 0.31
Campesterol 16.19 18.41 16.14 17.51 15.73 16.47 16.84 15.81 16.85 16.04 17.39 16.81 1.79 2.87
Campestanol 0.06 0.04 0.07 0.11 0.09 0.07 0.07 0.07 0.04 0.04 0.08 0.11 0.14 0.09
Stigmasterol 16.51 16.45 17.45 16.83 15.96 18.14 18.37 17.73 16.96 18.33 17.12 17.94 8.74 3.68
Clerosterol 0.58 0.6 0.49 0.52 0.45 0.49 0.49 0.49 0.52 0.49 0.26 0.52 0.58 0.06
β-Sitosterol 56.08 55.23 55.24 56.07 56.6 56.28 55.05 57.53 57.37 56.09 54.97 55.69 4.74 4.76
∆5- Avenasterol 6.46 5.78 7.01 5.71 7.63 5.5 5.67 5.04 5.6 5.59 6.66 5.88 7.17 2.52
∆5-24-Stigmastadienol 0.52 0.51 0.45 0.49 0.5 0.34 0.43 0.38 0.34 0.37 0.5 0.4 0.35 0.25
∆7-Stigmastadienol 1.34 1.22 1.18 1.17 1.31 1.22 1.14 1.17 0.92 1.26 1.29 1.22 0.60 0.56
∆7- Avenasterol 1.82 1.43 1.56 1.25 1.45 1.12 1.51 1.34 1.05 1.34 1.34 1.08 2.05 0.88
Table 4.1.19: Sterol Composition (Percent) of wild grown moringa tree landraces in selected locations of Punjab
59
60
Experiment 2
Seed yield and yield attributes
Moringa cultivation through stem cuttings at Khanewal collected from locations (FSD, RYK,
BWP, LAY, MUL, and KWL) produced flowers during first year except FSD. The data
recorded for seed yield and yield attributes are as under;
Pod length
Pod length was significantly (P<0.05) different among landraces during both years (Table
4.2.1). Maximum pod length was recorded in RYK trees during both years while the minimum
was noted in MUL during year I and in FSD during year II.
Pod weight
Pod weight was significantly (P<0.05) different among landraces during the years 2014 and
2015 (Table 4.2.2). Maximum pod weight was achieved by RYK trees during both years while
the minimum was noted in MUL during year 2014 and in FSD during year 2015.
Number of seeds per pod
Number of seeds per pod was significantly (P<0.05) different among landraces during both
years (Table 4.2.3). RYK produced significantly more seeds per pod during both years of
study, which was statically at par with BWP and KWL during year 2014 and with FSD and
KWL during year 2015 while minimum value was observed LAY during both years of study.
Seed weight per pod
Seeds weight per pod was statically (P<0.05) different among landraces during the years 2014
and 2015 (Table 4.2.4). Maximum seed weight per pod was noted in RYK during both years
(2014 and 2015) while minimum in LAY during year I and in FSD during year II.
The data in Table 4.2.5 showed that the number of pods per tree was statically ( P<0.05)
different in selected landraces during both years of experiment. Maximum pods per tree were
observed in KWL which was statically at par with RYK BWP and MUL while minimum in
LAY during year 2014. Similar trend was seen during year 2015.
61
Seeds weight per tree
Data in Table 4.2.6 exhibit that seed weight per tree was significantly (P<0.05) different during
both years of study. Maximum seeds weight per tree was recorded in RYK trees during year
2014 and 2015, while minimum in LAY during year I and in FSD during year II.
1000 seeds weight
1000 seeds weight was significantly (P<0.05) different among landraces during both years
(2014 and 2015) (Table 4.2.7). Maximum 1000 seeds weight was observed in RYK during
both seasons while minimum in LAY during year 2014 and in FSD during year 2015.
Oil yield per tree
Oil yield per tree was significantly (P<0.05) different between landraces during the years 2014
and 2015 (Table 4.2.8). Maximum oil yield was recorded in RYK trees during both growing
seasons (2014 and 2015) while least in LAY and FSD trees during year 2014 and 2015
respectively (Table 4.2.8).
Oil yield per hectare
Oil yield per hectare was significantly (P<0.05) different between landraces during the years
2014 and 2015 (Table 4.2.8). Maximum oil yield per hectare was achieved in RYK trees during
both years (2014 and 2015) while least in LAY and FSD trees during year 2014 and 2015
respectively (Table 4.2.9).
Physio-chemical parameters
Seed oil content
The hexane-extracted oil content was significantly (P<0.05) different among landraces during
both years of study (Table 4.2.10). Maximum seed oil recovery was recorded in KWL tree,
which was statically at par with RYK, BWP and MUL while the minimum in LAY during year
I. During year 2015 maximum seed oil content was observed in KWL trees, which were
statically at par with RYK, BWP, LAY and MUL while minimum value was observed in FSD.
Refractive index
Refractive index was non-significant (P>0.05) among landraces during both years of study
(Table 4.2.11) however; maximum value was recorded in RYK during years 2014 and 2015.
Saponification value
62
Saponification value was significantly (P<0.05) different among landraces during both years
of study (Table 4.2.10). Maximum saponification value was observed in RYK tree, which was
statically at par with LAY while minimum in BWP during year 2014. During year 2015,
highest saponification value was noticed in FSD, which was statically at par with RYK, BWP
and LAY while minimum MUL.
Iodine value
Iodine value of oil was significantly (P<0.05) different harvested from landraces during year I
while non-significant during year II (Table 4.2.13). Maximum iodine value was noted in MUL
tree, which was statically at par with RYK and BWP while minimum in LAY during year 2014
however, maximum value was recorded in RYK during years 2015.
Free fatty acid
Free fatty acid value of the moringa oil from the different landraces was not statically
significant (P>0.05) during both harvests in years 2014 and 2015.
Oxidative state of oil
Peroxide value
Peroxide value of oil was significantly (P<0.05) different among landraces during year 2014
while non-significant during year 2015 (Table 4.2.15). Maximum peroxide value was recorded
in LAY tree which was statically at par with BWP while minimum in KWL, which was
statically similar with MUL and RYK during year 2014. Maximum peroxide value was noticed
in RYK trees during year 2015.
p-anisidine value
p-anisidine value of moringa oil was significantly (P<0.05) different harvested from landraces
during first growing season while non-significant during second growing season (Table
4.2.16). Maximum p-anisidine value was recorded in LAY tree which was statically at par with
BWP while minimum in BWP, which was statically at par with KWL during year 2014.
Maximum peroxide value was observed in MUL trees during year 2015.
63
Specific extinctions
Specific extinction of seed oil at K232 and K268 was non-significant (P>0.05) among
landraces during both growing seasons (2014 and 2015) (Table 4.2.17) however, maximum
specific extinction at K232 was recorded in BWP during years 2014 and 2015. Similar trend
was observed in K268 values during both years of study.
Fatty acids composition
During both years of study (2014 and 2015), all landraces of different locations trees produce
oil with almost identical fatty acid composition (Table 4.2.19). Monounsaturated ω-9 oleic
(18:1) acid was the main fatty acid (more than 71% of total) in seed oil of all landraces, while
polyunsaturated ω-6 linoleic (18:1) and linolenic fatty acids were less than 1%. The content of
saturated fatty acids, palmitic (16:0), stearic (18:0), srachidic (20:0) and behenic (22:0) acids
were ranged from 9.22-11.01, 3.63-5.37, 2.51-3.34 and 3.66-4.85% respectively during both
years of study (Table 4.2.19).
Sterols Composition
Sterols compositions of all moringa landraces are given in Table 4.2.20. All moringa trees of
six landraces produced oil with nearly similar sterols composition. The β-sitosterol was the
main sterol (more than 55%). The content of other major sterols compounds, campesterol,
stigmasterol and ∆5 -Avenasterol were ranged from 16.42-17.80, 15.94-18.74 and 4.97-6.09%
respectively during both years of study.
Discussion
A sexually propagated moringa tree through stem cuttings can produce flowers and seeds
within a year. Animashaun et al. (2013) reported that trees propagated through seed, develops
longer roots then grown by stem cuttings. So trees grown by seeds are recommended in area
where the water table is deep. However, Ramachandran et al. (1980) reported plants
propagated through seeds produce poor fruits (pods) due to genetic diversity, as moringa highly
cross pollinated specie. Moringa is commonly cultivated in Southern Punjab, Pakistan (Anwar
et al., 2006). There was significant variation in pod yield of moringa landraces of different
regions (Rajangam et al., 2001; Ndubuaku et al., 2014). Six moringa landraces propagated
through stem cuttings, produced flowers and pods within one year of plantation except FSD.
However, during second year all landraces produced seeds. Ramachandran et al. (1980)
64
reported that moringa tree propagated through stem cutting produce seeds yield within one
year. Landrace selected from Rahimyar Khan (RYK) performed best during both years of study
and attained maximum pod length, pod weight, number of seeds per pod, seed weight per pod,
number of pods per tree and ultimately seed weight per tree. Values of these seed yield and
yield parameters were higher than the reported value of Ferrao and Ferrao (1970) and Proyecto
Biomasa (1996). Ayerza (2011) reported that significant differences in seed yield and yield
attributes and oil yield were recorded in genotypes of moringa.
Percent oil contents in seeds of both harvests were similar in all landraces. Seed oil
contents were in range of 36.80-38.43%. Ayerza (2011) reported that soil and environmental
conditions may affect seed oil content more than genotype. Muhl et al. (2014) also reported
that inadequate water supply during reproductive and flowering stage ensures better fruit set
and ultimately greater yield. Seed oil content was within the range of previously reported work
in Pakistan and other countries (Anwar et al., 2006; Anwar and Rashid, 2007; Orhevba et al.,
2013) and even higher as reported in African moringa variety (Mbololo) (Tsaknis et al., 1999)
and conventional oilseed crops like cotton (15-24%), soybean (17-21%), safflower (25-40%)
and mustard (24-40%)(Mcginely, 1991). Maximum oil yields per tree and oil yield per hectare
were recorded in RYK trees during both years of study. This indicates that might be RYK
landrace has more genetic potential than other landraces. Ayerza (2011) reported that seed and
oil yield potential of two varieties (Indian PKM-1 and African Mbololo) under same conditions
were significantly different. Rajangam et al. (2011) reported tremendous variation in seed yield
potential of moringa. Ndubuaku et al. (2014) also reported, in Nigeria seed yield ranges
between 4-24 tons per hectare depending upon climate, soil, available nutrients and
precipitation.
However, physio-chemical characteristics of seed oil had little variability and
comparable with the earlier reports of moringa seed oil characteristics. Refractive index at
40°C (1.4622-1.4627), saponification value (176.74-181.28 KOH/g of oil), iodine value
(68.80-71-81 g of iodine/100 g of oil) and free fatty acids (0.82-0.87% oleic acid) in this study
are within same range as of earlier reported work in Pakistan, Kenya, Malawi and India
(Tsaknis et al., 1998; Tsaknis et al., 1999; Lalas and Tsaknis, 2002; Anwar and Bhanger, 2003;
Anwar et al., 2006).
65
Peroxide value (PV), specific extinction and p-anisdine value (p-AV) are indicators of
oxidative state of oil. These values (Table 4.2.15-4.2.18) depict that moringa trees of different
landraces can produce oil of same quality regarding oxidative stability. . However, peroxide
value of different landraces oil was higher than earlier reported value in Pakistan and wild trees
of Malawi (Anwar and Bhanger, 2003; Tsaknis et al., 1998; Anwar et al., 2006) and less than
Indian variety PKM-1(Lalas and Tsaknis 2002). Specific extinction indicates oxidative
deterioration of oil. During both years of experiment, specific extinction values of oil were
similar in all landraces (Table 4.2.18 and 4.2.18). Values are within range of earlier reported
work in Pakistan (Anwar and Bhanger, 2003; Anwar et al., 2006) however, less than Indian
variety PKM-1 and African variety Mbololo (Tsakins et al., 1998; Lalas and Tsaknis 2002).
Different landraces of moringa propagated through stem cuttings also produced seeds
having similar in fatty acids composition. The contents of saturated fatty acids ranged between
21.40-22.50%. High behenic acid content in oil is the reason why moringa oil commercially
known as “Ben oil” or “Behen oil.” Moringa oil contains high level of monounsaturated fatty
acid. In seed oil of all landraces, oleic acid (ω-9) was major fatty acid (>71%), while
polyunsaturated fatty acids (linoleic and linolenic acid) were less than 1% (Table 4.2.19).
Amount of monounsaturated fatty acid (oleic acid) in moringa oil was higher than values of
conventional oil seed crop like Palm oil (45.6%), Canola oil (57.4%) and Soybean oil (24.8%)
(Abdulkarim et al., 2007). Current study revealed that moringa oil fall in the category of high
oleic acid oil, and contains high monounsaturated to the saturated fatty acid ratio
(MUFA/SFA). High MUFA/SFA is associated with health benefits and decreases chances of
heart diseases (Mensink and Katan, 1990; Corbett, 2003; Schwingshackl et al., 2014). Moringa
seed oil also resembles with olive oil regarding fatty acid composition (Ramachandran et al,.
1980). Warner and Knowlton (1997) reported that moringa oil is more stable than other edible
oils due to less oxidation both at ambient and high temperatures as it contains more
monounsaturated and less polyunsaturated fatty acids. Anwar et al. (2007) reported that proper
blending of moringa oil in high linoleic oils not only improves oil nutritional value but also
improves oil stability to oxidation during cooking and deep frying. The content of PUFA
(polyunsaturated fatty acid) in moringa oil was less than 1% in all landraces. Cosmetic industry
use moringa oil over other high oleic acid source due to very low content of PUFA, oil is less
prone to oxidation (Kleiman et al., 2008).
66
There was no significant difference in sterol composition of oil from selected landraces.
The sterol fractions of the oil from cultivated landraces mainly comprised of campesterol,
stigmasterol, β-sitosterol and delta 5-avenasterol. These four plant sterols comprised of 96%
total sterols in all selected landraces. β-sitosterol was most important and predominant sterol
(>55%) compound in oil of all landraces. It is reported to be involved in metabolism of
cholesterol and reduces the LDL cholesterol level in blood and ultimately decreases risk of
coronary heart diseases (Abumweis et al., 2008; Ros et al., 2014). Gupta et al. (2011) also
reported that β-sitosterol had an antidiabetic potential.
Sterol composition of moringa landraces of current study is different than earlier
reported work in Pakistan, India (PKM-1) and Kenya (Tsaknis et al., 1999; Lalas and Tsaknis,
2002; Anwar et al., 2006; Anwar and Bhanger, 2003). Sterol composition of selected landraces
comprises of more than 55% β-sitosterol, which are higher than earlier reported work.
Conclusion
Selected moringa landraces propagated through stem cuttings produced flowers and seed
within one year except FSD (which produced seed yield during second year). Significant
variabilities in seed production and oil yield were observed during both years of experiment.
Maximum seed and oil yield were recorded in RYK landrace. However, physio-chemical
characteristics, oxidative state and oil quality had little variability between landraces and their
values are much comparable with reported work in Pakistan and other countries.
67
Table 4.9: Mean sum of squares of data of pod length, pod weight, number of seeds pod -
1
and seed weight pod -1 of cultivated moringa landraces during year 2014

Treatment 4 17.01* 10.39* 10.21* 2.34*
Replication 3 2.00 0.11 1.18 0.012
Error 12 3.41 0.96 1.25 0.015
Table 4.10: Mean sum of squares of data of pod length, pod weight, number of seeds pod-
1 and Seed weight pod-1 of cultivated moringa landraces during year 2015

Treatment 5 12.76* 14.89* 0.98* 2.92*
Replication 3 0.12 0.046 0.25 0.02
Error 15 0.45 0.11 0.09 0.016
68
Table 4.11: Mean sum of squares of data of number of pods tree -1, seed weight tree-1,
1000 seeds weight, oil yield tree -1 and oil yield hectare-1 of cultivated moringa landraces
during year 2014
Number of Seed weight 1000 seed Oil yield per Oil yield per
SOV df
pods per Tree per tree weight tree hectare
Treatment 4 575.45* 0.13* 1196.91* 0020* 4806.82*
Replication 3 50.73 0.001 107.47 0.0001 17.71
Error 12 75.31 0.001 52.31 0.0001 36.52
Table 4.12: Mean sum of squares of data of number of pods tree -1, seed weight tree-1,
1000 seeds weight, oil yield tree -1 and oil yield hectare-1 of cultivated moringa landraces
during year 2015
Seed Oil yield per

Number of 1000 seed Oil yield
SOV df weight per hectare
pods per Tree weight per tree
tree
Treatment 5 994.66* 0.27* 7343.42* 0.0436* 9825.03*
Replication 3 273.39 0.0007 1.01 0.0002 12.10
Error 15
157.35 0.0003 18.58 0.0009 20.43
69
value, iodine value and free fatty acids of cultivated moringa landraces during year 2014

SOV df
Treatment 4 2.69ns 5.93ns 24.43* 0.88* 0.0040ns
Replication 3 2.36 2.81 1.79 0.26 0.0027
Error 12 1.39 1.83 5.43 0.19 0.0088
value, iodine value and free fatty acids of cultivated moringa landraces during year 2015

SOV df
Treatment 5 3.58ns 5.27ns 12.76* 5.66ns 0.0078ns
Replication 3 2.32 4.05 2.18 22.55 0.0027
Error 15 1.37 1.82 2.35 14.23 0.0053
70
Table 4.15: Mean sum of squares of data of oil peroxide value, p-anisdine value, K232,
and K268 of cultivated moringa landraces during year 2014
p-anisdine
1cm) K268(K1%
1cm)
value
Treatment 4 0.25* 0.29* 0.0044ns 0.0018 ns
Replication 3 0.0085 0.0046 0.0030 0.0022
Error 12 0.31 0.0095 0.0026 0.0061
Table 4.16: Mean sum of squares of data of oil peroxide value, p-anisdine value, K232,
and K268 of cultivated moringa landraces during year 2015
p-anisdine
1cm) K268(K1%
1cm)
value
Treatment 5 0.14ns 0.0647ns 0.0005 ns 0.0004ns
Replication 3 0.11 0.0025 0.0001 0.0003
Error 15 0.12 0.0269 0.0001 0.0002
71
Table 4.2.1: Effect of different landraces on pod length (cm) of cultivated moringa trees

FSD n/a* 40.67 d
RYK 46.16 a 45.95 a
BWP 41.26 b 42.71c
LAY 42.89 ab 43.39bc
MUL 41.17 b 43.45 bc
KWL 41.94 b 44.63 ab
*No seed
CVC 4.17 1.54
production
during year
2014 in FSD
Table 4.2.2: Locations Year 2014 Year 2015 Effect of

different landraces on
pod weight (g) FSD n/a* 5.57 c of cultivated
moringa trees RYK 11.32 a 11.55 a
BWP 8.27 b 8.22 b
LAY 8.22 b 7.56 b
MUL 7.56 b 8.23 b
KWL 7.99 b 7.98 b
CVC 0.70 0.75
*No seed production during year 2014 in FSD

72
Table 4.2.3: FSD n/a* 17.25 ab Effect of

different RYK 18.75 a 17.96 a landraces on
number of seeds per pod
of cultivated BWP 16.96 ab 16.64 b moringa
LAY 14.42 c 16.75 b
MUL 16.00 bc 16.79 b
KWL 17.20 ab 17.29 ab
CVC 2.53 0.69

FSD n/a* 2.11 d
RYK 4.90 a 4.77 a
BWP 3.17 b 3.28 bc
LAY 3.10 b 3.07 c
MUL 3.21 b 3.36 bc
KWL 3.34 b 3.45 b
CVC 0.28 0.29

Table 4.2.4: Effect of different landraces on seeds weight per pod (g) of cultivated
moringa
73
FSD n/a* 292 a
RYK 235 ab 293 a
BWP 239 ab 268 ab
*No seed
LAY 223 b 256 b production
during year 2014
MUL 250 a 283 ab in FSD
KWL 253 a 295 a Means sharing
same letters are
CVC 19.59 28.82 statically similar
CVC = Critical
value of comparison
Table 4.2.5: Effect of different landraces on number of pods per tree of cultivated
moringa

74
Table 4.2.6: Effect of different landraces on seeds weight per tree (kg) of cultivated
moringa trees


FSD n/a* 0.63 f
RYK 1.15 a 1.39 a
BWP 0.76 cd 0.87 d
LAY 0.69 d 0.79 e
MUL 0.80 bc 0.95 c
KWL 0.85 b 1.00 b
CVC 0.08 0.04
75
Table 4.2.7: Effect of different landraces on 1000 seeds weight (g) of cultivated moringa
trees
FSD n/a* 130.65 e

RYK 258.19 a 254.10 a
BWP 222.93 bc 224.47 c
LAY 213.27 c 210.03 d
MUL 221.27 bc 216.55 cd
KWL 229.98 b 236.67 b *No seed
production during
CVC 16.32 9.90 year 2014 in FSD
Means sharing
Table 4.2.8: Effect of different landraces on oil yield per tree (kg) of cultivated moringa
trees
FSD n/a* 0.23 f
RYK 0.44 a 0.53 a
BWP 0.29 cd 0.32 d
LAY 0.26 d 0.28 e
MUL 0.30 bc 0.36 c
KWL 0.33 b 0.39 b
CVC 0.03 0.02
*No seed
production during
year 2014 in FSD
76
Table 4.2.9: Effect of different landraces on oil yield ha -1 (kg) of cultivated moringa
trees
FSD n/a* 107.84 f
RYK 210.98 a 253.01 a
BWP 135.22 c 154.60 d
LAY 120.77 d 135.69 e
MUL 143.77 bc 168.54 c
KWL 155.30 b 182.76 b
CVC 13.64 10.38 *No seed
production during
year 2014 in FSD
Table 4.2.10: Effect of different

landraces on seed oil content (%) of
cultivated FSD n/a* 36.80 moringa trees
RYK 38.36 37.96

BWP 37.47 36.91
LAY 37.44 37.11
MUL 37.51 37.13
KWL 38.43 38.14
CVC 1.39 1.41
*No seed
production during year 2014 in FSD
77

landraces on Locations Year 2014 Year 2015 refractive index
(40°C) of cultivated moringa trees
FSD n/a* 1.4627
RYK 1.4627 1.4632
BWP 1.4621 1.4623
LAY 1.4623 1.4625
MUL 1.4626 1.4626
KWL 1.4621 1.4623
CVC 0.017 0.017

Table 4.2.12: Effect of different landraces on saponification value (mg of KOH/g of oil)
of cultivated moringa trees
FSD n/a* 179.66 a
RYK 183.33 a 179.24 ab
BWP 177.59 b 176.30 abc
LAY 180.83 ab 176.86 abc
MUL 178.02 b 175.45 c
KWL 178.02 b 175.88 bc
CVC 5.26 3.52
*No seed production
during year 2014 in FSD
78
Table 4.2.13: Effect of different landraces on iodine value (g of I/100 g of oil) of

FSD n/a 69.14
RYK 68.65 ab 68.80
BWP 68.2 ab 69.65
LAY 68.03 b 71.20
MUL 69.14 a 69.71
KWL 68.06 b 71.81
CVC 0.99 8.66 *No seed production
during year 2014 in
FSD
landraces on free Locations Year 2014 Year 2015 fatty acid (% oleic
acid) of cultivated moringa trees
FSD n/a* 0.45
RYK 0.52 0.48
BWP 0.45 0.45
LAY 0.44 0.42
MUL 0.49 0.43
KWL 0.45 0.41
CVC 0.21 0.16

79
80
Table 4.2.15: Effect of different landraces on peroxide value (meq of O 2 kg -1 of oil) of

FSD n/a* 1.25
RYK 1.43 bc 1.45
BWP 1.60 ab 1.45
LAY 1.88 a 1.05
MUL 1.48 bc 1.05
KWL 1.20 c 1.40
*No seed
CVC 0.37 0.80 production during
year 2014 in FSD
Table 4.2.16: Effect of different landraces on p-anisidine value of cultivated moringa

trees

FSD n/a* 1.37
RYK 1.30 c 1.49
BWP 1.75 ab 1.38
LAY 1.94 a 1.48
MUL 1.71 b 1.67
KWL 1.38 c 1.37
CVC 0.22 0.38 *No seed production
during year 2014 in
FSD
81
Table 4.2.17: Effect of different landraces on K232(𝐊 𝟏%
𝟏𝐜𝐦 ) of cultivated moringa trees

FSD n/a* 1.65
RYK 1.67 1.65
BWP 1.75 1.78
LAY 1.67 1.69
MUL 1.72 1.65
KWL 1.66 1.65
*No seed
CVC 0.11 0.03 production during
year 2014 in FSD
Means sharing

landraces on Locations Year 2014 Year 2015 K268(𝐊 𝟏%
𝟏𝐜𝐦 ) of
cultivated FSD n/a* 0.40 moringa trees
RYK 0.42 0.40

BWP 0.44 0.43
LAY 0.42 0.42
MUL 0.43 0.41
KWL 0.42 0.39
CVC 0.17 0.03

82
Table 4.2.19: Effect of different landraces of on fatty acids composition (g per 100 g of fatty acids) of cultivated moringa trees
during year 2014-15
Locations Year Fatty acids

16:0 16:1 18:0 18:1 18:2 18:3 20:0 20:1 22:0 24:0
2014 n/a* n/a n/a n/a n/a n/a n/a n/a n/a n/a
FSD
2015 9.87 1.98 4.63 71.90 0.65 0.09 3.12 2.35 4.29 1.11
2014 9.27 1.7 5.37 71.79 0.62 0.1 3.34 2.2 4.47 1.13
RYK
2015 10.85 1.94 3.9 72.27 0.74 0.09 2.68 2.37 4 1.16
2014 10.1 1.85 4.61 72.7 0.73 0.1 2.89 2.2 3.74 1.08
BWP
2015 10.01 2.06 4.31 71.79 0.66 0.1 3.04 2.41 4.46 1.16
2014 9.22 1.85 4.32 72.53 0.71 0.09 2.87 2.4 4.85 1.16
LAY
2015 10.23 2 4.13 72.06 0.68 0.09 2.91 2.38 4.34 1.17
2014 9.48 1.61 5.16 72.36 0.67 0.1 3.2 2.15 4.15 1.13
MUL
2015 10.58 1.97 3.83 72.84 0.73 0.09 2.65 2.43 3.78 1.09
2014 9.48 1.72 5.16 71.73 0.64 0.11 3.28 2.21 4.51 1.17
KWL
2015 11.01 1.94 3.63 72.85 0.76 0.09 2.51 2.45 3.66 1.1
2014 2.61 1.64 3.91 2.95 0.58 0.05 3.29 0.40 1.37 0.69
CVC
2015 2.51 0.50 1.19 1.39 0.20 0.04 0.77 0.19 1.41 0.18
83
FSD RYK BWP LAY MUL KWL CVC
Sterol compositon
2014 2015 2014 2015 2014 2015 2014 2015 2014 2015 2014 2015 2014 2015
Cholesterol n/a* 0.14 0.13 0.13 0.13 0.14 0.14 0.13 0.15 0.13 0.13 0.12 0.04 0.09
24-Methylenecholesterol n/a 0.21 0.30 0.25 0.36 0.27 0.26 0.27 0.21 0.25 0.31 0.25 0.32 0.16
Campesterol n/a 16.62 17.35 17.73 16.69 16.76 16.63 16.81 16.42 17.74 16.41 17.80 1.92 3.06
Campestanol n/a 0.07 0.05 0.07 0.06 0.10 0.10 0.10 0.07 0.07 0.05 0.07 0.13 0.14
Stigmasterol n/a 17.74 18.07 17.09 18.74 16.66 17.98 16.82 18.22 16.68 18.31 15.94 1.82 4.27
Clerosterol n/a 0.56 0.46 0.58 0.46 0.53 0.50 0.53 0.50 0.56 0.45 0.57 0.15 0.19
β-Sitosterol n/a 55.83 55.51 55.15 55.48 57.14 56.70 56.78 57.26 55.80 55.94 56.44 4.33 2.18
∆5- Avenasterol n/a 6.00 6.09 5.70 4.97 5.53 5.35 5.65 5.76 5.57 5.62 5.65 3.03 1.27
∆5-24-Stigmastadienol n/a 0.47 0.36 0.52 0.37 0.47 0.45 0.48 0.34 0.50 0.35 0.51 0.26 0.12
∆7-Stigmastadienol n/a 1.14 0.54 1.26 1.23 1.16 0.71 1.17 0.30 1.23 1.10 1.25 1.89 0.26
∆7- Avenasterol n/a 1.21 1.13 1.50 1.46 1.26 1.18 1.28 0.76 1.47 1.33 1.49 0.57 0.66
Table 4.2.20: Effect of different landraces of on sterol composition (%) of cultivated moringa trees during year 2014-15
84
85
Summary
This study was designed to explore qualitative and quantitative oil yield potential of wild and
vegetatively propagated moringa trees. Two experiments were performed to determine seed, oil
yield and oil quality of wild and cultivated moringa trees. In first experiment six locations in
Punjab {Faisalabad (FSD), Rahimyar Khan (RYK), Bahawalpur (BWP), Layyah (LAY), Mulan
(MUL) and Khanewal (KWL)} were selected where moringa is wildly grown. Five trees (at least
five years old) were tagged in each location for study. Mature dry pods were harvested from each
tagged tree by hand and data regarding seed and oil yield and quality were taken for two growing
seasons (2014 and 2015). During both years of experiment pods and oil yield of wild moringa
landraces were significantly different in all six locations. RYK landrace perform best among all
landraces and produced highest seed yield during both harvests (4.11 and 3.77 kg tree-1) while
least seeds weight per tree was recorded from FSD. Maximum seed oil contents (35.95 and
37.02%) and oil yield per tree (1.48 and 1.40 kg tree -1) were recorded in RYK landrace during
both years. However, oil physio-chemical characteristics, oxidative state, fatty acid and sterol
composition were almost similar in all selected landraces. Seed oil of all landraces possesses good
oxidative state. Peroxide and p-anisidine values of seed oil of selected landraces were ranged
between 0.95-1.68 meq of O2 kg-1 and 3.27-5.07 respectively during both growing seasons. The
monounsaturated fatty acid oleic acid (18:1) was dominate fatty acid (contribute more than 71%
of total fatty acid) in seed oil of all landraces while, polyunsaturated fatty acids were observed less
than 1%. The content of saturated fatty acids, Palmitic (16:0), Stearic (18:0), Arachidic (20:0) and
Behenic (22:0) acid were ranged from 8.99-10.99, 3.98-5.91, 2.67-3.53 and 4.01-4.99%
respectively during both years of study. All moringa landraces produced oil with nearly similar
sterols composition. The β-sitosterol was the main sterols (more than 55% of total sterol
composition). The content of other major sterols compounds; campesterol, stigmasterol and ∆5 -
Avenasterol were ranged from 15.81-18.41, 15.96-18.37 and 5.04-7.63% respectively during both
growing seasons.
In second experiment, stem cuttings of selected landraces were propagated at Farmer’s field
Khanewal to study seed yield response and to determine oil yield and quality. All landraces
propagation through stem cuttings produced flowers and pods within one year of plantation except
FSD, however, during second year all landraces produced seed yield. Seed and oil yield of
cultivated trees were significantly different among all landraces. Maximum seed yield (1.15 and
1.39 kg tree-1) and oil yield (210.98 and 253.01 kg ha -1) were noticed in RYK landrace during both
growing seasons. However, seed oil content, oil physio-chemical characteristics, oxidative
86
stability, fatty acid and sterol composition had little variability among all landraces. Oleic acid was
main fatty acid in seed oil of all landraces (>71% of total fatty acid). The content of saturated fatty
acids, Palmitic (16:0), Stearic (18:0), Arachidic (20:0) and Behenic (22:0) acid were ranged from
9.22-11.01, 3.63-5.37, 2.51-3.34 and 3.66-4.85% respectively during both year of study. The β-
sitosterol was the main plant sterol (more than 55% of total sterol composition). The content of
other major sterols compounds, campesterol, stigmasterol and ∆ 5 -Avenasterol were ranged from
16.42-17.80, 15.94-18.74 and 4.97-6.09% respectively during both year of study.
In conclusion, RYK landrace was appeared as high seed and oil yielder in both cases either wild
grown or cultivated. More importantly, seed oil quality was observed similar in all landraces in
all growing conditions. On basis of finding, it is concluded that RYK landrace has potential and
can be propagated as oil seed tree in Pakistan.
87
5. Literature Cited
Abdulkarim, S.M., K. Long, O.M. Lai, S.K.S. Muhammad and H.M. Ghazali. 2007. Some
physico-chemical properties of Moringa oleifera seed oil extracted using solvent and
aqueous enzymatic method. Food Chem. 93: 253-263.
Abumweis, S.S., R. Barake and P.J. Jones. 2008. Plant sterols/stanols as cholesterol lowering
agents: A meta-analysis of randomized controlled trials. Food Nutr. Res. 52: 4.
Agrawal, A.K., A.P. Joshi, S.K. Kandwal and R. Dhasmana. 1986. An ecological analysis of Malin
riverain forest of outer Garhwal Himalaya (western Himalaya). Indian J. Ecol. 13: 15-21.
American Oils Chemists Society (AOCS). 2009. Determination of Specific Extinction of oils and
Fats, Ultraviolet Absorption; Official Method Ch 5–91. New York, NY, USA, 2009.
Anhwange, B., V. Ajibola and S. Oniye. 2004. Amino acid composition of the seeds of Moringa
oleifera (Lam), Detarium microcarpum (Guill & Sperr) and Bauhinia monandra (Linn.).
Chem. Class J. 2004: 9-13.
Animashaun, J. 2013. Prospects of Agriculture Enterprise for Sustainable Economic Development:
Success Story of University of Ilorin Moringa Value-Addition Activities. In: Proceedings
of the 4th International Conference of the African Association of Agricultural Economists,
Hammamet, Tunisia.
Anwar, F. and M. Bhanger. 2003. Analytical characterization of Moringa oleifera seed oil grown
in temperate regions of Pakistan. J. Agric. Food Chem. 51: 6558-6563.
Anwar, F., M. Ashraf and M.I. Bhanger. 2005. Interprovenance variation in the composition of
Moringa oleifera oilseeds from Pakistan. J. Am. Oil Chem. Soc. 82: 45-51.
Anwar, F., N.S. Zafar and U. Rashid. 2006. Characterization of Moringa oleifera seed oil from
drought and irrigated regions of Punjab, Pakistan. Grasas Y Aceites, 7: 160-168.
Anwar, F., S. Latif, M. Ashraf and A.H. Gilani. 2007. Moringa oleifera: a food plant with multiple
bio-chemical and medicinal uses- a review. Phytother. Res. 21: 17-25.
Atta-Karh, A.N. 1990. Availability and use of fodder shrubs and trees in tropical Africa. In: Shrubs
and Tree Fodders for Farm Animals. Proceedings of a Workshop held in Denpasar,
Indonesia (Ed. Devendra C). IDRC, Ottawa, Canada, pp. 140-162.
Ayerza, R. 2011. Seed yield components, oil content, and fatty acid composition of two cultivars
of moringa (Moringa oleifera Lam.) growing in the Arid Chaco of Argentina. Ind. Crops
Prod. 33: 389-394.
Ayerza, R. 2012. Seed and oil yields of Moringa oleifera variety Periyakalum-1 introduced for oil
production in four ecosystems of South America. Ind. Crops Prod. 36: 70 -73.
88
Azam, M.M., A. Waris and N.M. Nahar. 2005. Properties and potential of fatty acid methyl esters
of some non-traditional seed oils for use as biodiesel in India. Biomass Bioenerg. 29: 293-
302.
Banerji, R., A. Bajpai and S.C. Verma. 2009. Oil and fatty acid diversity in genetically variable
clones of Moringa oleifera from India. J. Oloe. Sci. 58: 9-16
Banerji, R., S.C. Verma and P. Pushpangadan. 2003. Oil potential of moringa. Nat. Prod. Radiance
2: 68-69.
Basra, S.M.A., M.N. Irfan, I. Afzal, and M. Ashfaq. 2011. Potential of moringa leaf extract as seed
priming agent in hybrid maize. Int. J. Agric. Biol. 13: 1006-1010.
Basra, S.M.A., Z. Iqbal, K. Rehman, H. Rehman, M.F. Ejaz. 2014. Time Course Changes in pH,
Electrical Conductivity and Heavy Metals (Pb, Cr) of Wastewater Using Moringa oleifera
Lam. Seed and Alum, a Comparative Evaluation. J. Appl. Res. Technol. 12: 560-567
Benavides, J. 1994. La investigación en árboles forrajeros. Árboles y arbustos forrajeros de
América Central (CATIE), Turrialba, Costa Rica
Berger, M.R., M. Habs, S.A.A. Jahn, and D. Schmalhl. 1984. Toxicological assessment of seeds
from Moringa oleifera and Moringa stenopetala, two highly efficient primary coagulants
for domestic water treatment of tropical waters. East. African. Med. J. 61: 712 -717
Booth, F.E.M. and G.E. Wickens. 1988. Non-timber uses of selected arid zone trees and shrubs in
Africa. FAO Conservation Guide 19, Food and Agriculture Organization of the United
Nations, Rome
Boskou, D. 2011. Olive oil. In Vegetable oils in Food Technology; Wiley-Blackwell: Hoboken,
NJ, USA, pp. 243–271
Brault, M. and R. Maldiney. 1999. Mechanisms of cytokinin action. Plant Physiol. Biochem. 37:
403-412
Brockman, H. 2008. Production of biodiesel from perennials. Government of Western Australia:
Department of Agric. and Food.
Carvalho, I.S., M. Chaves and C.P. Ricardo. 2005. Influence of water stress on the chemical
composition of seeds of two Lupins (Lupinus albus and Lupinus mutabilis). J. Agr. Crop
Sci. 191: 95-98
Champion, H. G. 1936. Indian forest records: part 1 - a preliminary survey of forest types of India
and Burma. Government of India Press, New Delhi
Chandan, R.C. 2006. History and consumption trends. In: Chandan R.C. (ed.). Manufacturing
Yogurt and Fermented Milks. Blackwell Publishing, Ames, IA, USA. pp. 3-15.
89
Choudhary, S.K., S.K. Gupta, M.K. Singh and Sushant, 2016. Drumstick tree’ (Moringa oleifera
Lam.) is multipurpose potential crop in rural area of India. Int. J. Agric. Sci., 12: 115-122
Corbett, P. 2003. It is time for an oil change! Opportunities for high-oleic vegetables oils. Inform.
14: 480-1.
Crosby, G.W. 2007. Soilless culture of moringa (Moringa oleifera Lam.) for the production of
fresh biomass - Udini. Available at: http://udini.proquest.com/view/soillessculture-of
moringa-moringa-goid: 304844775/ [Accessed September 24, 2012].
CSIR .1962. The Wealth of India. A Dictionary of Indian Raw Materials and Industrial Products.
Raw Materials, Volume 6, L–M. CSIR, New Delhi, India.
Davies, P.J. 1995. Plant hormones: physiology, biochemistry and molecular biology. Kluwer
Academic Publishers, Dordrecht.
Demonty, I., R.T. Ras, H.C. van der Knaap, G.S. Duchateau, L. Meijer, P.L. Zock, J.M. Geleijnse
and E.A. Trautwein. 2009. Continuous dose-response relationship of the LDL-cholesterol-
lowering effect of phytosterol intake. J. Nutr. 139: 271-284.
Eilert, U., B. Wolters and A. Nahrstedt. 1980. Antibiotic principle of seeds of Moringa oleifera.
Planta Med. 39: 235.
Fahey, J.W. 2005. Moringa oleifera: a review of the medical evidence for its nutritional,
therapeutic, and prophylactic properties: part 1. Trees Life J. 1: 1-15.
FAO. 1982: Fruit-bearing forest trees: technical notes. Food and Agriculture Organization of the
United Nations, Rome
Ferrao, A.M.B.C. and J.E. Ferrao. 1970. Acidos gordos em oleo de Moringueiro (Moringa oleifera
Lam.). Agronomia Angolana. 8: 3-16.
Foidl, N., H.P.S. Makkar and K. Becker. 2001. The potential of Moringa oleifera for agricultural
and industrial Uses p. 45-76. In J. Lowell and C.T.A. Fuglie (Eds.). The miracle tree: The
multiple uses of Moringa. Wageningen, The Netherlands.
Fuglie, L.J., 2001. The miracle tree: moringa oleifera: natural nutrition for the tropics. Training
manual. Dakar: Church World Service.
Gbeassor, M., A.Y. Kedjagni, K. Koumaglo, C. de Souza, K. Agbo, K. Aklikokou and K.A.
Amegbo. 1990. In vitro antimalarial activity of six medicinal plants. Phytother. Res. 4:
115-117.
Gigon A., A.R. Matos, D. Laffray, Y.Z. Fodil and A.T. Pham-Thi. 2004. Effect of drought stress
on lipid metabolism in the leaves of Arabidopsis thaliana (Ecotype Columbia). Ann. Bot.
94:345-351.
90
Government of Pakistan. 2014-15. Agricultural Statistics of Pakistan. Ministry of Food,
Agriculture and Livestock, Islamabad
Gupta, R., A.K. Sharma, M.P. Dobhal, M.C. Sharma and R.S. Gupta. 2011. Antidiabetic and
antioxidant potential of β-sitosterol in streptozotocin-induced experimental
hyperglycemia. J. Diabetes. 3: 29-37.
Hartwell, J.L. 1971. Plants used against cancer. A survey. Lloydia 34: 386-425.
International Olive Council (IOC). Determination of the Composition and Content of Sterols and
Triterpene Dialcohols by Capillary Column Gas Chromatography; COI/T.20/Doc. No.
30/Rev.1 2013; International Olive Council (IOC): Madrid, Spain, 2013
International Olive Council (IOC). Determination of Trans-Unsaturated Fatty Acids by Capillary
Column Gas Chromatography; COI/T.20/Doc. no. 33-2015; International Olive Council
(IOC): Madrid, Spain, 2015
Jahn, S.A.A., H.A. Musnad and H. Burgstaller. 1986. The tree that purifies water: Cultivating
multipurpose moringaceae in the Sudan. Unasylva. 38: 23-28.
Jyothi, P.V., J.B. Atlura and C.S. Reddi. 1990. Pollination ecology of Moringa oleifera
(Moringaceae). Proc. Indian Academy of Sciences, Plant Sci. 100: 33-42.
Kale, S. and G. Megha. 2011. Formulation and in vitro evaluation for sun protection factor of
Moringa oleifera Lam (family-Moringaceae) oil sunscreen cream. Int. J. Pharm. Pharm.
Sci. 3: 371-375.
Kleiman, Robert, J. Brown and J. Hill. 2006. Moringa oil: a “new yet old” unique cosmetic
emollient. Information 17:739.
Kshirsagar, C.R. and T.F. D’souza. 1989. A new disease of drumstick. J. Maharashtra Agric. Univ.
14: 241-242.
Kumar, A.R., M. Prabhu, V. Ponnuswami, V. Lakshmanan and A. Nithyadevi. 2014. Scientific
seed production techniques in Moringa. Agric. Rev. 35: 69-73.
Lahjie, A.M. and B. Siebert. 1987. Kelor or horse radish tree (Moringa oleifera Lam.). A report
from East Kalimantan. German Forestry Group, Mulawarman Univ., GFG report. 6: 41 -43.
Lalas, S. and J. Tsaknis. 2002. Characterization of Moringa oleifera seed oil variety “periyakulam
L”. J. Food Compos. Anal. 15: 65077.
Lee, Y.W., J.D. Kim, J. Zheng and K.H. Row. 2007. Comparisons of isoflavones from Korean and
Chinese soybean and processed products. J Biochem Eng. 36: 49-53.
Lefroy E.C., P.R. Dann, J.H. Wildin, R.N. Wesley, Smith and A.A. McGowan. 1992. Trees and
shrubs as sources of fodder in Australia. Agrofores. Sys. 20: 117-139.
91
Little, E.L. and F.H. Wadsworth. 1964. Common trees of Puerto Rico and the Virgin Islands.
Agric. Handb. 249. U.S. Dep. Agric., Washington, D.C.
Lokuruka, M.N.I. 2007. Role of fatty acids of milk and dairy products in cardiovascular diseases:
a review. Afr. J. Food Agric. Nutr. Dev. 7: 45-59.
Lowell, J.F. 1999. Moringa oleifera: Natural nutrition for the tropics. Dakar Senegal: Church
World Service.
Mabberley, D.J. 1997. The Plant Book. 2 nd ed.. Cambridge Univ. Press, Cambridge, UK, 467.
Mahajan, S. and Y.K. Sharma. 1984. Production of rayon grade pulp from Moringa oleifera. Indian
For. 110: 303-306.
Makkar, H.P.S. and K. Becker. 1996. Nutrional value and antinutritional components of whole
and ethanol extracted Moringa oleifera Leaves. Anim. Feed Sci. Technol. 63: 211-228.
Mandloi, M., S. Chaudhari and G.K. Folkard. 2004. Evaluation of natural coagulants for direct
filtration. Environ. Technol. 25: 481-489.
Manzoor, M., F. Anwar, T. Iqbal and M.I. Bhanger. 2007. Physico-chemical characterization of
Moringa concanensis seeds and seed oil. J. Am. Oil Chem. Soc. 84: 413-419.
Martin, F.W. and R.M. Ruperté. 1979. Edible leaves of the tropics. 2 nd ed., U.S. Dep. Agric.,
Science and Education Admin., Agric. Res., Southern Region, Mayagüez, PR.
Mathur, B. 2006. Moringa for cattle fodder and plant growth. Treesfor Life J [online]. Available
at http://www.tfljournal.org/staticpages/index.php?page=call-for-studies-cattle-fodder.
Mcginely, L. 1991. Analysis and quality control for processing and processed fats. In J.B. Rossell
and J.L.R. Pritchard (Eds.), Analysis of oilseeds, fats and fatty foods New York, Elsevier
Applied Science, pp. 460-470
Mendieta-Araica, B., R. Sporndly, N.R. Sanchez and E. Sporndly. 2011. Moringa (Moringa
oleifera) leaf meal as a source of protein in locally produced concentrates for dairy cows
fed low protein diets in tropical areas. Livestock Sci. 137: 10-17.
Mensink, R.P. and M.B. Katan. 1990. Effect of dietary trans-fatty acids on high-density and low-
density lipoprotein cholesterol levels in healthy subjects. J. Clinical Nutr. 323: 439-445.
Morton, J.F. 1991. The horseradish tree, Moringa pterigosperma (Moringaceae). A boon to arid
lands. Econ. Bot. 45:318–333.
Mossa, J.S. 1985. A study on the crude antidiabetic drugs used in Arabian folk medicine. Int. J.
Crude Drug Res. 23: 137-145.
Mughal MH, Ali G, Srivastava PS, Iqbal M. 1999. Improvement of drumstick (Moringa
pterygosperma Gaertn.) – a unique source of food and medicine through tissue culture.
Hamdard Med 42: 37–42.
92
Munyanziza, E. and S.V. Sarwatt. 2003. The evaluation of Moringa oleifera for food security and
environmental rehabilitation in Tanzanian rural areas. J. Trop. For. Sci. 15: 450-456.
Nautiyal, B.P. and K.G. Venhataraman. 1987. Moringa (Drumstick) – an ideal tree for social
forestry. My Forest. 23: 53-58.
Ndubuaku, U.M., T.C.N. Ndubuaku and N.E. Ndubuaku. 2014. Yield characteristics of Moringa
oleifera across different ecologies in nigeria as an index of its adaptation to climate change.
Sustain. Agric. Res. 2014, 3, 95.
Negi, S.S. 1977. Fodder trees of Himachel Pradesh. Indian Forester 103: 616-622
Nel, A.A. 2001. Determination of sunflower seed quality for processing. Ph.D. Thesis. Deptt. of
Plant Production and Soil Sciences. University of Pretoria, Pretoria. pp. 40-56.
Newton, K.A., R.N. Bennett, R.B.L. Curto, E.A.S. Rosa, V.L. Turc, A. Giuffrida, A.L. Curto, F.
Crea and G.M. Timpo. 2010. Profling selected phytochemicals and nutrients in different
tissues of the multipurpose tree Moringa oleifera L., grown in Ghana. Food Chem. 122:
1047-1064.
Nordstrom, A., P. Tarkowsky, D. Tarkowská, R. Norbaek, C. Astot, K. Dolezal and G. Sandberg.
2004. Auxin regulation of cytokinin biosynthesis in Arabidopsis thaliana: a factor of
potential importance for auxin–cytokinin-regulated development. Proc. Natl. Acad. Sci.
101: 8039-8044
Nouman, W., M.T. Siddiqui, S.M.A. Basra, I. Afzal and H. Rehman. 2012. Enhancement of
emergence potential and stand establishment of Moringa oleifera Lam. by seed priming.
Turk. J. Agric. For. 36: 227-235.
Nouman, W., S.M.A. Basra, M.T. Siddiqui, A. Yasmeen, T. Gull, and M.A.C. Alcayde. 2014.
Potential of Moringa oleifera L. as livestock fodder crop: A review. Turk. J. Agric. For.
38: 1-14.
Nyein, M.M. and T. AYE. 1997: The use of Moringa oleifera (dan-da-lun) seed for the
sedimentation and decontamination of household water. Part II: community-based study.
Myanmar Health Sci. Res. J. 9: 163-166
Odee, D. 1998. Forest biotechnology research in drylands of Kenya: the development of moringa
species. Dryland Biodivers. 2: 7 - 8.
Ogunsina, B.S., T.N. Indira, A.S. Bhatnagar, C. Radha, S. Debnath, A.G.G. Krishna. 2014.
Quality characteristics and stability of Moringa oleifera seed oil of Indian origin. J. Food
Sci. Technol. 51: 5030510.
93
Oliveira, J.T.A., B.S. Silvana, M.V. Ilka, S.C. Benildo and A.M. Renato. 1999. Compositional and
nutritional attributes of seeds from the multiple purpose tree Moringa oleifera Lamarck. J.
Sci. Food Agric. 79: 815-820.
Olsen, A. 1987. Low technology water purification by bentonite clay and Moringa oleifera seeds
flocculation as performed in sudanese village: effects of Schistosoma Mansoni cericariae.
Water Res. 21: 81-92.
Orhevba, B.A., M.O. Sunmonu and H.I. Iwunze. 2013. Extraction and characterization of Moringa
oleifera seed oil. Res. Rev. J. Food Dairy Technol. 1: 22-27
Otsyina R. and B. Dzowela. 1995. Importance of Leucaena in Africa. In: Leucaena psyllid: A Treat
to Agroforestry in Africa (Eds. Ciesla WM, Nshubemuki L). FAO, Rome.
Palada, M.C. 1996. Moringa (Moringa oleifera Lam.): A versatile tree crop with horticultural
potential in the subtropical United States. HortScience. 31: 794-797.
Palada, M.C. and L.C. Chang. 2003. Suggested cultivation practice for Moringa AVRDC
publication.
Palada, M.C., L.C. Chang, R.Y. Yang and L.M. Engle. 2007. Introduction and varietal screening
of drumstick tree (Moringa spp.) for horticultural traits and adaptation in Taiwan. Acta.
Hort. 752: 249-253.
Palanisamy, V., K. Kumaresan, M. Jayabharathi and T.V. Karivaratharaju. 1985. Studies on seed
development and maturation in annual moringa. Vegetable Sci. 12: 74-78.
Paliwal, R. and V. Sharma. 2011. A review on horse radish tree (Moringa oleifera): A
multipurpose tree with high economic and commercial importance. Asian J. Biotechnol. 3:
317-328.
Parawira, W. 2010. Biodiesel production from Jatropha curcas: a review. Sci. Res. Essay. 5: 1796-
1808
Parrotta, J.A. 2001. Healing plants of peninsular India. Cabl Publishing, Wallingford, New York.
Pauwels, E.K. 2011. The protective effect of the mediterranean diet: focus on cancer and
cardiovascular risk. Med. Princ. Pract. 20:103-111.
Qaiser, M. 1973. Moringaceae. p. 1-4. In: E. Nasir and S.I. Ali. (Eds.). Flora of West Pakistan.
38, 1973, Dept. of Bot. Univ. Karachi, Karachi, Pakistan. 1973; pp 1-4.
Rahman, I.M.M., S. Barua, M. Nazimuddin, Z.A. Begum, M.A. Rahman and H. Hasegawa. 2009.
Physicochemical properties of Moringa oleifera Lam. Seed oil of the indigenous-cultivar
of bangladesh. J. Food Lipids, 16: 540-553.
94
Raja, S., B.G. Bagle and T.A. More. 2013. Drumstick (Moringa oleifera Lamk.) improvement for
semiarid and arid ecosystem: analysis of environmental stability for yield. J. Plant Breed.
Crop Sci. 5: 164-170.
Rajangam, J., R.S.A. Manavalan, T. Thangaraj, A. Vijayakumar and N. Muthukrishan. 2001.
Status of Production and Utilisation of Moringa in Southern India. In Proceedings of the
International Conference on Development Potential for Moringa Products, Dar es Salaam,
Tanzania, 29 October–2 November 2001.
Ramachandran, C., K.V. Peter and P.K. Gopalakrishnan. 1980. Drumstick (Moringa oleifera): a
multipurpose Indian vegetable. Econ. Bot. 34: 276-283.
Ras, R.T., J.M. Geleijnse and E.A. Trautwein. 2014. LDL-cholesterol-lowering effect of plant
sterols and stanols across different dose ranges: A meta-analysis of randomised controlled
studies. Br. J. Nutr. 112: 214-219.
Rashid, U., F. Anwar, B.R. Moser and G. Knothe. 2008. Moringa oleifera oil: A possible source
of biodiesel. Bioresour. Technol. 99: 8175-8179.
Richter, N., S. Perumal and B. Klaus. 2003. Evaluation of nutritional quality of moringa (Moringa
oleifera Lam.) leaves as an alternative protein source for Nile tilapia (Oreochromis
niloticus L.). Aquacul. 217: 599-611.
Roloff, A., H. Weisgerber, U. Lang and B. Stimm. 2009. Enzyklopädie der Holzgewächse,
Handbuch und Atlas der Dendrologie; WILEY-VCH: Weinheim, Germany.
Saint Sauveur, A. 2001 Moringa exploitation in the world: State of knowledge and challenges. In
Proceedings of the Development Potential for Moringa Products, Dar Es Salam,
Tanzania.
Sanchez, N.R., L. Stig and L. Inger. 2006. Biomass production and chemical composition of
Moringa oleifera under different management regimes in Nicaragua. Agrofores. Sys. 66:
231-242.
Santos, A.F., A.C. Argolo, L.C. Coelho and P.M Paiva. 2005. Detection of water soluble lectin
and antioxidant component from Moringa oleifera seeds. Water Res. 39: 975-980.
Schwingshackl, L. and G. Hoffmann. 2014. Monounsaturated fatty acids, olive oil and health
status: A systematic review and meta-analysis of cohort studies. Lipids Health Dis. 13:
154.
Sengupta, A. and M.P. Gupta. 1970. Studies on the seed fat composition of moringaceae family.
Fette Seifen Anstrichm. 72: 6-10.
95
Siddhuraju, P. and K. Becker. 2003. Antioxidant properties of various solvent extracts of total
phenolic constituents from three different agro climatic origins of drumstick tree (Moringa
oleifera Lam.). J. Agric. Food Chem. 51: 2144-2155
Singha, R. Advantages of moringa oil. Retrieved from www.buzzle.com/articles/moringa oil.html.
Accessed on 30 Dec 2010.
Statista, 2017. Available at: https://www.statista.com/statistics/263937/vegetable-oils-global-
consumption/ (Accessed: 20 March 2017)
Steel, R.G.D., J. H. Torrie and D. A. Dickey. 1997. Principles and procedures of statistics: A
biometrical approach. 3rd ed. McGraw Hill Book Co. Inc. New York: 400-428.
Suthanthirapandian, I.R., S. Sambandamurthy and I. Irulappan. 1989. Variations in seedling
populations of annual moringa (Moringa pterygosperma Gaertn.). South Indian Hortic. 37:
301-302.
Swarup, R., G. Parry, N. Graham, T. Allen and M. Bennett. 2002. Auxin cross-talk: integration of
signaling pathways to control plant development. Plant Mol. Biol. 49: 411-426
Taiz, L. and E. Zeiger. 2002. Plant physiology.3rd edition. Sinauer Associates, Inc. U.S.A.
Thurber, M.D. and J.W. Fahey. 2009. Adoption of Moringa oleifera tocombat under-nutrition
viewed through the lens of the “Diffusion of Innovations” theory. Ecol. Food Nutr. 48:
212-225.
Tsaknis J., S. Lalas, V. Gergis, V. Dourtoglou and V. Spiliotis. 1998. Characterization of Moringa
oleifera variety Mbololo seed oil of Kenya. J. Agric. Food Chem. 47:4495–4499
Tsaknis, J., V. Spiliotis, S.Lalas, V. Gergis and V. Dourtoglou. 1999. Quality changes of Moringa
oleifera, variety Mbololo of Kenya seed oil during frying. J. Agric. Food Chem. 47: 4495-
4499.
Umer, R., F. Anwar, B.R. Moser and G. Knothe. 2008. Moringa oleifera oil: a possible source of
biodiesel. Bioresource Technol. 99: 8175-8179.
Uzama, D., S.A. Thomas, A.T. Orishadipe and O.A. Clement. 2011. The development of a blend
of Moringa oleifera oil with diesel for diesel engines. J. Emerging Trends Eng. Appl. Sci.
2: 999-1001.
Warner, K. and S. Knowlton. 1997. Frying quality and oxidative stability of high-oleic corn oils.
J. Am. oil Chem. Soc. 74: 1317-1322
World Agroforestry Center. Agroforestry database.
http://www.worldagroforestry.org/Sites/TreeDBS/AFT/ SpeciesInfo.cfm?SpID=1169
96
Yasmeen, A, S.M.A. Basra, R. Ahmad and A. Wahid. 2012. Performance of late sown wheat in
response to foliar application of Moringa oleifera Lam. leaf extract. Chil. J. Agric. Res.
72:92-97
Yasmeen, A., S.M.A. Basra, M. Farooq, H. Rehman, N. Hussain and H.R. Athar. 2013. Exogenous
application of moringa leaf extract modulates the antioxidant enzyme system to improve
wheat performance under saline conditions. Plant Growth Reg. 69, 225–233
Yasmeen, A., W. Nouman, S.M.A. Basra, A. Wahid, H. Rehman, N. Hussain and I. Afzal. 2014.
Morphological and physiological response of tomato (Solanumly copersicum L.) to natural
and synthetic cytokinin sources: a comparative study. Acta Physiol. Plant. 36: 3147-3155.
97

Monringa

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Monringa

Uploaded by

Copyright:

Available Formats

“EXPLORING THE POTENTIAL OF MORINGA OLEIFERA L.

AS AN OIL SEED TREE”

MUHAMMAD IDREES FAISAL

A thesis submitted in partial fulfillment of requirements for the degree of

Muhammad Idrees Faisal

Muhammad Idrees Faisal

2.1 Origin and distribution 5

2.2 Morphology and physical characters 5

2.2.3 Leaves and young shoots 6

2.2.4 Flowers, fruits and seeds 6

2.2.5 Wood and bark 6

2.2.6 Root type 6

2.3 Genetic diversity and taxonomy 7

2.4 Growth and development 7

2.5 Cultivation and production 7

2.6 Genetic and breeding 8

2.8 Insect, pests and pathogens 10

2.9 Nutritional value of moringa 10

2.10 Economic importance 10

2.10.2 Human consumption 11

2.10.3 Water purification 11

2.10.4 Medicinal uses 12

2.10.5 Moringa as fodder for livestock 12

2.10.6 Crop growth enhancer 13

2.11 Moringa seeds oil (Ben oil) 13

2.13 Climatic factors and seed composition 15

2.14 Seed composition 15

2.15 Physio-chemical characteristics of moringa oil 16

2.16 Oxidative stability of oil 16

2.17 Fatty acid composition 17

2.18 Sterol composition 17

2.19 Moringa oil benefits 18

2.19.1 Health benefits 18

2.19.2 Medicinal and nutraceutical action 18

2.19.3 Biodiesel production 18

2.19.4 Cosmetic industry 19

3 Materials and Methods 20

3.1 Experiment duration 20

3.2 Wild tree selection 20

3.3 Pod harvest and seed yield data collection 20

3.4 Oil extraction 20

3.5 Degumming of oils 21

3.6.1 Pod length (cm) 21

3.6.2 Pod weight (g) 21

3.6.3 Number of seeds per pod 21

3.6.4 Seed weight per pod (g) 22

3.6.5 Number of pods per tree 22

3.6.6 Seed weight per tree (kg) 22

3.6.8 Refractive Index 22

3.6.9 Saponification value 23

3.6.10 Iodine Value 24

3.6.11 Free fatty acid 25

3.6.12 Peroxide value 27

3.6.13 p-anisidine value 28

3.6.14 Specific extension of oil 29

3.6.15 Fatty Acid Profile (FAP) 30

3.6.16 Sterol Composition 31

3.7 Experiment II: Exploring the potential of vegetatively propagated Moringa 35

3.7.1 Experimental Detail 35

3.7.2 Stem cutting propagation: 35

3.7.3 Soil analysis: 36

3.7.4 Pod harvest and seed yield data collection: 36

3.7.5 Oil extraction: 36