Morphological and Physiological responses of Moringa

(Moringa stenopetala L.) accessions seedlings to

water stress under greenhouse condition at Hawassa,
Southern Ethiopia, Ethiopia

M.Sc. Thesis

By; Gebre Garmame

Advisor: Hussien Mohammed B. (PhD)

Co-Advisor: Amsalu Gobena R. (PhD)

June, 2019
Hawassa, Ethioipia
 Presentation Outline

.
• Introduction

• Materials and Methods

. .

.
• Results and Discussions

• Summary, conclusion and

.
Recommendations
 1. INTRODUCTION

 The interest of food, nutrition and medicinal plant is putting

intense pressure in the world (Jane, 2015; FAO, 2018).

 Therefore, focusing on nutrient full, medicinal, multi-

purpose and drought tolerant plants is necessary.

 …Introduction…

 Surprisingly, Moringa  Due to its massive purpose

(Moringa stenopetala L.) is several people call Moringa

as:
one of the world’s most
 “The Tree of Life”,
Multipurpose, wide range  “The
10*Never
15*
12* the Die plant”,
thepotassium
vitamin A of
C of
of
25*
9* theprotein
17*the iron ofofspinach
calcium of
yogurt
milk
of agro- ecological  “The Magic carrot
banana
orange
plant”,
 “The Tree of Paradise”,
adaptation plants (Daba,
(Ahmad et al., 2014)
 “The Miracle plant”.
(Kumssa et al., 2017)
2016).  “Marvelous plant”
(Nina Glærum, 2012)
 …Introduction…

 The annual production of Moringa could succeed 1.1 to

1.3 million tons from 38000ha of land (Rajangam et al.,

2001).

 In India, small scale farmers get a net income of about

USD 1500 per hectare annually under adverse agro

ecology (Rajangam et al., 2001).

 …Introduction...

 Moringa can be affected and reduced its potential by a

number biotic and abiotic factors.

 While water does an essential component for the

plants life owes to the crucial role that water plays in

biochemical and physiological processes (Majken et

al., 2005).
 …Introduction...

 To meet sustainable development goals (SDGs) like

 Ending hunger,  improving nutrition and

 promoting sustainable

 Achieving food security, agriculture (FAO, 2018)

 Massive important and wide range agro ecology adaptive

plants such as Moringa can be focused and established

on water shortage conditions (Mall and Tripathi, 2017).

 …Introduction...

 But before establishing seedlings for commercial

production it is necessary to investigate if the selected

accessions will positively:

 Response,

 Survive and

 Grow under the water stressful environment,

 …Introduction...

 Moringa is highly heterozygous due to the highly cross

pollinated (Amoatey et al., 2012)

 Accessions could probably have a wide variations

in traits to water stress (Baiyeri et al., 2015;

Jacob, 2016).
 …Introduction...

 Several studies revealed that variations of Moringa

accessions are controlled by epigenetics (Kenneth, 2012;

Jacob, 2016).

 But no much is known about those traits of Moringa

stenopetala and its mechanisms coping against water

stress (Badran, 2016).

 …Introduction...

 Rather, the previous research carried out so far has

focused mainly on survey works:

 To assess the traditional uses of M. stenopetala,

 Most exp’ts also shown evidences on human and plant

nutrition, water purification, livestock feed, dyes, medicine

and oil production (Mall and Tripathi, 2017).

 …Introduction...

 These constraints directed us to evaluate different

Moringa accessions under water stress conditions,

regarding to each organ of the seedlings.

 …Introduction…
 The main objective of this study was therefore, to
investigate morphological and physiological responses of
Moringa stenopetala accessions seedlings to water
stress.
Specific objectives were:

 To study morphological responses of Moringa

stenopetala accessions seedlings to different
water stress,
 …..Introduction

 To study physiological responses of Moringa

stenopetala accessions seedlings to different water
stress,

 To identify adaptive mechanisms of Moringa

stenopetala accessions seedlings to water stress,

 To determine the optimum watering interval for

Moringa stenopetala accessions seedlings.

 2. MATERIALS AND METHODS
2.1. Description of the Study Area

 During October 2018 –January 2018 at HUCA campus.

 Located in the Hawassa, that is 275 km far from AA capital

city of Ethiopia.

 Lies at of altitude of 1750 m above sea level.

 The average annual rain fall is 971.9 mm

 Temperature(maxi & mini) is 27.9°c and 13.8°c, respectively

 2.2. Experimental Materials
Table 1. Description of Moringa stenopetala accessions
used for the current experiment.

Accession Accession Growing

No code Origin altitude Source

1 Konso Konso 1320 masl AARC

2 Arbaminch Zuria Arbaminch Zuria 1222 masl AARC

3 Humbo Humbo 1432masl AARC

 3.3. Treatments and Exp’tal Design

 3*4 ( 3 accessions and 4 water stress levels imposed as

irrigation interval) with 3 replications,

 Totally 36 experimental units

 Each consisting of 15 Perforated black polyethylene

tubes with size 22 cm x 16 cm per treatments.

 Factorial CRD
 3. 4. Experimental Procedure

 The seed was obtained from AARC

 Top soil (3): Compost (2): Sand (1) (Ede et al., 2015).

 Experimental media was sampled and analyzed for most

important:

 physical and

 chemical properties .
 … Experimental Procedure…

 Filled in polyethylene tube and arranged in CRD,

 The amount of water= (ƟFC - ƟAMC)* depth of the

Perforated black polythene tubes (cm) ˣ Perforated black
polythene tubes area (cm2),

 Then, two seeds were sown during October/ 09/2018 at

2cm depth per each polyethylene tubes, then thinned,
 … Experimental Procedure…

 All Experimental units were watered well to field

capacity up to October/29/2018,

 Thereafter, during October/30/2018, the seedlings were

subjected to water stress treatment.
3.5. Data Collection and Measurement

3.5.1. Morphological data

 Seedling height(cm)

 Stem collar girth(cm)

 Root length and

 Root diameter (cm)

 Leaf area (cm2)

 leaf, root & total seedling fresh
& dry weights
3.5. Data Collection and Measurement
3.5.2. Physiological data
 Stomata traits(Torre et al., 2003)

 chlorophylls concentration

(μg/ml) (Porra et al., 1989).

 Gas exchange traits
Chl a = 16.29*A665.2_8.54*A652.0
 Leaf water related traits

 IWUE=
Chl Ps 652.0
/ E _13.58*A665.2
b = 30.66*A
 LRWC%  = ˣ100

Chl a+b=22.12A652.0_2.71 A665.2

 3.6. Data Analysis

 Data's subjected to ANOVA using proc GLM

procedure of SAS version 9.3,

Tukey’s HSD test was used to separate means

at 5% level of significance.
4. RESULTS AND DISCUSSIONS

4.1. Morphological Response of Moringa

stenopetala Accessions Seedlings to Water
Stress
Table 4. Correlation
Table 3. Pearson'scoefficients
correlation among coefficients the morphological and physiological
among the morphological traits of traits
ofMoringa
Moringa stenopetala
Table 2. Responses
stenopetala accessions
of SH(cm), SG(cm), LN(count) andseedlings
accessions seedlings under
LA of three Moringa
grown water
stenopetala
under stress
accessions levels
seedlings
different to water stress.
water stress

Treatments
SH SG RL RD LN Parameters
LA TFW RFW LFW TDW RDW LDW
SH __SN SL Seedling
SW chl height Stem
a chl b girth
ch(a +b) RWC (%) Ps LeafEarea IWUE
SHAccessions
-0.68 __0.87 (cm)
0.81 0.82 0.92 (cm) 0.87 Leaf
0.88number 0.83(cm 0.87
2
) 0.76
SG 0.93 Baiyeri etb al. (2015); a
SGKonso-0.72 15.41± 0.26 0.69±0.010.83 22.88±1.14 0.89123.81 ±5.470.81
b b
0.83 0.79 0.76 0.88 0.89 0.89
RL 0.89 0.89 ___
Arbaminch Zuria 17.07±0.26a Sale et al.a (2015);
0.75±0.01 25.11±1.14a 149.50±5.47a
RL -0.68 0.82 0.77 0.82 0.87 0.86 0.85 0.81 0.8 0.82
RDHumbo0.91 0.97 0.91 __ c
RD -0.74 0.86
 Rajangam
13.87±0.26
0.81 0.78 Daba 0.88 andet
0.63±0.01 c
al. (2001);
18.47±1.14b
Adisu0.87
0.84 (2017) 97.56±5.47c
0.90 0.88 0.82
LN 0.9
Tukey/HSD 0.93 0.88 0.67
0.93 __ 0.03 2.85 13.66
LN -0.72 0.88 0.82 0.78 0.88 0.83 0.87 0.89 0.91 0.78
LAWater stress
0.96 0.95  Amoatey
0.91 0.94 0.94 __ et al. (2012);
LA -0.68 0.85 0.84 0.81 0.92 0.88 0.88 0.88 0.91 0.78
Daily (control)
TFW 0.9 0.97 0.86 0.95 0.93 0.94 ___
TFW -0.67 0.83 0.770.69
Baiyeri
20.39±0.30 a et al. 0.76
0.83
0.98±0.01 (2015)
a 0.86 ±1.32a
35.37 0.89215.13
0.91
±6.310.79
a
RFW 0.86 0.95 0.84 0.94 0.92 0.92 0.99 ___
RFW
5 days-0.65
interval 0.80 0.74 0.64b
17.29±0.30 0.80
0.70±0.010.72
b 0.82
21.96±1.32 b 0.88134.95±6.31
0.89 0.77
b

LFW 0.85
LFW -0.65 0.95 0.83
0.74 0.94 0.920.8 0.92 0.72
0.98 0.99 ___ c
10 days interval 0.79 12.34± 0.65
0.30c 0.56±0.01c 0.82 1.32
17.77± 0.8981.06±6.31
0.90 c0.77
TDW
TDW 0.95
15 days
-0.72 0.94
interval 0.83 0.93
0.82 0.92
0.83c 0.910.92
11.78±0.30 0.96 0.88
0.51±0.01 0.91
d 0.88
13.52± 0.87c
0.89 1.32 ___63.35±6.31
0.89 0.89 d0.83
Tukey/HSD
RDW
RDW 0.95
-0.74 0.94 0.84 0.85
0.87 0.91 0.83 0.920.930.940.040.89
0.94 0.92 3.64 0.88
0.89
0.92 0.9617.43
0.89 ___ 0.86
0.87
CV (%)
LDW
LDW 0.96
-0.67 0.94 0.82 4.25
0.90 0.86 0.79 0.920.910.945.210.86
0.92 0.92 12.64 0.89
0.89
0.93 0.9310.84
0.84 0.88
0.95 0.76
___
Table 4. Correlation coefficients among the morphological and physiological traits
of Moringa
Table stenopetala
5. Response accessions seedlings
of leaf fresh under
and dry waterof
weight stress
threelevels
Moringa
stenpetala accessions seedling to different water stress
SN SL SW chl a chl b ch(a +b)
WaterRWC
stress(%) Ps E IWUE
SH -0.68 0.87 0.81 0.82 0.92 0.87 0.88 0.83 0.87 0.76
Accessions Daily 5 Days interval 10 Days interval 15 Days interval
SG -0.72 0.83 0.79
 0.76
Majken et0.88
al. 0.83
(2005); 0.89
Wafa 0.89
(2006); 0.89 0.81
Leaf fresh weight (gram per plant)
RL -0.68
Konso
0.82 0.77 0.82
4.88±0.11b
0.87 0.86 0.85
2.25±0.11cd 1.93±0.11defg
0.81 0.8
1.52±0.11g
0.82
RD -0.74 0.86  Jacob
0.81 0.78 (2013)
5.33±0.11 0.88
a and0.84Badran
2.48±0.11 0.87 et al.
c
1.96±0.11 def 0.90 0.88 efg 0.82
1.63±0.11
Arbaminch Zuria
LN -0.72
Humbo 0.88 0.82 0.78 b
(2016)
4.56±0.11 0.88 0.83
2.03±0.11 d
0.87efg
1.70±0.11 0.89 0.91 fg 0.78
1.58±0.11

LA -0.68
Tukey/HSD0.85 0.84
= 0.42 0.81 0.92
CV (%) = 5.43 0.88 0.88 0.88 0.91 0.78
TFW -0.67 0.83 0.77 0.69 0.83dry weight
Leaf 0.76 (gram0.86
per plant) 0.89 0.91 0.79
b cd g h
RFW -0.65
Konso 0.80 0.74 0.64
2.56±0.04 0.80 0.72 0.75±0.04
1.84±0.04 0.82 0.88 0.89
0.50±0.04 0.77
a d f
2.80±0.04 1.70 ±0.04 0.43±0.04hi
LFW Arbaminch Zuria
-0.65 0.79 0.74 0.65 0.8 0.72 0.97±0.04
0.82 0.89 0.90 0.77
Humbo 1.88±0.04c 1.39± 0.04e 0.55±0.04h 0.32±0.04i
TDW -0.72 0.83 0.82 0.83 0.92 0.88 0.89 0.89 0.89 0.83
Tukey/HSD = 0.15 CV (%) =3.92
RDW -0.74 0.87 0.84 0.83 0.93 0.89 0.92 0.89 0.87 0.86
LDW P < 0.01
-0.67 0.90and 0.001
0.82 0.79for leaf fresh 0.86
0.91 and dry 0.93
weights, respectively.
0.84 0.88 0.76
 Table 6. Response of root length, root diameter, root fresh weight
and root dry weight of three Moringa stenopetala accessions
seedling to water stress.
Treatments Parameters
Root Root dry weight
Root length diameter Root fresh weight (gram plant-1)
Accessions (cm) (cm) (gram plant-1)
Konso 14.23±0.47b 1.71±0.05a 21.41±0.72b 10.29±0.56b

 Dunford
Arbaminch Zuria and
16.18±0.47 a
Vazquez
1.72±0.05 (2005);
a
24.33±0.72 a
13.64±0.56a
Humbo 13.08±0.47b 1.50±0.05b 19.23± 0.72c 7.90±0.56c
Tukey/HSD  Wafa (2006);
1.19 Sale 0.12
(2015); 1.81 1.41
Water stress

Daily (control) Badran et al. (2016);
18.296±0.55 2.38±0.05Daba
a and
51.60±0.83
a a
19.39±0.65a
5 days interval 14.96±0.55b 1.68±0.05b 17.34±0.83b 12.53±0.65b
Adisu (2017)
10 days interval 13.35±0.55c 1.41±0.05c 10.26±0.83c 6.92±0.65c
15 days interval 11.39±0.55d 1.24±0.05d 7.40±0.83d 3.61±0.65d
Tukey/HSD 1.52 0.16 2.31 1.80
CV (%) 8.06 7.45 8.2 13.07
 Table 7. Response of total seedling fresh weight and total dry
weight of Moringa stenopetala accessions seedling to water stress

Treatments Parameters
Total fresh weight Total dry weight
Similar pattern
Accessions on Moringa oleifera -1and Moringa peregrina
(gram plant ) (gram plant-1)
(Wafa,
Konso 2006; Badran et al., 27.80±0.77
2016). b 15.61±0.69b
Arbaminch Zuria 32.09±0.77a 23.08±0.69a
Humbo 24.45±0.77c 13.07±0.69c
Turkey/HSD 1.93 1.74
Water stress
Daily (control) 61.80 ±0.89a 31.05±0.80a
5 days interval 25.63±0.89b 20.00± 0.80b
10 days interval 14.92±0.89c 11.62±0.80c
15 days interval 10.11±0.89d 6.34± 0.80d
Tukey/HSD 2.47 2.22
CV (%) 6.76 9.91
… RESULTS AND DISCUSSIONS…

4.2. Physiological Responses of Moringa

stenopetala Accessions Seedlings to
Water Stress
 Table 8. Response of stomata traits of different Moringa
stenopetala accessions seedling to varied water stress

Treatments Parameters

Stomata Stomata Stomata

Accessions number length (μm) width (μm)
Konso 16.50± 0.85ab 0.39±0.01a 0.22±0.01a
Arbaminch Zuria 15.25±0.85b 0.40±0.01a 0.25±0.01b
Humbo  Bosabalidis andaKofidis
17.83±0.85 (2002);
0.33±0.01 b
0.18±0.01c
Turkey/HSD  Hetherington
2.12 0.04
and Woodward 0.03
(2003)
Water stress and
Daily (control) 13.88±0.98b 0.49±0.01a 0.29±0.01a
 Belhadj et al. (2011)
5 days interval 16.44±0.98ab 0.40±0.01b 0.23±0.01b
10 days interval 17.11±0.98a 0.32±0.01c 0.19±0.01bc
15 days interval 18.66± 0.98a 0.28±0.01c 0.16±0.01c
Tukey/HSD 2.71 0.05 0.04
CV (%) 12.63 10.96 14.67
 Table 9. Response of chlorophylls concentration of different
Moringa stenopetala accessions seedling to different water stress.
Treatments Parameters
Accessions chl a(μgml-1) chl b(μgml-1) chl a+b(μgml-1)
Konso 5.49±0.08b 2.85±0.07b 8.34±0.15b
Arbaminch Zuria 6.00±0.08a 3.15±0.07a 9.15±0.15a
Humbo 4.48±0.08c
2.45±0.07
 Manivannan c
6.93±0.15c
et al. (2007b);
Turkey/HSD 0.21 0.18 0.39
 Kiani et al. (2008) and
Water stress
 Alireza et al. (2011)
Daily (control) 6.05±0.09a 3.55±0.08a 9.60 ±0.18a
5 days interval 5.60±0.09b 3.10±0.08b 8.70±0.18b
10 days interval 5.06±0.09c 2.47±0.08c 7.53±0.18c
15 days interval 4.53± 0.09d 2.19±0.08d 6.72±0.18d
Tukey/HSD 0.26 0.24 0.50
CV (%) 3.89 6.53 4.74
 Responses of gas exchanges & RWC of Moringa
stenopetala accessions seedlings to water stress.

 Wafa (2006)
Rivas et al. (2013)
Badran (2016) and
Claudiana et al. (2017)
IWUE of three Moringa stenpetala accessions seedlings
under different water stress.

 (Rivas
Farooq et (2009);
et al. al., 2013)
Hanafey et
 (2018)
al. (Andreia, 2014)

Means with different letter at each bar of each factor is

statistically significant at P-values ≤ 0.1%.
 5. SUMMARY, CONCLUSION AND
RECOMMENDATIONS
5.1. Summary
 M. stenopetala L. is one of the most multi-purpose medicinal
plants which adapt wide range of agro ecology.

 This study tested the responses of some morphological and

physiological parameters to different water stress level and

M.stenopetala accessions seedlings.

 Morph-physiology of M. stenopetal was influenced by

accessions and imposed water stress.

 5.2. Conclusion

 Variations in morphological and physiological responses of

M. stenoptala accessions seedlings to water stress levels
were observed.

 Accordingly, all the accessions of M. stenopetla seedlings

produced more values in morphological and physiological
traits except stomata no, when the seedlings watered daily.
 5.2. Conclusion

 But “Arbaminch Zuria” produced the largest values.

Contrarily, the lowest value was observed “Humbo”, when
subjected to fifteen days interval watering.
 5.3. Recommendations

 Accession “Arbaminch Zuria” of M. Stenopetala

 Daily watering.

 Further research on (indigenous and exotic) and water

stress for long period of time (due to its perennial character)
to follow its field performance and adaptation.
Have a Moringa Tea!!

With Many Thanks!!