Nutritional Status of Some Aromatic Plants Grown to Produce Volatile Oils Under Treated Municipal Wastewater Irrigation

Abstract

To avoid any contamination risk of edible crops, the safety use of treated municipal wastewater is growing industrial non-food crops such as aromatic plants to produce volatile oils for manufacturing soaps, cosmetics and perfumes, etc. Therefore, two field experiments were conducted in a sandy soil to investigate the influence of treated domestic sewage effluents or freshwater (control) on the essential oils of geranium, peppermint, fennel, marjoram, and chamomile plants. In the first experiment, the plants were irrigated with treated domestic sewage effluents without mineral fertilizers addition, while in the second the plants irrigated with fresh water and received the mineral fertilizers. Heavy metals accumulation in plant organs and the nutritional status of plants were studied. Results indicated that the treated municipal waste water contain obvious amount of nutrients sufficiently for growing the tested crops and the plants had higher herb yield than that irrigated with fresh water. Irrigation of geranium and fennel plants with wastewater caused a significant increment in the essential oils concentration, but lowered it in peppermint and sweet marjoram plants. The oil yields of five crops were higher under wastewater irrigation and with geranium plants exceeded than that irrigated with fresh water. Uptake and accumulation of heavy metals in plant parts was varied. No detectable amount of the potential toxic elements was recorded in the essential oils of the tested aromatic plants. From this standpoint, treated municipal waste water can be used for growing aromatic plants in the arid area without mineral fertilizers addition to produce volatile oils without causing any reduction in quantity and quality of volatile oils.

Full text

ISSN: 2087-3948
Vol. 5, No. 1, pp. 22-29 E-ISSN: 2087-3956
May 2013

Response of Silybum marianum plant to irrigation intervals combined
with fertilization
SABER F. HENDAWY1,♥, MOHAMED S. HUSSEIN1, ABD-ELGHANI A. YOUSSEF2,
REYAD A. EL-MERGAWI3
1Medicinal and Aromatic Plants Research Department, National Research Centre, Dokki 12311, Giza, Egypt. Tel. +202-3366-9948, +202-33669955,
Fax: +202-3337-0931, ♥email: hendawysaber@yahoo.com
2Departement of Chemistry, Faculty of Science, Jazan University, Saudi Arabia
3 Department of Plant Production and Protection, College of Agriculture and Veterinary Medicine, Qassim University,P.O.Box 6622, Buhrida 51452, Al-
Qassim; Saudi Arabia
Manuscript received: 11 April 2013. Revision accepted: 5 May 2013.
Abstract. Hendawy SF, Hussein MS, Youssef AA, El-Mergawi RA. 2013. Response of Silybum marianum plant to irrigation intervals
combined with fertilization. Nusantara Bioscience 5: 22-29. This study was investigated to evaluate the influence of different kinds of
organic and bio fertilization under different irrigation intervals on the growth, production and chemical constituents of Sylibium
marianum plant. Data indicated that all studied growth and yield characters were significantly affected by the duration of irrigation
intervals. Fertilizer treatments had a primitive effect on growth and yield characters. The interaction between irrigation intervals and
fertilizer treatments has a clear considerable effect on growth and yield characters. The obtained results indicated the favorable effect of
organic and bio fertilizers which reduce the harmful effect of water stress. Different treatments had a pronounced effect on silymarin content.
Key words: Sylibium marianum, silymarin, bio fertilization and irrigation intervals.
Abstrak. Hendawy SF, Hussein MS, Youssef AA, El-Mergawi RA. 2013. Respons tanaman Silybum marianum terhadap interval irigasi
yang dikombinasi dengan pemupukan. Nusantara Bioscience 5: 22-29. Penelitian ini bertujuan untuk mengevaluasi pengaruh berbagai
jenis pupuk organik dan hayati dengan interval irigasi yang berbeda terhadap pertumbuhan, produksi dan kandungan kimia tanaman
Sylibium marianum. Data menunjukkan bahwa semua sifat pertumbuhan dan produksi yang diteliti secara signifikan dipengaruhi oleh
durasi interval irigasi. Perlakuan pemupukan berpengaruh nyata terhadap karakter pertumbuhan dan hasil panen. Interaksi antara interval
irigasi dan perlakuan pemupukan berpengaruh besar pada karakter pertumbuhan dan hasil panen. Hasil yang diperoleh menunjukkan
efek menguntungkan dari pupuk organik dan hayati yang dapat mengurangi efek berbahaya cekaman air. Perlakuan yang berbeda
berpengaruh kuat terhadap kandungan silymarin.
Kata kunci: Fagus orientalis, serat, sifat biometri, pohon unggul.
INTRODUCTION
Milk thistle (Silybum marianum L. Gaertn.), a member
of the Mediterranean Basin, as a crop and weed on
agricultural plantations, it occurs in many European
countries, North Africa, South and North America, Central
and Western Asia and southern Australia (Carrier et al.
2002).The pharmaceutical compound of milk thistle is
derived from its fruits, which are achenes (Fructus silybi
mariani). In their dry pericarp and seed coat the plant
accumulates a group of flavonolignans commonly referred
to as silymarin (Cappelletti and Caniato 1984). Taxifolin is
their precursor. The main flavonolignans of milk thistle are
silybinin, isosilybinin, silydianin and silychristin. Several
other compounds of that type have also been identified, but
their importance in the silymarin complex is insignificant
(Kurkin et al. 2001). Silymarin, derived from the seeds of
milk thistle plant has been used widely for the treatment of
toxic liver damage (Dewick 1998). Silymarin primarily
consists of an isomeric mixture of six phenolic compounds:
silydianin, silychristin, diastereoisomers of silybin (silybin
A and B), and diastereoisomers of isosilybin (isosilybin A
and B) (Lee et al. 2007).
The compost must be added to conventional NPK
fertilizer to improve soil structure, making the soil easier to
cultivate, encouraging root development, providing plant
nutrients and enabling their increased uptake by plants.
Moreover, compost aids water absorption and retention by
the soil, reducing erosion and run-off and thereby
protecting surface waters from sedimentation, help binding
agricultural chemicals, keeping them out of water ways and
protecting ground water from contamination (leaMaster et
al. 1998). Compost has already been established as a
recommended fertilizer for improving the productivity of
several medicinal and aromatic plants, as peppermint
(O’Brien and Barker 1996), Tagetes erecta (Khalil et al.
2002), Sideritis montana (El-Sherbeny et al. 2005), Ruta
graveolens (Naguib et al. 2007) and Dracocephalum
moldavica L. ( Amer 2008). Compost tea is a highly
concentrated microbial solution produced by extracting

HENDAWY et al. – Response of Silybum marianum to irrigation and fertilizer 23
beneficial microbes from vermicompost and or compost.
Compost tea provides direct nutrition as a source of foliar
and soil organic nutrient and as chelated micronutrients for
easy plant absorption. Also, compost tea provide microbial
functions, that compete with disease causing microbes,
degrade toxic pesticides, produce plant growth hormones,
mineralize plant available nutrients and fix nitrogen
(Hendawy 2008).
Arbuscular mycorrhiza (AM) fungi (Endogonaceae)
form a mutualistic relationship with the roots of most plant
species. This plant-fungus association involves the
translocation of carbon from the plant to the fungus and
enhanced uptake and transport of soil nutrients, primarily
phosphorus, to the plant via the fungus (Newman and
Reddel 1987). Other potential benefits of AM fungal
colonization to host plants include improved uptake of
poorly mobile nutrients such as zinc (Gildon and Tinker
1983), improved plant water relations (Allen and Allen
1986) and reduced pathogenic infections (Newsham, et al.
1995). AMF can also benefit plants by stimulating the
production of growth regulating substances, increasing
photosynthesis, improving osmotic adjustment under
drought and salinity stresses and increasing resistance to
pests and soil borne diseases (Al-Karaki 2006).
However, water deficit is a limiting factor in
production of many field crops (Kafi and Mahdavi
Damghani 2001; Munns 2002) and water stress causes
different morphological, physiological and biochemical
changes including: leaf area reduction, leaf senescence and
reduction in cell development (Kafi and Mahdavi
Damghani 2001), stomatal closure (Safar-Nezhad 2003)
and photosynthetic limitation (Kafi and Mahdavi
Damghani 2001). It appears that the effect
of water stress on economic yields of
medicinal plants which are mainly
secondary metabolites, are somehow
positive (Baher et al. 2002). In many cases,
a moderate stress could enhance the content
of secondary metabolites.
This current experiment targeted the
evaluation of the influence of different kinds
of organic and bio fertilization under
different irrigation intervals on the growth,
production and chemical constituents of
Sylibium marianum plant.
MATERIALS AND METHODS
Field experiment
Location
The field experiment was carried out at
El-Nubareia Research Station (El-Behira
Governorate, Egypt), National Research
Centre, to investigate the influence of
Chemical, organic and bio fertilizers under
different irrigation intervals on growth,
yield and chemical constituents of milk
thistle.
Soil
The experiment was set up on sand loam soil as shown
in Table 1.
Table 1. Main characteristics of soil
Characteristics Value
Mechanical analysis
Sand% 68.08
Silt% 16.00
Clay% 15.92
Texture Sandy loam
Chemical analysis
PH 1:2.5ext. 8.50
Ca Co3 21.70
Electrical conductivity 1:2.5ext 0.61
Soluble cations meq/l
Ca++ 3.38
Mg++ 3.62
Na+3.23
K+0.49
Soluble anions meq/l
HCO3-1.12
Cl-1.5
SO4-9.1
Macro-elements (ppm)
N 30.00
P 20.00
K 368.00
Micro-elements (ppm)
Zn 0.28
Mn 2.50
Fe 3.70
Cu 0.96
Figure 1. Inflorescense of milk thistle (Silybum marianum L. Gaertn.)

5 (1): 22-29, May 2013

24
Experiment design and agronomic practices
The fertilization factor experiment was set up in a
randomized design in three replicates.
Experimental treatments
A. Irrigation every 3 days
1. NPK (100 kg super phosphate+150 kg nitrate ammonium+50
kg potassium sulphate).
2. Compost 20m3/feddan
3. Compost 20m3/feddan+mycorrhiza
4. Compost 20m3/feddan+compost tea 20 L/feddan
5. Compost 20m3/feddan+compost tea 20 L/feddan+mycorrhiza
B. Irrigation every 6 days
6. NPK (100 kg super phosphate+150 kg nitrate ammonium+50
kg potassium sulphate) as control.
7. Compost 20m3/feddan
8. Compost 20m3/feddan+mycorrhiza
9. Compost 20m3/feddan+compost tea 20 L/feddan
10. Compost 20m3/feddan+compost tea 20 L/feddan+mycorrhiza
C. Irrigation every 9 days
11. NPK (100 kg super phosphate+150 kg nitrate ammonium+50
kg potassium sulphate) as control.
12. Compost 20m3/feddan
13. Compost 20m3/feddan+mycorrhiza
14. Compost 20m3/feddan+compost tea 20 L/feddan
15. Compost 20m3/feddan+compost tea 20 L/feddan+mycorrhiza
The seeds were directly sown in 20th of October 2010.
Each plot was 13.5 m2 consisting of 9 rows with a distance
of 50 cm between the rows and 30 cm between each
successive plant.. Weeding and thinning was done after 30
days of plantation. Recommended agronomic practices
were adopted.
Super phosphate or compost was added during
preparing soil. The other chemical fertilizers (Ammonium
nitrate and Potassium sulphate were divided into two equal
portions during the growing season, the 1st portion was
added after one month of sowing, while the second one was
applied after one month from the 1st. Tea compost (Table 2,
3) was sprayed after 60 days from sowing and repeated
after 15 days.Vesicular arbscular mycorrhiza (VAM) fungi
which contained 3 effective strains representing Glomus
etunicatum, Glomus fasciculatum and Glomus intraradices.
VAM fungi was used for soil inoculation. The VAM
inoculation was applied into sowing hills at a rate of 5
mL/hill. The amount contained about 200 VAM spores/hill.
The effect of the above treatments was measured by
plant height, branches number, capitula number/plant, seed
yield and silymarin content.
Table 2. Microbial population of organic compost tea
Constituent Value
Bacterial Plate Count (CFU/ml) 7.1 X 107
Bacterial Direct Count (Cell/ml) 6.4 X 108
Spore Forming Bacteria (CFU/ml) 7 X 104
Total Fungi (CFU/ml) 2.8 X 105
Table 3. Chemical analysis of organic compost tea
Constituent Value
Bulk Density kg/m3 510
Moisture Content% 18.2
Electrical conductivity dS/m 9.65
PH 7.6
Total Organic Carbon% 24.6
Total Organic Matter% 42.41
Total Nitrogen% 1.35
C/N Ratio 18.22
NH4-N, mg/kg 880
NO3-N, mg/kg 450
Total Phosphorus% 1.6
av. Phosphorus mg/kg 410
Total Potassium% 2.3
av. Potassium mg/kg 620
Trace Element (ppm)
Fe 960
Zn 280
Mn 320
Cu 140
Note: Nematodes (nil), Weeds germination (nil), Parasites (nil),
Pathogenic (nil), Humus value (5)
Extraction procedure
Silymarin content was extracted according to ( Cacho et
al. 1999). Gram of seeds were defated in a Soxhlet
apparatus with 50 mL of petroleum-ether at 40-60 oC for 12
h. The residue was extracted with 50 mL of methanol at 65-
70 oC over 8 h. The methanolic solution was concentrated
to a dry residue. The extract was dissolved in 10 mL of
methanol.
HPLC analysis
HPLC was carried out using an HPLC pump monitored
at 280 nm by a UV detector and quantified by an integrator.
A Shim-pack C18 ( 1250 x 4.6 mm ID) column was used,
eluting with MeOH-H2O-AcOH 40:60:5, at a flow rate of 2
mL/min. Mixture of flavonolignans obtained from Alex
Pharm, Egypt (specifications: Silychristin 25% Rt 2.94
min, silydianin 9.7% Rt 3.64 min, silybin A 21.3% Rt 7.84
min, silybin B 32% Rt 9.18 min, isosilbin A 8.7% Rt 13.61
min and isosilybin B 3% Rt 15.18 min).
RESULTS AND DISCUSSION
Vegetative growth and yield
Irrigation intervals
Data tabulated in Table 4 indicated that all studied
growth and yield characters were significantly affected by
the duration of irrigation intervals.
By increasing the severity and duration of drought from
3 days to 9 days, plant height (cm) showed significant
reduction. Such reduction in plant height in response to
drought may be due to blocking up of xylem and phloem
vessels thus hindering any translocation through (Lovisolo
and Schuber 1998). Similar results were obtained by Singh
et al. (2006) and Khalil et al. (2010).

HENDAWY et al. – Response of Silybum marianum to irrigation and fertilizer 25
Table 4. Effect of irrigation intervals on vegetative growth and
yield of Silybum marianum
Seed yield
(g/Plant)
Flowers
heads
no/plant
Branches
no/plant
Plant height
(cm)
Irrigation
intervals
20.9120.606.60179.003 days
17.3918.207.20169.406 days
14.8915.808.60166.2 9 days
0.4770.5820.5760.504LSD at 5%
Data on hand, illustrated also that, number of
branches/plant increased significantly with decreasing of
irrigation, this may be due to that drought reduced cycling-
dependent kinase activity results in slower cell division as
well as inhibition of growth (Schuppler et al. 1998). This
supported by the results of (Rahmani et al. 2008) on
Calendula officinalis L. and (Taheri et al. 2008) on
Cichorium intybw L.
Significant higher numbers of flowers head/plant and
seed yield (g/plant) were recorded with the shortest
irrigation interval (3 days) followed by (6 days). The
decrease in yield attributes under the longest irrigation
interval (9 days) may be due that water stress changing the
hormonal balance of mature leaves, thus enhancing leaf
senescence and hence the number of active leaves
decreased, as well as leaf area was reduced by water
shortage, which was attributed to its effect on cell division
and lamina expansion. When the number of active leaves
decreased the light attraction and CO2 diffusion inside the
leaf decreased and the total capacity of photosynthesis
decreased, therefore, the photosynthetic materials that
transferred to seeds will decreased (Ahmed and Mahmoud
2010; Moussavi et al. 2011).
Fertilizer treatments
Data tabulated in Table 5 show that fertilizer
treatments had a significant effect on growth and yield
characters of Silybum marianum plants. The mean values
of plant height were 174.33, 164.33, 168.33, 171.0 and
179.67 cm as a result of NPK, compost, compost+
mycorrhiza, compost+compost tea and compost+compost
tea+mycorrhiza treatments, respectively. So, the highest
value of plant height was obtained as a result of
compost+compost tea+mycorrhiza treatment.
Table 5. Effect of fertilizer treatment on vegetative growth and
yield of Silybum marianum
Seed
yield
(g/plant)
Flowers
heads
no/plant
Branches
No/plant
Plant
height
(cm)
Fertilizer treatments
18.1521.338.33174.33NPK
15.7417.006.33164.33Compost
18.9016.677.33168.33Compost+mycorrhiza
16.4917.007.33171Compost+compost tea
19.37198.00179.67Compost+compost
tea+mycorrhiza
1.0540.3180.4490.825LSD at 5% 
The results in Table 5 reveal that, fertilizer treatments
had a pronounced effect on branches number. It can be
noticed that, mean values of branches number recorded
8.33, 6.33, 7.33, 7.33 and 8.00/plant were obtained from
NPK, Compost, Compost+mycorrhiza, compost+compost
tea and compost+compost tea+mycorrhiza treatments,
respectively. Thus, the maximum mean value of branches
number/plant (8.33) was obtained as a result of NPK
treatment followed by compost+compost tea+mycorrhiza
treatment, which recorded 8.00/plant. There is no
significant difference between NPK treatment and
compost+compost tea+mycorrhiza treatment.
The averages of heads flowers number were 21.33,
17.00, 16.67, 17.00 and 19.00/plant as a result of NPK,
Compost, Compost+mycorrhiza, compost+compost tea and
compost+compost tea+mycorrhiza treatments, respectively.
Thus, the maximum mean value of flowers heads
number/plant (21.33) was obtained from NPK treatment
followed by compost+compost tea treatment, which
recorded 19.00/plant.
It is evident from data in Table 5 that fertilizer
treatments had a significant effect on seed yield (g/plant).In
this respect, mean values of seed yield (g/plant) were
18.15, 15.74, 18.90, 16.49 and 19.37 g/plant as a result of
as a result of NPK, compost, compost+mycorrhiza,
compost+compost tea and compost+compost tea+
mycorrhiza treatments, respectively. Therefore, compost+
compost tea+mycorrhiza treatment gave the highest mean
value of seed yield (19.37g/plant) followed by
compost+mycorrhiza treatment which recorded (18.90
g/plant).
The promotion effect of compost on the growth and
yield of plant could be explained through the role of
organic materials including composts in improving soil P
availability (Gichangi et al. 2009). Since during
composting, labile nutrients are converted into stabilized
organic material (Zucconi and De Bertoldi 1987), therefore
a large proportion of nutrients are labile. Composts provide
microbes not only with P but also C and N and are
therefore likely to induce changes in P pools that differ
from those of inorganic P addition (Hassan et al. 2012).
The favorable effects of the combination between
compost +compost tea+mycrohiza may be explained based
on the beneficial effects of them on the improvement soil
physical and biological properties and also, the chemical
characteristics resulting in more release of available
nutrient elements to be absorbed by plant root and its effect
on the physiological processes such as photosynthesis
activity as well as the utilization of carbohydrates. A
similar suggestion was made by Hanafy et al. (2002) on
rocket plants. Furthermore, this stimulative effect may be
related to the good equilibrium of nutrients and water in the
root medium (Abdelaziz and Balbaa 2007) or to the
beneficial effects of mycorrhiza on vital enzymes and
hormonal, stimulating effects on plant growth and yield.
Interaction treatments
The interaction between irrigation intervals and
fertilizer treatments has a clear considerable effect on
growth and yield characters (Table 6). It can be observed

5 (1): 22-29, May 2013

26
that the maximum mean value of plant height (190.00 cm)
was obtained from the combination treatment between
irrigation intervals every 3 days and fertilized with
compost+compost tea+mycorrhiza. On the other hand, the
lowest average of plant height (158.00 cm) was obtained
from the combination between irrigation intervals every 9
days and compost treatment. The variation in plant height
between maximum and the minimum values reached to
20.25%.
For branches number/plant, it can be observed that, the
highest mean value of branches number/plant (10.00/plant)
against the lowest value (5.00/plant) were obtained as a
result of the combination between irrigation intervals every
9 days and NPK treatment and the combination irrigation
intervals every 3 days with compost treatment,
respectively. The variation in branches number/plant
between maximum and the minimum values reached to
100%.
Data shown in Table 6 indicated that, the combination
between irrigation intervals every 3 days and NPK
treatment gave the highest mean value of flowers heads
number (25.00/plant),while the combination between
irrigation intervals every 9 days and compost+compost tea
treatment gave the lowest mean value (13.00/plant). The
variation in flowers heads number/plant between maximum
and the minimum values reached to 92.31%.
Concerning the interaction treatments, it can be noticed
that the combination between irrigation intervals every 3
days and compost+compost tea+mycorrhiza treatment
resulted in the maximum mean value of seed yield (23.40
g/plant) while the interaction between irrigation intervals
every 9 days and compost+compost tea treatment gave the
lowest one (13.00 g/plant). The variation in seed yield
(g/plant) between maximum and the minimum values
reached to 78.49%.
The obtained results indicated the favorable effect of
organic and bio fertilizers which reduce the harmful effect
of water stress through their effect on improving the soil
texture. The structural improvement can encourage the
plant to have a good root development by improving the
aeration in the soil. The favorable effects of these fertilizers
may be due to the role of organic material for continues
supply of nutrients, which improve some physical
properties of soil and increase water retention (Abd-
Elmoez et al. 1995; Fliessbach et al. 2000).
Table 6. Effect the interaction treatments between irrigation intervals and fertilization on growth and yield of Silybum marianum
Seed yield
(g/plant)
Flowers heads
no/plant
Branches
no/plant
Plant height
(cm) Fertilizer treatmentsIrrigation
intervals
19.5025.007.00183.00 NPK 3 days
18.7019.005.00170.00 Compost
22.6018.006.00175.00 Compost+mycorrhiza
20.3321.008.00177.00 Compost+compost tea
23.4020.007.00190.00 Compost+compost tea+mycorrhiza
18.5520.008.00174.00 NPK 6 days
15.4017.007.00165.00 Compost
19.6018.007.00167.00 Compost+mycorrhiza
15.5017.006.00166.00 Compost+compost tea
17.9019.008.00175.00 Compost+compost tea+mycorrhiza
16.4019.0010.00166.00 NPK 9 days
13.1115.007.00158.00 Compost
14.5014.009.00163.00 Compost+mycorrhiza
13.6513.008.00170.00 Compost+compost tea
16.8018.009.00174.00 Compost+compost tea+mycorrhiza
1.0671.3011.2881.126 LSD at 5%
Table 7. Effect irrigation intervals on silymarin content (mg/g seed) of Silybum marianum
Irrigation Intervals Silychristin Silydianin Silybin ASilybin BIsosilybin AIsosilybin BTotal
3 days 17.952 11.182 11.092 18.576 7.216 2.814 68.832
6 days 18.584 12.032 12.086 19.332 7.538 3.078 72.65
9 days 22.028 13.352 14.776 23.34 8.734 3.184 85.414
Table 8. Effect of fertilizer treatment on silymarin content (mg/g seed) of Silybum marianum
Fertilizer treatmentsSilychristin Silydianin Silybin ASilybin BIsosilybin AIsosilybin BTotal
NPK19.62 11.76 12.61 20.82 7.64 3.07 75.52
Compost19.49 13.10 12.35 20.00 7.92 3.09 75.95
Compost+mycorrhiza20.49 12.53 13.35 21.48 8.21 2.79 78.85
Compost+compost tea19.37 12.34 12.80 20.27 8.18 3.36 76.32
Compost+compost tea+ mycorrhiza 18.34 11.22 11.96 19.51 7.20 8.47 76.7

HENDAWY et al. – Response of Silybum marianum to irrigation and fertilizer 27
Table 9. Effect the interaction treatments between irrigation intervals and fertilization on silymarin content (mg/g seed) of Silybum marianum
Total Isosilybin
B Isosilybin
A Silybin
B Silybin
A Silydianin Silychristin Fertilizer treatments Irrigation
intervals
71.74 3.02 7.49 19.45 11.66 11.48 18.64 NPK 3 days
69.64 2.95 7.26 18.46 11.02 11.83 18.12 Compost
64.86 2.03 6.98 17.70 10.79 10.34 17.02 Compost+mycorrhiza
72.00 3.38 7.57 19.14 11.38 12.03 18.50 Compost+compost tea
65.92 2.69 6.78 18.13 10.61 10.23 17.48 Compost+compost tea+mycorrhiza
72.24 3.29 7.33 18.74 11.71 12.80 18.37 NPK 6 days
68.96 2.69 7.22 18.51 11.30 11.30 17.94 Compost
75.95 3.13 8.15 20.21 12.55 12.39 19.52 Compost+mycorrhiza
71.3 2.92 7.63 19.35 12.48 10.81 18.11 Compost+compost tea
74.8 3.36 7.36 19.85 12.39 12.86 18.98 Compost+compost tea+mycorrhiza
82.58 2.89 8.11 24.27 14.47 10.99 21.85 NPK 9 days
89.24 3.64 9.28 23.02 14.74 16.16 22.40 Compost
96.29 3.20 9.50 26.54 17.25 14.87 24.93 Compost+mycorrhiza
85.64 3.77 9.33 22.32 14.54 14.17 21.51 Compost+compost tea
73.32 2.42 7.45 20.55 12.88 10.57 19.45 Compost+compost tea+mycorrhiza
Silymarin content
Data tabulated in Tables 7, 8 and 9 indicated that total
silymarin content (mg/g seed) ranged from 64.86 to 96.29
mg/g. The main constituent of silymarin were Silybin B
(17.70-26.54 mg/g) followed by Silychristin (17.48-24.93
mg/g). In this connection, dried extracts of milk thistle
seeds contain approximately 60% silymarin, where
silymarin consists of four flavonolignans of silybinin (~ 50
to 60%), isosilybinin (~ 5%), silychristin (~ 20%) and
silydianin (~ 10%) (Burgess, 2003). (Ibrahim et al. 2007)
found that the concentration and total yield of six silymarin
compounds showed wide variations between lines, varieties
and generations ranged from 11.92 to 62.85 mg/g seed and
between 329.8 to 2121.3 mg/plant, respectively. Six
silymarin compounds: silychristin, silydinin, silybin A,
silybin B, isosilybin A and isosilybin B were detected in
the extract of all tested treatments. These results were in
agreement with (Ibrahim et al. 2007).
Irrigation intervals
Data tabulated in Table 7 show that, the mean values of
total Silymarin content (mg/g seed) were 68.83, 72.65 and
85.41 mg/g were obtained as a result of irrigation intervals
at 3, 6 and 9 days, respectively.
Silybin B followed by silychristin were the main
components of silymarin. The maximum mean values of
Silybin B (23.34 mg/g) and Silychristin (22.03 mg/g) were
observed as a result of irrigation intervals every 9 days.
Drought stress increases the secondary products
percentage of more medicinal and aromatic plants, because
in case of stress, more metabolites are produce in the plants
and substances prevent from oxidization in the cells, but
secondary products content reduce under drought stress,
because the interaction between the amount of the
secondary products percentage and mass production is
consider important as two components of the secondary
products content and by exerting stress, increases the
secondary products percentage but mass production
decreases by the drought stress, therefore secondary
products content reduces. The data from (de Abreu and
Mazzafera 2005) showed that also the total amount of some
secondary plant products per plant indeed is significantly
higher in plants grown under drought stress than in those
cultivated under normal conditions. Although stressed
plants had been quite smaller, the product of biomass and
substance concentration yields in a 10% higher amount of
phenolic compounds; however, the total content of
betulinic acid was nearly the same in plants when grown
under drought stress or under standard conditions. Also the
studies published by Nogues et al. (1998), who found a
massive increase of phenolic compounds in stressed peas,
allow calculating the overall yield of the related substances.
Despite the fact that the total biomass of pea plants grown
under drought stress is just about one third of those
cultivated under standard condition, the overall amount of
anthocyanins (product of biomass and anthocyanin
concentration) is about 25% higher in the stressed plants.
Apart from that, the overall yield of total flavanoids was
nearly the same in Pisum sativum plants grown under
drought stress or under non-stress conditions.
Fertilizer treatments
Data tabulated in Table 8 indicated the effect of
different fertilizer treatments on silymarin content (mg/g).
Total silymarin content ranged from 75.52 to 78.85 mg/g.
Compost+mycorrhiza treatment gave the maximum mean
values of total silymarin content (78.85 mg/g) followed by
Compost+compost tea+mycorrhiza treatment which gave
76.70 mg/g. The highest mean values of Silybin B (21.48
mg/g) and Silychristin (20.49 mg/g) were obtained as a
result of compost+mycorrhiza treatment compared with
other treatments.
As for the favorable effect of applying organic and/or
bio fertilizers on silymarin content may be due to effect of
these fertilizers on accelerating metabolism reactions as
well as stimulating enzymes. Application of bio fertilizers
and compost significantly improved secondary products
such as essential oil, rutin and coumarin (El-Sherbeny et al.
2007 a, b). Variations in plant growth and active principles
in mycorrhizae inoculated plants have been reported for

5 (1): 22-29, May 2013

28
many other medicinal plants (Sailo and Bagyara 2005;
Copetta et. al. 2006).
Interaction treatments
It can be noticed that compost+ mycorrhiza treatment
under 9 days irrigation intervals gave the maximum value
of total silymarin content (96.29 mg/g) followed by
compost treatment under the same irrigation intervals
which gave 89.24 mg/g (Table 9). The lowest value of
Sylimarin content (64.86 mg/g) was obtained as a result of
compost+mycorrhiza treatment under 3 days irrigation
intervals.
Moreover, the highest values of Silybin B (26.54 mg/g)
and Silychristin (24.93 mg/g) were observed as a result of
compost+ mycorrhiza treatment under 9 days irrigation
intervals. In this respect, mycorrhiza fungi play a critical
role in interest cycling and ecosystem function. They
improve plant growth and survival through a mutuality
relationship in which photosynthates are exchanged for
increased access to water and nutrients (Kernaghan 2004).
These effects may be played an important role to increase
the secondary metabolites accumulation.
CONCLUSION
All presented data indicated that all studied growth and
yield characters were significantly affected by the duration
of irrigation intervals also organic and bio fertilizer showed
a primitive effect on growth and yield characters. The
interaction between irrigation intervals and fertilizer
treatments has a clear considerable effect on growth and
yield characters. Organic and bio fertilizers can reduce the
harmful effect of water stress.
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