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Journal of Applied Sciences Research, 4(3): 270-277, 2008
© 2008, INSInet Publication
Allelopathic Effect of Acacia Raddiana Leaf Extract on the Phytochemical
Contents of Germinated Lupinus termis Seeds
1
Samia El-Sayed Saffan and 2Amani M.D. El-Mousallamy
1
2
Botany Department, Faculty of Science, Zagazig University.
Chemistry Department, Faculty of Science, Zagazig University.
Abstract: There are some foods that contain mutagenic or carcinogenic agents, some of which occur
naturally and other that may be formed during preparation or cooking. Several foods, such as Legumes,
also contain natural antimutagens and / or anticarcinogens. Lupine is one such legume that contains high
amouts of protein (40%) and oils (14%). However, the use of this crop as a source of food has been
limited by the presence of antinutritional agents such as phenolic compounds and alkaloids. It has also
been suggested that consuming these compounds can affect human health and may even reduce the risk
of disease. The objective of this work was to determine the effect of Acacia raddiana leaf extract on
alkaloid, flavoniod, oligosaccharides, total tocopherol and phytosterol contents of Lupinus termis. The
present study aims at investigation of the response of germinated seeds of Lupinus termis to effect
allelopathic of Acacia leaf extract (control, 2% and 4%) for a period of 2 days. â-OH- lupanine and D-isolupanine were more abundant of alkaloids in germinated seeds of lupinus under two different
concentrations of leaf extract of Acacia. Moreover, X-iso- sparteine and sparteine were decreased under
both concentrations, also, OH- sparteine and termisine were decreased only at 2% concentration, while
increased at 4% concentration. The treatment of Acacia leaves resulted in accumulated amount of
lupinisoflavone, parvisoflavone, kaempferol, quercetin and epigenin isoflavonoid contents of germinated
seeds of lupinus, but declined 2- hydroxygenisteine and wighteone at both concentrations. Oligosaccharide
and total tocopherol contents of germinated lupinus were accumulated at 2% but reduced at 4%
concentration.
Meanwhile, total phytosterol of germinated seeds of lupinus were increased under both concentrations
of Acacia leaves extract.
Key words: Lupinus – Acacia – alkaloid – isoflavonoid – oligosaccharide – tocopherol – phytosterol
fungi and viruses [1 0 ]. Phenolic compounds can interact
with human salivary proline-rich protein to produce an
astringent sensation [4 ]. Their ability to interact with
proteins has led to their importance in the food
industry, herbal medicine and agriculture [4 ]. Flavonoids
are the major group of phenolic compounds, thus
biologically active and may be potent antioxidants,
scavengers of active oxygen species and electrophiles,
blockers of nitration, or chelators of metals. They can
undergo autoxidation to produce hydrogen peroxide in
the presence of metals and are capable of modulating
the activity of certain cellular enzymes [1 7 ]. The principal
role of alkaloids and other secondary compounds in
plant tissues appears to be one of chemical defence
against predators. A few crop plants contain alkaloids,
which are bitter tasting compounds to mammalian
herbivores. Alkaloids can be pharmacologically active
and may have narcotic, analgesic, antitussive,
chemotherapeutic, antiarrythmic (cardiotonic), diuretic,
hypoglycemic or uterotonic properties [2 3 ]. It has been
INTRODUCTION
In recent years, much emphasis has been placed on
dietary habits as being significant factors affecting
human health. Epidemiological studies have shown a
good relationship between good dietary habits and a
low incidence of cancer. N aturally occuring
antimutagens and anticarcinogens can be found in
fresh fruits and vegetables, including legumes [2 0 ].
The lupinus genus is widely distributed approximately
300 species are found in the M editerranean countries,
Africa and North & South America [1 5 ]. Lupin is a
legume with a rich source of plant protein and amino
acid [4 1 ]. Similar to other legumes, lupine contain
phenolic compounds and carbohydrates that may affect
human health or results in a reduced risk of
disease [4 1 ,5 1 ]. Phenolic compounds are secondary plant
metabolites commonly found in fruits, vegetables and
legumes, and which seen to be involved in the defence
of plants against invading pathogens, insects, bacteria,
Corresponding Author: Samia El-Sayed Saffan, Botany Department, Faculty of Science, Zagazig University.
270
J. Appl. Sci. Res., 4(3): 270-277, 2008
shown that oral administration of Lupinus termis
reduces high blood pressure and hyperglycemia in
rabbits, rats and mice [3 5 ]. Indeed, the addition of lupin
seeds to the food of diabetic-hypercholesterolemic
rabbits decreases cholesterol levels and postprandial
hyperglycemia [1 ]. O n the other hand, alkaloids sparteine
sulfate administered by intravenous in fusion to normal
men increases either basalor glucose-induced insulin
secretion [1 1 ].
M oreover, Pereira et al.,[3 7 ] have demonstrated that
aqueous lupine extract enhances insulin release
from isolated rat pancreatic islets, and sparteine
has
been shown to increase insulin secretion in
vitro [3 6 ]. However, a number of health benefits
results from ingestion of oligosaccharides such as
proliferation of bifidobacteria and reduction of
detrimental bactaria, reduction of toxic metabolites
and detrimental enzymes, prevention of pathogenic
and
autogenous
diarrhoea,
prevention
of
constipation protection of liver function, reduction
of serum cholesterol, reduction of blood pressure
and they may have anticancer effects [3 4 ]. But recently
it
has
been
throughly investigated taking into
account especially the phytochemical representing the
minor compounds like tocopherols and phytosterols [4 3 ].
This interest is connected with the activity of such
compounds against cardiovascular diseases [7 ]. Most of
these beneficial effects are due to antioxidant activity
especially when the presence of phenolic compounds
and tocopherols are involved in the stability of oils and
thus its toxicological safety [2 2 ].
In general, phytosterol supplementation tends to
decrease serum levels of total and LDLs (low-density
lipoproteins) cholesterol and has little effect on serum
levels of HDL (high-density lipoproteins) cholesterol
and triglycerides. The trees of Acacia spp. are
generally planted along the field bordes as windbreaksshelterbelts or are scattered within the fields with no
systematic design [3 8 ]. The tree is used for a variety of
purposes (fuel, fudder, timbers, gums, etc.) and its
nitrogen fixing ability makes it one of the most
preferred species for agricultural fields. Tree-plant
interactions are of p aram o unt im p o rtance in
agroforestry system [4 4 ]. W oody plants produce a large
number of chemical compounds which vary in their
chemical composition and concentration [4 0 ]. There is a
general belief among farmers that when trees are grown
in combination with agricultural crops the latters are
adversely affected. Acacia species have been reported
to suppress the germination and seedling growth of
many agricultural crops [4 2 ]. Such effect was related to
the presence of allelochemicals in various parts of
Acacia spp., including tannis, wax, several flavonoids
and phenolic acids [9 ]. Biosynthesis and accumulation of
phenolic compounds are one of the common responses
of plants to various biotic and abiotic stress conditions,
indicating their ability to participate in the process of
stress acclimation [1 9 ].
M ATERIAL AND M ETHODS
M aterial: Seeds of lupinus termis were obtained from
the Agriculture Research Center, Giza, Cairo. Samples
of Acacia raddiana leaves were collected from Zagazig
area, during February 2002 and identified by Dr. ElHaddidy, professor of Botany, Cairo University, Egypt.
M ethods: Four different weights (one, two, three and
four grams) of dry Acacia raddiana leaves, each was
separately soaked in 100 mls of double distilled water
for 24 hrs. at room temperature. The mixture was
filtered through whatman No.1 filter paper to obtain
1%, 2% , 3% and 4% w/v. Four levels besides the
control were chosen for germination experiments.
Twenty seeds were soaked in fifty mls of each level of
the Acacia extract for 24 hours, then washed with
distilled water. Twenty seeds of Lupine were arranged
in petri-dishes on W hatman N o. 1 filter paper under
laboratory condition at 25 o C. Double distilled water
was applied to all samples. The germination percentage
was recorded at 12 hours intervals up to 2 days, [4 1 ].
The biochemical analysis were determined after 2 days
from germination for 2% and 4% concentration of
Acacia leaves. W hereas the alkaloids, flavonoids,
oligosaccharides, tocopherol and phytosterols were
determined in germinated seeds (2 days) for 2% and
4% concentrations of Acacia leaves.
C
C
271
Alkaloids were extracted as described by Muzquiz
et al.,[3 1 ] and they are estimated by HPLC column
C 1 8 with flow rate 0.5 ml min -1 of mobile phase
was mixture of MeOH: H 2 O (1:1) and detected by
UV detectors at ë 2 5 6 . 1 ml extract of plant with
chloroform was dissolved in 2 ml of dilute
hydrochloride, if precipitate is obtained on addition
of few drops of Mayer`s reagent, W agner`s
reagent, when few drops of the solution were
applied to filter paper and sprayed with
Dragendroff`s reagent, orange colour is observed
this indicates alkaloids may be present.
Extraction of flavonoids estimated by Mabry et
at.,[2 8 ]. Dry ground seeds (20 gm) were extracted
with 85% methanol / H 2 O by soaking over-night.
The filterate was evaporated using a rotary
evaporator. The extract was redissolved in water,
and was spotted on two dimensional paper
chromatography using (n- Butanol – acetic acid –
water, 3:1:1) in the long direction, and 15% acetic
acid in the second direction using W hatmann No.3
mm paper. Flavonoids were identified using
methods described by [1 6 ], including R f value, colour
under long UV light (366nm) on cellulose plates,
the changes of colour with ammonia were
observed under long UV light, HPLC column C 1 8
J. Appl. Sci. Res., 4(3): 270-277, 2008
C
C
Table 1:
with flow rate of 1.0 ml/min. was used with two
solvents (A+B) 0.1% formic acid in water (A) and
(B) consisted of 80% acetonitrite and 20% acetic
acid. Isoflavonoid contents were detected by UV
detectors at ë 3 1 0 . About 5 g of each of the dried
powder was separately maccerated overnight in 50
ml of 1% hydrochloric acid and filtered, each
filtrate was separately tested for flavonoid
compounds as follow: 10 ml of each filtrate was
rendered alkaline with sodium hydroxide faint
yellow colour was obtained, this indicates the
probable presence of flavonoids.
For extraction of oligosaccharides in germinated
seeds of Lupinus samples with aqueous ethanol
(50% ) using HPLC column C 1 8 (250*4.6mm) with
flow rate of 1 ml/min. and acetonitrite: water
(65:35 v/v) as the mobile phase as method of
Muzquiz et al.,[3 2 ]. Fifty microliters of the sample
was injected and quantified by the external
standard material using sucrose, raffinose,
melibiose, stachyose, verbascose and ciceritol
purchased from Sigma Chemical Co..
Saponification for sterol and tocopherol analysis
were estimated by HPLC according to standard
procedure [2 9 ]. For phytosterol analysis, 20 ìl sample
was injected into C 8 column (250*4.6 mm) with
flow rate of 1.6 ml/min. at 50ºC and mobile phase
was 80% acetonitrite and 20% water at ë 2 0 5 . Total
tocopherol were detected by absorbance at 305 nm
after separation on column, (250*4.6 mm) at flow
rate of 1.2 ml/min. at 25ºC at ë 2 9 2 with a mobile
phase of 2- propanol (1% v/v) in hexane and
peaks were identified by retention time relative to
the standard.
Influence of A cacia raddiana leaves extract on the
alkaloids contents of germ inated seeds of lupinus term is
(2 days old) (m g/g dry wt.)
Treatm ent
-------------------Contents
L-oxo-Sparteine
17-oxo-Lupanine
D -Lupanine
13-O H -Lupanine
D -iso-Lupanine
x-iso-Sparteine
O H -Sparteine
Sparteine
Term isine
Control
2%
4%
Rf
35.5
84.4
50.9
350.4
498.1
12.9
25.5
15.6
80.1
32.5
88.6
48.4
355.6
499.3
8.81
22.53
12.7
83.6
29.65
89.78
57.5
362.7
505.6
7.76
31.6
10.4
85.8
0.88
0.81
0.71
0.75
0.38
0.2
0.12
0.9
0.35
M ass spectral
248
247
248
246
248
234
232
234
264
that quantification of quinolizidinic alkaloids (Qas)
have several pharmacological activities such as
nicotine-like activity, antiarrythmic (cardiotonic),
uterotonic properties and they have a hypoglycemic
effect [2 1 ] . T his result is in agreement with Saffan and
Salama [4 1 ], found that aqueous leaves extract of Acacia
contained the phenolic compounds and flavonoids that
might be implicated as allelochemicals agent which has
been attributed to inhibited of enzymes of biosynthesis
of Guinolizidine alkaloids. In this respect, Takamatsu
et al.,[4 7 ], indicated that certain members of the genus
lupinus are rich in esters of bicyclic quinolizidine
alkaloids with the derivatives of cinnamic acid. Also,
Takamatsu et al., [4 7 ] revealed that two new lupine
alkaloids (-) 13 á-tigloyloxymultiflorine and epilupinine
were isolated and their concentration increased rapidly
during the frist 3-10 days of growth of seedling
lupinus. So far, these enzymatic activities for
biotransformation of alkaloids tend to increase
temporarily during the first 3 to 6 days after
germination. Garcia-Lopez et al., [1 1 ], found the
quinolizidine alkaloids lupanine, 13-á-OH-lupanine, 17oxo-lupanine and 2-thionosparteine enchanced insulin
secretion of rat islets at different concentration of
glucose. On the other hand, Banuelos et al.,[3 ],
indicated that the large concentration of quinolizidine
alkaloids extracts from lupines caused of brain damage
of adult rat. Meanwhile, Gay et al., [1 3 ] revealed that
concentration of alkaloid lupinus leucophyllus was
modrately high in young leaf materials, decreased in
the more mature leaf growth, high in flower buds and
highest in seeds. There was also marked year-to-year
variation in the proportion of the total alkaloids. Also,
Gay et al.,[1 3 ] studied that , the total alkaloid of lupinus
effect by shading was left in place for 7 days. On the
other hand, lupinus leucophyllus is one of many lupine
species known to contain toxic and/or teratogenic
alkaloids that can cause congenital birth defects to
calves and cows and induced crooked calf diseases and
challenge in cattle. These alkaloids distributed in the
different tissues, where anagyrine levels were highest
in the floral tissue, lupanine and unknown F
RESULTS AND DISCUSSION
Table (1) revealed that the percentage of the most
important known components of the total alkaloid
content of the germinated seeds of lupinus termis under
the different concentration of leaves extract of Acacia.
It appears that neither L-oxo- sparteine, x-iso- sparteine
nor sparteine percentage was affected by various
concentration. Treatment of leaves extract of Acacia
(4%) was highly accumulated of D-lupanine and OHsparteine in germinated lupinus. However, these were
decreased under concentration (2%). D-iso- lupanine
was the most abundant alkaloid ranging from 499.3 to
505.6 mg/g respectively. 17-oxo-lupanine and termisine
were present at similar concentration in germinated
seeds of lupinus table (1). On the other hand, sparteine
and x-iso-sparteine alkaloids were highly lower
p erc e n ta g e in g e rm in a te d lup inus under all
concentrations. These results are important from a
toxicological point of view because it is well known
272
J. Appl. Sci. Res., 4(3): 270-277, 2008
Table 2:
accumulated to the greatest level in the vegetative
tissue and 5-6-dehydrolupanine accumulated to the
highest level in the stem. Anagyrine is, thus, the
phenological stage poising the greatest danger to
grazing livestock [3 9 ,2 4 ,2 5 ]. In this respect Saffan and
Salama,[4 1 ] revealed that accumulation of specific amino
acid such as arginine, glutamic acid, lysine and
pyruvate family were clearly observed in germinated
seeds of lupinus at different concentrations of Acacia
leaves extract (2% and 4%) for 2 days from
germination. Besides being incorporated into proteins,
free amino acids can serve as precursor for polyamines.
A large number of reports have demonstrated
accumulation of polyamines under a variety of
enviromental stress [2 6 ] . Serine and arginine are
precursors for the synthesis of glycinebetaine,
nicotinamide and putrescine that are commonly
encountered osmolytes that accumulates in plants under
stresses [2 7 ]. Besides, glutamic acid was considered as
precursor for glutathione that is involved in oxidative
defense [4 5 ]. Also, Saffan and Salama [4 1 ], showed that,
accumulation of amino acid lysine and pyruvate family
at different concentrations of Acacia leaves extract (2%
and 4%) lead to increase in Quinolizidine alkaloids of
germinated lupinus, which it was a precursor of
biosysnthesis of lupanine and sparteine W ink and
Hartmann [5 3 ].
Table (2) showed that flavonoids of lupinus termis
contained flavones, isoflavones and tentatively
flavonones. The flavones identified are apigenin, cglycoflavones and luteolin. These compounds exist in
the plant as free a glycones as well as in several
glycosylated forms. C-glycoflavones occur widely in
lupinus plants. The isoflavone genistein, kaempferol
and quercetin were observed as a glycones as well as
glycosides bound through positions 3 and 7.
Treatm ent
---------------Contents
Genistein
2-hydroxygenistein
W ighteone
Luteone
Luteolin
Lupinisoflavone
Parvisoflavone
Kaem pferol
Q uercetin
Apigenin
C
C
C
C
C
C
C
C
C
Control
2%
4%
30.9
31
25
35
25
54.5
29.6
42.6
44.8
30.6
28.6
29.8
21.5
36.1
24.0
57.3
30.9
45.5
46.8
31.3
35.6
27.8
19.2
38.1
27.8
59.9
33.7
46.1
48.75
33.8
M ass spectral
132
137
311
410
386
353
326
286
302
270
Compound Lupinisaflavone A colour UV bright
yellow, NH 3 yellow-green PCR f 0.731 UV m max
in MeOH, 255, 265.
Compound Parvisoflavone colour UV faint blue,
NH 3 intense blue, PCR f: 0.58 in B AW , UV nm
max 250, 270.
Compound kaempferol colour UV yellow, NH 3
yellow, PCR f: 0.8, UV nm max in MeOH: 266,
322, 367.
Compound Quercetin colour UV yellow, NH 3
yellow, NH 3 orange PCR f: 0.57, UV nm max in
MeOH, 255, 370.
Compound Epigenin colour UV blue grey, NH 3
dull ochre, PCR f: 0.83 U V nm max in MeOH,
269, 336.
Also, table (2) revealed that mostly flavonoids
patterns of germinated seeds of lupinus which treated
with (4%) of Acacia leaves extract were higher than
treated with (2%). W hereas, Acacia raddiana contains
high amount of phenolic compound [4 1 ]. Therefore, the
germinated seeds of lupinus after 48 hours treated with
aqueous extract of Acacia (4%) were increased in
amount of flavono id com pound s. T he major
isoflavonoids in lupinus termis are week natural
oestrogens, power natural insecticides, while related
coumestans such as pisatin are phytoalexins, protective
substances formed in plants in response to disease
attack [2 ]. These results are in accordance with
previously reported findings, Nicholls and Bohm,[3 3 ]
found that flavonoids types of American lupines
e n c o u n te r e d w e r e fla v o n es, C -g lyc o fla v o n e s ,
isoflavones, flavonols and tentatively flavonones. Also,
Stobiecki and W ojtaszek,[4 6 ] showed that isoflavonoids
in lupine root extracts are genistein, luteone, 2
hydroxygenistein, wighteone, lupinisoflavone and
parvisoflavone by using HPLC and (GC-MS) for the
identification of isoflavonoids. On the other hand,
Chukwumah et al.,[6 ] found that peanut had the highest
total flavonoids and polyphenol content such as
isoflavone glycosides daidzin, genistein, sissostrin
(biochanin A glycoside), luteolin and kaempferol.
Identification of Flavonoids:
C
Influence of Acacia raddiana leaves extract on the
isoflavonoid contents of germ inated seeds of lupinus
term is (2 days old) (m g/g dry wt.)
Compound Genistein: colour UV dull purple, NH 3
dull brown, visible colour, None. PCR f (cellulose
plate): 0.94 B AW , UV nm max in ETOH 250,
260, 302, 263, 325.
Compound Hydroxygenistein: dark mauve, NH 3 ,
faint brown, R f: 0.92 BAW , UV nm max 250,
260, 302, 263, 325.
Compound W ighteone colour UV dark mauve,
NH 3 pale yellow, PCR f 0.59 UV nm max 330,
350.
Compound Luteone: colour UV yellow-green, NH 3
yellow, PCR f 0.72 BAW , UV nm max in MeOH,
248, 269, 355.
Compound Luteolin: colour UV dull ochre, NH 3
dull ochre bright. PCR f: 0.66, UV nm max in
MeOH, 255, 268, 350.
273
J. Appl. Sci. Res., 4(3): 270-277, 2008
Table 3:
Table (3) showed that decreases of total
oligosaccharides compounds of lupinus germinated the
4% concentration of Acacia leaves while, increases at
2% concentration of Acacia. But, stachyose, verbascose
and raffinose enhanced under 2% concentration of
Acacia than the other one. These results are in
accordance with previously reported findings, Saffan
and Salama [4 1 ], showed that the aqueous extract of
Acacia leaves inhibited the amylase activity, an affect
which decreased at the increase in the level of
treatments (4%). This reduction caused a decrease in
the level of the total solube sugars (reducing and non
reducing) and a increase relatively in the level of
polysaccharides of germinated seeds of lupinus. On the
other hand, Einhelling [8 ], found that the most phenolic
compound in Acacia leaves extract such as tannins or
juglone inhibited the activity of amylase and most
hydrolytic enzymes. These allelochemical substances
might be present in leaves extract of Acacia and
retarded the activity of amylase and decreased the
soluble sugar contents of germinated seeds of lupinus
after 2 days old. Table (3) represent that the amount of
carbohydrates in germinated lupinus was 114.23 mg g -1
which included mostly mono, di and oligosaccharides.
The presence of these oligosaccharides are quite
characteristic of lupinus spp. Stachyose was the most
abundant oligosaccharides present in the lupinus termis
(55.33 and 57.37 mg g -1 control and 2% concentration
of Acacia leaves extract respectivly). This result is
agreement with other lupinus spp, such as L. albus, L.
angustifolius, L. atlanticus and L. campestris [1 8 ]. The
amount of stachyose observed in the present study was
higher than in other legumes such as chick pea,
soybean and red lentil[5 ]. It is worthy to point that, the
main advantages of oligosaccharides intake in humans
are gaining attention since the daily requirment is small
(usually 3 g day-1 ) and the health benefits are
significant (or great). Tomomatsu,[4 8 ] noted that
oligosaccharides are do not bind minerals, easy to
incorporate into processed foods and drinks. In
addition, they do not cause diarrhoea in recommended
doses and the average d aily intake of
fructooligosaccharides from natural sources in the USA
is estimated to be approximately 0.8 g day -1 / 130 Ib
body weight. Thus, the ingestion of supplemental
oligosaccharides may be beneficial for mature adults
who desire to maintain a healthy digestive system.
Table (4) revealed that the levels of total tocopherol of
germinated seeds of lupinus were increased under 2%
aqueous extract of Acacia. while, decreased under 4%
concentration. Tocopherols, act as antioxidants by
trapping the hydroperoxide intermediates and stopping
the autoxidation chain reaction. But the ã-tocophenol
roles was the most abundant tocophenol than other.
Differences in relative amounts of total tocophenol are
Influence of A cacia raddiana leaves extract on the
carbohydrate contents of germ inated seeds of lupinus
term is (2 days old) (m g/g dry wt.)
Treatm ent
------------Contents
Sucrose
M elibiose
Stachyose
Verbascose
Ciceritol
Raffinose
Table 4:
Control
2%
4%
18.87
5.8
55.33
19.03
4.23
10.97
19.2
4.5
57.37
22.33
3.07
12.67
14.1
3.37
48.97
17.97
2.3
11.83
Influence of A cacia raddiana leaves extract on total
tocopherol and phytosterol contents of germ inated seeds
of lupinus term is (2 days old) (µg/g oil).
Treatm ent
--------------Control
á-Tocopherol
ã-Tocopherol
ä-Tocopherol
Cam pesterol
Stigm asterol
â-Sitosterol
Control
2%
4%
710.1
950.6
371.3
66.3
38.1
991.2
721.4
970.8
375.8
70.8
40.8
993.7
705.3
880.5
350.7
67.1
38.3
996.4
important, á-tocopherol affects human nutrition and
health aspects, while ã-tocopherol shows a strong
activity in the seed protecting compounds like fatty
acids. The amount of tocophenols are very high in
maize and soybean seed oils (1618.4 and 1797.6mg/kg,
respectively, [4 9 ].
Also, the order of increasing total phytosterols
content illustrated in germinated seeds of lupinus under
2% extract of Acacia, while these were decreasing
under 4% concentration. â-sitosterol level was the most
abundant phytosterols. Campesterol and stigmasterol
were present at similar concentrations (table 4). But,
the â-sitosterol leve was the highest in germinated
lupinus under 2% and 4% aqueous extract of Acacia
leaves. This results were satisfactory, especially when
considering that the highest polyunsaturated and ratio
between oleic / linoleic fatty acids in germinated seeds
of lupinus [4 1 ]. Phytosterols are found in plant foods and
are structurally and functionally analogous to
cholesterol in vertebrate animals. â-sitostero l,
campesterol and stigmasterol are the most commonly
occurring phytosterols and constitute 95% of total
phytosterols in the diet. In addition to nuts,
phytosterols are found in a range of seeds, legumes,
vegetables and unrefined vegetable oils [5 2 ]. The effects
of dietary supplementation with phytosterols on serum
cholesterol levels in humans have been reviewed
comprehensively by[3 0 ].
In conclution, the results of the present work can
suggest a mechanism by which allelopathic effect of
Acacia leaves extract may be increase the major
Quinolizidine alkaloid levels in germinated seeds of
lupinus termis. This suggestion is based on the
accumulation of amino acid (e.g. Lysine). The
274
J. Appl. Sci. Res., 4(3): 270-277, 2008
increased amounts of arginine amino acid under
allelopathic effects could derive the biosynthesis of
polyamins (e.g, putrescine). The polyamines were found
to induce nitrite oxide biosynthesis [ 5 0 ] . Also,
accumulation of isoflavonoids of germinated seeds of
lupinus were observed at different concentrations of
Acaica leaves extract. Total tocopherol and phytosterol
contents were reduced at the highest concentration of
polyphenolic compounds or allelochemicals of Acacia
leaves extract.
10. Friedman, M . and H. Jürgens, 2000. Effect of PH
on the stability of plant phenolic compounds.
Journal of Agriculture and Food Chemistry., 48:
2102-2110.
11. Garcia-Lopez, P.M., P. Garzon dela M ora, W .
W ysocka, B. Maiztegui, M .E. Alzugaray, H.D.
Zoto and M.I. Boralli, 2004. Guinolizidine
alkaloids isolated from lupinus species enhance
insulin secretion. European J. of Pharmacology. ,
504: 139-142.
12. Gay, C.C., E.S. Motteram, K .E. Panter and T.
W ierenga, 2007. year to year variation in alkaloid
in Lupinus leucophyllus growing on the Scablands
of central W ashington. Poisonous-plants: globalresearch-and-solutions., 2007: 414-417.
13. Gay, C.C., K.E. Panter, E.S. Motteram, J.M. Gay,
T. W ierenga and T. Plalt, 2007. Effect of shade on
alkaloid content of Lup inus leu co phyllus.
Poisonous plants: global-research and solutions.,
411-413.
14. Gay, C.C., A. Tibary, K.E. Panter, K.L. Mealey,
J.M. Gay, S.W . Hjartarson, E. Motteram, T.
W ierenga and L.F. James, 2007. Lupine-induced
crooked call diseases: plasma disposition of lupine
alkaloids following repeated challenge in cattle that
have given birth to affected calves or those that
have borne normal calves. Poisonous plants:
global–research and solutions., 2007: 151-155.
15. Gland-stones, J.S., 1998. Distribution origin,
taxonomy, history and importance In: Glandsrones,
J.S., Atkin, J.S., Hamblin, C. (Eds), lupinus as
crop plants. Biology, Production and Utilization.
Cab International, New York, pp: 1-39.
16. G rayer, R.J., 1989. “M ethods in
plant
Biochemistry” (Harborne, J. B., ed.) Vol, 1.
Academic Press. London.
17. Huang, M. and T. Ferraro, 1992. Phenolic
compounds in food and cancer prevention. C Y
phenolic compounds in Food and Their effect on
health II: Antioxidants and Cancer
Chemoprevention edited by M. Huang, C.HO and
H. S. Lee (W ashington. DC: American Chemical
Society), pp: 8-34.
18. Jimènaz Martinez, C., G. Loarca-Pina and G.
Davila, 2003. Antimutagenic activity of phenolic
compounds, oligosaccharides and quinolizidinic
alkaloids from Lupinus campestris seeds. Food
Additives and Contaminants, 20(10): 940-948.
19. Juszczuk, I.M ., A. W iktorowska, E. Malusa and
A.M. Rychter, 2004. Changes in the concentration
of phenolic compounds and exudation induced by
phosphate deficiency in bean plants (Phseolus
vulgaris L.). Plant Soil., 267: 41-49.
20. Karakaya, S. and A. Kavas, 1999. Antimutagenic
activities of some foods. Journal of the Science of
food and Agriculture, 79: 237-242.
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
Ahmed, R.T. and E.E. Esmail, 1993. Influence of
dietary lupin seed mixed diet on serum cholesterol
level in normal and diabetic rabbits. Egypt J.
Pharm Sci., 34: 565-576.
Bailey, J.A. and J.W . Mansfield, (eds), 1982.
phytoalexins, Blackie, Glasgow.
Banuelos-Pineda, J., G. Nolasco-Rodriguez, J.A.
Monteon, P.M. Garcia-Lopez, M.A. Ruiz-Lopez
and J. G arcia-E strad a, 20 0 5 . H istological
evaluation of brain damage caused by crude
quinolizidine alkaloid extracts from lupines,
Histology and Histopathology., 20(4): 1147-1153.
Bartolomè, B., L. Estrella and M.T. Hernandez,
2000. Interaction of low molecular weight
phenolics with protein (BSA). Food Chemistry and
Toxicology., 65: 617-621.
Carlsson, N.G., H. Karlons and A. Sardhberg,
1992. Determination of oligosaccharides in foods,
diets and intestinal contents by high-temperature
gas chromatography and gas chromatography /
mass spectrometry. Journal of Agriculture and
Food Chemistry, 40: 2404-2412.
Chukwumah, Y., L. W alker, B. Vogler and M .
Verghese, 2007. Changes in the phtochemical
composition and profile of row, bailed and roasted
peanuts. J. Agric. Food. Chem. 55(22): 9266-9273.
Delplanque, B., B. Le Roy, F. Mendy, E. Fenart,
A. Thaminy-Dekar and F. Syeda, et al., 2002.
Oleic, linoleic and alphalinoleic acids from
vegetable oils: where are the limits for benefical
effects on lipemia and atherothrombotic parameters
in humans OCL- Oleagineu Corps Gras Lipides,
9: 237-244.
Einhellig, F.A., 1995. Mechanism of action of
allelochemicals in alelopathy. In “Allelopthy:
organisms, processes and applications” (F. A.
Einhellig. Inderjet and K . M. M. Dakshinni, eds.)
p p : 9 6 -1 1 6 . A m eaican chem ical Society:
W ashington: USA.
Fagg, C.W . and J.L. Stewart, 1994. The value of
Acacia and Prosopis in arid and semi-arid
enviroments. J. Arid Environ., 27: 3-25.
275
J. Appl. Sci. Res., 4(3): 270-277, 2008
21. Kinghorn, A.D. and M.F. Balandrin, 1984.
Guinolizidine alkaloids of the leguminosae:
structural types, analysis chematoxonomy and
biological activities. Alkaloids Chemical and
Biological perspectives, edited by W .S. Pelletier
(New York: W iley)., pp: 105-148.
22. Koski, A., E. Psomiadou, M. Tsimidou, A. Hopia,
P. Kefalas and K. W ähälä, 2002. Oxidative
stability and minor constituents of virgin olive oil
and coldpressed rapeseed oil. European. J. of Lipid
Science and Technology, 214: 294-298.
23. Kutchan, T.M., 1995. Alkaloid biosynthesis-the
basis for metabolic engineering of medicinal
plants. Plant Cell., 7: 1059-1070.
24. Launchbaugh, K., J. Pfister, S. Lopez-Ortiz and R.
Frost, 2007. Bodycondition affects blood alkaloid
and monoterpene kinetics and voluntary in take of
c h e m ic a lly-d efen d ed p lan ts b y live rstoc k.
Poisonous-plants: global-research and solutions.,
2007: 394-400.
25. Lee, S.T. M.H. Ralphs, K.E. Panter, D. Cook and
D . R . G a r d n e r , 2 0 0 7 . A lk a lo id p r o file s ,
concentration and pools in velvet lupine (Lupinus
leucophyllus) over the growing season. J.
of
chemical Ecology., 33(1): 75-84.
26. Legocka, J. and A. Kluk, 2005. Effect of salt and
osmotic stress on changes in polyamine content
and arginine decarboxylase activity in Lupinus
luteus seedlings. J. Plant Physiol., 162: 662-668.
27. Liu, J.H., K . Nada, C. Honda, H. Kitashiba, X.P.
W en, X.M. Pang, Moriguchi, T. (2006): Polyamine
biosynthesis of apple callus under salt stress:
Importance of the arginine decarboxylase pathway
in stress response. J. Exp. Bot., 57: 2589-2599.
28. Mabry, T.J., K.R. Markham and M.B. Tiiomas,
1 9 7 0 . “ T h e S yste m atic Id e ntific atio n o f
Flavonoids”. Springer, New York.
29. Marini, D. and F. Balestricri, 1996. Metodi di
analisi chimica dei prodotti alimentari. Monolite,
Roma, Italy., 1: 214-218.
30. Moroeau, R.L., B.D. W hitaker and K.B. Hicks,
2002. Phytosterols, phytostanols and their
c o n ju g a tes in fo o d s: structural diversity,
quantitative analysis and health-promoting uses.
Prog. Lipid Res., 41: 457-500.
31. M uzquiz, M., C. Cuadrado, G. Ayet, C. De La
Cuadra, C.C. Burbano and A. Osagie, 1994.
Variation of alkaloid components of lupin seeds in
49 genotypes of Lupinus albus L. from different
countries and location. Journal of Agriculture and
Food Chemistry, 42: 1447-1450.
32. Muzquiz, M., C. Rey and C. Cuadrado, 1992.
Effect of germination on the oligosaccharides
c o nte nts o f lupin sp e cie s. Jo urna l o f
chromatography, 607: 349-352.
33. Nicholls, K.W . and B.A. Bahm, 1982. flavonoids
and affinities of some north american lupines. Can.
J. Bot., 61: 708-730.
34. Oku, T., 1994. Special physiological functions of
newly development mono- and oligosacchrides.
Functional Foods. Designer Foods. Pharmafoods
and Nutraceutical. Edited by l. Golbery (London:
Chapmanx Hill), pp: 202-218.
35. Omran, A.A., 1996. Farmakologiczna aktywnose
wyeiagow znasion wybranych gatunkow Lubinu
(L. angustifolius L., L. albus L., L. Luteus L.,)
ph.D thesis Department of the pharmacology Karol
Marcinkowski University of Medical Sciences.
36. Paolisso, G., M. Nenquin, F. Schmeer, H. Mathat,
H.P. Meissner and J.C. Henquin, 1985. Sparteine
increases insulin release by decreasing the K +
permeability of the B-cell membrane. Biochem.
Pharmacol., 34: 2355-2361.
37. Periera, F., R. Gluedrasgo, P. Lebrum, R. Barbosa,
A. Cunha, R. Santos and L. Rosario, 2001.
Insulinotrophic action of white lupine seeds
(Lupinus albus L.): effects on ion fluxes and
insulin secretion from isolated pancreatic islets.
Biomed. Res., 22: 103-109.
38. Puri, S., S. Singh and A. Kumar, 1994. Growth
and productivity of crops in association with
Acacia nilotica tree belt. J. Arid Environ. 27:
17-18.
39. Ralphs, M.H., K.E. Panter, C. Gay, E. Motteram
and S.T. Lee, 2007. Cattle grazing velvet lupine
(Lupinus leucophyllus): Influence of associated
forages, alkaloid levles and population cycles.
Poisonous plants: global research and solutions.
2007: 401-406.
40. Rice, E., 1995. Allelopathy in forestry. In
“Biological Control of W eeds and Plant Diseases:
Advances in Applied Allelopathy”. (E. Rice, ed.)
pp. 317-378. Univ. of Oklahoma Press, USA.
41. Saffan, S.E. and H.M. Salma, 2005. Influence of
allelopathic of Acacia raddiana leaf extract on
germination and some metabolites seedling of
Lupinus termis. Egypt. J. Biotechnol., 21: 32-43.
42. Schumann, A. W . ; Little, K. M. and Eccles, N. S.
(1995): Suppression of seed germination and early
seedling growth by planatation harvest residues.
South-Africa J. Plant Soil, 12: 170-172.
43. Shahidi, F., 2004a. Quality characteristics of edible
oils. In quality of fresh and processed foods
(pp: 239-249). New York, USA: Kluwer Academic
/ Plenum Publishers.
44. Sharma, K.D., K.L. Sidana and N.R. Singhvi,
1982. Allelochemic effect of Peganum harmala L.
On Pennisetum typhoides L. (Pajra). Indian J.
Bot., 5: 15-119.
45. Sharma, S.S. and K.J. Dietz, 2006. The
276
J. Appl. Sci. Res., 4(3): 270-277, 2008
46.
47.
48.
49.
50.
significance of amino acids and amino acid-derived
molecules in plant responses and adaptation to
heavy metal stress. J. Exp. Bot, 57: 711-726.
Stobiecki, M. and P. W ojtaszek, 1990. Application
of gas chromatography mass spectrometry to the
identification of isoflavonoids in lupine root
extracts. J. of chromatography, 508: 391-398.
Takamatsu, S., K. Saito and I. Murakoshi, 1991.
New lupine alkaloids from the seedlings of lupinus
and change of alkaloid pattern with germination. J.
of Natural Products., 54(2): 477-482.
Tomomatsu, H., 1994. Health effects of
oligosaccharides. Food Technology, 61-65.
Tuberoso, C.I.G., A. Kowalezyk, E. Sarritzu and P.
Cabras, 2007. Determination of antioxidant
compounds and antioxidant activity in commercial
oil seeds for food use. Food Chemical 103:
1494-1501.
Tun, N.N., C. Santa-Catarine, T. Begum, V.
Silveira, W .H. EnyIochevet, S. Folh and G.F.E.
S c h e re r, 2 0 0 6 . P o lyam ine s in d u c e ra p id
biosynthesis of nitric oxide (NO) in Arabidopsis
thaliana seedlings. Plant Cell Physiol., 47:
346-354.
51. Tzyh-Chyuan, H., I. Yu-Chih, C. Iou-Sen and L.
Jen-Kun, 1999. Inhibition of eleven mutagens by
various tea extracts: (-) epilogallocatechin- 3
–gallate, gallic acid abd caffeine. Food and
Chemical Toxicology. 37: 569-579.
52. W eihrauch, J.L. and J.M. Gardner, 1978.s Sterol
content of foods of plant origin. J. Am. Diet.
Assoc., 73: 39-47.
53. W ink, M. and T. Hartmann, 1985. Natural
products chemistry”. Ed. by Zalewski, R. I. and
Skolik, J. J., pp: 511.
277