World Journal of Agricultural Sciences 11 (2): 89-93, 2015

ISSN 1817-3047
IDOSI Publications, 2015
DOI: 10.5829/idosi.wjas.2015.11.2.12526

Effect of Alfalfa (Medicago sativa) Extract on Yield and Yield Components of

Lettuce (Lactucasativa), Beetroot (Beta vulgaris) and Pepper (Capsicum annum)

Tafesse Kibatu Shikur

Dilla University, College of Agriculture and Natural Resources, P.O.Box: 419, Dilla, Ethiopia

Abstract: Alfalfa extract has an autotoxic or growth regulatory effect which is determined by the way the extract
is prepared and applied to plants. Three separate studies were undertaken to determine the effect of hot and
cold water crude extract of alfalfa leaf on growth parameters and yield of lettuce, beetroot and pepper with an
attempt to promote the crops growth and yield. To prepare the extract, dried alfalfa leaf (15g) was soaked
separately in 1 litre distilled hot and cold water for 24 hours. To study the extract effect, 2 levels of cold water
extracts (50% diluted and without dilution), 2 levels of hot water extracts (50% diluted and without dilution) and
a control were considered as an experimental treatment. These treatments were applied for all crops in
randomized complete block design (RCBD) with three replications. Each plant was sprayed with 200ml diluted
alfalfa extracts. Spraying alfalfa extract on lettuce and pepper resulted in non-significant difference in growth
parameters and in all yield traits as compared to untreated treatment. However, in case of beetroot, the study
showed that the aqueous extracts have a significant effect on root length and yield.

Key words: Autotoxicity Lucerne Growth Regulators Aqueous Extract Plant Extract

INTRODUCTION hay or watering plants with extract made by soaking

Vegetables are cash crops and grown for income
generation. In order to achieve high quantity and quality
products, appropriate and improved production practices
must be applied. Adaptation of these practices requires
investment, time, labour and availability of resources. It is
clear that the longer the crop stays in the field the more
difficult the required field activities. In addition to escape
diseases and other unfavourable conditions, rapid growth
rate can reduce the cost of field operations and enhance
the rate of marketing products. 1Rapid growth rate can
be achieved with plant growth regulators. Triacontanol,
a common constituent of plant waxes, was shown to be an
active growth regulator, which increased the growth and
yield of crops [1].
Synthetic growth regulators are expensive and not
available in many countries like Ethiopia, particularly
when Ethiopian farmers are considered. Hence, alternative
and relatively cheaper practices such as use of plant
extracts could be solution for enhancing growth of
vegetables. This can be achieved by mulching with alfalfa
alfalfa in water [2, 3, 4]. Alfalfa is well known for its ability
to improve soil structure and, as a legume, is an effective
source of biological nitrogen (N2). It can be grown in
almost all agro-ecologic zones of Ethiopia [5].
Growth stimulating effect of alfalfa on several crop
plants has interest of researchers and nursery men [4]. A
growth promoter crystalline substance isolated from the
chloroform-soluble fraction of the alfalfa was identified as
triacontanol [1, 4, 6]. Growth stimulation of alfalfa is
brought about by mulching with alfalfa or watering plants
with an extract made by soaking alfalfa in water [1, 4].
In order to get the promoting effect of alfalfa in potted
plants, which are just starting their growth cycle,
finely chopped alfalfa with some mulch material can be
used. Alternatively, alfalfa aqueous extract can be used as
growth stimulant [4]. However, in the other hand,
alfalfa is claimed to be autotoxic and hetrotoxic [7].
Whatever the case it is, in order to get positive response
from alfalfa mulch and/or extract, optimum methods of
application must be established, such as rate and time of
application to crops in the field [8].

Corresponding Author: Tafesse Kibatu Shikur, Dilla University, College of Agriculture and Natural Resources,
P.O. Box: 419, Dilla, Ethiopia.
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 World J. Agric. Sci., 11 (2): 89-93, 2015
Previously, pot experiments showed a promising
result on troyer Citrange and pencil cedar seedlings [2, 3].
During the experiments different concentrations showed
different responses. However, in all experiments, almost
constant effect was observed with increasing of soaking
time from 24 to 72hrs. In addition, from these studies and
other previous results, the application of the alfalfa
extracted triacontanol is recommended to be at optimum
response time during the early growth stage of plants.
Based on these results, for this study, different level of
24hrs soaked alfalfa leaf which applied at the seedling
stage of plants was considered to bring positive effect on
crop growth. Therefore, the present study was planned to
determine the extracts effect on growth of beet as
representative of root crops, lettuce as representative of
leafy vegetables and pepper as representative of fruit
vegetables.
MATERIALS AND METHODS
Experimental Site: Dilla is located at 5.84-6.43 latitudes
and 38.08-38.44 longitudes. The altitude of the
experimental sites is about 1470 masl. The area is
characterized by Wet/Moist Kolla and highly recognized
by its agro-forestry cropping system.
Alfalfa Preparation: The seed of alfalfa cultivar 6984 was
collected from International Livestock Research Institute
and grown in Dilla agriculture farm and cut at bud stage
(about 10% flower). The leaves were separated and air
dried until constant weight was achieved. Then, it was
stored in air ventilated dry place, until it was used for the
experiment.
Alfalfa Extracts Preparation and Application: 15g dried
alfalfa leaf was soaked separately in 1 litre hot and cold
water for 24 hours. The residue was removed using
four-fold cheese cloth and the desired amount was diluted
by water. Each plant was sprayed with 200ml treatment.
Accordingly, for each of the experimental plots the
amount was calculated based on number of plants and the
treatment application was started 20 days after emergency
for lettuce and beetroot and 20 days after transplanting
for pepper.
Treatment and Design: To study the extract effect,
2 levels of cold water extracts (50% diluted and without
dilution), 2 levels of hot water extracts (50% diluted and
90
without dilution) and a control were considered as
experimental treatments. No treatment was applied for the
control plot. These treatments were applied randomly for
all crops in randomized complete block design (RCBD)
with three replications. Beetroot as representative of root
crops, lettuce as representative of leafy vegetable and
pepper as representative of fruit vegetables are selected
for the experiments. To achieve uniform stand at the
beginning of the establishment and before the application
of treatments, within the block transplanted pepper was
assigned based on uniformity of leaf number and plant
height. In case of direct sowed lettuce and beet crops the
stand uniformity was maintained by thinning non-uniform
stands.
Data Collection: Four randomly selected central
plants per experimental plot were tagged and used
to measure the parameters. Plant height was measured
in cm from the ground level to the top of the shoot.
To count the number of leaves, true leaves were
considered. Fresh weights, immediately after harvest, were
measured with the help of an analytical balance and
average weight was taken for analysis. The root length
was measured using a ruler and expressed in cm. Branch
at blooming stage was counted and recorded as the
number of branches. The total numbers of fruits per
plant were counted for pepper. Total weight of edible
portion was taken as yield for each crops and converted
in ton per hectare.
Chemical Analysis: Soil chemical analysis of the
two sites was made by the standard methods.
Total organic carbon was measured by the method of
Walkley and Black [9]. Total Kjeldahl nitrogen was
determined by the method of Bremner and Mulvaney
[10]. The pH was determined by potentiometric method.
Electrical conductivity was measured by using
hydrometer and APHA 2510 Conductivity. The
phosphorous was determined by Olsen et al. [11] method
(Table 1).
Statistical Analysis: The data were processed with
Microsoft Excel 2007. Analysis of variance (ANOVA)
was done using SPSS for Randomized Complete Block
Design (RCBD) to determine differences between
treatment means. The Means of treatments were
distinguished using Duncans Multiple Range Test at p
0.05.
 World J. Agric. Sci., 11 (2): 89-93, 2015

Table 1: Soil chemical analysis.

Parameters Pepper crop research Site lettuce and beet crops research Site
pH water 6.45 6.27
Electrical conductivity (ds/m) 0.11 0.10
Available phosphorus (mg/kg) 14.4 1.86
Organic carbon 2.54 1.06
Total nitrogen 0.20 0.10
Available potassium(cmol(+)/kg 1.85 2.52

Table 2: Mean square for pepper parameters subjected to alfalfa extract.

Source of variation df Total Biomass Number of leaf Number of Branch Number of Pod yield Seedling Length Root length
Treatment 4 2015ns 13193ns 33ns 9.8ns 278ns 20.5ns 3.4ns
Block 2 6173 8987 208 15.7 1061 48.6 3.5
Error 8 9894 9644 70 62.8 1523 30.3 10.5

Table 3: Mean square for lettuce parameters subjected to alfalfa extract.

Source of variation df Seedling length Root length Total Biomass Leaf dry weight Number of leave Root Weight Yield
Treat 4 1.4ns 0.65ns 15639ns 47ns 286ns 77ns 14198ns
Rep 2 4.9 2.47 34925 95 338 178 30182
Error 8 4.9 0.80 48873 112 452 105 45341

Table 4: Mean square for beet parameters subjected to alfalfa extract.

Source of variation df Root length Leaf number Yield Shoot length Total biomass
Rep 2 12.7 229.3 6780.3 7.4 46577
Treat 4 28.5** 46.2ns 118300.5* 2.6ns 88875ns
Error 8 3.5 123.0 20670.4 6.5 59488

RESULT AND DISCUSSION

None of the parameters of pepper plant and lettuce

were affected by sprays of alfalfa extract (Table 2 and 3).
In case of beetroot, parameters that are related to shoot
part did not show any significant difference. However,
root length and yield of beetroot were significantly
affected by the extract at p<0.05 and p<0.01 respectively
(Table 4).
The tallest beetroot plants were recorded in
50% diluted cold extract, which is similar in effect with the
control and undiluted hot mextract treatments (Fig. 1). Fig. 1: Root length of beetroot subjected to alfalfa
The least length was recorded from undiluted cold water extracts.
and hot water soaked treatments. The highest amount of
yield was shown by the control and 50% diluted cold
extract. The least yield was recorded from undiluted hot
water and cold water experiments and 50% diluted hot
water extracts (Fig. 2).
When alfalfa water extract is applied in high
concentration, the growth response of plants may decline.
Ries et al. [12] showed that when 0.1 and 0.3 g/l alfalfa
extracted triacontanol was applied as a colloidal
dispersion in 374 l/ha, it increased a pot grown maize dry
weight by 10% within 7 days. When the amount was
further increased to 0.5 g/l, 13.7% increase was attained.
However, when the amount was increased into 0.7, 0.9 and Fig. 2: Yield of beet subjected to alfalfa extracts.

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 World J. Agric. Sci., 11 (2): 89-93, 2015
1.1 g/l, the percentage increases over the control were
decreased to 8.6, 6.9 and 4.0 %, respectively. Similarly,
0.001 g/l extracted triacontanol was applied in tomato and
rice plant the dry weight increased by 20 and 27 %,
respectively. When the concentration further increased
to 0.01g/l, the result of both plant increased to
32%. However, when the concentration increased to
1 g/l the result decreased to 23% for tomato and 12% for
rice. As much as 400 kg/ha band (2.5cm to the side and
2.5cm below seed) application of alfalfa mulch shown no
growth inhibiting effect on field corn [13].
Laboratory evaluations continued to point to
autotoxic substances, despite the mixed field results
[7, 14, 15, 16, 17]. The water soluble phytotoxic compound
in alfalfa has been characterized as a phenolic compound
[14]. In earlier work, Guenzi et al. [18] had implicated
saponins as being a possible water-soluble substance
released by alfalfa forage. While various studies indicated
saponins are phytotoxic to cotton, Marchaim et al. [19]
found no effect on alfalfa. Medicarpin was implicated as
the allelochemical that causes autotoxicity in seed
germination and seedling growth [20]. Dornbos et al. [21]
identified medicarpin from aqueous extracts and found it
inhibited germination and seedling growth. Hall and
Henderlong [14] had identified nine distinct compounds,
some of which exhibited promoter effects.
The toxic effect of alfalfa depends on the extract
concentration, stage of growth and genetic tolerance to
alfalfa toxicity. Most of the alfalfa plant parts were toxic to
plant growth when they were applied in high
concentration [22]. In allelopathic experiments less
concentrated alfalfa water extract showed no effect or
increasing tendency on growth. The highest growth
retarding chemicals were showed to exist in leaf and the
least extract was observed to be in stem compared to
other parts of the alfalfa plant [7]. Chung and Miller
[7] further observed that aqueous extracts from various
plant parts did not equally affect germination, radicle
length, or hypocotyl length. Extracts from alfalfa stems
and roots were generally much less inhibitory than those
from leaf and reproductive tissues.
There is occasional discrepancy between field and
laboratory results. Differences in soil characteristics, time
after alfalfa, increased rainfall/leaching, the varying
autotoxic degrees of the different varieties used and the
interaction between these factors may likely have led to
the discrepancies [7, 17, 22, 23]. Jennings and Nelson
[17] found that the latency of autotoxic substances was
longer in a clay soil than in a sandy soil. The common
field recommendation to avoid autotoxicity is to delay
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seeding for at least 2 weeks and in some cases up to two
years [23, 24]. The potential leaching of the autotoxic
compounds from lighter soil types may explain the
absence of autotoxicity after only 2 weeks in the Michigan
field trials of Tesar [23]. Findings that alfalfa varieties vary
in their autotoxicity also indicate that the field test
conditions vary considerably [18, 7]. In allelopathic
experiments less concentrated alfalfa water extract showed
no effect or increasing tendency on growth [23, 25].
CONCLUSIONS
This field experiment was performed based on the
response of tree plant seedlings to alfalfa extract on
greenhouse and plastic house condition. With some
methodological modification attempts was made to see the
effect of alfalfa extract in field condition. However,
from the begging, discrepancy between field and the
previous pot experiments were expected especially due to
the seasonal rainfall. It seems one time application during
seedling stage did not work for growth promotion, but can
be considered for retarding effects, which might be further
explored for weed control. To point out starting point
for field experiments, pot experiments on the selected
vegetables are recommended with a focus on application
interval and extraction method. In case of extract
preparation, in addition to hot and cold water soaking
methods, the consideration of affordable chemical
approach is also recommended to get better results.
ACKNOWLEDGEMENTS
The financial and materials support from Dilla
University is gratefully appreciated. The author also
acknowledges International Livestock Research Institute
(ILRI) for providing alfalfa seed.
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