INTRODUCTION

Allelopathy is a common biological phenomenon by which one organism produces

biochemicals that influence the growth, survival, development, and reproduction of other

organisms. These biochemicals are known as allelochemicals and have beneficial or detrimental

effects on target organisms. Cited from Penn State Extension entitled “Phytotoxicity”,

phytotoxicity is a toxic effect by a compound on plant growth, which may occur when

chemicals are employed to protect plants from pests, fertilize plants, regulate plant

growth, and more (Moorman, 2011). Plants with allelopathic properties give off substances

that are toxic to other plants. These toxins inhibit growth or keep seeds from germinating. These

toxic substances either vaporize and settle to earth or are washed there by rains. They can also be

released through the roots or as the allelopathic plant decomposes.

Herbicide, an agent, usually chemical, for killing or inhibiting the growth of unwanted

plants—i.e., weeds. Selective herbicides control specific weed species, while leaving the desired

crop relatively unharmed, while non-selective herbicides can be used to clear waste ground,

industrial and construction sites, railways and railway embankments as they kill all plant material

with which they come into contact.

Mangifera indica L.belonging to the family Anacardiaceae is one of the best known and

most evergreen trees, native to Indian subcontinent and found widely distributed in tropical and

subtropical regions. According to some studies from Keio Research Information System of Keio

University, entitled “Allelopathic Potential and an Allelopathic Substance in Mango Leaves”,

mango leaves contain allelopathic substances and have potential as soil additive material for

weed management. Mango tree is one of the most important multipurpose trees growing with

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wide range of cultivated species and forms a component of agroforestry practices like home

garden and silvipastoral system, however, in recent years decline in crop yield under this tree is

reportedly attributed mostly to its allelopathic effects. (Sahoo et al, 2010)

Mango leaves are potential bioresources due to their capability to produce allelochemical

compound that can inhibit weed development (Yulifrianti et al., 2015). Mango leaf contains

phenol, flavonoid and terpenoid (El-Rokiek et al., 2010).

Several studies have indicated that the allelochemicals, such as Phenolics, terpenoids,

alkaloids and their derivatives are toxics which may inhibit shoot/ root growth, nutrient uptake,

or may attack a naturally occurring symbiotic relationship thereby destroying the usable source

of plants of a nutrient. Some of these compounds which are known as allelochemicals alter the

growth or physiological functions of receiving species. The most commonly found

allelochemicals, cinnamic and benzoic acids, flavonoids, and various terpenes (Singh et al.,

2003), these compounds are known to be phytotoxic (Einhellig, 2002).

Capsicum annuum is a species of the plant genus Capsicum native to southern North

America and northern South America. This species is the most common and extensively

cultivated of the five domesticated capsicums. This food crop can be easily grown and cultivated

at home and is one of the few home garden food crops in the country early 20th century through

south Florida.

Cyperus rotundus, commonly known as purple nutsedge, or nut grass is a slender, erect,

glabrous, perennial grasslike plant, found throughout the Philippines in open areas at low and

medium altitudes, common weed in gardens, and has been called the world's worst weed. It is

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one of the most extensively researched non-cultivated plant species on the planet, yet the

complexities of its life cycle.

The chemicals with allelopathy activity are present in many plants and various organs

including leaves and fruits and have potential inhibitory effect on crops (Seigler et al., 1996).

According to the study “Allelopathic Effects of Leaf Leachate of Mangifera indica on Initial

Growth Parameters of Few Homegarden Food Crops”,which was cited from Meissner and

Antonio, other researchers have evaluated the allelopathic effects grasses/trees on the

germination, shoot length, root length, bioassay on different crop species. For instance, pure

tuber extract of Cyperus rotundus impede the cucumber, onion, radish and tomato’s radical

elongation at time, the effect was confined to only certain parts of the plant, as it was evident

that, the elongation of forage grasses radicle was more responsive than seed germination upon

treatment with Leucaena (Sahoo et al., 2010).

A recent study in 2010 was found relevant and related to the current study of the

researcher. According to Sahoo et al, of the same study, aqueous leaf extract of different

concentrations of Mangifera indica L., was used to investigate their effects on germination, shoot

and root lengths and dry matter yield of 5 food crops viz. Capsicum annum L. (Chilli), Glycine

max (L.) Merr. (Soybean), Zea mays L. (Maize), Oryza sativa L. (Rice) and Abelmoschus

esculentus (L.) Moench (Lady’s finger). Both bioassays and pot culture indicated that the

inhibitory effect was much more pronounced at higher concentrations, while the lowest

concentration showed stimulatory effect in some cases. The most affected crop was lady’s finger

among the test crops.

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 Cited from a study of James et al., entitled “Allelopathy: How plants suppress other

plants”, the dried mango leaf powder was reported to significantly inhibit sprouting of purple

nutsedge tubers and its aqueous extract inhibiting germination and growth of some crops.

Cyperus rotundus and Cynodon dactylon could be controlled by phytochemical compounds

derived from extract of mango leaves (El-Rokiek et al., 2010).

The procedure for the preparation of the extracts was based on the study of El-Rokiek et

al., in 2010, “Inhibitory effect of mango (Mangifera indica L.) leaf extracts on the germination of

Cassia occidentalis seeds”, wherein 10 g of both dry and fresh pulverized leaf materials were

carefully transferred into cleaned, sterilized and labeled jars, and 200, 100, 66.5 and 50 ml of

sterile, deionised distilled water was added to the corresponding labeled jars. The mixture was

shaken and the jars were covered and left for 24 h at room temperature and filtered through a

cheese cloth to remove debris and finally filtered using Whatman No.1 filter paper to give 5, 10,

15 and 20% concentrations, respectively.

In the continual search for natural herbicides from plant products, leaf extracts from

Mangifera indica were investigated for inhibitory effect on the seed germination of the popular

weed, that is, Cyperus rotundus. Earlier reports have also shown that water soluble leachate from

the matured fresh leaves of M. indica has the allelopathic potential to reduce the development of

food crops, like Capsicum annuum. However, such studies are conducted separately between

weeds and food crops, therefore still unclear to know whether which among the kind of plants

will be more affected by the phytotoxicity potential of M. indica.

In accordance to this, the researcher aimed to investigate the phytotoxicity or inhibitory

effect of the aqueous extracts of Mangifera indica leaves to the germination of a weed and food

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crop, such as Cyperus rotundus and Capsicum anuum, to evaluate allelopathic potential of

mangoes.

STATEMENT OF THE PROBLEM

The study aimed to investigate inhibitory effect of the leaves aqueous extracts of M.

indica to the germination of Cyperus rotundus tubers and Capsicum anuum seeds.

Specifically, the objectives of this study are:

1. To measure the growth of the Capsicum annuum in terms of:

a. Radicle Length

b. Coleoptile Length

2. To measure the growth of Cyperus rotundus tubers in terms of:

a. Shoot Length

b. Root Length

3. To determine which plant (weed or food crop) would show more inhibition than

the other.

4. To compare the treatments of extract to the positive (distilled water) and negative

(commercial herbicide) controls.

5. To compare which of the treatments would give the best result for the inhibition

of the germination of the seeds.

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 HYPOTHESIS

Null Hypothesis - The mango leaves will not have a significant effect to the Cyperus

rotundus and Capsicum annuum

Alternative Hypothesis - The mango leaves will have a significant effect to the Cyperus

rotundus and Capsicum annuum

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 SIGNIFICANCE OF THE STUDY

DEPARTMENT OF AGRICULTURE. Allelopathy from mango leaves could help them

determine the important role of its effect not just in weed control, but to crop productivity.

DEPARTMENT OF EDUCATION. The knowledge of the allelopathic potential of

mango leaves in weeds and home garden food crops.

BUSINESS PROFESSIONALS. They can come up with new products using the

knowledge about the allelopathic effect of mango leaves.

GARDENERS. An idea would be presented to know what effect does mango leaves have

in food crop germination, especially at home gardens.

STUDENTS. They would gain knowledge about plant allelopathy and how plants

suppress other plants.

FUTURE RESEARCHERS. The study would help them gain knowledge that will be of

aid to have future studies about plant allelopathy.

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SCOPE AND LIMITATIONS

The study mainly focused on determining the inhibitory effect of the extracts of dry

mango leaves to the germination of the seeds of chili peppers and tubers of purple nutsedge. The

materials for the experiment were only collected from Centro Occidental, Polangui, Albay. The

growth parameters that were measured were the radicle and coleoptile lengths for the chili

peppers, and the shoot and root lengths for the purple nutsedge.

Experimentation was conducted in a controlled environmental setup, and the extraction

used for the experimentation was maceration of the ‘Haden’ mango leaves. The study did not

utilize a chemical analysis for the experiment.

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 METHODOLOGY

This study used an experimental research design, specifically the Two-Group Design,

wherein there are two groups present. The presence of controlled groups, such as the negative

control, distilled water, and the positive control, commercial herbicide, served as control for

comparison of the results. ANOVA was used to determine if there is significance effect in using

mango leaves in inhibiting the germination of Cyperus rotundus tubers and Capsicum annuum

seeds.

Collection and preparation of materials

Fresh mango leaves were selected from one specific mango tree in a residence grown in

Centro Occidental, Polangui, Albay. The leaves were washed thoroughly, air-dried, then

pulverized before extraction. The dried leaf materials were pulverized into powder using mortar

and pestle then stored in a container until extraction. Capsicum annuum seeds were purchased

from Harvest Plus, Polangui, Albay. Cyperus rotundus seeds were collected from a single

population in Centro Occidental, Polangui, Albay. The mango leaves were authenticated in Bicol

University College of Agriculture and Forestry in Guinobatan, Polangui, Albay.

Extract preparation

Fresh mature mango leaves were solar dried for 48 hours before extraction. After solar

drying, the leaves were pulverized into fine pieces using mortar and pestle. 10 g of dry

pulverized leaf materials were carefully transferred into cleaned, sterilized and labeled sample

containers. Using a graduated cylinder to measure the volume, 200, 66.5and 40 ml of sterile,

distilled water was added to the corresponding labeled sample containers. The mixture were

9
shaken and the containers were left covered for 24 hours. Then the mixtures were filtered using

filter paper to make extracts of 5%, 15%, and 25% concentration, respectively.

Experimental setup preparation and treatment application

Table 1. Effect of Mangifera indica leaf extracts on root lengths (RTL) and shoot lengths

(SL)of Cyperus rotundus (Purple Nutsedge) tubers and radicle length (RL) and coleoptile length

(CL) of Capsicum annuum (Chili Pepper) seeds.

Cyperus rotundus Capsicum annuum

RL CL RTL SL
T0
Distilled water
T1
5%
T2
15%
T3
25%
T4
Commercial herbicide

Petri dishes were sterilized before the experiment, then a filter paper was put inside of

each petri dish. 10 seeds were placed in each petri dish containing the filter papers. Each petri

dish were initially moistened or saturated with 5mL of each treatment and as for the negative

control, 5mL of distilled water was used, and herbicide, Spitfire, for positive control.

Three sets of treatments were prepared for the experimentation: In Set A, the mango

leaves extracts were applied to Cyperus rotundus, and control; In Set B, the mango leaves

extracts were applied to Capsicum annuum; and in Set C, the extracts were applied to both

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Cyperus rotundus and Capsicum annuum. For Set C, 5 seeds of Cyperus rotundus and Capsicum

annuum were placed on each petri dish. Each treatment was replicated three times.

Monitoring of experimental set-up

Measurements were taken such as the emergence of radicle and coleoptile for the seeds,

and root and shoot for the tubers in each experimental treatment. Radicle length was measured

for it is the first part of a seedling to emerge from the seed during the process of germination, and

was commonly used as a growth parameter on different studies about seed germination. The

coleoptile length showed the same importance as a growth parameter because it is the first leaf

above the ground, forming a sheath around the stem tip. Same procedure was done to measure

the root and shoot lengths of tubers, the root length is the same as the radicle length of seeds,

and the shoot length as the coleoptile length.

The given parameters were measured by picking the seed using forceps and a centimeter-

scaled ruler were used in reading the measurements. Readings were recorded after 9 days of

germination.

Data Analysis

After the time allotted for the set-up, the researcher analyzed the data gathered, in terms

of the length of the radicles and coleoptiles of the chili peppers and the length of the roots and

shoots of the purple nutsedge, with the use of statistical tests such as ANOVA. The researcher

evaluated the data after being exposed to each treatment and determined the effect of the

extracts. Comparison between the concentrations of extracts was first evaluated, then comparison

between the parameters of the plants afterwards.

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 Collection and
preparation of materials

Extract preparation

Experimental setup
preparation and
treatment application

Monitoring of
experimental set-up

Gathering of Data

Data Analysis

Formulation of
Conclusion

Figure 1. Experimental Flowchart

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 RESULTS AND DISCUSSIONS

The results of the radicle length and coleoptile lengths (cm) of chili pepper seeds after 9

days from sowing are shown in Table 1, 2, 3 and 4. These results revealed that the radicle and

coleoptile lengths were affected significantly by each concentrations and treatments 9 days after,

compared with the respective control.

In determining the inhibitory effect of the leaf extracts to the germination of the seeds

and tubers, the radicle and coleoptile lengths of the seeds, and root and shoot lengths of the

tubers were measured, wherein the lower the measurements of the given parameters in respect to

the negative controlled treatment, the higher the inhibitory effect of the leaf extracts to its

germination. While the higher the measurements of the given parameters compared to the

negative controlled treatment, the lower the inhibitory effect and showed stimulatory effect on

the seeds.

To further show the difference between the treatments for each set, a graph was

illustrated for each table given.

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 1. SET A: CYPERUS ROTUNDUS

T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 2.245 3.255 4.155 0.660 0
Replicate 2 2.855 3.000 3.150 0.715 0
Replicate 3 4.150 4.045 3.255 0.440 0

AVERAGE 3.083 3.520 0.605 0

3.433333
v
Table 1.1 Root Length (in cm) of Cyperus rotundus tubers on mango leaves extract (Set A)

The radicle lengths of the germinating Cyperus rotundus was lowest in the T3 with an

average of 0.605 cm. Stimulatory effect was seen in Treatment 1 and 2, compared to the negative

control, while Treatment 3 showed inhibitory effect and relatively near the result of the positive

control. The results showed stimulatory effects at lower concentration.

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 T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 2.125 1.820 1.725 0.14 0

Replicate 2 1.485 1.765 1.425 0.367 0

Replicate 3 2.925 3.305 1.44 0.330 0

AVERAGE 2.178 2.297 1.530 0.279 0

Table 1.2. Shoot Length (in cm) of Cyperus rotundus tubers on mango leaves extract (Set A)

The results on the shoot lengths of the tubers showed both stimulatory and inhibitory

effects on the different treatments, The inhibitory effect on the radicle length of the seeds was

more evident in T3 at an average of 0.279 cm, and the germination of the Cyperus rotundus seeds

was found to be highest at T0, with an average of 2.178 cm. T1 showed stimulation while the other

treatments inhibited the germination.

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 2. SET B. Capsicum annuum

Table 2.1. Radicle Length (in cm) of Capsicum annuum seeds on mango leaves extract (Set
B)

T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 1.235 1.375 0.39 0.18 0

Replicate 2 1.225 0.910 0.465 0.26 0

Replicate 3 2.200 1.385 0.480 0.145 0

AVERAGE 1.553 1.223 0.445 0.195 0

Table 2.1 shows the radicle lengths of the seeds after 9 days of receiving the treatment in

the mango leaves extract, wherein T0 has the highest elongation of the radicles, with an average

of 1.553 cm. Inhibitory effect was also seen on the results, for the average lengths for the three

treatments was lower than that of the positive control.

Table 2.2. Coleoptile length (in cm) of Capsicum annuum seeds on mango leaves extract
(Set B)

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 T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 2.180 0.780 0.190 0.040 0

Replicate 2 1.505 0.515 0.275 0.030 0

Replicate 3 1.295 0.715 0.280 0.010 0

AVERAGE 1.660 0.670 0.248 0.027 0

Coleoptile lengths of the seeds were found highest on the control, as shown on table 2.2,

with an average length of 1.660 cm while all treatments showed inhibitory effects, however, it

was much pronounced at T3 with an average of 0.027 cm.

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 Graph 1.1 Average Root Length in cm of Cyperus rotundus tubers and Radicle Length of

Capsicum anuum seeds on mango leaves (Set A and Set B)

4
3.5
3
2.5
2 Cyperus rotundus
Capsicum annuum
1.5
1
0.5
0
T0 T1 T2 T3 T4

Graph 1.2. Average Shoot Length in cm of Cyperus rotundus tubers and Coleoptile Length

of Capsicum anuum seeds on mango leaves (Set A and Set B)

3. SET C: Cyperus rotundus and Capsicum annuum

2.5

1.5
Cyperus rotundus
1 Capsicum annuum

0.5

0
T0 T1 T2 T3 T4

Table 3.1.1 Root Length (in cm) of Cyperus rotundus tubers on mango leaves extract (Set C)

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 T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 6.650 6.600 1.800 0.370 0

Replicate 2 5.760 2.360 3.140 0 0

Replicate 3 5.120 3.350 3.690 0.890 0

AVERAGE 5.843 4.103 2.877 0.420 0

The negative control had an average length of the roots at 5.843 cm, while T3 had an

average length of 0.420 cm, which showed great inhibition and a close result to the positive

control.

Table 3.1.2. Shoot Length (in cm) of Cyperus rotundus tubers on mango leaves extract (Set
C)

T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 3.680 4.900 0.960 0.210 0

Replicate 2 1.760 1.140 0.690 0 0

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Replicate 3 3.520 2.140 0.730 0.900 0

AVERAGE 2.987 2.727 0.793 0.370 0

The results for the shoot lengths of the tubers in Set C showed inhibitory effects

on all treatments, however, T3 showed the most inhibition compared to the negative control, with

an average of 0.370 cm, which is near that of the positive control. T 2 also showed great inhibition

with an average of 0.793 cm.

Table 3.2.1. Radicle Length (in cm) of Capsicum annuum seeds on mango leaves extract
(Set C)

T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 3.940 2.250 1.520 0.530 0

Replicate 2 3.000 1.550 1.720 0.400 0

Replicate 3 2.020 2.360 1.550 0.570 0

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AVERAGE 2.987 2.053 1.597 0.500 0

The growth of the chili pepper seeds on the treatments were inhibited compared to the

negative control with average 2.987 cm. T3 showed the most inhibition with an average of 0.500

cm, which is close to the results of the positive control.

Table 3.2.2. Coleoptile length (in cm) of Capsicum annuum seeds on mango leaves extract
(Set C)

T0 T1 T2 T3 T4
(5%) (15%) (25%)
Replicate 1 2.280 0.890 0.260 0.460 0

Replicate 2 2.280 0.390 1.330 0.320 0

Replicate 3 1.140 0.880 0.630 0.950 0

AVERAGE 1.900 0.720 0.740 0.577 0

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 The results for the coleoptile lengths of the seeds on Set C also showed inhibitory effects

on all treatments, with lowest average on T3, , with an average of 0.577 cm.

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 Graph 1.1 Average Root Length in cm of Cyperus rotundus tubers and Radicle Length of

Capsicum anuum seeds on mango leaves (Set C)

6
5
4
3 Cyperus rotundus
Capsicum annuum
2
1
0
T0 T1 T2 T3 T4

Graph 1.2. Average Shoot Length in cm of Cyperus rotundus tubers and Coleoptile Length

of Capsicum anuum seeds on mango leaves (Set C)

2.5

1.5 Cyperus rotundus

Capsicum annuum
1

0.5

0
T0 T1 T2 T3 T4

CONCLUSION

The researcher therefore concluded that the best treatment was Treatment 3,

which is the 25% concentration. Given the best treatment, the researcher evaluated that the best

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results of the study was of the Cyperus rotundus tubers and Capsicum annuum seeds germinated

with the 25% aqueous extracts of the mango leaves . It shows that phytotoxicity of mango leaf

extract showed pronounced variation among different concentrations on the growth of chili

pepper. The effect was highly depending on concentration of the extract.

Based on the calculations using Analysis of Variance, all sets showed a significance

effect on the germination of the tubers and seeds in terms of the radicle and coleoptile lengths,

and root and shoot lengths. The critical value for determining the area of rejection of the null

hypothesis is 3.478, and the calculated values for the given parameters for each table, are all

greater than the critical value. Therefore, there was a significant effect of using the extract of

mango leaves to inhibit the growth of Cyperus rotundus and Caspsicum annuum, based on the

computations using the ANOVA.

RECOMMENDATIONS

After evaluating the results and finishing the experiment, several recommendations were

useful for future purposes. Other studies could be formulated by other researchers, such as

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 testing Mangifera indica leaf extracts to other food crops or weeds, using a different method of

extraction, and extending the growth parameters of the plants to be tested for further more

significant information and data. Other researchers could also conduct comparative studies using

other parts of Mangifera indica, such as the fruit’s peels. This study served as a reference for

other researchers to study the same field.

Allelopathic potential and an allelopathic substance in mango leaves. (n.d.). Retrieved from

https://www.tandfonline.com/doi/abs/10.1080/09064710.2016.1215517?journalCode=sagb20

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