Efficacy of Garlic Extract on the Reproductive Parameters of Root Knot Nematode, Meloidogyne incognita (Kofoid and White) Chitwood
Ayesha Javed1, Huma Abbas1*, Sajid Aleem Khan1, Maham Batool1, Aqsa Ashfaq1, Muhammad Kamran2,4, Akhtar Hameed3, Azhar Iqbal2 and Muhammad Ehetisham-ul-Haq2
1Department of Plant Pathology, University of Agriculture, Faisalabad, Pakistan
2Plant Pathology Research Institute, Ayub Agricultural Research institute, Faisalabad, Pakistan
3Institute of Plant Protection, MNS University of Agriculture, Multan, Pakistan
4Sugarcane Research Institute, Ayub Agricultural Research Institute, Iaisalabad
ABSTRACT
The present study aimed to assess the efficacy of garlic aqueous extract (GAE) on the reproduction of the root knot nematode (RKN), Meloidogyne incognita, in tomato plants. Four experiments were conducted during this investigation using different concentrations of GAE (50%, 75%, and 100%). The effects of GAE were evaluated on the mortality and egg hatching of M. incognita under in vitro conditions. In field conditions, GAE was applied using both root dipping and soil drenching methods. In the soil drench method, 20ml of GAE was applied around the root zone, and 1000 freshly hatched juveniles of M. incognita were inoculated. In the root dipping method, tomato roots were immersed in GAE for 20 min before transplantation, and then the plants were inoculated with 1000 freshly hatched juveniles. Each treatment was replicated five times, and various parameters including the number of galls, egg masses, the number of juveniles per 100g of soil, shoot and root length, weight, number of branches, leaves, and stem girth were recorded. The results showed significant juvenile mortality under in vitro conditions, with the highest mortality and the lowest egg hatching rates observed in the 100% solution of GAE. Furthermore, a significant reduction in RKN reproduction parameters was observed when using the soil drench application method compared to the root dipping method. The active substance in garlic extract, diallyl polysulfide, acts on cellular targets. Overall, soil drench treatment with garlic extract resulted in reduced root galling in tomato plants compared to root dipping treatment. Therefore, garlic extract could be successfully used for the management of the root knot nematode, M. incognita in field conditions.
Article Information
Received 29 September 2022
Revised 10 July 2024
Accepted 23 July 2024
Available online 23 January 2025
(early access)
Published 27 December 2025
Authors’ Contribution
AJ, HA and SAK planed, designed and executed experimental work. MB and AA assisted in processing of samples. MK and AH reviewed, edited and analysed the data. AI and ME helped in writing the manuscript. All authors have read and approved the final version of the manuscript.
Key words
Root knot, Nematode, Meloidogyne incognita, Diallyl polysulfide
DOI: https://dx.doi.org/10.17582/journal.pjz/20220929050919
* Corresponding author: [email protected]
0030-9923/2026/0001-0299 $ 9.00/00
Copyright 2026 by the authors. Licensee Zoological Society of Pakistan.
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
INTRODUCTION
Tomato (Solanum lycopersicum L.) is a significant crop, yielding approximately 182.3 million tons of fruit on 4.85 million hectares annually, making it the second most important fruit and vegetable crop after potato (Solanum tuberosum L). These are a valuable source of nutrients, including vitamin A, carotenoids, β-carotene, and lycopene, and are beneficial for reducing cardiovascular risks associated with type-2 diabetes (Shifdar et al., 2011). However, tomatoes are susceptible to a wide range of diseases, with approximately 200 diseases identified, each exhibiting unique symptoms and economic impacts. Among these, root knot nematodes (RKN) are common plant-parasites, found in soil. In England, the Meloidogyne species of RKN was initially observed in the roots of cucumber plants by Berkeley in 1855. RKNs disrupt the plant’s ability to absorb nutrients and water, leading to stunted growth, yellowing foliage, and reduced yields.
The life cycle of RKN begins with the deposition of eggs into a gelatinous matrix that protects and holds them together (Maggenti and Fortuner, 1987). The first-stage RKN larvae (J1) shed their skins before hatching into eggs, often independently of plant root stimuli. The freshly hatched J2s move over the soil in search of roots to feed on, heading straight to the root tissue tips and entering the roots (Jones et al., 2013). Once a J2 locates a root, it secretes enzymes and uses its stylet to penetrate the plant’s roots. The larvae’s bodies undergo two additional moults, with the third and fourth juvenile stages possessing a functional stylet but not feeding. In the final moult, mature females form, comprising a functional stylet, median bulb, uterus, and vagina (Perry et al., 2009; Jones et al., 2013). Under ideal conditions, RKNs can complete their life cycle in three to four weeks, with five to eight generations reproducing in a single growing season (Noling, 2014).
In Pakistan, estimates of crop loss due to nematodes range from 5 to 20% (Maqbool et al., 1988), with disease occurrence in Punjab reaching between 75 and 100% (Shahid et al., 2007; Khan, 2009). In some areas, such as Okara, yield losses due to RKNs can be as high as 85.2%, with an incidence frequency of 38.89% (Hussain et al., 2012). Globally, the economic impact of nematodes is substantial, with assessments reaching up to US $78 billion (Mokrini, 2016).
To combat nematodes, various plant extracts have been used as effective methods for control. These extracts in combination with fertilizers and biopesticides are cost-effective, easy to use, environmentally friendly, and capable of improving soil health (Sultana et al., 2010; Abbas et al., 2023). Garlic (Allium sativum L.), a member of the Allium family, has been found to contain diallyl polysulfide as its active substance, which targets cellular mechanisms. This action, observed particularly in the roots, can lead to the development of resistance against RKNs. GAE has been found to have nematicidal and nematode-hatching inhibitory effects, reducing nematode infestations such as gall formation, egg masses, and hatched juveniles on tomato roots, as well as the number of juveniles in the soil. In view of above the current investigation was designed to evaluate the effectiveness of GAE in reducing juvenile mortality and egg hatching of M. incognita, and to assess its efficacy through soil drenching and root dipping treatments against M. incognita in tomato plants.
MATERIALS AND METHODS
Collection and culturing of nematodes
Diseased samples of eggplant roots and soil infested with RKNs were collected and processed for mass culturing of nematodes. Nursery raising of the tomato cultivar “Money Maker” was carried out, and after three weeks of transplantation into pots, approximately 1000 freshly hatched juveniles were inoculated around the plant rhizosphere by creating holes in each pot. For each experiment, five replications were conducted.
Preparation of garlic aqueous extract (GAE)
GAE was prepared by soaking 25g of crushed garlic cloves in 1000ml of distilled water for 72 h and then filtered through Whatman no.1filter paper. The prepared GAE was kept in sterilized vessel at 4°C temperature till needed. The crude extract and diluted extract were used for experiment purposes at different concentrations of 50 %( v/v), 75 %( v/v), 100 %( v/v).
Effect of GAE on egg hatching of RKN
Five ml of GAE was poured into Petri dishes. In separate plates having different concentration of (50%, 75%, 100%) treatments were inoculated with 50 M. incognita eggs. Control treatment was kept with distilled water. Data were recorded after the 3rd, 5th, and 7th days of interval after treatments applications and observed the no. of hatched eggs and calculated.
Effect of GAE on juvenile mortality of RKN
Five ml of GAE was poured into each Petri plate. In separate plates having different concentration of treatments (50%, 75%,100%) were inoculated with 100 freshly hatched juveniles of RKN. Control treatment was kept with distilled water. The total juveniles mortality was detected 24, 48 and 72 h after the treatment using the formula:
Root dipping of tomatoes seedling in GAE
The test plant roots of tomato seedlings were dipped into 20ml of solutions of different concentrations of (50%, 75%,100%) in GAE for 15-20 min before transplanting them into pots, filled with sterilized soil. The control plants were dipped in equal volume of water for the same period. After three weeks of transplantation, tomato seedlings were inoculated with 1000 J2 of M. incognita. Around the plant root rhizosphere three holes was made at a distance of 2cm away from tomato plant. Inoculation was done by adding water suspension containing juveniles into these holes and covering it with soil to avoid desiccation.
Soil drenching of tomato seedlings by GAE
The soil around the root zone was drenched with 20 ml of the extract at various concentrations. After one week, inoculation of 1000 freshly hatched juveniles into the tomato plants was conducted by creating three holes around the plant stem. The roots were then covered with topsoil. The same process was performed for the control plants using water. Plant growth parameters and nematode reproduction parameters were recorded.
After 15 days of treatment application, a second application of garlic extract was administered to the soil-drenched plants. Experiments were repeated to confirm the results.
Data recorded
After 60 days, harvesting was done. Following parameters were measured: shoot and root length, stem girth, fresh shoot and root weight, number of leaves per plant, number of branches per plant, number of flowers per plant, number of fruits per plant, egg mass of M. incognita, number of root galls, number of juveniles, and number of females.
RESULTS
Effect of GAE on egg hatching and juvenile mortality
The efficacy of GAE was observed on egg hatching 3rd, 5th, and 7th days under in vitro conditions at different concentrations (Table I). The highest percentage of egg hatching was observed in the control plate with water, while the lowest percentage of egg hatching was observed in the 100% crude extract of garlic after 3 days.
Table I. Effect of garlic extract (GAE) on egg hatching and juveniles mortality of M. incognita under in vitro condition.
|
GAE treatment |
Egg hatching |
Juveniles mortality after |
||||
|
3rd day |
5th day |
7th day |
24 h |
48 h |
72 h |
|
|
Control |
26.24d |
35.14b |
48.03a |
0j |
0j |
0j |
|
50% GAE |
16.50f |
25.88d |
31.67c |
7i |
19g |
46d |
|
75% GAE |
11.03g |
20.33e |
28.73cd |
12h |
22f |
64b |
|
100% GAE |
5.09h |
12.11g |
19.03ef |
24e |
48c |
84a |
*Mean values sharing similar letters do not differ significantly α = 0.05
The efficacy of GAE was examined regarding juvenile mortality under in vitro conditions at different concentrations (Table I). Juvenile mortality was recorded after 24 h, 48 h, and 72 h. An increase in juvenile mortality was observed with 100% crude garlic extract. At lower concentrations of GAE, specifically 50%, the minimum number of juvenile mortalities were observed. No mortality was observed in the control plate containing only water.
Effect of GAE on RKN reproduction and plant growth parameters by soil drench method
First application
Table II shows the effect of GAE on RKN reproduction parameters and plant growth parameters by the soil drench method. A significant reduction in the number of galls, the number of juveniles (J2), the number of egg masses, and the number of females was observed due to the application of GAE at different concentrations. The maximum number of galls was observed in diseased control plants where no treatment was applied. In healthy control plants, no galling was observed. A significant reduction in the number of juveniles (J2) was observed due to the application of GAE at different concentrations. At higher concentrations of GAE, the number of juveniles per 100g of soil was reduced compared to diseased control plants where the maximum number of juveniles per 100g of soil was present. At a higher concentration, the minimum number of egg masses were obtained compared to diseased plants where the maximum number of egg masses were observed. A higher concentration of GAE minimized the number of nematode females compared to diseased plants where a higher number of females were observed. Therefore, a higher concentration of GAE was effective in controlling nematode reproduction parameters.
Table II. Effect of first and second application of aqueous garlic extract (GAE) on root knot nematode reproduction parameters by soil drench method.
|
GAE treatment |
No. of galls |
No. of J2/ 100g of soil |
No. of egg masses |
No. of females |
|
First application |
||||
|
Control (healthy) |
0e |
0e |
0e |
0e |
|
Control (diseased) |
180.39a |
1047.43a |
167.49a |
186.31a |
|
50% GAE |
73.3b |
487.18b |
68.11b |
77.37b |
|
75% GAE |
54.51c |
321.45c |
42.18c |
58.43c |
|
100% GAE |
43.56d |
202.33d |
32.35d |
47.38d |
|
Second application |
||||
|
Control (healthy) |
0e |
0e |
0e |
0e |
|
Control (diseased) |
175.33a |
1040.2a |
159.27a |
177.25a |
|
50% GAE |
69.12b |
481.15b |
61.01b |
70.46b |
|
75% GAE |
46.39c |
318.37c |
38.24c |
52.31c |
|
100% GAE |
37.13d |
198.52d |
27.41d |
40.35c |
*Mean values sharing similar letters do not differ significantly α = 0.05
There was a significant effect on the plant growth parameters of tomato plants due to the application of GAE as a soil drench method at various concentrations (Table III). A higher shoot length of tomato plants was observed in healthy control plants compared to diseased control plants where the minimum shoot length was recorded. Significantly, the shoot weight of tomato plants was recorded as the maximum in healthy control plants compared to disease control plants where the minimum shoot weight was observed. The root length of tomato plants was higher in healthy control plants compared to diseased control plants where the minimum root length was observed. As the concentrations of GAE were increased, the root weight of tomato plants decreased because the number of galls was reducing due to the nematicidal activity of GAE. The maximum number of
Table III. Effect of first and second application of GAE on plant growth parameters by soil drench method
|
GAE treatment |
Shoot length |
Shoot weight |
Root length |
Root weight |
Number of leaves |
Number of branches |
Stem girth |
|
First application |
|||||||
|
50% GAE |
31.13c |
9.63cd |
9.75b |
6.17ab |
7.58c |
5.56bc |
1.46c |
|
75% GAE |
33.13bc |
11.40bc |
10.39b |
5.15bc |
8.04bc |
6.27abc |
1.55c |
|
100% GAE |
34.32ab |
12.62ab |
11.62ab |
4.48c |
9.51b |
7.47ab |
1.82b |
|
Control (healthy) |
36.5a |
13.82a |
12.79a |
4.92bc |
11.63a |
8.12a |
2.16a |
|
Control (diseased) |
19.52d |
8.16d |
6.67c |
7.0a |
4.34d |
5.34c |
0.92d |
|
Second application |
|||||||
|
50% GAE |
32.03b |
10.14c |
10.53c |
7.17ab |
8.55b |
4.47b |
1.17bc |
|
75% GAE |
34.02ab |
12.02b |
11.21bc |
6.31bc |
9.34b |
5.76ab |
1.36b |
|
100% GAE |
35.04ab |
13.41ab |
12.39ab |
5.39c |
10.47ab |
6.49ab |
1.54b |
|
Control (healthy) |
37.03a |
14.55a |
13.52a |
5.61c |
12.46a |
7.46a |
2.18a |
|
Control (diseased) |
19.09c |
8.10d |
7.30d |
8.29a |
5.69c |
4.38b |
0.96c |
*Mean values sharing similar letters do not differ significantly α = 0.05
leaves on tomato plants was observed in healthy control plants compared to diseased control plants where the minimum number of branches were recorded. The stem girth of tomato plants was maximum in healthy control plants compared to diseased control plants where the minimum stem girth was observed. As the concentrations of GAE were increased, the plant growth parameters, including shoot length and weight, root length and weight, stem girth, number of leaves, and branches, also increased due to the plant growth-promoting factors present in garlic extract.
Second application
A significant reduction in the number of galls, the number of juveniles (J2), the number of egg masses, and the number of females was observed due to the 2nd application of GAE at different concentrations (Table II). The maximum number of galls was observed in diseased control plants. The number of egg masses was significantly reduced by the 2nd application of GAE, as a higher concentration of GAE resulted in a minimum number of egg masses compared to diseased plants where the maximum number of egg masses was observed. A significant reduction in the number of females was also observed after the 2nd application of GAE, especially at a higher concentration.
There was a significant effect on the plant growth parameters of tomato plants due to the 2nd application of GAE as a soil drench method at various concentrations (Table III). The maximum shoot length was observed in healthy control plants compared to diseased control plants, where the minimum shoot length was recorded. The maximum number of leaves on tomato plants was observed in healthy control plants compared to diseased control plants, where the minimum number of branches was recorded. The stem girth of tomato plants was maximum in healthy control plants compared to diseased control plants, where the minimum stem girth was observed. As the concentrations of the 2nd application of GAE were increased, the plant growth parameters also increased.
Table IV. Effect of GAE on root RKNs reproduction parameters by root dipping method.
|
GAE treatment |
No. of galls |
No. of J2/ 100g of soil |
No. of egg masses |
No. of females |
|
Control (healthy) |
0e |
0e |
0e |
0e |
|
Control (diseased) |
191.08a |
1124.41a |
183.19a |
193.08a |
|
50% GAE |
93.94b |
494.13b |
89.25b |
98.02b |
|
75% GAE |
57.16c |
375.13c |
51.44c |
64.49c |
|
100% GAE |
44.25d |
221.55d |
40.22d |
51.12d |
*Mean values sharing similar letters do not differ significantly α = 0.05
Effect of GAE on RKN reproduction and plant growth parameters by root dipping method
Table IV shows the effect of GAE on RKN reproduction parameters by the root dipping method. A significant reduction in the number of galls was observed due to the application of GAE at different concentrations. The maximum number of galls was observed in diseased control plants. There was a significant reduction in the number of juveniles (J2) due to the application of GAE
Table V. Effect of GAE on plant growth parameters by root dipping method.
|
GAE treatment |
Shoot length |
Shoot weight |
Root length |
Root weight |
No. of leaves |
Number of branches |
Stem girth |
|
Control (healthy) |
38.35a |
14.0a |
13.17a |
4.43c |
11.99a |
9.78a |
2.12a |
|
Control (disease) |
21.39e |
10.56b |
8.01d |
7.77a |
5.38d |
6.23c |
0.89c |
|
50% GAE |
25.08d |
7.11c |
9.26cd |
6.64ab |
8.1c |
6.65bc |
1.41b |
|
75% GAE |
30.35c |
8.26bc |
11.07bc |
5.21bc |
9.05bc |
7.52bc |
1.66ab |
|
100% GAE |
35.0b |
9.15bc |
12.13ab |
3.47c |
10.74ab |
8.35ab |
1.83ab |
*Mean values sharing similar letters do not differ significantly α = 0.05
at higher concentrations. The number of egg masses was significantly reduced by the application of aqueous garlic extract at higher concentrations, with the minimum number of egg masses obtained compared to diseased plants where the maximum number of egg masses was observed. Higher concentrations of GAE also reduced the number of nematode females compared to diseased plants, where a higher number of females were observed.
There was a significant effect on plant growth parameters in tomato plants due to the application of aqueous garlic extract by the root dipping method at different concentrations (Table V). The shoot weight of tomato plants was significantly higher in healthy control plants compared to diseased control plants, where the minimum shoot weight was observed. The root length of tomato plants was greater in healthy control plants compared to diseased control plants, where the minimum root length was observed. The maximum number of leaves on tomato plants was observed in healthy control plants compared to diseased control plants, where the minimum number of branches was recorded. The stem girth of tomato plants was maximum in healthy control plants compared to diseased control plants, where the minimum stem girth was observed. As the concentrations of aqueous garlic extract increased, the stem girth also increased.
DISCUSSION
The present study was conducted to assess the efficacy of GAE on the plant growth parameters and reproduction parameters of RKNs, specifically Meloidogyne incognita, in tomato plants. Severe infections resulting from nematode attacks lead to a significant decrease in plant height, crop losses, and reduced product quality. Due to their broad host range, RKN are challenging to manage, causing worldwide losses estimated at up to US $78 billion (Mokrini, 2016). RKNs are known to cause damage ranging from 50% to 85% in tomato production.
In this study GAE with laboratory test at different concentrations (50%, 75%, and 100%) inhibited the egg hatching of M. incognita compared to the control treatment. These findings are in line with those of Adomako and Kwoseh (2013), who observed increased egg hatching in control treatments compared to 100% crude garlic extract, where less egg hatching occurred. Lower concentrations of the treatments were less effective compared to higher concentrations in all treatments examined. These results align with those of Adegbite and Adesiyan (2005) suggesting that the inhibitory impact of botanicals may be attributed to the presence of compounds in the extract with larvicidal and ovicidal characteristics. The results revealed that the maximum juvenile mortality was observed at a 100% concentration of GAE, while the minimum egg hatching was observed at the same concentration.
Under field conditions, GAE showed a significant impact on nematode reproduction, and plant parameters indicated increased growth soil drenching and root dipping methods. GAE has been reported to contain complex growth-promoting substances such as vitamins, saponins, carbohydrates, proteins, alkaloids, and sugars like fructose (Martins et al., 2016). A well-maintained dose of these compounds actively participates in promoting the growth of recipient plants.
The shoot length of tomato plants was maximized in the 100% GAE, which proved to be effective. Maximum shoot weight was observed at higher concentrations of GAE. Increased root length was observed at higher concentrations of GAE, while minimum root weight was detected at the 100% concentration. These nematodes deprive plants of nutrition by forming galls on roots and colonizing root tissue (Bird, 1974). The minimum number of galls was observed in the 100% GAE treatment, along with the minimum number of egg masses. Additionally, the minimum number of juveniles per 100g of soil was observed in the 100% GAE treatment. In contrast to nematode-infested plants, soil treatment with GAE reduced the nematode root gall index and increased the activity of catalase and B-1,3 glucanase enzymes in tomato plants, as stated by Abd-Elgawad et al. (2009). The active substance in garlic extract is diallyl polysulfide, and its mode of action targets cells. This pattern was consistent in the roots, indicating the development of resistance to RKN. GAE caused no phytotoxicity in plants (Sukul et al., 1974).
Conclusion
The most beneficial impact of GAE was a considerable reduction in the galling index and other reproductive factors, such as egg mass count, number of females, and number of juveniles. The results of this investigation demonstrate that GAE has great potential for nematode control and significantly enhances the growth of tomato plants.
The soil drenching application method was more effective compared to the root dipping method. Therefore, the crude aqueous garlic extract has proven to be effective for the management of root-knot nematodes under both in vitro and field conditions.
Declarations
Funding
The study received no external funding.
Statement of conflict of interest
The authors have declared no conflict of interest.
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