with the collaboration of Iranian Scientific Association for Landscape (ISAL)

Evaluation of the Allelopathic Effects of some Plant Species on the Germination and Growth of Lettuce Seedlings (Lactuca sativa var. Great Lakes 366) using the Sandwich Method

Document Type : Research Article

Authors

1 1- Badiee Saffron Production Agricultural Company, Mashhad, Iran 2- Department of Horticulture, Faculty of Agriculture, Birjand University, Birjand, Iran

2 Department of Plant Genetics and Production Engineering, Faculty of Agriculture, Birjand University, Birjand, Iran

Abstract
Introduction
Invasion of weeds is one of the major factors in the reduction of yield of agricultural crops in the field conditions. The use of allelopathic plants is one of the methods for weed control. To date, numerous laboratory and field techniques have been developed to evaluate allelopathy in various plant species. Recently, advancements in allelopathy research have emerged with the introduction of innovative techniques and more effective bioassays. Researchers have shown that many challenges in allelopathy studies can be addressed through these improved methods. Traditional approaches have increasingly been replaced by newer techniques, such as the Sandwich method, which allows the allelopathic properties of different plants to be evaluated in a very short time using only a minimal amount of plant material.
 
Materials and Methods
Considering that there is a significant evidence of the presence of allelopathic compounds in invasive weeds as well as medicinal plants, therefore, in the current research, the main focus was on these two groups of plants for the selection. In this research, the allelopathic activity of 57 plant species (60 samples including leaf, stem, seed, rhizome, flower, gum) of 18 families was investigated using the Sandwich method. In this method, plant samples were first dried at 40 degrees Celsius and then weighed in three replicates with amounts of 10 and 50 mg. Special containers with 6 well were used for cultivation. The diameter of each well was 3 cm. In one row of the wells, 10 mg and 50 mg of plant sample powder were placed in the other row. Then 5 ml of 0.5% agar solution was added to the samples and after the agar cooled and coagulated, another 5 ml of agar solution was added to each well. In this way, the plant sample was placed between two layers of agar. Then, in each well, 5 seeds of lettuce (Lactuca sativa) variety Great Lakes 366 were planted vertically on the agar surface. This variety of lettuce has been selected as an indicator due to its high sensitivity to metabolites. Then the containers were sealed and placed in an incubator with a temperature of 25 degrees Celsius in the dark conditions. After 3 days, the percentage of seed germination was checked by measuring the length of lettuce radicle and hypocotyl. To make it easier to compare plants, data clustering was performed using Minitab software.
 
 
 
Results and Discussion
Using this method, 59 plant samples were placed into 10 distinct clusters. The similarity of plants in each cluster was 85%. The findings revealed that the leaf residues of Anethum graveolens and a Cynara cardunculus clustered together based on their allelopathic activity, demonstrating the most significant growth-inhibitory effect at a concentration of 10 mg of the plant sample. The average percentage of inhibition of radicle and hypocotyl growth of lettuce seedlings caused by these two plants was reported to be 94.95 and 87.65 percent, respectively. Also, 13 plants including leaves of Chenopodium album, Prangos ferulacea, Lepidium draba, Petroselinum crispum, Dianthus  barbatus, Convolvulus arvensis, Narcissus tazetta, Heracleum persicum, Onosma dichroantha, Stellaria media, Brassica cretica, Ipomoea  purpurea and Apium graveolense Seeds were placed in one cluster. This group also had significant growth inhibitory effects. Although their inhibitory effects were always less than Anethum graveolens and a Cynara cardunculus. The average percentage of radicle and hypocotyl growth inhibition in these plants with 10 mg of dry plant sample was 79.76 and 62.27, respectively. This study demonstrated that both the leaf residue of Hypericum perforatum and Hypericum scabrum significantly enhanced the growth of lettuce seedlings. Even minimal amounts (10 mg of the dried plant sample) resulted in a remarkable increase in radicle growth by 14.65% and hypocotyl growth by 45.15%. Additionally, at amount of 50 mg, Pistacia lentiscus gum demonstrated exceptional efficacy, enhancing radicle growth by an impressive 90%. Among the plants studied in this study, Cupressus sempervirens leaf residues had an inhibitory effect on radicle growth but stimulated hypocotyl growth. Centaurea behen leaf residues only produced a growth-stimulating effect (43.3%) on hypocotyl growth at amount of 10 mg, and in other cases, they showed allelopathic effects in the form of growth inhibition.
 
Conclusion
This study confirms the reliability of the sandwich method for identifying allelopathic plants and assessing their properties. It efficiently evaluates a variety of plants with fewer samples and time. The results suggest that plant residues could be used in organic farming to control weeds. Further research is necessary to identify the specific allelopathic compounds involved.
 
Acknowledgement
The authors of this article would like to thank Mr. Mohammad Reza Joharchi, a faculty member of the Herbarium of Ferdowsi University of Mashhad, for his sincere cooperation in identifying the plants used in this study from a botanical perspective, confirming the Persian names, and matching them with scientific names.

Keywords

Subjects

Authors retain the copyright. This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0).

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  • Receive Date 08 September 2024
  • Revise Date 06 March 2025
  • Accept Date 22 March 2025
  • First Publish Date 22 March 2025