Dish Pack Method Research Articles

Evaluation of Allelopathic Activity Interactions of Some Medicinal Plants Using Fractional Inhibitory Concentration and Isobologram

Allelopathy is a physiological process with an ecological concept and application. Allelopathy is the result of the production of biologically active molecules by growing plants or their remains, which may have a direct effect on the growth and development of individuals of the same species or other species after changing their shape and entering the environment. As regards, the use of natural compounds in the control of weeds and pests is a priority. In this research, the allelopathic activity of 123 specimens of medicinal and aromatic plants were investigated individually by the dish-pack method using lettuce seeds as a model. Then, the strongest inhibitory ones were selected and their allelopathic interaction effects were investigated for the first time by interacting them together. Two methods were used to evaluate allelopathic interaction effects: calculating Fractional Inhibitory Concentration (FIC) and drawing Isobologram diagrams. Lettuce hypocotyl length, root length, germination percentage, and germination rate were investigated. Pelargonium graveolens (leaf) had the greatest inhibitory effect on lettuce radicle growth (EC50 = 5.31 mg/well) and Echinophora platyloba (stem) had the greatest effect on hypocotyl growth inhibition (EC50 = 7.91 mg/well). Also, the lowest lettuce germination percentages were observed in the treatments Lavandula officinalis (flower) and Nepeta binaloudensis (leaf), respectively (23.61, 22.85%). The highest inhibitory effect by considering lettuce germination rate was detected in Salvia ceratophylla (leaf), (12.86 seed/day) and the lowest belonged to Nepeta binaloudensis (leaf) and Lavandula officinalis (flower), respectively (3.60, 3.32 seed/day). According to FIC calculations and isobolograms, two types of interaction, including synergist (Nepeta binaloudensis (leaf) with Trachyspermum ammi (fruit) and Nepeta binaloudensis (leaf) with Lavandula officinalis (flower) and antagonist (Pelargonium graveolens (leaf) with Lavandula officinalis (flower)), were observed significantly among the plants tested in this research. These interactions can be used to prepare more effective natural herbicides and decrease the use of herbicides.

Relationship between the Antioxidant Activity and Allelopathic Activities of 55 Chinese Pharmaceutical Plants.

Pharmaceutical plants contain several phytochemicals that are sources of myriad biological activities. These biological activities can be explored in multiple fields for the benefit of mankind. Pharmaceutical plants with high ethnobotanical indices (i.e., use value and relative frequency of citation) were reported with the potential to inhibit lettuce elongation through leachates and volatiles. The focus of the study was to assess Chinese pharmaceutical plants for both antioxidants, as well as allelopathic potentials to explore any underlying relationship. The estimation of antioxidative capacity and content of total phenolics (TPC) for the 55 Chinese pharmaceutical plants was conducted by the assays of DPPH radical scavenging activity (DPPH-RSA), oxygen radical absorbance capacity (ORAC) and the means of Folin–Ciocalteu. The estimation of the activity of allelopathy for collected medicinal plants was done by adopting the sandwich method for plant leachates and the dishpack method for volatile constituents, respectively. The fruits of sea buckthorn (Hippophae rhamnoides) had the most remarkable ORAC value (168 ± 7.04 μmol TE/g) and DPPH radical scavenging activity (440 ± 7.32 μmol TE/g) and contained the highest contents of total phenolic compounds (236 ± 7.62 mg GAE/g) in the 55 pharmaceutical plant species according to the results. In addition, sea buckthorn showed dominant allelopathic potential through plant leachates evaluated by using the sandwich method. Star anise (Illicium verum Hook. f.) showed conspicuous allelopathic activity through plant volatiles assessed by the dishpack bioassay method. Among the same plant species, antioxidative ability and total phenolics, in comparison with potential allelopathy of medicinal herbs indicated that volatile allelochemical had a weak active effect (r = 0.407 to 0.472, p < 0.01), with antioxidant capacity by the dishpack method. However, the evaluation by the sandwich method showed a significant positive correlation (r = 0.718 to 0.809, p < 0.001) with antioxidant capacity. Based on these results, a new hypothesis is that the antioxidant activity of plants may have an involvement with the potential allelopathic activity.

Comparative response of Malaysian weedy rice (Oryza sativa) initial growth towards the allelopathic potential of Leucaena leucocephala (Lam.) de Wit and Dicranopteris linearis (Burm. f.) Underw

The infestation of weedy rice has becoming a threat which leads to competition between weedy rice and commercial rice for nutrient uptake. Weedy rice management is more dependent on the herbicides application that can contribute to various environmental problems. Thus, this study was conducted to assess the allelopathic potential of two invasive plants namely L. leucocephala and D. linearis in controlling the growth of weedy rice. The allelopathic potential was evaluated through laboratory bioassay namely the dish pack method, sandwich method and plant box method to determine the allelopathic potential on the seedling growth of two weedy rice variants collected in Peninsular Malaysia (designated as WRE and WRN) and MR220 CL2 rice variety. Highest concentration of L. leucocephala leaf litter (50 mg) inhibited the radicle elongation of MR220 CL2, WRN and WRE by 49 %, 51 % and 83 % compared to control, respectively. Through the dish pack method, the volatile compound from D. linearis leaf were found to disrupt the radicle growth of MR220 CL2, WRN and WRE by 21 %, 29 % and 25 % of the control at the nearest distance (41 mm) of the bioassay plants with the D. linearis leaf. Compared with D. linearis, the root exudate from L. leucocephala exhibited higher inhibitory effect towards the bioassay species. Overall, the findings from this study showed that the allelopathic potential of L. leucocephala was more preferable to be investigated further for sustainable weedy rice management in the future by infusing the biological approach.

Determination of the Allelopathic Potential of Cambodia’s Medicinal Plants Using the Dish Pack Method

Plants produce several chemically diverse bioactive substances that may influence the growth and development of other organisms when released into the environment in a phenomenon called allelopathy. Several of these allelopathic species also have reported medicinal properties. In this study, the potential allelopathic effects of more than a hundred medicinal plants from Cambodia were tested using the dish pack method. The dish pack bioassay method specifically targets volatile allelochemicals. Twenty-five species were found to have significant inhibitory effects on lettuce radicle growth. Eleven different plant families, including Iridaceae (2), Apocynaceae (2), Poaceae (2), Sapindaceae, Araceae, Combretaceae, Orchidaceae, Clusiaceae, Zingiberaceae, Rutaceae and Asparagaceae had the plant species with high inhibitory effects. Allophyllus serrulatus had the highest growth inhibitory effect on lettuce radicles more than 60%, followed by Alocasia macrorrhiza, Iris pallida, Terminalia triptera, Wrightia tomentosa, Cymbidium aloifolium, Garcinia villersiana and Kaempferia parviflora. The candidate species were subjected to further studies to identify the volatile allelochemicals in the volatile constituents.

Allelopathic screening of Malaysian noxious weeds and several medicinal plants as potential alleloherbicides

Aim: This research was conducted to assess the allelopathic potential of 100 plants species, including noxious weeds and medicinal plants of Malaysia as donor plant on the growth of Lactuca sativa (lettuce) as the recipient plant. This study was also carried out to justify common weeds found in Malaysia to act as a natural herbicide for weed management through allelopathy phenomenon. Methodology: Weed and medicinal plant species commonly found in Malaysia were identified according to their morphological characteristics. Sandwich method was carried out to determine the allelopathic potential of weed species through leaf litter leachate mechanism. On the other hand, dish pack method was conducted to discover the allelopathic potential of weed species through volatilization pathway. Results: 100 weed species and medicinal plants commonly found in Malaysia inhibited the radicle and hypocotyl growth of L. sativa by both sandwich method and dish pack method. Among 100 weed species, Passiflora foetida showed the maximum inhibitory effects on L. sativa growth in both sandwich and dish pack method. The highest amount of P. foetida leaf litter (50 mg), completely inhibited the growth of radicle and hypocotyl of L. sativa. Volatilization of weed and medicinal plants leaf litter inhibited the growth of L. sativa with increasing distances of lettuce seed with the weed and medicinal plants leaf litter. Interpretation: The findings of this study provides a base line for identifying plants with allelopathic properties for controlling weed propagation.

Potential of Octanol and Octanal from Heracleum sosnowskyi Fruits for the Control of Fusarium oxysporum f. sp. lycopersici

The antifungal activity of volatile compounds from the fruit, leaf, rhizome and root of 109 plant species was evaluated against Fusarium oxysporum f. sp. lycopersici (FOL) race 1—the tomato wilt pathogen—by using the modified dish pack method. Eighty-eight plant samples inhibited mycelial growth, including volatiles from fruits of Heracleum sosnowskyi, which exhibited the strongest antifungal activity, showing 67% inhibition. Two volatile compounds from the fruits of H. sosnowskyi (octanol and octanal) and trans-2-hexenal as a control were tested for their antifungal activities against FOL race 1 and race 2. In terms of half-maximal effective concentration (EC50) values, octanol was found to be the most inhibitory compound for both pathogenic races, with the smallest EC50 values of 8.1 and 9.3 ng/mL for race 1 and race 2, respectively. In the biofumigation experiment, the lowest disease severity of tomato plants and smallest conidial population of race 1 and race 2 were found in trans-2-hexenal and octanol treated soil, while octanal had an inhibitory effect only on race 2. Therefore, our study demonstrated the effectiveness of volatile octanol and trans-2-hexenal on the control of the mycelial growth of two races of Fusarium oxysporum f. sp. lycopersici and may have potential for the future development of novel biofumigants.

Plant Growth Inhibitory Activities and Volatile Active Compounds of 53 Spices and Herbs.

The inhibitory activities of the leachates and volatiles from 53 plant species (spices and herbs) were evaluated against lettuce (Lactuca sativa “Great Lakes 366”) seedling growth using the sandwich and dish pack methods, respectively. With the sandwich method, parsley (Petroselinum sativum) showed the strongest inhibitory effect on lettuce radicle growth (77%), followed by tarragon (Artemisia dracunculus) (72%). However, caraway (Carum carvi), dill (Anethum graveolens) (seed), laurel (Laurus nobilis), rosemary (Rosmarinus officinalis), and sage (Salvia officinalis) were the most inhibitory species (100% inhibition of lettuce radicle and hypocotyl growth inhibition at all distance wells) in the dish pack method. Cardamom (Elettaria cardamomum) and thyme (Thymus vulgaris) also showed strong inhibitory activity (100% for radicle and hypocotyl growth inhibition at all 41 and 58 mm distance wells). The headspace sampling and gas chromatography-mass spectrometry (GC-MS) analysis identified the main inhibitory active compounds as carvone in caraway and dill (seeds), 1,8-cineole in laurel and cardamom, and borneol in thyme. Both camphor and 1,8-cineole were detected in rosemary and sage, and the total activity evaluation showed that camphor was the major inhibitory compound in rosemary, although both compounds played equal roles in sage.

Allelopathic effects of the revegetation species Juniperus sabina L. in semiarid areas of China

Juniperus sabina L. is a native evergreen conifer of the Mu Us Sandy Land. It has prostrating stems that can prevent sand shifting and is an important revegetation species. Allelopathy is a phenomenon in which the chemical substances released from one individual have effects, both inhibitory and promotive, on other individuals and it has a considerable impact on revegetation. In this study, to effectively use J. sabina for revegetation, we clarified the allelopathic effects of J. sabina on the germination and initial growth of the native species Artemisia ordosica Krasch. in comparison with a non-native cultivated lettuce (Lactuca sativa L. ‘Great Lakes 366’) in semiarid areas of China, and identified volatile allelochemicals. To evaluate the allelopathic effects of J. sabina, the dish pack method, which measures the effects of volatile compounds on seed germination and initial growth, was applied and gas chromatography–mass spectrometry was used to identify the volatile allelochemicals. The volatile compounds from J. sabina had almost no effects on the germination of lettuce seeds, while the germination ratio of A. ordosica seeds was extremely low. The major volatile compounds were sabinene, α-pinene, and β-myrcene, which are monoterpenes and species selective. J. sabina has several advantages as a revegetation woody species; however, our results suggest that J. sabina could have strong inhibitory effects on some specific native species of the Mu Us Sandy Land. Therefore, to effectively use J. sabina for revegetation, it is important to consider both its negative and positive effects.

Evaluation of the Allelopathic Potential of Fifteen Common Malaysian Weeds

The use of allelopathy concept in weed management has received attentions to minimize extensively the reliance on herbicide applications on the agriculture industry in Malaysia. A laboratory study was conducted to evaluate the allelopathic potential of 15 Malaysian common weed species of different morphological characteristics (broadleaves, sedges and grasses). They were screened using the Sandwich method (from leaf litter leachate) and the Dish pack method (for testing the presence and content of volatile compounds in weeds). Among the 15 weed species tested, the leaf litter leachate of Centrosema pubescens was observed to be the most sensitive plant material inhibiting the growth of lettuce radicle (84%) and hypocotyl (55%) in the Sandwich bioassay compared to the control. This was followed by Asystasia gangentica (81%) and Cynodon dactylon (80%) inhibiting the lettuce radicle growth. In the Dish pack bioassay, Rhynchelytrum repens demonstrated maximum inhibition on the radicle and hypocotyl elongations by 44% and 29%, respectively, (over control) at 41 mm distance from the source well. Meanwhile, at the same distance, Cynodon dactylon was observed to have the least inhibitory effect on lettuce radicle growth by 12%. The results presented can be utilized as benchmark information for further research on the identification and isolation of allelochemicals for weed control strategies.

Determination of allelopathic potentials in plant species in Sino-Japanese floristic region by sandwich method and dish pack method

&lt;p&gt;The Sino-Japanese Floristic Region appears as one of the major centers of development of higher plants. This region have been relevant for the study of evolution and systematics of many flowering plants. The taxonomic richness of endemic plant species in this region have survived several years of extreme climate conditions. Endemic mountainous plant species that have survived extreme climate conditions are of allelopathic and medicinal interest. For this reason, 251 plant species collected from the Sino-Japanese Floristic Region were screened for allelopathic plant species. Sandwich method and dish pack method were respectively used to screen plant leaf leachates and volatile materials with lettuce (&lt;em&gt;Lactuca sativa&lt;/em&gt; CV. Great Lakes 366) as receptor plant. Among the 84 species that showed inhibitory effect on lettuce radicle elongation in our sandwich bioassay, &lt;em&gt;Photinia glabra &lt;/em&gt;showed complete inhibition of lettuce radicle elongation (0% radicle elongation). In the dish pack bioassay, &lt;em&gt;Photinia glabra&lt;/em&gt;, &lt;em&gt;Liquidambar styraciflua&lt;/em&gt;, and &lt;em&gt;Cinnamomum camphora&lt;/em&gt; (90.6%, 61.4%, and 50.2% respectively) were among the nine species that were observed with strong inhibitory effect on lettuce radicle growth. On the other hand, nine other species promoted lettuce radicle growth when compared to the control. &lt;em&gt;Aesculus turbinata&lt;/em&gt; and &lt;em&gt;Quercus gilva&lt;/em&gt; were the species with the highest growth stimulatory effect (33.0% and 16.1% respectively). We hereby present &lt;em&gt;Photinia glabra &lt;/em&gt;as an allelopathic candidate species for both leachate and volatile compounds.&lt;/p&gt;

Identification of Safranal as the Main Allelochemical from Saffron (Crocus sativus)

Dried parts of 75 medicinal plant species collected from different regions in Iran were assayed by the Dish Pack Method for volatile allelopathic activity, using Lactuca sativa (lettuce) as the test plant. The highest (60%) inhibition was observed for saffron (stigma of Crocus sativus), followed by Dracocephalum kotschyi, Solanum nigrum and Artemisia aucheri. Safranal was identified as the main chemical by Headspace Gas Chromatography-Mass Spectrometry (HS- GC-MS) analyses of saffron. Moreover, the EC50 of safranal was evaluated as 1.2 μg/L (ppb). This is the first report on allelopathic activity of safranal as a bioactive compound identified from saffron.

Determination of allelopathic potential in some medicinal and wild plant species of Iran by dish pack method

Plants have the ability to interfere with other organisms in their immediate environment, a phenomenon called allelopathy. Allelopathy is a common phenomenon in nature (Qian Chemosphere 75(368):375, 2009). The use of allelopathic compounds for the development of new agrochemicals has several benefits as opposed to common synthetic products. Cognizant of reports on the allelopathic action of medicinal and invasive plants, 118 Iranian plants from 104 species which belongs to 34 families have been screened for allelopathic volatiles by the dish pack method. A total of 118 plants (104 species) belonging to 34 families were collected in 2009 from the tropical, subtropical and temperate regions of Iran. The Dish pack method as described by Fujii et al. (Weed Sci Technol 45(80):81, 2000) utilized for screening the presence of allelopathic volatiles released from different parts of the plant specimen. This method can be used to recognize allelopathic properties very easy and quickly. Results of bioassay with lettuce seed (Lactuca sativa cv. Great Lakes 366) revealed that 23 species elicited growth inhibitory action with the greatest (≥96% reduction) observed in the inflorescences of Achillea wilhelmsii and Achillea filipendula. Furthermore, a high suppression (83–95 %) in radicle elongation was observed in lettuce exposed to: flowers of Achillea nobilis, Lavandula vera, and Perovskia abrotanoides; fruit of Ruta graveolens; seeds of Bunium persicum and Trachyspermum copticum; leaves of Achillea biebersteinii, Pulicaria gnaphalodes, Ziziphora clinopodioides, Zataria multiflora; gum of Ferula foetida; and stigma of Crocus sativus. On the other hand, 14 species instead exhibited growth-promoting properties. Of these, the greatest promoting effect were exhibited by the leaves of Cardaria draba, Verbascum speciosum and Urtica dioica eliciting 69 %, 56 %, and 53 %, respectively, which had longer radicles compare to the control. The results hereby presented provide insight into the allelopathic action of the medicinal and wild plants of Iran. Data herein reported serves as benchmark information for further research on the characterization of the allelochemicals in these plants.

Allelopathy of Teucrium Royleanum Wall. Ex Benth. from Pakistan

Teucrium royleanum was tested by four variant assays using lettuce seeds, showing excellent inhibition. The plants leaf leachates were assayed by Sandwich and Homogenated Sandwich Methods while Dish Pack Method was used for the evaluation of essential oils. The above stated methods confirm the presence of allelochemicals in T. royleanum and were further evaluated in bioassay guided way by total activity approach. The methanol subsequent fractions; hexane, chloroform, ethylacetate, butanol and water fractions were assayed by the Total Activity Method. The essential oils, hexane and chloroform fractions played a decisive role in our findings. The results from this study suggested that the secondary metabolites from T. royleanum can be potential candidates for the phytotoxicity (Allelopathy) and can be utilized on commercial scale for the development of new herbicides or weedicides. Key words: Allelopathy, Teucrium royleanum, lettuce seeds, leaf leachates, extracts essential oils.

Antifungal Effects of Volatile Compounds from Black Zira (Bunium persicum) and Other Spices and Herbs

The dish pack method, which measures growth inhibition or promotion effects of volatile compounds on germinating seeds, was applied to measure the antifungal effects of 52 dried samples of spices and herbs against a soil-borne phytopathogenic fungus, Fusarium oxysporum. Black zira showed the strongest effect, followed by cumin and cardamom. Headspace sampling and gas chromatography-mass spectrometry analysis of black zira identified seven volatile compounds, gamma-terpinene, limonene, p-cymene, beta-pinene, alpha-pinene, cuminaldehyde, and myrcene. Among these, cuminaldehyde and p-cymene showed the strongest antifungal activities against F. oxysporum, suggesting roles in the antifungal activity of black zira. The same compounds also showed antifungal activities against another soil-borne phytopathogenic fungus, Verticillium dahliae, and foliar phytopathogenic fungi, Botrytis cinerea and Alternaria mali. The total activity calculated from the concentration of cuminaldehyde contained in black zira and its EC(50) against F. oxysporum demonstrated that cuminaldehyde is the main antifungal compound detected in black zira.

炭によるアレロパシー軽減効果の改良プラントボックス法、ディッシュパック法による評価

炭によるアレロパシー軽減効果の改良プラントボックス法、ディッシュパック法による評価

植物から放出される植物生育阻害作用が強い揮発性物質の分析と cis-3-hexenal の同定

植物から放出される植物生育阻害作用が強い揮発性物質の分析と cis-3-hexenal の同定

ディッシュパック法による植物体から放出される揮発性他感物質の分析・同定

ディッシュパック法による植物体から放出される揮発性他感物質の分析・同定

揮発性他感物質の検定法と作用成分の分析法の開発

揮発性他感物質の検定法と作用成分の分析法の開発