ABSTRACTObjective: Tagetes minuta (Family Asteraceae) is an aromatic plant possessing volatile essential oil.Â  Its oil finds extensive use in flavor and perfume industry. T. minuta oil also possess medicinal and insecticidal properties as antiheminthic, nematicidal, bactericidal, antiviral, fngicidal and insecticidal. The plant has been explored for herbicidal potential in very few reports. The aim of present study is to find out its allelopathic potential against common wasteland and agricultural weeds.Material &amp; Methods: Various agricultural and wasteland weeds were selected for laboratory growth studies. Oil was applied in solution form using an emulsifier.Results: Growth of all test weeds was inhibited by T. minuta oil; however, effect was maximum in Amaranthus tricolor with complete inhibition at 1Âµl/ml and E. cruss-galli was least affected with complete inhibition at 5Âµl/ml concentration of T. minuta oil.Conclusion: T. minuta oil offers great potential for effective weed management in agricultural as well as wasteland areas.Â Keyowrds: Tagetes minuta, essential oil, allelopathy, solution form, wasteland, agricultural, weed

Arora, Komal

BATISH, DAIZY

KOHLI, RAVINDER K

SINGH, HARMINDER P

English

Innovare Academic Sciences: E-Journals

Vol 5, Issue 1, 2017 ISSN - 2321-6832ALLELOPATHIC IMPACT OF ESSENTIAL OIL OF TAGETES MINUTA ON COMMON AGRICULTURAL AND WASTELAND WEEDSKOMAL ARORA1*, DAIZY BATISH2, RAVINDER K KOHLI3, HARMINDER P SINGH41Department of Botany, DAV College, Jalandhar, Punjab, India. 2Department of Botany, Panjab University, Chandigarh, India. 3Central University of Punjab, Bathinda, Punjab, India. 4Department of Environment Studies, Panjab University, Chandigarh, India. Email: komal.swami@yahoo.comReceived: 30 November 2016, Revised and Accepted: 02 December 2016ABSTRACTObjective: Tagetes minuta (Family Asteraceae) is an aromatic plant possessing volatile essential oil. T. minuta oil possesses medicinal and insecticidal properties as antiheminthic, nematicidal, bactericidal, antiviral, fungicidal, and insecticidal. However, it has been explored for herbicidal potential in very few reports. The aim of this study is to find out its allelopathic potential against common wasteland and agricultural weeds.Methods: The various agricultural and wasteland weeds were selected for laboratory growth studies. Oil was applied in solution form using an emulsifier.Results: Growth of all test weeds was inhibited by T. minuta oil; however, the effect was maximum in Amaranthus tricolor with complete inhibition at 1 μl/ml, and Echinochloa crus-galli was least affected with complete inhibition at 5 μl/ml concentration of T. minuta oil.Conclusion: T. minuta oil offers great potential for effective weed management in agricultural as well as wasteland areas.Keywords: Tagetes minuta, Essential oil, Allelopathy, Solution form, Wasteland, Agricultural Weeds.INTRODUCTIONTagetes minuta L. is an aromatic plant commonly known as stinking roger, Khakibush, wild marigold, Khali weed, Mexican marigold [1], etc. It is one of the most important species of genus Tagetes known to yield highest amount of essential oil [2]. T. minuta is large annual herb growing over 1 m with erect branched stems; green pungent, glabrous, and pinnately compound leaves with serrated leaflets. Inflorescence is terminal shortly stalked corymb having yellowish green flower heads. Fruits are black flattened spindle-shaped achenes [3,4]. The plant is native to South America and is often found growing in disturbed areas during succession [5] and in areas with loose soil [6]. T. minuta has made alien introduction to many parts of the world including Asia, Africa, United States, Europe, and New Zealand as a weed [7]. In India, it was introduced for its essential oil [8]; however, it escaped cultivation and started growing abundantly at high altitudes and disturbed sites of deforestation or forest fires [9]. The plant has been known to acquire weedy habit competing with many economically important crops, viz., Zea mays, Oryza sativa, and beans [3].An exhaustive study on the antibacterial, antifungal, insecticidal, acaricidal, nematicidal, and insecticidal properties of T. minuta essential oil has been documented by number of researchers [10-18]. T. minuta oil also possesses therapeutic and antioxidant properties [19,20]; however, very few researchers have tested essential oil of this plant for weed management. Recently, we have reported allelopathic potential of T. minuta oil against Chenopodium murale L., Phalaris minor Retz., Amaranthus viridis L., and Cassia occidentalis L. [21,22] where oil was used in volatile form. However, for foliar or soil application, it has to be applied in solution form. In view of this objective, this study was conducted to explore allelopathic potential of T. minuta oil on few agricultural (Amaranthus tricolor, A. viridis, Echinochloa crus-galli, and P. minor) and wasteland (Bidens pilosa and Chenopodium murale) weeds.METHODSCollection of plant material and extraction of oilAbove-ground flowering shoots of T. minuta were collected from various locations around Solan (Himachal Pradesh, India). The plant specimen was identified from herbarium of IHBT, Palampur (Voucher PLP2537). These were hydrodistilled in Clevengers apparatus for 2 hrs. Oil was collected in glass vials and kept in refrigerator in dark till used.Procurement of weed seedsThe seeds of agricultural and wasteland weeds, viz., green amaranth (A. viridis L.), vegetable amaranth (A. tricolor L.), hairy beggars tick (B. pilosa L.), little seed canary grass (P. minor Retz.), nettle leaf goosefoot (C. murale L.), and barnyard grass (E. crus-galli [L.] Beauv.) were collected from wildly growing stands of these weeds in Panjab University Campus, Chandigarh and agricultural fields in adjoining areas.Bioassay studiesThe effect of oil on weeds was studied through laboratory bioassay using oil in solution form. Various concentrations of T. minuta oil (0.25, 0.50, 1, 2, 3, 4, and 5 μl/ml, v/v) were prepared by dissolving requisite amount of oil in distilled water with the help of emulsifier Tween-20 (0.1%, and v/v). About 15 seeds each of E. crus-galli, B. pilosa, 20 seeds each of C. murale and P. minor, and 50 seeds each of A. viridis and A. tricolor were used per Petri dish. Seeds of P. minor were pre-treated with concentrated sulfuric acid for 1 minute and washed several times with tap water before imbibition to remove acid completely. Imbibed seeds of all test plants except A. viridis and A. tricolor (used without imbibition) were spaced equidistantly on double layer of Whatman filter paper No. 1 moistened with 8 ml of respective concentration of oil solution. The Petri dishes were sealed immediately with double layer of adhesive tape and kept in growth chamber under standard conditions of 16/8 hrs, light/dark photoperiod, 25±2°C temperature, 75±2% relative humidity, Research Article2Innovare Journal Of Agri. Sci, Vol 5, Issue 1, 1-4 Arora et al. and 150 μmol/m2/seconds photon flux density for summer plants (B. pilosa, E. crus-galli, A. viridis, and A. tricolor), whereas winter plants (C. murale and P. minor) were kept at temperature 10±2°C maintaining rest of the conditions similar to summer plants. A parallel control was kept without oil treatment and 5 replicates were maintained for each concentration. After 7 days, observations/measurements were made in terms of germination percentage, radical length, and plumule length.Statistical analysisThe experiment was conducted in completely randomized block design and repeated twice. All the data were presented as mean values of both the experiments. Data were subjected to one-way analysis of variance followed by separation of treatment means from the control at p≤0.01 and 0.05 applying post-hoc Dunnett’s Test using SPSS PC software.RESULTS AND DISCUSSIONThe effect of T. minuta oil on growth of test weeds was studied in the form of germination percentage, radical length and plumule length of seedlings grown under lab conditions. The % germination of control samples was recorded as 94.44±1.93, 97.78±1.92, 100±0, 96.67±5.77, 83.33±2.89, and 100±0% in A. tricolor, A. viridis, B. pilosa, C. murale, E. crus-galli, and P. minor, respectively (Fig. 1a). At lower concentrations, a dose-dependent decrease in germination was noticed in all test plants, which was statistically significant too except P. minor. None of the seeds of A. tricolor germinated at 1 μl/ml treatment whereas among other weeds maximum inhibition was observed in C. murale (65%) and B. pilosa (53.3%). At higher concentrations (2 μl/ml), the % germination declined to zero in C. murale, A. viridis, and B. pilosa. Least allelopathic effect on germination was observed in E. crus-galli where a reduction of 65% was observed at 4 μl/ml followed by complete inhibition at 5 μl/ml concentration. From the dose-response curves, inhibitory threshold values were calculated. Among agricultural weeds, half maximal inhibitory concentration value (IC50) was maximum for E. crus-galli (3.36 μl/ml) and minimum for A. tricolor (0.47 μl/ml) (Table 1).Similarly, reduction in radicle length was also observed in response to concentration of oil (Fig. 1b). Radicle length in control seedlings was 5.64±0.04, 3.04±0.02, 1.87±0.08, 3.49±0.06, 6.75±0.28, and 6.5±0.12 cm in A. tricolor, A. viridis, B. pilosa, C. murale, E. crus-galli, and P. minor, respectively. With treatment of 0.25 μl/ml, reduction in radicle length was nearly 23, 24, 26, and 16% in A. tricolor, A. viridis, C. murale, and E. crus-galli, respectively. At 0.5 μl/ml, radicle length in weeds A. tricolor, A. viridis, B. pilosa, C. murale, E. crus-galli, and P. minor was 1.89±0.04, 2.05±0.03, 2.13±0.03, 1.83±0.06, 5.39±0.05, and 5.21±0.23, respectively. At 1 and 2 μl/ml concentrations, most sensitive plant was P. minor (nearly 65% reduction) and least was B. pilosa (nearly 4% reduction). At 3 μl/ml, radicle length in E. crus-galli and P. minor was 2.42±0.08 and 0.67±0.03 cm, respectively. At 5 μl/ml concentration, none of the weeds germinated at all.In general, with an increase in concentration, plumule length also decreased (Fig. 2). In control, seedlings of broad-leaved weeds, viz., A. tricolor, A. viridis, B. pilosa, and C. murale and it was 1.61±0.16, 1.29±0.02, 2.69±0.20, and 2.03±0.21, respectively. With the treatment of 0.25 μl/ml, the % reduction observed in A. tricolor, A. viridis, C. murale, E. crus-galli, and P. minor was nearly 28, 16, 38, 3, and 11%, respectively. Promotory effect of oil was noticed in B. pilosa, with increase of 2% in plumule length at this concentration. In response to 0.5 μl/ml concentration of oil, maximum reduction was observed in B. pilosa and A. tricolor (nearly 67 and 65%, respectively). Similarly at Table 1: IT and IC50 values for weeds and crops using solution form of oil treatmentTest plant IT (μl/ml) IC50 (μl/ml)Agricultural weedsA. tricolor 0.25 0.47A. viridis 0.25 1.25E. crus-galli 0.25 3.36P. minor 1 3.15Wasteland WeedsB. pilosa 0.25 0.98C. murale 0.25 0.49B. pilosa: Bidens pilosa, C. murale: Chenopodium murale, P. minor: Phalaris minor, E. crus-galli: Echinochloa crus-galli, A. tricolor: Amaranthus tricolor, IT: Inhibitory threshold, IC50: Inhibitory concentrationFig. 1: (a) Effect of Tagetes minuta oil on germination and (b) radicle length of agricultural and wasteland weeds. Results are expressed as mean ± standard error (Y error bar); *,**along each symbol in each figure represent significant difference at p≤0.05 and 0.01, respectively, applying Dunnett’s testa b3Innovare Journal Of Agri. Sci, Vol 5, Issue 1, 1-4 Arora et al. 1 μl/ml concentration, among all the target plants, maximum inhibition of plumule length was observed in C. murale (nearly 52%). At 2 μl/ml, B. pilosa, A. viridis, A. tricolor, C. murale were completely inhibited, and plumule length in E. crus-galli and P. minor was reduced by nearly 25% and 58%, respectively. At 4 μl/ml concentration, it was 3.28±0.07 cm in E. crus-galli leading to approximately 51.3% reduction.Thus, allelopathic impact of T. minuta essential oil resulted in overall reduction of growth of test weeds, in general.Germination and growth bioassays are primary tools for determining phytotoxic activity [23], and may detect potential allelopathic effects under controlled laboratory conditions. In this study, it is clear that T. minuta oil exerts allelopathic effect on various weeds. In general, a dose-dependent type of relationship between oil concentrations and germinating parameters of all test plants was apparent, indicating greater inhibition with increase in concentration. Similar observations that essential oil exerts inhibitory effect were also made by several other workers [21-29]. Since essential oils are composed of monoterpenes and/or sesquiterpenes, thus their inhibitory effect may be contributed by all or few major constituents either synergistically or additively. In this study, effect of volatile essential oil of Tagetes was species specific. Some weeds were affected to greater extent (A. tricolor and A. viridis) than others (E. crus-galli). The differential susceptibility of different plants may be attributed to their seed size [30] or due to their different genetic constitution. In relevance to observations made in this study, we may infer that T. minuta oil inhibits germination and growth of other plants, however, detailed study for determining the mode of action of oil with regards to biochemical, anatomical, and morphological changes needs to be done.CONCLUSIONT. minuta oil possesses phytotoxicity toward test weeds. Radicle growth was affected more compared to the plumule. The effect of T. minuta oil varied from species to species with maximum allelopathic inhibition of A. viridis and A. tricolor.REFERENCES1. Randall R, Kessal O. National list of naturalized invasive and potentially invasive garden plants. Australia, Sydney: WWF; 2004. p. 98.2. Singh V, Singh B, Kaul VK. Domestication of wild marigold (Tagetes minuta L.) as a potential economic crop in western Himalaya and north Indian plains. Econ Bot 2003;57:535-44.3. Holm LG, Doll J, Holm E, Pancho JV, Herberger JP. World Weeds: Natural Histories and Distribution. New York: John Wiley and Sons Publishers; 1997.4. Wang CM, Chen CH. Tagetes minuta L. (Asteraceae), a newly naturalized plant in Taiwan. Taiwania 2006;51:32-5.5. Soule J. Tagetes minuta: A potential new herb from South America. In: Janick J, Simon JE, editors. New Crops. New York: Wiley; 1993. p. 649-54.6. Khatri S, Phougat N, Chaudhary R, Singh B, Chillar AK. 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ALLELOPATHIC IMPACT OF ESSENTIAL OIL OF TAGETES MINUTA ON COMMON AGRICULTURAL AND WASTELAND WEEDS

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ALLELOPATHIC IMPACT OF ESSENTIAL OIL OF TAGETES MINUTA ON COMMON AGRICULTURAL AND WASTELAND WEEDS

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