Leaf functional traits and resource use strategies facilitate the spread of invasive plant Parthenium hysterophorus across an elevational gradient in western Himalayas.

Parthenium hysterophorus L. (Asteraceae) is a highly prevalent invasive species in subtropical regions across the world. It has recently been seen to shift from low (subtropical) to high (sub-temperate) elevations. Nevertheless, there is a dearth of research investigating the adaptive responses and the significance of leaf functional traits in promoting the expansion to high elevations. The current study investigated the variations and trade-offs among 14 leaf traits (structural, photosynthetic, and nutrient content) of P. hysterophorus across different elevations in the western Himalayas, India. Plots measuring 20 × 40 m were established at different elevations (700 m, 1100 m, 1400 m, and 1800 m) to collect leaf trait data for P. hysterophorus. Along the elevational gradient, significant variations were noticed in leaf morphological parameters, leaf nutrient content, and leaf photosynthetic parameters. Significant increases were observed in the specific leaf area, leaf thickness, and chlorophyll a, total chlorophyll and carotenoid content, as well as leaf nitrogen and phosphorus content with elevation. On the other hand, there were reductions in the amount of chlorophyll b, photosynthetic efficiency, leaf dry matter content, leaf mass per area, and leaf water content. The trait-trait relationships between leaf water content and dry weight and between leaf area and dry weight were stronger at higher elevations. The results show that leaf trait variability and trait-trait correlations are very important for sustaining plant fitness and growth rates in low-temperature, high-irradiance, resource-limited environments at relatively high elevations. To summarise, the findings suggest that P. hysterophorus can expand its range to higher elevations by broadening its functional niche through changes in leaf traits and resource utilisation strategies.

Comparative assessment of two biodegradable chelants, S,S-ethylenediamine disuccinic acid and nitrilotriacetic acid, in facilitating Cd remediation by lesser swine cress (Coronopus didymus, Brassicaceae).

Chemically assisted phytoremediation is suggested as an effective approach to amplify the metal-remediating potential of hyperaccumulators. The current study assessed the efficiency of two biodegradable chelants (S,S-ethylenediamine disuccinic acid, EDDS; nitrilotriacetic acid, NTA) in enhancing the remediation of Cd by Coronopus didymus (Brassicaceae). C. didymus growing in Cd-contaminated soil (35-175 mg kg-1 soil) showed increased growth and biomass due to the hormesis effect, and chelant supplementation further increased growth, biomass, and Cd accumulation. A significant interaction with chelants and different Cd concentrations was observed, except for Cd content in roots and Cd content in leaves, which exhibited a non-significant interaction with chelant addition. The effect of the NTA amendment on the root dry biomass and shoot dry biomass was more pronounced than EDDS at all the Cd treatments. Upon addition of EDDS and NTA, bio-concentration factor values were enhanced by ~184-205 and ~ 199-208, respectively. The tolerance index of root and shoot increased over the control upon the addition of chelants, with NTA being better than EDDS. With chelant supplementation, bio-accumulation coefficient values were in the order Cd35 + NTA (~163%) > Cd105 + NTA (~137%) > Cd35 + EDDS (~89%) > Cd175 + NTA (~85%) > Cd105 + EDDS (~62%) > Cd175 + EDDS (~40%). The translocation factor correlated positively (r ≥ 0.8) with tolerance index and Cd accumulation in different plant parts. The study demonstrated that chelant supplementation enhanced Cd-remediation efficiency in C. didymus as depicted by improved plant growth and metal accumulation, and NTA was more effective than EDDS in reclaiming Cd.

Nature of phytotoxic interference of alien weed 'Calyptocarpus vialis' against some crop plants.

Calyptocarpus vialis (syn. Synedrella vialis; Asteraceae), a native of the tropical Americas, has acquired an invasive status in the eastern Asia and Africa and, of late, in India. It is an annual herbaceous weed that forms a dominant ground cover due to its prostrate expansion and interferes with the growth of other plant species. However, the reasons for this interference are largely unknown. Therefore, we examined the allelopathic interference of C. vialis via leachation and residue degradation on the emergence, growth, and development of three crop species (Brassica nigra, Triticum aestivum, and Avena sativa). In a laboratory bioassay, the leachates (0.5-4%) of C. vialis exhibited a dose-dependent inhibitory effect on various growth parameters of the test plants. Similarly, under screenhouse, C. vialis-amended soil (1-4%) affected the growth of test species in a dose-dependent manner. Further, the phytotoxicity of the residues of C. vialis was examined using rhizospheric soil (RS) and residue-amended soil (RAS). It was observed that RAS exerted the maximum allelopathic effect on the test species accompanied by significant changes in pH, electrical conductivity, and total water-soluble phenolic content, as compared with the control soil (CS) and RS. Liquid chromatography and mass spectroscopy analyses confirmed the presence of eleven allelochemicals as the major phytotoxins. The study demonstrated that C. vialis exerts strong phytotoxic effects on other plants through the release of potent allelochemicals, both via leachation and residue degradation.

Projected Impacts of Climate Change on the Range Expansion of the Invasive Straggler Daisy (Calyptocarpus vialis) in the Northwestern Indian Himalayan Region.

Human-induced climate change modifies plant species distribution, reorganizing ecologically suitable habitats for invasive species. In this study, we identified the environmental factors that are important for the spread of Calyptocarpus vialis, an emerging invasive weed in the northwestern Indian Himalayan Region (IHR), along with possible habitats of the weed under current climatic scenarios and potential range expansion under several representative concentration pathways (RCPs) using MaxEnt niche modeling. The prediction had a high AUC (area under the curve) value of 0.894 ± 0.010 and a remarkable correlation between the test and expected omission rates. BIO15 (precipitation seasonality; 38.8%) and BIO1 (annual mean temperature; 35.7%) had the greatest impact on the probable distribution of C. vialis, followed by elevation (11.7%) and landcover (6.3%). The findings show that, unlike the current situation, "high" and "very high" suitability areas would rise while less-suited habitats would disappear. All RCPs (2.6, 4.5, 6.0, and 8.5) indicate the expansion of C. vialis in "high" suitability areas, but RCP 4.5 predicts contraction, and RCPs 2.6, 6.0, and 8.5 predict expansion in "very high" probability areas. The current distribution of C. vialis is 21.59% of the total area of the state, with "medium" to "high" invasion suitability, but under the RCP 8.5 scenario, it might grow by 10% by 2070. The study also reveals that C. vialis may expand its niche at both lower and higher elevations. This study clarifies how bioclimatic and topographic factors affect the dispersion of invasive species in the biodiverse IHR. Policymakers and land-use managers can utilize the data to monitor C. vialis hotspots and develop scientifically sound management methods.

Biodegradable chelant-metal complexes enhance cadmium phytoextraction efficiency of Solanum americanum.

Toxic contaminants (metals and metal-containing compounds) are accumulating in the environment at an astonishing rate and jeopardize human health. Remarkable industrial revolution and the spectacular economic growth are the prime causes for the release of such toxic contaminants in the environment. Cadmium (Cd) is ranked the 7th most toxic compound by the Agency for Toxic Substances and Disease Registry (USA), owing to its high carcinogenicity and non-biodegradability even at miniscule concentration. The present study assessed the efficiency of four biodegradable chelants [nitrilotriacetic acid (NTA), ethylenediamine disuccinate (EDDS), ethylene glycol tetraacetic acid (EGTA), and citric acid (CA)] and their dose (5 mM and 10 mM) in enhancing metal accumulation in Solanum americanum Mill. (grown under 24 mg Cd kg-1 soil) through morpho-physiological and metal extraction parameters. Significant variations were observed for most of the studied parameters in response to chelants and their doses. However, ratio of root and shoot length, and plant height stress tolerance index differed non-significantly. The potential of chelants to enhance Cd removal efficiency was in the order - EGTA (7.44%) > EDDS (6.05%) > NTA (4.12%) > CA (2.75%). EGTA and EDDS exhibited dose-dependent behavior for Cd extraction with 10 mM dose being more efficient than 5 mM dose. Structural equation model (SEM) depicted strong positive interaction of metal extraction parameters with chelants (Z-value = 11.61, p = 0.001). This study provides insights into the importance of selecting appropriate dose of biodegradable chelants for Cd extraction, as high chelant concentration might also result in phytotoxicity. In the future, phytoextraction potential of these chelants needs to be examined through field studies under natural environmental conditions.

Nitric oxide alleviates arsenic toxicity by reducing oxidative damage in the roots of Oryza sativa (rice).

Nitric oxide (NO) is a bioactive gaseous, multifunctional molecule playing a central role and mediating a variety of physiological processes and responses to biotic and abiotic stresses including heavy metals. The present study investigated whether NO applied exogenously as sodium nitroprusside (SNP) has any protective role against arsenic (As) toxicity in Oryza sativa (rice). Treatment with 50 microM SNP (a NO donor) significantly ameliorated the As-induced (25 or 50 microM) decrease in root and coleoptile length of rice. Further, As-induced oxidative stress measured in terms of malondialdehyde (MDA), superoxide ion (O(2)(-)), root oxidizability and H2O2 content was lesser upon supplementation of NO. It indicated a reactive oxygen species (ROS) scavenging activity of NO. NO addition reversed (only partially) the As-induced increase in activities of antioxidant enzymes - superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and catalase. The study concludes that exogenous NO provides resistance to rice against As-toxicity and has an ameliorating effect against As-induced stress.

Phytotoxicity of major constituents of the volatile oil from leaves of Artemisia scoparia Waldst. & Kit.

The phytotoxicity of the three major monoterpene constituents of the essential oil from leaves of Artemisia scoparia Waldst. & Kit. (redstem wormwood) was investigated. GC/GC-MS analysis revealed that the essential oil (yield 0.84%) is a complex mixture containing 19 monoterpenes, 7 sesquiterpenes and 15 other compounds--aliphatic alcohols, ketones, aromatic hydrocarbons and esters. The three major monoterpenes were beta-myrcene (30.2%), p-cymene (12.8%) and dl-limonene (12.4%). The essential oil and the three monoterpenes exhibited phytotoxicity and reduced germination, seedling growth, chlorophyll content and percent respiration of Avena sativa and Triticum aestivum in a dose-response manner. The inhibitory effect of monoterpenes was comparatively smaller than of the crude essential oil and beta-myrcene was most toxic followed by p-cymene, whereas limonene was least toxic. The study suggests that A. scoparia oil and beta-myrcene can be explored for phytotoxicity against weeds.

Assessment of phytotoxicity of parthenin.

Phytotoxicity of parthenin, a sesquiterpene lactone, was evaluated against four weedy species (Amaranthus viridis, Cassia occidentalis, Echinochloa crus-galli, and Phalaris minor) through a series of experiments conducted under laboratory or greenhouse conditions to assess its herbicidal potential. Under laboratory conditions, parthenin (0.5-2 mM) severely reduced seedling growth (root and shoot) and dry weight of test weeds. However, the effect was greater on root growth. Parthenin (1 mM) suppressed the mitotic activity in the onion root tip cells that could possibly be responsible for the reduction in seedling growth. Both pre- and post-emergent application of parthenin caused a significant loss of chlorophyll pigments and affected photosynthesis. Parthenin ( > or =1 mM) caused an excessive electrolyte leakage in the plant tissues which was light-dependent. The root inhibition was associated with swelling and blackening of the root tip, shriveling and damage to the epidermal tissue and non-formation of root hairs. The study concludes that parthenin possesses weed-suppressing potential (both pre- and post-).

Phytotoxicity of the volatile monoterpene citronellal against some weeds.

A study was undertaken to assess the phytotoxicity of citronellal, an oxygenated monoterpenoid with an aldehyde group, towards some weedy species [Ageratum conyzoides L., Chenopodium album L., Parthenium hysterophorus L., Malvastrum coromandelianum (L.), Garcke, Cassia occidentalis L. and Phalaris minor Retz.]. A significant effect on weed emergence and early seedling growth was observed in a dose-response based laboratory bioassay in a sand culture. Emergence of all test weeds was completely inhibited at 100 micro/g sand content of citronellal. Seeds of A. conyzoides and P. hysterophorus failed to emerge even at 50 microg/g content. Root length was inhibited more compared to shoot length. The failure of root growth was attributed to the effect of citronellal on the mitotic activity of growing root tips cells as ascertained by the onion root tip bioassay. At 2.5 mM treatment of citronellal, mitosis was completely suppressed and at higher concentrations cells showed various degrees of distortion and were even enucleated. The post-emergent application of citronellal also caused visible injury in the form of chlorosis and necrosis, leading to wilting and even death of test weeds. Among the test weeds, the effect was severe on C. album and P. hysterophorus. There was loss of chlorophyll pigment and reduction in cellular respiration upon citronellal treatment indicating the impairment of photosynthetic and respiratory metabolism. Scanning electron microscopic studies in C. occidentalis leaves upon treatment of citronellal revealed disruption of cuticular wax, clogging of stomata and shrinkage of epidermal cells at many places. There was a rapid electrolyte leakage in the leaf tissue upon exposure to citronellal during the initial few hours. In P. minor electrolyte leakage in response to 2 mM citronellal was closer to the maximum leakage that was obtained upon boiling the tissue. The rapid ion leakage is indicative of the severe effect of citronellal on the membrane structure and loss of membrane integrity. In all, the study concludes that citronellal causes a severe phytotoxicity on the weeds.

Effect of 2-benzoxazolinone (BOA) on seedling growth and associated biochemical changes in mung bean (Phaseolus aureus).

BOA (2-benzoxazolinone) is a potent phytotoxin present in several graminaceous crops such as rye, maize and wheat. Due to its wide range of phytotoxicity, it is considered as a potential pesticide. A study was conducted to explore the impact of BOA on the radicle and plumule elongation of mung bean (Phaseolus aureus) and associated changes in the macromolecular content - proteins and carbohydrates - and activities of enzymes like amylases, proteases, polyphenol oxidases and peroxidases. BOA significantly reduced the radicle and plumule length of P. aureus, and the contents of proteins and carbohydrates in both root and leaf tissue. On the other hand, activities of hydrolytic enzymes - proteases, amylases, polyphenol oxidases and peroxidases - increased substantially in both root and leaf tissue of P. aureus upon BOA exposure. This indicated that BOA treatment induced stress in P. aureus and enhanced enzyme activities to counter the induced stress and continue the growth. In other words, BOA-induced stress altered the plant biochemical status and related enzyme activities resulting in increased metabolism that serves to provide protection against cellular injury. Such studies providing information about the biomolecular content and enzymatic activities in response to natural products serve as clues for furtherance of knowledge about the modes of action of natural compounds of commercial interest.

Chemical composition and phytotoxicity of volatile essential oil from intact and fallen leaves of Eucalyptus citriodora.

A total of 23 volatile constituents was identified and characterized by GC and GC-MS in the volatile essential oil extracted from intact (juvenile and adult) and fallen (senescent and leaf litter) leaves of lemon-scented eucalyptus (Eucalyptus citriodora Hook.). The leaves differed in their pigment, water and protein content, and C/N ratio. The oils were, in general, monoterpenoid in nature with 18 monoterpenes and 5 sesquiterpenes. However, a great variability in the amount of essential oils and their individual constituents was observed in different leaf tissues. The amount was maximum in the senescent leaves collected from the floor of the tree closely followed by that from juvenile leaves. In all, 19 constituents were identified in oil from juvenile and senescent leaves compared to 23 in adult leaves and 20 in leaf litter, respectively. Citronellal, a characteristic monoterpene of the oil reported hitherto was found to be more (77-78%) in the juvenile and senescent leaves compared to 48 and 54%, respectively, in the adult leaves and leaf litter. In the adult leaves, however, the content of citronellol--another important monoterpene-- was very high (21.9%) compared to other leaf types (7.8-12.2%). Essential oil and its two major monoterpenes viz. citronellal and citronellol were tested for their phytotoxicity against two weeds (Amaranthus viridis and Echinochloa crus-galli) and two crops (Triticum aestivum and Oryza sativa) under laboratory conditions. A difference in the phytotoxicity, measured in terms of seedling length and dry weight, of oil from different leaves and major monoterpenes was observed. Oil from adult leaves was found to be most phytotoxic although it occurs in smaller amount (on unit weight basis). The different toxicity of different oil types was due to the relative amount of individual monoterpenes present in the oil, their solubility and interactive action. The study concludes that oil from senescent and juvenile leaves being rich in citronellal could be used as commercial source of citronellal whereas that from adult leaves for weed management programmes as it was the most phytotoxic.

Chemical composition and inhibitory activity of essential oil from decaying leaves of Eucalyptus citriodora.

A study was undertaken to explore the content and composition of volatile oil from decaying leaves of lemon-scented eucalypt (Eucalyptus citriodora Hook.) not analyzed earlier. GC and GC-MS analysis of the oil (yield 0.6%) revealed the monoterpenoid nature with citronellal (52.2%), citronellol (12.3%) and isoisopulegol (11.9%) as the major constituents. Overall, 17 components were identified that accounted for over 94% of the decaying leaf oil. Surprisingly, the decaying leaf oil contained nearly 1.8% of trans-rose oxide, which is generally absent in eucalypt essential oil. Decaying leaf oil and its major 2 components (citronellal and citronellol) inhibited the germination and root elongation of two weeds--Cassia occidentalis (broad-leaved) and Echinochloa crus-galli (grassy weed). Based on the dose-response studies, I50 values were determined for decaying leaf oil and the effect was more on germination only of broad-leaved weed (C. occidentalis), whereas that of citronellal and citronellol were on germination as well as root length of E. crus-galli (grassy weed). Based on I50 values it was observed that citronellal was more phytotoxic and germination inhibiting in nature, whereas citronellol was a more potent root inhibitor, thereby indicating a possible different mode of action. The study concludes that decaying leaf oil hold a good commercial value for exploitation as weed management agent.

ß-Pinene inhibited germination and early growth involves membrane peroxidation.

ß-Pinene, an oxygenated monoterpene, is abundantly found in the environment and widely occurring in plants as a constituent of essential oils. We investigated the phytotoxicity of ß-pinene against two grassy (Phalaris minor, Echinochloa crus-galli) and one broad-leaved (Cassia occidentalis) weeds in terms of germination and root and shoot growth. ß-Pinene (0.02-0.80 mg/ml) inhibited the germination, root length, and shoot length of test weeds in a dose-response manner. The inhibitory effect of ß-pinene was greater in grassy weeds and on root growth than on shoot growth. ß-Pinene (0.04-0.80 mg/ml) reduced the root length in P. minor, E. crus-galli, and C. occidentalis over that in the control by 58-60, 44-92, and 26-85 %, respectively. In contrast, shoot length was reduced over the control by 45-97 % in P. minor, 48-78 % in E. crus-galli, and 11-75 % in C. occidentalis at similar concentrations. Further, we examined the impact of ß-pinene on membrane integrity in P. minor as one of the possible mechanisms of action. Membrane integrity was evaluated in terms of lipid peroxidation, conjugated diene content, electrolyte leakage, and the activity of lipoxygenases (LOX). ß-Pinene (≥0.04 mg/ml) enhanced electrolyte leakage by 23-80 %, malondialdehyde content by 15-67 %, hydrogen peroxide content by 9-39 %, and lipoxygenases activity by 38-383 % over that in the control. It indicated membrane peroxidation and loss of membrane integrity that could be the primary target of ß-pinene. Even the enhanced (9-62 %) activity of protecting enzymes, peroxidases (POX), was not able to protect the membranes from ß-pinene (0.04-0.20 mg/ml)-induced toxicity. In conclusion, our results show that ß-pinene inhibits root growth of the tested weed species through disruption of membrane integrity as indicated by enhanced peroxidation, electrolyte leakage, and LOX activity despite the upregulation of POX activity.

Cr(VI) imposed toxicity in maize seedlings assessed in terms of disruption in carbohydrate metabolism.

The present study examined the toxic effects of Cr(VI; 100, 250 and 500 µM) in maize seedlings by investigating the changes in carbohydrate metabolism after 48, 96, and 144 h of exposure. Cr-stress results in severe alterations in the contents of carbohydrates and reducing sugars and the activities of carbohydrate metabolizing enzymes, amylases, phosphatases and phosphorylases, and invertases in maize seedlings. Under Cr stress, the contents of carbohydrates and reducing sugars declined in roots, whereas an increase was noticed in leaves. The catalytic activity of carbohydrate metabolizing enzymes, except invertases, in roots declined in the presence of Cr(VI) in a concentration- and exposure time-dependent manner. In contrast, the activities of these enzymes were enhanced in leaves under Cr(VI) stress. The activity of invertases increased with increasing amount of Cr(VI) but declined with an increase in the time interval. In conclusion, our results show that carbohydrate metabolism is severely affected under Cr(VI) toxicity. The study suggests that Cr-induced perturbations in the carbohydrate metabolism are one of the factors resulting in growth inhibition under Cr(VI) stress.

Lead (Pb)-inhibited radicle emergence in Brassica campestris involves alterations in starch-metabolizing enzymes.

Lead (Pb) is a toxic heavy metal released into the natural environment and known to cause oxidative damage and alter antioxidant mechanism in plants. However, not much is known about the interference of Pb with the biochemical processes and carbohydrate metabolism during seed germination. We, therefore, investigated the effect of Pb (50-500 µM) upon biochemical alterations in germinating seeds (at 24-h stage) of Brassica campestris L. Pb treatment significantly enhanced protein and carbohydrate contents that increased by ~43% and 200%, respectively, at 500-µM Pb over control. In contrast, the activities of starch/carbohydrate-metabolizing enzymes--α-amylases, ß-amylases, acid invertases, and acid phosphatases--decreased by ~54%, 60%, 74%, and 52%, respectively, over control. Activities of peroxidases and polyphenol oxidases, involved in stress acclimation, however, increased by ~1.2- to 3.9-folds and 0.4- to 1.4-folds upon 50-500-µM Pb treatment. Pb enhanced oxidizing ability by 10 to 16.7 times over control suggesting interference with emerging root's oxidizing capacity. The study concludes that Pb exposure inhibits radicle emergence from B. campestris by interfering with the biochemical processes linked to protein and starch metabolism.

In vitro screening of essential oil from young and mature leaves of Artemisia scoparia compared to its major constituents for free radical scavenging activity.

The present study investigated the chemical characterization, and antioxidant activity of essential oil hydrodistilled from young and mature leaves of Artemisia scoparia. GC-MS analyses revealed a monoterpenoid nature (64-67%) with 44 and 31 constituents in young and mature leaves oil, respectively. The oil from young leaf contained greater amount of oxygenated compounds. Beta-myrcene (24.13%) and p-cymene (27.06%) were the major constituents in young and mature leaves oil, respectively. A. scoparia leaf oils (25-200 microg/ml) exhibited a strong 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity and antioxidant activity against hydroxyl radical and hydrogen peroxide. However, the activities of major constituent monoterpenes, beta-myrcene and p-cymene, were less. In general, the DPPH radical scavenging and antioxidant activity was in the order: mature leaf oil > young leaf oil > beta-myrcene > p-cymene.

Characterization and antioxidant activity of essential oils from fresh and decaying leaves of Eucalyptus tereticornis.

The composition of essential oils hydrodistilled from fresh and decaying leaves of Eucalyptus tereticornis was analyzed by means of gas chromatography and mass spectrometry, and a total of 68 constituents were identified. The essential oils were assayed for antioxidant activity in terms of scavenging of 2,2-diphenyl-1-picrylhydrazil (DPPH) and hydroxyl (OH(*)) radical, and superoxide anion (O2(-*)).The major constituents of the fresh leaf oil were alpha-pinene (28.53%) and 1,8-cineole (19.48%), whereas in the decaying leaf oil, beta-citronellal (14.15%), (-)-isopulegol (13.35%), and (+)-beta-citronellol (10.73%) were the major components. Both essential oils exhibited a strong radical scavenging activity against DPPH radical with IC50 values of 110 and 139.8 microg/mL for fresh and decaying leaf oil, respectively (IC50 of BHT = 164.2 microg/mL). Further, the essential oils (at 400 microg/mL) also exhibited OH(*) (56-62%) and O2(-*) (65-69%) scavenging activity parallel to the commercial antioxidant BHT/ascorbic acid. However, unlike the essential oils, the major monoterpene constituents exhibited significantly less scavenging activity (<35% DPPH or OH(*); at 400 microg/mL). The study concluded that fresh and decaying leaves of E. tereticornis are a source of monoterpenoid rich oil exhibiting antioxidant activity.

alpha-Pinene inhibits growth and induces oxidative stress in roots.

BACKGROUND AND AIMS: Determining the mode of action of allelochemicals is one of the challenging aspects in allelopathic studies. Recently, allelochemicals have been proposed to cause oxidative stress in target tissue and induce an antioxidant mechanism. alpha-Pinene, one of the common monoterpenoids emitted from several aromatic plants including forest trees, is known for its growth-inhibitory activity. However, its mechanism of action remains unexplored. The aim of the present study was to determine the inhibitory effect of alpha-pinene on root growth and generation of reactive oxygen species, as indicators of oxidative stress and changes in activities of antioxidant enzymes. METHODS: Effects of alpha-pinene on early root growth were studied in five test species, Cassia occidentalis, Amaranthus viridis, Triticum aestivum, Pisum sativum and Cicer arietinum. Electrolyte leakage, lipid peroxidation, hydrogen peroxide generation, proline accumulation, and activities of the enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT) and glutathione reductase (GR) were studied in roots of C. occidentalis. KEY RESULTS: alpha-Pinene inhibited the radicle growth of all the test species. Exposure of C. occidentalis roots to alpha-pinene enhanced solute leakage, and increased levels of malondialdehyde, proline and hydrogen peroxide, indicating lipid peroxidation and induction of oxidative stress. Activities of the antioxidant enzymes SOD, CAT, GPX, APX and GR were significantly elevated, thereby indicating the enhanced generation of reactive oxygen species (ROS) upon alpha-pinene exposure. Increased levels of scavenging enzymes indicates their induction as a secondary defence mechanism in response to alpha-pinene. CONCLUSIONS: It is concluded that alpha-pinene inhibits early root growth and causes oxidative damage in root tissue through enhanced generation of ROS, as indicated by increased lipid peroxidation, disruption of membrane integrity and elevated antioxidant enzyme levels.