Repurposing spent biomass of vetiver grass used for stormwater treatment to generate biochar and ethanol.

Stormwater pollution is a key factor contributing to water quality degradation, posing substantial environmental and human health risks. Although stormwater retention ponds, also referred to as wet ponds, are commonly implemented to alleviate stormwater challenges by reducing peak flow and removing suspended solids, their effectiveness in removing heavy metals and nutrients is limited. This study evaluated the performance of floating treatment platforms (FTPs) featuring vetiver grass (Chrysopogon zizanioides), a non-invasive, nutrient- and metal-accumulating perennial grass, in removing heavy metals (Cu, Pb, and Zn) and nutrients (P and N) in stormwater retention ponds. Furthermore, the potential for utilizing the spent vetiver biomass for generating biochar and bioethanol was investigated. The study was conducted in a greenhouse setup under simulated wet and dry weather conditions using pond water collected from a retention pond in Stafford Township, New Jersey, USA. Two FTPs with vetiver (vegetated FTPs) were compared with two FTPs without vetiver (non-vegetated FTPs), which served as controls. Results showed that the removal of heavy metals and nutrients by the FTPs with vetiver was significantly higher (p < 0.05) than the FTPs without vetiver. Notably, vetiver showed resilience to stormwater pollutants and hydroponic conditions, displaying no visible stress symptoms. The biochar and bioethanol generated from the spent vetiver exhibited desirable yield and quality, without raising concerns regarding pollutant leaching, indicated by very low TCLP and SPLP concentrations. This study provides compelling evidence that the implementation of vetiver-based FTPs offers a cost-effective and environment-friendly solution for mitigating stormwater pollution in retention ponds. Furthermore, the utilization of vetiver biomass for biofuel and biochar production supports clean production and fostering circular economy efforts.

Vetiver grass cleans up arsenic contaminated field for subsequent safe cultivation of rice with low arsenic in grains: A two year field study.

The presence of high concentrations of arsenic (As) in agricultural soils and its subsequent accumulation in rice crop is a serious issue threatening sustainability of agriculture and human health. In the present work, remediation of As contaminated field in Nadia, West Bengal, India was done through the cultivation of Vetiver (Vetiveria zizanoides L. Nash) and the same field was subsequently used for rice (Oryza sativa L.) cultivation. The results showed that V. zizanoides could reduce As concentrations in the field to bring it lower than the maximum permissible limit (20 mg kg-1) in 11 months' time. The rice plants grown in remediated field showed improvement in growth and photosynthesis parameters as compared to that of contaminated field. Importantly, yield related parameters (filled seed, 1000 grain weight, number of panicles etc.) were also significantly higher in remediated field than that in contaminated field. Arsenic concentration in roots, shoot, husk and grains of rice was found to be significantly lower in remediated field than in contaminated field. Grain As decreased from 0.75 to 0.77 µg g-1 dw in contaminated field to 0.15-0.18 µg g-1 dw. In conclusion, replacing rice for single year with V. zizanoides crop can significantly remediate the field and can be a viable option.

Co-cropping vetiver grass and legume for the phytoremediation of an acid mine drainage (AMD) impacted soil.

Acid mine drainage (AMD) is a form of environmental pollution from mining activity that can negatively affect soil environments by acidification, salinisation, and metal(loid) contamination. The use of plants to remediate (phytoremediation) these impacted environments while generating plant-based value is a promising approach to more accessible and cost-benefiting restoration of post-mining, marginal lands. In this study, a 3-month growth-chamber pot experiment was conducted to investigate the influence of co-cropping two plant species, Chrysopogon zizanioides (vetiver grass) and the legume Medicago truncatula (barrel clover) with a wheat straw biochar amendment on the phytostabilisation of metal(loid)s Cr, Zn, and As and the phytoextraction of rare earth element (REE) in an AMD impacted soil from a gold mining region in South Africa. The results showed that co-cropping with vetiver significantly lowered the legume's Cr, Zn, and As root contents by 80%, 32% and 54%, respectively, and improved the plant's overall metal(loid) tolerance by increasing its translocation from root to shoot tissue. The biochar further inhibited root uptake of Cr and Zn, by 71% and 36%, and increased the legume biomass by 40%. Both plant species and cropping treatments exhibited low REE extraction capabilities by shoot tissue, which accounted for less than 0.2% of total soil REE contents. The study shows that co-cropping with vetiver and biochar amendment are effective tools for the phytoremediation of AMD impacted soil mainly by lowering plant uptake and improving plant metal(loid) tolerance. Likely mechanisms at play include the alteration of rhizosphere chemistry and species-specific physiological and molecular responses. These effects offer support for the phytostabilisation of AMD impacted soil with the generation of plant-based value through dual (and safe) cultivation (phytoprotection) rather than through REE recovery from plant biomass (phytoextraction). These techniques could allow for the simultaneous restoration of post-mining, mining-impacted and marginal lands with agricultural production.

Stenocarpella chrysopogonis, a New Species Causing Leaf Streak Disease of Chrysopogon zizanioides in Southern China.

Vetiver grass (Chrysopogon zizanioides) has been widely used in recent years for ecological environment management, restoration of degraded ecosystems, and essential oil extraction. In 2019, a leaf streak disease of C. zizanioides was observed in Zhanjiang, Guangdong Province, China. The disease appeared as large streak lesions on the leaves, on which conidiomata were formed. A pathogenicity test with the fungus isolated from these lesions confirmed Koch's postulates and thus the fungus as the causal agent of this disease. A morphological resemblance of the pathogen to Stenocarpella was noted upon microscopic examination. Phylogenetic trees inferred from both individual and combined ITS, LSU, and tef1 sequences confirmed the pathogen as a species of the Diaporthaceae and revealed it to be closely related to Phaeocytostroma and Stenocarpella species. As morphological characters clearly placed the pathogen in the genus Stenocarpella, it was described as S. chrysopogonis.

Improving capacity for phytoremediation of Vetiver grass and Indian mustard in heavy metal (Al and Mn) contaminated water through the application of clay minerals.

One of the consequences of mining is the release of heavy metals into the environment, especially water bodies. Phytoremediation of areas contaminated by heavy metals using Vetiver grass and Indian mustard is cost-effective and environmentally friendly. This study aimed at enhancing remediation of heavy metal contaminated water through the simultaneous hybrid application of clay minerals (attapulgite and bentonite) and Vetiver grass or Indian mustard. A 21-day greenhouse experiment was carried out to investigate the effectiveness of the clay minerals to improve heavy metal phytoremediation. The highest accumulation of aluminium (Al) by Vetiver grass was 371.8 mg/kg in the BT2.5VT treatment, while for Mn, the highest accumulation of 34.71 mg/kg was observed in the AT1VT treatment. However, Indian mustard showed no significant uptake of heavy metals, but suffered heavy metal toxicity despite the addition of clay minerals. From this study, it was evident that bentonite added at 2.5% (w/v) could improve the phytoremediation capacity of Vetiver grass for Al and Mn polluted water. The current laboratory-scale findings provided a basis for field trials earmarked for remediation in a post-mining coal environment in South Africa. This remediation approach can also be adopted in other places.

Effects of biochar on the physiology and heavy metal enrichment of Vetiveria zizanioides in contaminated soil in mining areas.

The effects of biochar addition on the physiological and biochemical characteristics of Vetiveria zizanioides, and the enrichment of heavy metals, were studied herein. The aim was to provide a theoretical reference for biochar to regulate the growth of V. zizanioides in the heavy metal-contaminated soil of mining areas and the enrichment capacity of Cu, Cd, and Pb. The results showed that the addition of biochar significantly increased the contents of various pigments in the middle and late growth stages of V. zizanioides, reduced the contents of malondialdehyde (MDA) and proline (Pro) in each growth period, weakened the peroxidase (POD) activity during the entire growth period; superoxide dismutase (SOD) activity decreased in the initial stages and substantially increased in the middle and late stages. The addition of biochar reduced the enrichment of Cu in the roots and leaves of V. zizanioides, while the enrichment of Cd and Pb increased. In conclusion, it was found that biochar could reduce the toxicity of heavy metals in contaminated soil in the mining area, affect the growth of V. zizanioides and its accumulation of Cd and Pb, and is, therefore, beneficial to the restoration of contaminated soil and the overall ecological restoration of the mining area.

Integrated biochemical and transcriptomic analysis reveals the effects of Burkholderia sp. SRB-1 on cadmium accumulating in Chrysopogon zizanioides L. under Cd stress.

Application of plant growth-promoting rhizobacteria plays a vital role in enhancing phytoremediation efficiency. In this study, multiple approaches were employed to investigate the underlying mechanisms of Burkholderia sp. SRB-1 (SRB-1) on elevating Cd uptake and accumulation. Inoculation experiment indicated that SRB-1 could facilitate plant growth and Cd tolerance, as evidenced by the enhanced plant biomass and antioxidative enzymes activities. Cd content in plant shoots and roots increased about 36.56%-39.66% and 25.97%-130.47% assisted with SRB-1 when compared with control. Transcriptomics analysis revealed that SRB-1 upregulated expression of amiE, AAO1-2 and GA2-ox related to auxin and gibberellin biosynthesis in roots. Auxin and gibberellin, as hormone signals, regulated plant Cd tolerance and growth through activating hormone signal transduction pathways, which might also contribute to 67.94% increase of dry weight. The higher expression levels of ATP-binding cassette transporter subfamilies (ABCB, ABCC, ABCD and ABCG) in Chrysopogon zizanioides roots contributed to higher Cd uptake in Cd15 B (323.83 mg kg-1) than Cd15 (136.28 mg kg-1). Further, SRB-1 facilitated Cd migration from roots to shoots via upregulating the expression of Nramp, ZIP and HMA families. Our integrative analysis provided a molecular-scale perspective on Burkholderia sp. SRB-1 contributing to C. zizanioides performance.

Essential Oils Composition and Biological Activity of Chamaecyparis obtusa, Chrysopogon nigritanus and Lavandula coronopifolia Grown Wild in Sudan.

Generally, there are scant data about the constituents and eventually the biological activity of essential oils (EOs) from aromatic plants that grow naturally in Sudan. The present study aimed to determine the chemical composition, and antioxidant and enzyme inhibitory activities of EO extracted from the fruit of Chamaecyparis obtusa (Siebold and Zucc.) Endl. (family Cupressaceae), root of Chrysopogon nigritanus (Benth.) Veldkampis (family Poaceae) and aerial part of Lavandula coronopifolia Poir (family Lamiaceae). The fruit of C. obtusa contained only monoterpenes, mainly hydrogenated ones, with α-pinene (69.07%) as the major component. Oxygenated sesquiterpenes comprised the highest content of the C. nigritanus root EO with cedr-8-en-15-ol (28.69%) as the major constituent while aerial parts of L. coronopifolia contained both monoterpenes and sesquiterpenes and the oxygenated monoterpene lavandulol (26.56%) as dominant compounds. The EO of the root of C. nigritanus significantly displayed (p < 0.05) the highest anti-DPPH radical, Fe3+- and Cu2+-reducing and metal-chelating activities, while that of C. obtusa fruit significantly exerted (p < 0.05) the best anti-ABTS radical and total antioxidant activity. The two EOs significantly exhibited (p < 0.05) the highest anti-acetylcholinesterase and -butyrylcholinesterase activities, respectively, while EO of L. coronopifolia was the only oil to show a considerable inhibitory effect against the tyrosinase and α-glucosidase enzymes. In conclusion, EOs from these three plants could be natural agents with promising functional properties for food, cosmetics, and pharmaceutical applications.

Enhanced phytoremediation of atrazine-contaminated soil by vetiver (Chrysopogon zizanioides L.) and associated bacteria.

The intensive and long-term use of atrazine (ATZ) has led to the contamination of agricultural soils and non-target organisms, posing a series of threats to human health through the transmission of the food chain. In this study, a 60-day greenhouse pot experiment was carried out to explore the phytoremediation by Chrysopogon zizanioides L. (vetiver). The uptake, accumulation, distribution, and removal of ATZ were investigated, and the degradation mechanisms were elucidated. The results showed that the growth of vetiver was inhibited in the first 10 days of the incubation; subsequently, the plant recovered rapidly with time going. Vetiver grass was capable of taking up ATZ from the soil, with root concentration factor ranging from 2.36 to 15.55, and translocating to the shoots, with shoot concentration factor ranging from 7.51 to 17.52. The dissipation of ATZ in the rhizosphere soil (97.51%) was significantly higher than that in the vetiver-unplanted soil (85.14%) at day 60. Metabolites were identified as hydroxyatrazine (HA), deethylatrazine (DEA), deisopropylatrazine (DIA), and didealkylatrazine (DDA) in the samples of the shoots and roots of vetiver as well as the soils treated with ATZ. HA, DEA, DIA, and DDA were reported first time as metabolites of ATZ in shoots and roots of vetiver grown in soil. The presence of vetiver changed the formation and distribution of the dealkylated products in the rhizosphere soil, which remarkably enhanced the occurrence of DEA, DIA, and DDA. Arthrobacter, Bradyrhizobium, Nocardioides, and Rhodococcus were the major atrazine-degrading bacterial genera, which might be responsible for ATZ degradation in the rhizosphere soil. Our findings suggested that vetiver grass can significantly promote ATZ degradation in the soil, and it could be a strategy for remediation of the atrazine-contaminated agricultural soil.

Review of Antimicrobial and Other Health Effects in 5 Essential Oil Producing Grass Species.

The warm season essential oil producing grass species including lemongrass (Cymbopogon citratus), palmarosa grass (C. martini), geranium grass (C. schoenanthus), vetiver grass (Chrysopogon zizanioides), and scented top grass (Capillipedium parviflorum) are used worldwide for their cosmetic and health properties. A discussion providing evidence from literature reviews about the potential uses of these grass species for antimicrobial and other health uses are presented. These species could be used as new therapies for treating microbial infections. The purpose of this study is to discuss in detail, evidence from literature reviews supporting potential health uses and to provide some discussion regarding some agronomic traits for these essential oil producing species.

Biological pilot treatment reduces physicochemical and microbiological parameters of dairy cattle wastewater.

The objectives of the present study were to characterize and evaluate a pilot treatment unit (PTU) for dairy cattle wastewater (DCW) in relation to its efficiency in reducing the physicochemical and microbiological parameters and possible application of this fertilizer in organic production. A PTU was set up, composed of the following elements: a dung pit of 7.8 m3, already in place; a septic tank; a set of anaerobic biological filters comprising an upflow filter and a downward-flow filter filled with fragments PVC corrugated conduit; and two constructed wetland systems (CWSs) of horizontal subsurface flow in two parallel routes (Routes 1 and 2), controlled by means of a flow rate divider box. Route 1 passed through CWS 1 cultivated with cattail (Typha domingensis) and Route 2 passed through CWS 2 cultivated with vetiver grass (Chrysopogon zizanioides). To evaluate the treatment stages, biweekly investigations were carried out to collect effluent samples. The results of monitoring, in absolute values, were evaluated by means of the medians and variation coefficients and compared by means of Kruskal-Wallis non-parametric test followed by the Student Newman Keuls test. The treatment efficiencies of Routes 1 and 2 were calculated. The influence of vetiver on the removal of nutrients from the DCW was analyzed and the productivity estimate (t.ha-1) was performed. CWS 1 was not able to reduce the organic load indices, but it was able to retain fatty material and sodium. CWS 2 showed a reduction in nitrogenous forms and also for other nutrients, achieving the greatest removal of sodium and greatest decay of fecal contamination indicators, thermotolerant coliforms (56.13%), and E. coli (46.82%).

Enzyme-based green synthesis, characterisation, and toxicity studies of cellulose nanocrystals/fibres produced from the Vetiveria zizanioides roots agro-waste.

Vetiver zizanioides roots are considered the most useful part of the plant. It is widely used to extract oil. The aromatic oil is used in perfumery, food-flavouring and cosmetic industries. However, presently, there are no reports available for the usage of vetiver roots agro-waste after oil extraction in nano-based products. Considering the concept of value-added products and green-chemistry approaches, synthesising cellulose nanoparticles (CNPs) using enzymatic treatment from agro-waste has emerged as a viable option. CNP's non-toxicity, biodegradability, and biocompatibility have sparked the industry's interest in its production. Therefore, in the present study, 3 enzymes, cellulase, pectinase, and viscozymes, were used for the green synthesis of CNP. The characterisation of CNP was done using techniques like DLS, FTIR, TEM, SEM, AFM, and TG/DTG, and cytotoxicity of CNP was studied in human skin cell-line (HaCaT) using MTT assay. Results show that CNPs synthesised using viscozyme and pectinase were of crystalline nature (2.0-3.0 nm) and cellulase were of fibres (40-60 nm). The FTIR confirmed that CNPs were devoid of lignin/hemicellulose. The AFM pictures revealed thick and thin nanoparticles with a variety of morphologies. The thermal stability of cellulose was higher compared to CNP. All the synthesised CNPs were crystaline, with a 60-70% crystallinity index. Furthermore, CNP did not show cytotoxic effect on HaCaT cells upto 500 µg/mL concentrations. In conclusion, pectinase and viscosyme may be used for synthesing cellulose-nanocrystals and cellulase enzyme for cellulose-nanofibers from the vetiver roots agro-waste. The findings revealed that Vetiveria zizanioides agro-waste-derived CNP is a sustainable material that can be used as a reinforcing agent/nanocarrier in textile and drug-delivery systems.

Optimistic influence of multi-metal tolerant Bacillus species on phytoremediation potential of Chrysopogon zizanioides on metal contaminated soil.

The current study investigated the plant growth promoting (PGP) characteristics of multi-metal-tolerant Bacillus cereus and their positive effect on the physiology, biomolecule substance, and phytoremediation ability of Chrysopogon zizanioides in metal-contaminated soil. The test soil sample was detrimentally contaminated by metals including Cd (31 mg kg-1), Zn (7696 mg kg-1), Pb (326 mg kg-1), Mn (2519 mg kg-1) and Cr (302 mg kg-1) that exceeded Indian standards. The multi-metal-tolerant B. cereus seemed to have superb PGP activities including fabrication of hydrogen cyanide, siderophore, Indole Acetic Acid, N2 fixation, as well as P solubilisation. Such multi-metal-tolerant B. cereus attributes can dramatically reduce or decontaminate metals in contaminated soils, and their PGP attributes significantly improve plant growth in contaminated soils. Hence, without (study I) and with (study II) the blending of B. cereus, this strain vastly enhances the growth and phytoremediation potency of C. zizanioides on metal contaminated soil. The results revealed that the physiological data, biomolecule components, and phytoremediation efficiency of C. zizanioides (Cr: 7.74, Cd: 12.15, Zn: 16.72, Pb: 11.47, and Mn: 14.52 mg g-1) seem to have been greatly effective in study II due to the metal solubilizing and PGP characteristics of B. cereus. This is a one-of-a-kind report on the effect of B. cereus's multi-metal tolerance and PGP characteristics on the development and phytoextraction effectiveness of C. zizanioides in metal-polluted soil.

Removal of atrazine from submerged soil using vetiver grass (Chrysopogon zizanioides L.).

The long-term widespread application of atrazine poses significant threats to the eco-environment and human health. To investigate the potential of vetiver (Chrysopogon zizanioides L.) for phytoremediation of the environmental media contaminated by atrazine, an indoor incubation experiment was conducted in submerged soil over 30 days. Results showed that the chlorophyll level of the vetiver was not significantly affected by exposure to atrazine. Vetiver could take up and accumulate atrazine from submerged soil and peaked around the 20th day with a concentration of 1.0 mg kg-1 in leaf. The metabolites Hydroxyatrazine (HA), deethylatrazine (DEA), Deisopropylatrazine (DIA), and didealkylatrazine (DDA) were detected in the leaf on the 30th day, indicating vetiver could degrade atrazine inside the leaf tissue. The atrazine removal rate in the vetiver planted and unplanted jars were 69.72 and 60.29%, respectively, indicating that 9.43% higher atrazine removal was achieved in the presence of vetiver (p < 0.05). The atrazine dissipation in the submerged soil followed first-order kinetics, the degradation constant was 0.066, and the half-life of atrazine dissipation was shortened by 6.86 days in the presence of vetiver. The present study suggests that vetiver can take up atrazine from submerged soil and accumulate in the leaf, which could then degrade in the leaf.Novelty statement: Although the fate of atrazine in agricultural soils has been extensively investigated through various experiments, little is known about the effect of vetiver grass on atrazine dissipation from submerged soil. With the identification of soil-leaf transportation and four metabolites in vetiver leaf and soils, significantly accelerated atrazine dissipation from the submerged soil was achieved in the presence of vetiver. Particularly, the formation of less toxic dealkylated products in the leaf indicated vetiver is a promising grass for atrazine removal from submerged soil.

Two P450 genes, CYP6SN3 and CYP306A1, involved in the growth and development of Chilo suppressalis and the lethal effect caused by vetiver grass.

Chilo suppressalis is a widely distributed pest occurring in nearly all paddy fields, which has developed high level resistance to different classes of insecticides. Vetiver grass has been identified as a dead-end trap plant for the alternative control of C. suppressalis. In this study, two cytochrome P450 monooxygenase (P450) genes, CsCYP6SN3 and CsCYP306A1, were identified and characterized, which are expressed at all developmental stages, with the highest expression in the midguts and fat bodies of 3rd instar larvae. Vetiver significantly inhibited the expression levels of CsCYP6SN3 and CsCYP306A1 in 3rd larvae after feeding. RNA interference showed that silencing CsCYP6SN3 and CsCYP306A1 genes dramatically reduced the pupation rate and pupa weight. Feeding on vetiver after silencing CsCYP6SN3 and CsCYP306A1 led to higher mortality compared with feeding on rice. In conclusion, these findings indicated that the expression levels of CsCYP6SN3 and CsCYP306A1 were associated with the lethal effect of vetiver against C. suppressalis larvae and functional knowledge about these two detoxification genes could provide new targets for agricultural pest control.

Combined effect of humic acid and vetiver grass on remediation of cadmium-polluted water.

Effective treatment of water pollution is an economic and social requirement globally. Humic acid (HA) is a popular mitigator for such waters. However, the combined effect of HA and restorative plants on cadmium (Cd) remediation is not well understood. Therefore, we experimented on Cd remediation using HA along with vetiver grass and HA-vetiver grass. We observed that vetiver grass effectively removed Cd at 15～30 mg/L. The accumulation capacity of the root was significantly higher than the shoots (P < 0.05), and Cd distribution followed the trend: cell wall > organelle > soluble substance (F1 > F2 > F3). The plant's accumulation capacity against 25 mg/L Cd was higher than for other treatments. The root accumulation capacity was much higher (702.3 mg/L) than those without added HA. However, upon adding 200 and 250 mg/L HA, the phytoremediation of Cd in the root and shoot significantly reduced (P < 0.05). Conversely, HA improved the Cd removal efficiency of the plants, notably at a lower HA concentration (150 mg/L). In addition, HA (especially at 150 mg/L) influences Cd distribution in vetiver cells (P < 0.05) and can significantly increase the proportion of Cd in the root cytoplasm. Consequently, a low HA concentration can significantly improve Cd accumulation in the vetiver, shorten the metal's bioremediation cycle, and improve the biological absorption efficiency.

Exploration of Cytotoxic Potential of Longifolene/Junipene Isolated from Chrysopogon zizanioides.

Since ancient times, Chrysopogon zizanioides has been utilized as a traditional medicinal plant for the treatment of numerous ailments, but neither its plant extract form nor its phytoconstituents have been fully explored. With this in mind, the present research was designed to isolate and structurally characterize one of its chemical constituents and evaluate its cytotoxic potential. Therefore, an ethanolic extract of roots was prepared and subjected to column chromatography using solvents of varying polarities. The obtained pure compound was characterized using various chromatographic and spectroscopic techniques such as high-performance liquid chromatography (HPLC), carbon and proton nuclear magnetic resonance (NMR), and liquid chromatography-mass spectroscopy (LC-MS) and identified as longifolene. This compound was evaluated for its cytotoxic potential using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on the prostate (DU-145), oral (SCC-29B) cancer cell line and normal kidney cell line (Vero cells), taking doxorubicin as a standard drug. The obtained outcomes revealed that longifolene possesses cytotoxic potential against both prostate (IC50 = 78.64 µg/mL) as well as oral (IC50 = 88.92 µg/mL) cancer cell lines with the least toxicity in healthy Vero cells (IC50 = 246.3 µg/mL) when compared to doxorubicin. Hence, this primary exploratory study of longifolene exhibited its cytotoxic potency along with wide safety margins in healthy cell lines, giving an idea that the compounds possess some ability to differentiate between cancerous cells and healthy cells.

Analysis of cell death in Bacillus subtilis caused by sesquiterpenes from Chrysopogon zizanioides (L.) Roberty.

Recently, the antibacterial effects of essential oils have been investigated in addition to their therapeutic purposes. Owing to their hydrophobic nature, they are thought to perturb the integrity of the bacterial cell membrane, leading to cell death. Against such antibiotic challenges, bacteria develop mechanisms for cell envelope stress responses (CESR). In Bacillus subtilis, a gram-positive sporulating soil bacterium, the extracytoplasmic function (ECF) sigma factor-mediated response system plays a pivotal role in CESR. Among them, σM is strongly involved in response to cell envelope stress, including a shortage of available bactoprenol. Vetiver essential oil, a product of Chrysopogon zizanioides (L.) Roberty root, is also known to possess bactericidal activity. σM was exclusively and strongly induced when the cells were exposed to Vetiver extract, and depletion of multi-ECF sigma factors (ΔsigM, ΔsigW, ΔsigX, and ΔsigV) enhanced sensitivity to it. From this quadruple mutant strain, the suppressor strains, which restored resistance to the bactericidal activity of Vetiver extract, emerged, although attempts to obtain resistant strains from the wild type did not succeed. Whole-genome resequencing of the suppressor strains and genetic analysis revealed inactivation of xseB or pnpA, which code for exodeoxyribonuclease or polynucleotide phosphorylase, respectively. This allowed the quadruple mutant strain to escape from cell death caused by Vetiver extract. Composition analysis suggested that the sesquiterpene, khusimol, might contribute to the bactericidal activity of the Vetiver extract.

Evaluation of Vetiver Volatile Compound Production under Aeroponic-Grown Conditions for the Perfume Industry.

Vetiver (Chrysopogon zizanioides (L.) Roberty) is a major tropical perfume crop. Access to its essential oil (EO)-filled roots is nevertheless cumbersome and land-damaging. This study, therefore, evaluated the potential of vetiver cultivation under soilless high-pressure aeroponics (HPA) for volatile organic compound (VOC) production. The VOC accumulation in the roots was investigated by transmission electron microscopy, and the composition of these VOCs was analyzed by gas chromatography coupled with mass spectrometry (GC/MS) after sampling by headspace solid-phase microextraction (HS-SPME). The HPA-grown plants were compared to plants that had been grown in potting soil and under axenic conditions. The HPA-grown plants were stunted, demonstrating less root biomass than the plants that had been grown in potting soil. The roots were slender, thinner, more tapered, and lacked the typical vetiver fragrance. HPA cultivation massively impaired the accumulation of the less-volatile hydrocarbon and oxygenated sesquiterpenes that normally form most of the VOCs. The axenic, tissue-cultured plants followed a similar and more exacerbated trend. Ultrastructural analyses revealed that the HPA conditions altered root ontogeny, whereby the roots contained fewer EO-accumulating cells and hosted fewer and more immature intracellular EO droplets. These preliminary results allowed to conclude that HPA-cultivated vetiver suffers from altered development and root ontology disorders that prevent EO accumulation.