Drought has serious effects on forests, especially semi-arid and arid forests, around the world. Zagros Forest in Iran has been severely affected by drought, which has led to the decline of the most common tree species, Persian oak (Quercus brantii). The objective of this study was to determine the effects of drought on the anatomical structure of Persian oak. Three healthy and three declined trees were sampled from each of two forest sites in Ilam Forest. Discs were cut at breast height, and three sapwood blocks were taken near the bark of each tree for sectioning. The anatomical characteristics measured included fiber length (FL), fiber wall thickness (FWT), number of axial parenchymal cells (NPC), ray number (RN), ray width (RW), and number of calcium oxalate crystals. Differences between healthy and declined trees were observed in the abundance of NPC and in RN, FL, and FWT, while no differences occurred in the number of oxalate crystals. The decline had uncertain effects on the FL of trees from sites A and B, which showed values of 700.5 and 837.3 μm compared with 592.7 and 919.6 μm in healthy trees. However, the decline resulted in an increase in the FWT of trees from sites A and B (9.33 and 11.53 μm) compared with healthy trees (5.23 and 9.56 μm). NPC, RN, and RW also increased in declined individuals from sites A and B (28.40 and 28.40 mm−1; 41.06 and 48.60 mm−1; 18.60 and 23.20 μm, respectively) compared with healthy trees (20.50 and 19.63 mm−2; 31.60 and 28.30 mm−2; 17.93 and 15.30 μm, respectively). Thus, drought caused measurable changes in the anatomical characteristics of declined trees compared with healthy trees.
Rigidoporus microporus, which causes white root rot disease (WRD) in Hevea brasiliensis, is a looming threat to rubber plantation in Malaysia. The current study was conducted to determine and evaluate the efficiency of fungal antagonists (Ascomycota) against R. microporus in rubber trees under laboratory and nursery conditions. A total of 35 fungal isolates established from the rubber tree rhizosphere soil were assessed for their antagonism against R. microporus by the dual culture technique. Trichoderma isolates can inhibit the radial growth of R. microporus by 75% or more in the dual culture test. Strains of T. asperellum, T. koningiopsis, T. spirale, and T. reesei were selected to assess the metabolites involved in their antifungal activity. Results indicated that T. asperellum exhibited an inhibitory effect against R. microporus in both volatile and non-volatile metabolite tests. All Trichoderma isolates were then tested for their ability in producing hydrolytic enzymes such as chitinase, cellulase and glucanase, indole acetic acid (IAA), siderophores production, and phosphate solubilization. From the positive results of the biochemical assays, T. asperellum and T. spirale were selected as the biocontrol candidates to be further tested in vivo against R. microporus. The nursery assessments revealed that rubber tree clone RRIM600 pretreated with only T. asperellum or with the combination of T. asperellum and T. spirale was able to reduce the disease severity index (DSI) and exert higher suppression of R. microporus compared to other pretreated samples, with the average DSI below 30%. Collectively, the present study demonstrates that T. asperellum represents a potential biocontrol agent that should be further explored to control R. microporus infection on rubber trees.
The application of miRNA mimic technology for silencing mature miRNA began in 2007. This technique originated from the discovery of the INDUCED BY PHOSPHATE STARVATION 1 (IPS1) gene, which was found to be a competitive mimic that prevents the cleavage of the targeted mRNA by miRNA inhibition at the post-transcriptional level. To date, various studies have been conducted to understand the molecular mimic mechanism and to improve the efficiency of this technology. As a result, several mimic tools have been developed: target mimicry (TM), short tandem target mimic (STTM), and molecular sponges (SPs). STTM is the most-developed tool due to its stability and effectiveness in decoying miRNA. This review discusses the application of STTM technology on the loss-of-function studies of miRNA and members from diverse plant species. A modified STTM approach for studying the function of miRNA with spatial-temporal expression under the control of specific promoters is further explored. STTM technology will enhance our understanding of the miRNA activity in plant-tissue-specific development and stress responses for applications in improving plant traits via miRNA regulation.
An aqueous extract of Syzygium cumini seeds was utilized to green synthesize titanium dioxide nanoparticles (TiO2 NPs). UV-Visible, DLS, FTIR, XRD, FESEM, TEM, SAED, EDAX, and photoluminescence spectroscopy techniques were employed to characterize the prepared TiO2 nanoparticles. The rutile crystal structure of TiO2 NPs was revealed by XRD study. The TEM and FESEM images of the TiO2 NPs revealed an average particle size of 50-100 nm. We employed EDAX to investigate the elemental compositions of TiO2 NPs. The O-Ti-O stretching bands appeared in the FTIR spectrum of TiO2 NPs at wavenumbers of 495 cm-1. The absorption edge peaks of TiO2 NPs were found in the UV-vis spectra at 397 nm. The MTT study revealed that TiO2 NPs effectively inhibited the growth of liver cancer Hep3 and Hep-G2 cells. The results of the corresponding fluorescent staining assays showed that TiO2 NPs significantly increased ROS generation, decreased MMP, and induced apoptosis in both liver cancer Hep3 and Hep-G2 cells. TiO2 nanoparticles lessened SOD, CAT, and GSH levels while augmenting MDA contents in Hep3 and Hep-G2 cells. In both Hep3 and Hep-G2 cells treated with TiO2 NPs, the Bax, CytC, p53, caspase-3, -8, and -9 expressions were remarkably augmented, while Bcl-2 expression was reduced. Overall, these findings revealed that formulated TiO2 NPs treatment considerably inhibited growth and triggered apoptosis in Hep3 and HepG2 cells.
Drought has been a major limiting factor for rice production. Drought yield QTLs (qDTYs; QTLs = quantitative trait loci) were pyramided into MRQ74 and MR219 to produce drought tolerant lines. In this study, new drought tolerant MRQ74 and MR219 pyramided lines (PLs) were evaluated under drought stress (RS) and non-stress (NS) conditions to evaluate the effects of different qDTYs combinations on morphological and agronomical traits. MRQ74 PLs having qDTY12.1 possessed the best root length (RL) under both RS and NS but the effect was only significant for MR219 PLs under RS. Some qDTYs combinations also found to have consistent effect on the same trait of both populations. PLs with only qDTY12.1 showed the highest grain yield (GY) under RS in both populations which means qDTY12.1 controlled RL and caused higher GY under drought condition. The interaction of major-effect qDTY12.1 with qDTY2.2 also shows significant effect on leaf rolling (LR) of both PL populations. These qDTYs proved to be beneficial in improving traits related to drought tolerance. Selected PLs with qDTY12.1 combinations also found to have better RL and root weight (RW) under RS. Improvement of morphological and agronomical traits led to higher GY of PLs. Therefore, qDTY12.1 either is present singly or in combination with other qDTYs was the best qDTY due to its consistent effect on morphological and agronomical traits and GY across populations under RS and NS.
Obesity has become a worldwide epidemic and its prevalence continues to increase at an alarming rate. It is considered a major risk factor for the development of several comorbidities, including type 2 diabetes, stroke, other cardiovascular diseases and even cancer. Conventional treatments for obesity, such as dietary interventions, exercise and pharmacotherapy, have proven to have limited effectiveness and are often associated with undesirable side effects. Therefore, there is a growing interest in exploring alternative therapeutic approaches. Nigella sativa (NS), a medicinal plant with multiple pharmacological properties, has gained attention due to its potential role in the treatment of obesity and its associated complications. The aim of this review is therefore to assess the effects of NS on obesity and its complications and to provide insights into the underlying mechanisms. From this review, NS appears to play a complementary or supportive role in the treatment of obesity and its complications. However, future studies are needed to verify the efficacy of NS in the treatment of obesity and its complications and to prove its safety so that it can be introduced in patients with obesity.
Cardiac diseases, such as myocardial infarction and heart failure, have become a major clinical problem globally. The accumulating data demonstrate that bioactive compounds with antioxidant and anti-inflammatory properties have favorable effects on clinical problems. Kaempferol is a flavonoid found in various plants; it has demonstrated cardioprotective properties in numerous cardiac injury models. This review aims to collate updated information regarding the effects of kaempferol on cardiac injury. Kaempferol improves cardiac function by alleviating myocardial apoptosis, fibrosis, oxidative stress, and inflammation while preserving mitochondrial function and calcium homeostasis. However, the mechanisms of action of its cardioprotective properties remain unclear; therefore, elucidating its action could provide insight into directions for future studies.
Impatiens glandulifera or Himalayan balsam is one of the most invasive weeds across Europe and can seriously reduce native plant diversity. It often forms continuous monocultures along river banks, but the mechanisms of this arrested succession are largely unknown. Here, we investigated the effect of arbuscular mycorrhizal (AM) fungi on balsam competitive ability with two native plant species, Plantago lanceolata and Holcus lanatus. We also studied how competition with Impatiens affects colonisation by foliar endophytes and mycorrhizas of two other co-occurring native species, Urtica dioica and Cirsium arvense. Mycorrhizal colonisation reduced balsam growth when the plants were grown singly, but appeared to have little effect when balsam experienced intra- or interspecific competition. Competition with balsam together with the addition of mycorrhizas had no effect on P. lanceolata biomass, suggesting that the fungi were beneficial to the latter, enabling it to compete effectively with balsam. However, this was not so with H. lanatus. Meanwhile, competition with Impatiens reduced endophyte numbers and mycorrhizal colonisation in U. dioica and C. arvense, leading to enhanced susceptibility of these plants to insect attack. Himalayan balsam is known to degrade soil fungal populations and can also reduce foliar beneficial fungi in neighbouring plants. This allows the plant to compete effectively with itself and other native species, thereby leading to the continuous monocultures.
Water deficit caused by drought is a significant threat to crop growth and production. Nitric oxide (NO), a water- and lipid-soluble free radical, plays an important role in cytoprotection. Apart from a few studies supporting the role of NO in drought responses, little is known about this pivotal molecular amendment in the regulation of abiotic stress signaling. In this review, we highlight the knowledge gaps in NO roles under drought stress and the technical challenges underlying NO detection and measurements, and we provide recommendations regarding potential avenues for future investigation. The modulation of NO production to alleviate abiotic stress disturbances in higher plants highlights the potential of genetic manipulation to influence NO metabolism as a tool with which plant fitness can be improved under adverse growth conditions.
Microorganisms are an important element in modeling sustainable agriculture. Their role in soil fertility and health is crucial in maintaining plants' growth, development, and yield. Further, microorganisms impact agriculture negatively through disease and emerging diseases. Deciphering the extensive functionality and structural diversity within the plant-soil microbiome is necessary to effectively deploy these organisms in sustainable agriculture. Although both the plant and soil microbiome have been studied over the decades, the efficiency of translating the laboratory and greenhouse findings to the field is largely dependent on the ability of the inoculants or beneficial microorganisms to colonize the soil and maintain stability in the ecosystem. Further, the plant and its environment are two variables that influence the plant and soil microbiome's diversity and structure. Thus, in recent years, researchers have looked into microbiome engineering that would enable them to modify the microbial communities in order to increase the efficiency and effectiveness of the inoculants. The engineering of environments is believed to support resistance to biotic and abiotic stressors, plant fitness, and productivity. Population characterization is crucial in microbiome manipulation, as well as in the identification of potential biofertilizers and biocontrol agents. Next-generation sequencing approaches that identify both culturable and non-culturable microbes associated with the soil and plant microbiome have expanded our knowledge in this area. Additionally, genome editing and multidisciplinary omics methods have provided scientists with a framework to engineer dependable and sustainable microbial communities that support high yield, disease resistance, nutrient cycling, and management of stressors. In this review, we present an overview of the role of beneficial microbes in sustainable agriculture, microbiome engineering, translation of this technology to the field, and the main approaches used by laboratories worldwide to study the plant-soil microbiome. These initiatives are important to the advancement of green technologies in agriculture.
Palm oil is a very important commodity which will be required well into the future. However, the consequences of growing oil palm (OP) are often detrimental to the environment and contribute to climate change. On the other hand, climate change stress will decrease the production of palm oil by causing mortality and ill health of OP, as well as reducing yields. Genetically modified OP (mOP) may be produced in the future to resist climate change stress, although it will take a long time to develop and introduce, if they are successfully produced at all. It is crucial to understand the benefits mOP may bring for resisting climate change and increasing the sustainability of the palm oil industry. This paper employs modeling of suitable climate for OP using the CLIMEX program in (a) Indonesia and Malaysia, which are the first and second largest growers of OP respectively, and (b) Thailand and Papua New Guinea, which are much smaller growers. It is useful to compare these countries in terms of future palm oil production and what benefits planting mOP may bring. Uniquely, narrative models are used in the current paper to determine how climate change will affect yields of conventional OP and mOP. The effect of climate change on the mortality of mOP is also determined for the first time. The gains from using mOP were moderate, but substantial, if compared to the current production of other continents or countries. This was especially the case for Indonesia and Malaysia. The development of mOP requires a realistic appreciation of what benefits may accrue.
Research has confirmed that the utilisation of Antarctic microorganisms, such as bacteria, yeasts and fungi, in the bioremediation of diesel may provide practical alternative approaches. However, to date there has been very little attention towards Antarctic microalgae as potential hydrocarbon degraders. Therefore, this study focused on the utilisation of an Antarctic microalga in the bioremediation of diesel. The studied microalgal strain was originally obtained from a freshwater ecosystem in Paradise Bay, western Antarctic Peninsula. When analysed in systems with and without aeration, this microalgal strain achieved a higher growth rate under aeration. To maintain the growth of this microalga optimally, a conventional one-factor-at a-time (OFAT) analysis was also conducted. Based on the optimized parameters, algal growth and diesel degradation performance was highest at pH 7.5 with 0.5 mg/L NaCl concentration and 0.5 g/L of NaNO3 as a nitrogen source. This currently unidentified microalga flourished in the presence of diesel, with maximum algal cell numbers on day 7 of incubation in the presence of 1% v/v diesel. Chlorophyll a, b and carotenoid contents of the culture were greatest on day 9 of incubation. The diesel degradation achieved was 64.5% of the original concentration after 9 days. Gas chromatography analysis showed the complete mineralisation of C7-C13 hydrocarbon chains. Fourier transform infrared spectroscopy analysis confirmed that strain WCY_AQ5_3 fully degraded the hydrocarbon with bioabsorption of the products. Morphological and molecular analyses suggested that this spherical, single-celled green microalga was a member of the genus Micractinium. The data obtained confirm that this microalga is a suitable candidate for further research into the degradation of diesel in Antarctica.
The bacterial soft rot and vascular wilt of sugar beet are the major diseases of sugar crops globally induced by Pectobacterium betavasculorum and P. carotovorum subsp. carotovorum (Pcc). The control of this bacterial disease is a severe problem, and only a few copper-based chemical bactericides are available for this disease. Because of the limitations of chemicals to control plant bacterial pathogens, the essential oils and extracts have been considered one of the best alternative strategies for their control. In this study, twenty-seven essential oils and twenty-nine plant extracts were extracted and evaluated for their antibacterial activities against Pectobacterium betavasculorum isolate C3, using the agar diffusion method at 0.01%, 0.1%, and 100% (v/v). Pure Pimpinella anisum L. oil exhibited the most anti-bacterial activity among three different concentrations of essential oils and extracts, followed by Thymus vulgaris L. oil and Rosa multiflora Thunb. extract. The efficacy of effective essential oils and extracts on Ic1 cultivar of sugar beet seeds germination and seedling growth in vivo also were tested. The seed germination of the Ic1 cultivar was inhibited at all the concentrations of essential oils used. Only extracts of Rosa multiflora Thunb., Brassica oleracea L., Lactuca serriola L., Salvia rosmarinus Spenn., Syzygium aromaticum (L.) Merr. and L.M.Perry, Eucalyptus globulus Labill., and essential oils of Ocmium basilicum L., Pimpinella anisum L., and Mentha× piperita L.L. in 0.1% concentration had no inhibition on seed germination and could improve seedling growth. This is the first report of the antibacterial activity of essential oils and extracts on Pectobacterium betavasculorum.
Eurycomanone and eurycomalactone are known quassinoids present in the roots and stems of Eurycoma longifolia. These compounds had been reported to have cytotoxic effects, however, their mechanism of action in a few cancer cell lines have yet to be elucidated. This study was aimed at investigating the anticancer effects and mechanisms of action of eurycomanone and eurycomalactone in cervical (HeLa), colorectal (HT29) and ovarian (A2780) cancer cell lines via Sulforhodamine B assay. Their mechanism of cell death was evaluated based on Hoechst 33342 assay and in silico molecular docking toward DHFR and TNF-α as putative protein targets. Eurycomanone and eurycomalactone exhibited in vitro anticancer effects manifesting IC50 values of 4.58 ± 0.090 µM and 1.60 ± 0.12 µM (HeLa), 1.22 ± 0.11 µM and 2.21 ± 0.049 µM (HT-29), and 1.37 ± 0.13 µM and 2.46 ± 0.081 µM (A2780), respectively. They induced apoptotic cancer cell death in dose- and time-dependent manners. Both eurycomanone and eurycomalactone were also predicted to have good inhibitory potential as demonstrated by the docking into TNF-α with binding affinity of -8.83 and -7.51 kcal/mol, respectively, as well as into DHFR with binding affinity results of -8.05 and -8.87 kcal/mol, respectively. These results support the evidence of eurycomanone and eurycomalactone as anticancer agents via apoptotic cell death mechanism that could be associated with TNF-α and DHFR inhibition as among possible protein targets.
The effect of fructose on γ-aminobutyric acid (GABA) content and its metabolic pathway in broccoli sprouts was investigated. The results demonstrated that the fructose treatment not only significantly increased the fresh weight, GABA, and glutamate contents in sprouts, but also promoted the activity of glutamic acid decarboxylase (GAD) and the expressions of BoGAD1 and BoGAD2. Meanwhile, fructose treatment inhibited the stem length of broccoli sprouts and enhanced the abscisic acid (ABA) production in comparison with the control. Ca2+, CaM contents, and BoCaM2 expression in broccoli sprouts were also stimulated after fructose treatment. Exogenous fructose increased inositol trisphosphate (IP3) content and activated the activity of phosphatidylinositol-specific phospholipase C (PI-PLC) and the expression of BoPLC2, contributing to Ca2+ influx into the cells. These results suggested that Ca2+ played an essential role in GABA enrichment under fructose treatment, which may be associated with GAD and PI-PLC.
Water scarcity negatively impacts oil palm production, necessitating the development of drought-tolerant varieties. This study aimed to develop molecular markers for oil palm breeding programs focused on drought tolerance. Genes associated with drought tolerance were selected, and single nucleotide polymorphism (SNP)-based markers were developed. Genomic DNA was successfully extracted from 17 oil palm varieties, and 20 primers out of 44 were effectively amplified. Screening with single-strand conformation polymorphism (SSCP) revealed an informative SNP marker from the choline monooxygenase (CMO) gene, exhibiting CC, CT, and TT genotypes. Notably, the oil palm variety La Mé showed the CT genotype, while Surat Thani 2 (Deli × La Mé) exhibited the CT and CC genotypes in a 1:1 ratio. Gene expression analysis confirmed the association of the CMO gene with drought tolerance in commercial oil palm varieties. The full-length CMO gene was 1308 bp long and shared sequence similarities with other plant species. However, amino acid sequence variations were observed compared with existing databases. These findings highlight the potential utility of the CMO marker for drought tolerance selection, specifically within the La Mé parent of oil palm Surat Thani 2 varieties, and strongly confirm the La Mé S5 population and Surat Thani 2 as drought-tolerant varieties.
The red palm weevil (RPW; Rhynchophorus ferrugineus Olivier (Coleoptera Curculionidae)) is an invasive insect pest that is difficult to manage due to its nature of infesting the host palm trees from within. A holistic, molecular-based approach to identify proteins that correlate with RPW infestation could give useful insights into the vital processes that are prevalent to the host's infestation response and identify the potential biomarkers for an early detection technique. Here, a shotgun proteomic analysis was performed on oil palm (Elaeis guineensis; OP) under untreated (control), wounding by drilling (wounded), and artificial larval infestation (infested) conditions at three different time points to characterise the RPW infestation response at three different stages. KEGG pathway enrichment analysis revealed many overlapping pathways between the control, wounded, and infested groups. Further analysis via literature searches narrowed down biologically relevant proteins into categories, which were photosynthesis, growth, and stress response. Overall, the patterns of protein expression suggested abscisic acid (ABA) hormone signalling to be the primary driver of insect herbivory response. Interspecies molecular docking analysis between RPW ligands and OP receptor proteins provided putative interactions that result in ABA signalling activation. Seven proteins were selected as candidate biomarkers for early infestation detection based on their relevance and association with ABA signalling. The MS data are available via ProteomeXchange with identifier PXD028986. This study provided a deeper insight into the mechanism of stress response in OP in order to develop a novel detection method or improve crop management.
Ficus is one of the largest genera in the plant kingdom that belongs to the Moraceae family. This review aimed to summarize the medicinal uses, phytochemistry, and pharmacological actions of two major species from this genus, namely Ficus benghalensis and Ficus religiosa. These species can be found abundantly in most Asian countries, including Malaysia. The chemical analysis report has shown that Ficus species contained a wide range of phytoconstituents, including phenols, flavonoids, alkaloids, tannins, saponins, terpenoids, glycosides, sugar, protein, essential and volatile oils, and steroids. Existing studies on the pharmacological functions have revealed that the observed Ficus species possessed a broad range of biological properties, including antioxidants, antidiabetic, anti-inflammatory, anticancer, antitumor and antiproliferative, antimutagenic, antimicrobial, anti-helminthic, hepatoprotective, wound healing, anticoagulant, immunomodulatory activities, antistress, toxicity studies, and mosquitocidal effects. Apart from the plant parts and their extracts, the endophytes residing in these host plants were discussed as well. This study also includes the recent applications of the Ficus species and their plant parts, mainly in the nanotechnology field. Various search engines and databases were used to obtain the scientific findings, including Google Scholar, ScienceDirect, PMC, Research Gate, and Scopus. Overall, the review discusses the therapeutic potentials discovered in recent times and highlights the research gaps for prospective research work.
Rice is a worldwide staple food and heavy metal contamination is often reported in rice production. Heavy metal can originate from natural sources or be present through anthropogenic contamination. Therefore, this review summarizes the current status of heavy metal contamination in paddy soil and plants, highlighting the mechanism of uptake, bioaccumulation, and health risk assessment. A scoping search employing Google Scholar, Science Direct, Research Gate, Scopus, and Wiley Online was carried out to build up the review using the following keywords: heavy metals, absorption, translocation, accumulation, uptake, biotransformation, rice, and human risk with no restrictions being placed on the year of study. Cadmium (Cd), arsenic (As), and lead (Pb) have been identified as the most prevalent metals in rice cultivation. Mining and irrigation activities are primary sources, but chemical fertilizer and pesticide usage also contribute to heavy metal contamination of paddy soil worldwide. Further to their adverse effect on the paddy ecosystem by reducing the soil fertility and grain yield, heavy metal contamination represents a risk to human health. An in-depth discussion is further offered on health risk assessments by quantitative measurement to identify potential risk towards heavy metal exposure via rice consumption, which consisted of in vitro digestion models through a vital ingestion portion of rice.
For centuries, Azadirachta indica or neem has been utilized as a primary source of medicine due to its antimicrobial, larvacidal, antimalarial and antifungal properties. Recently, its potential as an effective biopesticide has garnered attention, especially towards efficient and continuous production of its bioactive compounds. The present study investigated the effect of the plant growth regulators (PGRs) thiadiazuron (TDZ) and 2,4-dichlorophenoxyacetic acid (2,4-D) on the induction of colored callus formation and subsequent accumulation of azadirachtin (AZA) in A. indica. An efficient protocol was established for micropropagation and colored callus production of this species, followed by quantification of AZA (a mixture of azadirachtin A and B) and its safety assessment. For induction of the callus, leaf and petiole explants obtained from a young growing neem plant were excised and cultured on Murashige and Skoog (MS) medium supplemented with TDZ (0.2-0.6 mg L-1) and 2,4-D (0.2-0.6 mg L-1), either applied singly or in combination. Callus was successfully induced from both explant types at different rates, where media with 0.6 mg L-1 of TDZ resulted in the highest fresh weight (3.38 ± 0.08 g). In general, media with a single hormone (particularly TDZ) was more effective in producing a high mass of callus compared to combined PGRs. A culture duration of six weeks resulted in the production of green, brown and cream colored callus. The highest callus weight and accumulation of AZA was recorded in green callus (214.53 ± 33.63 mg g-1 dry weight (DW)) induced using TDZ. On the other hand, small amounts of AZA were detected in both brown and cream callus. Further experimentation indicated that the green callus with the highest AZA was found to be non-toxic (LC50 at 4606 µg mL-1) to the zebrafish animal model. These results suggested that the addition of different PGRs during in vitro culture could prominently affect callus and secondary metabolite production and can further be manipulated as a sustainable method for the production of a natural and environmentally friendly pesticide.