Unraveling the potential of hydrogen sulfide as a signaling molecule for plant development and environmental stress responses: A state-of-the-art review.

Over the past decade, a plethora of research has illuminated the multifaceted roles of hydrogen sulfide (H2S) in plant physiology. This gaseous molecule, endowed with signaling properties, plays a pivotal role in mitigating metal-induced oxidative stress and strengthening the plant's ability to withstand harsh environmental conditions. It fulfils several functions in regulating plant development while ameliorating the adverse impacts of environmental stressors. The intricate connections among nitric oxide (NO), hydrogen peroxide (H2O2), and hydrogen sulfide in plant signaling, along with their involvement in direct chemical processes, are contributory in facilitating post-translational modifications (PTMs) of proteins that target cysteine residues. Therefore, the present review offers a comprehensive overview of sulfur metabolic pathways regulated by hydrogen sulfide, alongside the advancements in understanding its biological activities in plant growth and development. Specifically, it centres on the physiological roles of H2S in responding to environmental stressors to explore the crucial significance of different exogenously administered hydrogen sulfide donors in mitigating the toxicity associated with heavy metals (HMs). These donors are of utmost importance in facilitating the plant development, stabilization of physiological and biochemical processes, and augmentation of anti-oxidative metabolic pathways. Furthermore, the review delves into the interaction between different growth regulators and endogenous hydrogen sulfide and their contributions to mitigating metal-induced phytotoxicity.

Hydrogen gas: A new fresh keeping agent of perishable horticultural products.

Hydrogen gas (H2), a gaseous signaling molecule, is involved in plant growth and development. This review collates emerging evidence to show that H2 regulates the postharvest senescence of horticultural products through critical biochemical processes, including the improvement of antioxidant systems, the activation of cell wall metabolism, the promotion of energy metabolism, the inhibition of ethylene biosynthesis and the regulation of bacterial communities. Additionally, the interactions between H2 and other signaling molecules are also discussed. This paper presents the current status of H2 research in terms of its biological effects and safety in postharvest products by combining the research results on the molecular mechanisms of biological effects and H2 signaling. The action mechanism of H2 for postharvest preservation is also proposed, and it reflects the complexity and diversity of the pathways involved. Furthermore, a growing body of evidence has found a large number of downstream pathways or targets for the medical effects of H2. Therefore, the scientific and practical aspects of H2 biology are proposed for the postharvest preservation of horticultural products.

Ethylene-induced potassium transporter AcKUP2 gene is involved in kiwifruit postharvest ripening.

BACKGROUND: Potassium (K) is important in the regulation of plant growth and development. It is the most abundant mineral element in kiwifruit, and its content increases during fruit ripening. However, how K+ transporter works in kiwifruit postharvest maturation is not yet clear. RESULTS: Here, 12 K+ transporter KT/HAK/KUP genes, AcKUP1 ~ AcKUP12, were isolated from kiwifruit, and their phylogeny, genomic structure, chromosomal location, protein properties, conserved motifs and cis-acting elements were analysed. Transcription analysis revealed that AcKUP2 expression increased rapidly and was maintained at a high level during postharvest maturation, consistent with the trend of K content; AcKUP2 expression was induced by ethylene, suggesting that AcKUP2 might play a role in ripening. Fluorescence microscopy showed that AcKUP2 is localised in the plasma membrane. Cis-elements, including DER or ethylene response element (ERE) responsive to ethylene, were found in the AcKUP2 promoter sequence, and ethylene significantly enhanced the AcKUP2 promoter activity. Furthermore, we verified that AcERF15, an ethylene response factor, directly binds to the AcKUP2 promoter to promote its expression. Thus, AcKUP2 may be an important potassium transporter gene which involved in ethylene-regulated kiwifruit postharvest ripening. CONCLUSIONS: Therefore, our study establishes the first genome-wide analysis of the kiwifruit KT/HAK/KUP gene family and provides valuable information for understanding the function of the KT/HAK/KUP genes in kiwifruit postharvest ripening.

Nitric oxide, gravity response, and a unified schematic of plant signaling.

Plant signaling components are often involved in numerous processes. Calcium, reactive oxygen species, and other signaling molecules are essential to normal biotic and abiotic responses. Yet, the summation of these components is integrated to produce a specific response despite their involvement in a myriad of response cascades. In the response to gravity, the role of many of these individual components has been studied, but a specific sequence of signals has not yet been assembled into a cohesive schematic of gravity response signaling. Herein, we provide a review of existing knowledge of gravity response and differential protein and gene regulation induced by the absence of gravity stimulus aboard the International Space Station and propose an integrated theoretical schematic of gravity response incorporating that information. Recent developments in the role of nitric oxide in gravity signaling provided some of the final contextual pillars for the assembly of the model, where nitric oxide and the role of cysteine S-nitrosation may be central to the gravity response. The proposed schematic accounts for the known responses to reorientation with respect to gravity in roots-the most well studied gravitropic plant tissue-and is supported by the extensive evolutionary conservation of regulatory amino acids within protein components of the signaling schematic. The identification of a role of nitric oxide in regulating the TIR1 auxin receptor is indicative of the broader relevance of the schematic in studying a multitude of environmental and stress responses. Finally, there are several experimental approaches that are highlighted as essential to the further study and validation of this schematic.

NB-LRR-encoding genes conferring susceptibility to organophosphate pesticides in sorghum.

Organophosphate is the commonly used pesticide to control pest outbreak, such as those by aphids in many crops. Despite its wide use, however, necrotic lesion and/or cell death following the application of organophosphate pesticides has been reported to occur in several species. To understand this phenomenon, called organophosphate pesticide sensitivity (OPS) in sorghum, we conducted QTL analysis in a recombinant inbred line derived from the Japanese cultivar NOG, which exhibits OPS. Mapping OPS in this population identified a prominent QTL on chromosome 5, which corresponded to Organophosphate-Sensitive Reaction (OSR) reported previously in other mapping populations. The OSR locus included a cluster of three genes potentially encoding nucleotide-binding leucine-rich repeat (NB-LRR, NLR) proteins, among which NLR-C was considered to be responsible for OPS in a dominant fashion. NLR-C was functional in NOG, whereas the other resistant parent, BTx623, had a null mutation caused by the deletion of promoter sequences. Our finding of OSR as a dominant trait is important not only in understanding the diversified role of NB-LRR proteins in cereals but also in securing sorghum breeding free from OPS.

Belowground fungal volatiles perception in okra (Abelmoschus esculentus) facilitates plant growth under biotic stress.

Microbial volatile organic compounds (mVOCs) have great potential in plant ecophysiology, yet the role of belowground VOCs in plant stress management remains largely obscure. Analysis of biocontrol producing VOCs into the soil allow detailed insight into their interaction with soil borne pathogens for plant disease management. A root interaction trial was set up to evaluate the effects of VOCs released from Trichoderma viride BHU-V2 on soil-inhabiting fungal pathogen and okra plant growth. VOCs released into soil by T. viride BHU-V2 inhibited the growth of collar rot pathogen, Sclerotium rolfsii. Okra plants responded to VOCs by increasing the root growth (lateral roots) and total biomass content. VOCs exposure increased defense mechanism in okra plants by inducing different enzyme activities i.e. chitinase (0.89 fold), ß-1,3-glucanase (0.42 fold), peroxidase (0.29 fold), polyphenol oxidase (0.33 fold) and phenylalanine lyase (0.7 fold) when inoculated with S. rolfsii. In addition, T. viride BHU-V2 secreted VOCs reduced lipid peroxidation and cell death in okra plants under pathogen inoculated condition. GC/MS analysis of VOCs blend revealed that T. viride BHU-V2 produced more number of antifungal compounds in soil medium as compared to standard medium. Based on the above observations it is concluded that okra plant roots perceive VOCs secreted by T. viride BHU-V2 into soil that involved in induction of plant defense system against S. rolfsii. In an ecological context, the findings reveal that belowground microbial VOCs may play an important role in stress signaling mechanism to interact with plants.

Optogenetic control of plant growth by a microbial rhodopsin.

While rhodopsin-based optogenetics has revolutionized neuroscience1,2, poor expression of opsins and the absence of the essential cofactor all-trans-retinal has complicated the application of rhodopsins in plants. Here, we demonstrate retinal production in plants and improved rhodopsin targeting for green light manipulation of plant cells using the Guillardia theta light-gated anion channelrhodopsin GtACR13. Green light induces a massive increase in anion permeability and pronounced membrane potential changes when GtACR1 is expressed, enabling non-invasive manipulation of plant growth and leaf development. Using light-driven anion loss, we could mimic drought conditions and bring about leaf wilting despite sufficient water supply. Expressed in pollen tubes, global GtACR1 activation triggers membrane potential depolarizations due to large anion currents. While global illumination was associated with a reversible growth arrest, local GtACR1 activation at the flanks of the apical dome steers growth direction away from the side with increased anion conductance. These results suggest a crucial role of anion permeability for the guidance of pollen tube tip growth. This plant optogenetic approach could be expanded to create an entire pallet of rhodopsin-based tools4, greatly facilitating dissection of plant ion-signalling pathways.

GENOMES UNCOUPLED1-independent retrograde signaling targets the ethylene pathway to repress photomorphogenesis.

When germinating in the light, Arabidopsis (Arabidopsis thaliana) seedlings undergo photomorphogenic development, characterized by short hypocotyls, greening, and expanded cotyledons. Stressed chloroplasts emit retrograde signals to the nucleus that induce developmental responses and repress photomorphogenesis. The nuclear targets of these retrograde signals are not yet fully known. Here, we show that lincomycin-treated seedlings (which lack developed chloroplasts) show strong phenotypic similarities to seedlings treated with ethylene (ET) precursor 1-aminocyclopropane-1-carboxylic acid, as both signals inhibit cotyledon separation in the light. We show that the lincomycin-induced phenotype partly requires a functioning ET signaling pathway, but could not detect increased ET emissions in response to the lincomycin treatment. The two treatments show overlap in upregulated gene transcripts, downstream of transcription factors ETHYLENE INSENSITIVE3 and EIN3-LIKE1. The induction of the ET signaling pathway is triggered by an unknown retrograde signal acting independently of GENOMES UNCOUPLED1. Our data show how two apparently different stress responses converge to optimize photomorphogenesis.

Synthesis and Biological Activity of Brassinosteroid Analogues with a Nitrogen-Containing Side Chain.

Brassinosteroids are a class of plant hormones that regulate a broad range of physiological processes such as plant growth, development and immunity, including the suppression of biotic and abiotic stresses. In this paper, we report the synthesis of new brassinosteroid analogues with a nitrogen-containing side chain and their biological activity on Arabidopis thaliana. Based on molecular docking experiments, two groups of brassinosteroid analogues were prepared with short and long side chains in order to study the impact of side chain length on plants. The derivatives with a short side chain were prepared with amide, amine and ammonium functional groups. The derivatives with a long side chain were synthesized using amide and ammonium functional groups. A total of 25 new brassinosteroid analogues were prepared. All 25 compounds were tested in an Arabidopsis root sensitivity bioassay and cytotoxicity screening. The synthesized substances showed no significant inhibitory activity compared to natural 24-epibrassinolide. In contrast, in low concentration, several compounds (8a, 8b, 8e, 16e, 22a and 22e) showed interesting growth-promoting activity. The cytotoxicity assay showed no toxicity of the prepared compounds on cancer and normal cell lines.

Chemical Compounds and Biologic Activities: A Review of Cedrela Genus.

The genus Cedrela P. Browne, which belongs to the Meliaceae family, has eighteen species. Trees of this genus are of economic interest due to wood quality, as well as being the focus of studies because of relevant biologic activities as in other Meliaceae species. These activities are mainly related to limonoids, a characteristic class of compounds in this family. Therefore, the aim of this review is to perform a survey of the citations in the literature on the Cedrela genus species. Articles were found on quantitative and qualitative phytochemical studies of the Cedrela species, revealing the chemical compounds identified, such as aliphatics acid and alcohol, flavonoids, tocopherol, monoterpenes, sesquiterpenes, triterpenes, cycloartanes, steroids, and limonoids. Although some activities were tested, the majority of studies focused on the insecticidal, antifeedant, or insect growth inhibitor activities of this genus. Nonetheless, the most promising activities were related to their antimalarial and antitripanocidal effects, although further investigations are still needed.

Plant-Derived Smoke Affects Biochemical Mechanism on Plant Growth and Seed Germination.

The role of plant-derived smoke, which is changed in mineral-nutrient status, in enhancing germination and post-germination was effectively established. The majority of plant species positively respond to plant-derived smoke in the enhancement of seed germination and plant growth. The stimulatory effect of plant-derived smoke on normally growing and stressed plants may help to reduce economic and human resources, which validates its candidature as a biostimulant. Plant-derived smoke potentially facilitates the early harvest and increases crop productivity. Karrikins and cyanohydrin are the active compound in plant-derived smoke. In this review, data from the latest research explaining the effect of plant-derived smoke on morphological, physiological, biochemical, and molecular responses of plants are presented. The pathway for reception and interaction of compounds of plant-derived smoke at the cellular and molecular level of plant is described and discussed.

Increased production of valuable secondary products in plants by leaf applied radiation-processed polysaccharides.

Oligosaccharides derived through irradiation of polysaccharides act as efficient plant elicitors and stimulate responses associated with primary as well as secondary metabolic pathways in plants. Reduced molecular weight together with the structural rearrangement, induce plant growth promotion activity in the polysaccharides after irradiation. In addition to the increased activities of different enzymes involved in photosynthesis and nutrient assimilation, various secondary metabolism enzymes are up-regulated by the leaf-applied oligomers. Oligosaccharide-induced elicitation of different signal transduction cascades leads to the increased biosynthesis of valuable secondary metabolism products in plants. The present review presents a comprehensive approach regarding the irradiation-induced structural changes and molecular weight reduction in polysaccharides and their role in increasing the production of economically valuable secondary products in various medicinally important plants. This review also encompasses the role of oligosaccharides in regulation of plant growth and enzyme activities as well as the signal transduction mechanism involved in the elicitation of secondary metabolites.

Phytoremediation effect of Medicago sativa colonized by Piriformospora indica in the phenanthrene and cadmium co-contaminated soil.

BACKGROUND: The coexistence of polycyclic aromatic hydrocarbons (PAHs) and heavy metals has deleterious effects on environmental quality. Few reports have studied the mechanisms of plant inoculation with Piriformospora indica to remediate PAH-metal co-contaminated soil by analyzing the chemical speciation of the contaminants. This study investigated the influence of the inoculation of Medicago sativa with P. indica to remediate soil co-contaminated with phenanthrene (a kind of PAH) and cadmium (a heavy metal) by analyzing plant growth, physiological parameters and chemical speciation in rhizosphere and nonrhizosphere soils. RESULTS: The presence of P. indica significantly increased plant tolerance, chlorophyll a, chlorophyll b, maximum quantum efficiency of PSII photochemistry and electron transport rate values in phenanthrene- and/or cadmium-contaminated soil. P. indica inoculation in M. sativa roots increased fluorescein diacetate activities in soils contaminated with phenanthrene, cadmium or both, especially in the nonrhizosphere. The presence of phenanthrene prevented the inoculated plant from accumulating cadmium to some extent, whereas the presence of cadmium did not prevent the degradation of phenanthrene in either the rhizosphere or the nonrhizosphere after P. indica colonization. Although the low bioavailability of cadmium in the rhizosphere restricted its transportation into the stem, P. indica colonization in plants effectively increased cadmium accumulation in roots in soil co-contaminated with cadmium and phenanthrene. CONCLUSIONS: In conclusion, this work provides a theoretical basis for the use of P. indica combined with M. sativa for the remediation of PAH-metal co-contaminated soil.

Size effect of iron (III) oxide nanomaterials on the growth, and their uptake and translocation in common wheat (Triticum aestivum L.).

Nanomaterials (NMs) have emerged in the last decades and are used in many disciplines such as industry, material sciences, biomedicine, biotechnology, bioenergy, and agriculture. The size of the NMs is a critical factor that affects NMs' integration and transfer into the biological systems. Therefore, this study aims at investigating the effect of NMs-size on i) plant growth and physiology, and ii) NMs uptake and translocation in plant tissues. For these purposes, iron (III) oxide (Fe2O3) NMs with varied sizes, 8-10, 20-40, and 30-50 nm, have been applied to wheat plants in a hydroponic system. Results showed that Fe2O3 NMs enhanced root length, plant height, biomass, and chlorophyll content of wheat. Confocal microscopy analysis indicated that Fe2O3 NMs cause injury in root-tip cells without a visible toxic symptom. Vibrating sample magnetometer (VSM), and inductively coupled plasma-mass spectroscopy (ICP-MS) analyses of leaf tissues revealed that all tested NMs were up taken by wheat plant and translocated to the leaves. Iron content was found to be dramatically increased in NMs-treated plant tissues, which possibly contributed to the growth enhancement. Experiments confirmed that Fe2O3 NMs with 20-40 nm size is much more efficient in plant growth compared to those with 8-10 and 30-50 nm size. Overall, Fe2O3 NMs with 20-40 nm in size could be proposed as a nano-fertilizer for agricultural applications. On the other hand, the translocation of NMs in the wheat plant requires further investigation of their effects on the end users.

CRK2 and C-terminal Phosphorylation of NADPH Oxidase RBOHD Regulate Reactive Oxygen Species Production in Arabidopsis.

Reactive oxygen species (ROS) are important messengers in eukaryotic organisms, and their production is tightly controlled. Active extracellular ROS production by NADPH oxidases in plants is triggered by receptor-like protein kinase-dependent signaling networks. Here, we show that CYSTEINE-RICH RLK2 (CRK2) kinase activity is required for plant growth and CRK2 exists in a preformed complex with the NADPH oxidase RESPIRATORY BURST OXIDASE HOMOLOG D (RBOHD) in Arabidopsis (Arabidopsis thaliana). Functional CRK2 is required for the full elicitor-induced ROS burst, and consequently the crk2 mutant is impaired in defense against the bacterial pathogen Pseudomonas syringae pv tomato DC3000. Our work demonstrates that CRK2 regulates plant innate immunity. We identified in vitro CRK2-dependent phosphorylation sites in the C-terminal region of RBOHD. Phosphorylation of S703 RBOHD is enhanced upon flg22 treatment, and substitution of S703 with Ala reduced ROS production in Arabidopsis. Phylogenetic analysis suggests that phospho-sites in the C-terminal region of RBOHD are conserved throughout the plant lineage and between animals and plants. We propose that regulation of NADPH oxidase activity by phosphorylation of the C-terminal region might be an ancient mechanism and that CRK2 is an important element in regulating microbe-associated molecular pattern-triggered ROS production.

Remediation of hexavalent chromium contaminated water through zero-valent iron nanoparticles and effects on tomato plant growth performance.

Contaminated water with hexavalent chromium Cr(VI) is a serious environmental problem. This study aimed to evaluate the Cr(VI) removal by zero valent iron nanoparticles (nZVI) reduction process and the impact of Cr(VI), nZVI and combined treatment with nZVI and Cr(VI) on tomato growth performance. To evaluate the Cr(VI) toxic effect on germination capability, seeds were exposed to increasing Cr(VI) concentrations up to 1000 mg L-1. The inhibition of seed germination and the decrease of hypocotyl and root length started from Cr(VI) 5 mg L-1. Under treatment with Cr(VI) + nZVI 5 mg L-1, seed germination, hypocotyl and root length resulted significantly higher compared to Cr(VI) 5 mg L-1 treatment. The impact of only nZVI was investigated on chlorophyll and carotenoid in leaves; iron levels in leaves, roots, fruits and soil; carotenoid, fat-soluble vitamin and nicotianamine in mature fruits. A significant increase of leaf chlorophyll and carotenoids was observed after nZVI 5 mg L-1 treatment compared to controls. No significant variations were observed in carotenoids, fat-soluble vitamins and nicotianamine levels after treatment with nZVI 5 mg L-1 in mature fruits. For their ability to reduce Cr(VI) and to stimulate tomato growth, nZVI might to be considered as alternative for remediation purposes.

HpaXpm, a novel harpin of Xanthomonas phaseoli pv. manihotis, acts as an elicitor with high thermal stability, reduces disease, and promotes plant growth.

BACKGROUND: Harpins are proteins secreted by the type III secretion system of Gram-negative bacteria during pathogen-plant interactions that can act as elicitors, stimulating defense and plant growth in many types of non-host plants. Harpin-treated plants have higher resistance, quality and yields and, therefore, harpin proteins may potentially have many valuable agricultural applications. Harpins are characterized by high thermal stability at 100 °C. However, it is unknown whether harpins are still active at temperatures above 100 °C or whether different temperatures affect the activity of the harpin protein in different ways. The mechanism responsible for the heat stability of harpins is also unknown. RESULTS: We identified a novel harpin, HpaXpm, from the cassava blight bacteria Xanthomonas phaseoli pv. manihotis HNHK. The predicted secondary structure and 3-D structure indicated that the HpaXpm protein has two ß-strand domains and two major α-helical domains located at the N- and C-terminal regions, respectively. A phylogenetic tree generated using the maximum likelihood method grouped HpaXpm in clade I of the Hpa1 group along with harpins produced by other Xanthomonas spp. (i.e., HpaG-Xag, HpaG-Xcm, Hpa1-Xac, and Hpa1Xm). Phenotypic assays showed that HpaXpm induced the hypersensitive response (HR), defense responses, and growth promotion in non-host plants more effectively than Hp1Xoo (X. oryzae pv. oryzae). Quantitative real-time PCR analysis indicated that HpaXpm proteins subjected to heat treatments at 100 °C, 150 °C, or 200 °C were still able to stimulate the expression of function-related genes (i.e., the HR marker genes Hin1 and Hsr203J, the defense-related gene NPR1, and the plant growth enhancement-related gene NtEXP6); however, the ability of heat-treated HpaXpm to induce HR was different at different temperatures. CONCLUSIONS: These findings add a new member to the harpin family. HpaXpm is heat-stable up to 200 °C and is able to stimulate powerful beneficial biological functions that could potentially be more valuable for agricultural applications than those stimulated by Hpa1Xoo. We hypothesize that the extreme heat resistance of HpaXpm is because the structure of harpin is very stable and, therefore, the HpaXpm structure is less affected by temperature.

Effect of plant-growth-promoting rhizobacteria on phytoremediation efficiency of Scirpus triqueter in pyrene-Ni co-contaminated soils.

The aim of this study was to investigate whether the plant-growth-promoting rhizobacteria (PGPR) could enhance phytoremediation efficiency of Scirpus triqueter (S.triqueter) in the pyrene-Ni co-contaminated soil. We also expected to reveal the possible mechanism for the affected phytoremediation efficiency induced by PGPR. We used three kinds of contaminated soils (Ni-contaminated soil, pyrene-contaminated soil and pyrene-Ni co-contaminated soil) to conduct this pot study. After harvest, plants growth indicators, polyphenol oxidase (PPO) activity and soil microbial community structure of each treatment were investigated to explain the different dissipation rates of pyrene and removal rates of Ni between treatments with and without PGPR. The results showed that PGPR-inoculated S. triqueter increased dissipation rates of pyrene and removal rates of Ni in all three contaminated soils, among which Ni removal rates in Ni single contaminated soil was elevated most significantly, from 0.895‰ to 8.8‰, increasing nearly 9 folds. However, Ni removal rate efficiency in co-contaminated soil was weakened because more toxic and complicated co-contaminated soil restrained plant growth and Ni absorption. We also observed that co-contamination harmed the soil microbial community more severely than that in single pyrene or Ni contaminated soil through phospholipid fatty acids analysis. Furthermore, dissipation rates of pyrene and removal rates of Ni were found positively correlated to the PPO activity and the abundance of branched and saturated fatty acids reflected by Pearson correlation analysis.

Detection of oil pollution impacts on vegetation using multifrequency SAR, multispectral images with fuzzy forest and random forest methods.

Oil pollution harms terrestrial ecosystems. There is an urgent requirement to improve on existing methods for detecting, mapping and establishing the precise extent of oil-impacted and oil-free vegetation. This is needed to quantify existing spill extents, formulate effective remediation strategies and to enable effective pipeline monitoring strategies to identify leakages at an early stage. An effective oil spill detection algorithm based on optical image spectral responses can benefit immensely from the inclusion of multi-frequency Synthetic Aperture Radar (SAR) data, especially when the effect of multi-collinearity is sufficiently reduced. This study compared the Fuzzy Forest (FF) and Random Forest (RF) methods in detecting and mapping oil-impacted vegetation from a post spill multispectral optical sentinel 2 image and multifrequency C and X Band Sentinel - 1, COSMO Skymed and TanDEM-X SAR images. FF and RF classifiers were employed to discriminate oil-spill impacted and oil-free vegetation in a study area in Nigeria. Fuzzy Forest uses specific functions for the selection and use of uncorrelated variables in the classification process to yield an improved result. This method proved an efficient variable selection technique addressing the effects of high dimensionality and multi-collinearity, as the optimization and use of different SAR and optical image variables generated more accurate results than the RF algorithm in densely vegetated areas. An Overall Accuracy (OA) of 75% was obtained for the dense (Tree Cover Area) vegetation, while cropland and grassland areas had 59.4% and 65% OA respectively. However, RF performed better in Cropland areas with OA = 75% when SAR-optical image variables were used for classification, while both methods performed equally well in Grassland areas with OA = 65%. Similarly, significant backscatter differences (P < 0.005) were observed in the C-Band backscatter sample mean of polluted and oil-free TCA, while strong linear associations existed between LAI and backscatter in grassland and TCA. This study demonstrates that SAR based monitoring of petroleum hydrocarbon impacts on vegetation is feasible and has high potential for establishing oil-impacted areas and oil pipeline monitoring.

Establishment of Actinorhizal Symbiosis in Response to Ethylene, Salicylic Acid, and Jasmonate.

Phytohormones play a crucial role in regulating plant developmental processes. Among them, ethylene and jasmonate are known to be involved in plant defense responses to a wide range of biotic stresses as their levels increase with pathogen infection. In addition, these two phytohormones have been shown to inhibit plant nodulation in legumes. Here, exogenous salicylic acid (SA), jasmonate acid (JA), and ethephon (ET) were applied to the root system of Casuarina glauca plants before Frankia inoculation, in order to analyze their effects on the establishment of actinorhizal symbiosis. This protocol further describes how to identify putative ortholog genes involved in ethylene and jasmonate biosynthesis and/or signaling pathways in plant, using the Arabidopsis Information Resource (TAIR), Legume Information System (LIS), and Genevestigator databases. The expression of these genes in response to the bacterium Frankia was analyzed using the gene atlas for Casuarina-Frankia symbiosis (SESAM web site).