Biocatalytic Production of Aldehydes: Exploring the Potential of Lathyrus cicera Amine Oxidase.

Aldehydes are a class of carbonyl compounds widely used as intermediates in the pharmaceutical, cosmetic and food industries. To date, there are few fully enzymatic methods for synthesizing these highly reactive chemicals. In the present work, we explore the biocatalytic potential of an amino oxidase extracted from the etiolated shoots of Lathyrus cicera for the synthesis of value-added aldehydes, starting from the corresponding primary amines. In this frame, we have developed a completely chromatography-free purification protocol based on crossflow ultrafiltration, which makes the production of this enzyme easily scalable. Furthermore, we determined the kinetic parameters of the amine oxidase toward 20 differently substituted aliphatic and aromatic primary amines, and we developed a biocatalytic process for their conversion into the corresponding aldehydes. The reaction occurs in aqueous media at neutral pH in the presence of catalase, which removes the hydrogen peroxide produced during the reaction itself, contributing to the recycling of oxygen. A high conversion (>95%) was achieved within 3 h for all the tested compounds.

Overexpression of SlGSNOR impairs in vitro shoot proliferation and developmental architecture in tomato but confers enhanced disease resistance.

The pervasive presence of nitric oxide (NO) in cells and its role in modifying cystein residues through protein S-nitrosylation is a remarkable redox based signalling mechanism regulating a variety of cellular processes. S-NITROSOGLUTATHIONE REDUCTASE (GSNOR) governs NO bioavailability by the breakdown of S-nitrosoglutathione (GSNO), fine-tunes NO signalling and controls total cellular S-nitrosylated proteins. Most of the published data on GSNOR functional analysis is based on the model plant Arabidopsis with no previous report for its effect on in vitro regeneration of tissue cultured plants. Moreover, the effect of GSNOR overexpression (O.E) on tomato growth, development and disease resistance remains enigmatic. Here we show that SlGSNOR O.E in tomato alters multiple developmental programs from in vitro culture establishment to plant growth and fruit set. Moreover, constitutive SlGSNOR O.E in tomato showed enhanced resistance against early blight (EB) disease caused by Alternaria solani and reduction in hypersensitive response (HR)-mediated cell death after Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) infiltrations. High GSNOR transcript levels led to the inhibition of in vitro shoot proliferation in transformed explants as revealed by the fluorescence microscopy after YFP labelling. Transgenic tomato lines overexpressing SlGSNOR showed defective phenotypes exhibiting stunted plant growth and bushy-type plants due to loss of apical dominance, along with reduced seed germination and delayed flowering. Furthermore, SlGSNOR O.E plants exhibited altered leaf arrangement, fruit shape and modified locules number in tomato fruit. These findings give a novel insight into a multifaceted regulatory role of SlGSNOR in tomato plant development, reproduction and response to pathogens.

OsUEV1B, an Ubc enzyme variant protein, is required for phosphate homeostasis in rice.

Phosphorus is a crucial macronutrient for plant growth and development. The mechanisms for maintaining inorganic phosphate (Pi) homeostasis in rice are not well understood. The ubiquitin-conjugating enzyme variant protein OsUEV1B was previously found to interact with OsUbc13 and mediate lysine63-linked polyubiquitination. In the present study, we found OsUEV1B was specifically inhibited by Pi deficiency, and was localized in the nucleus and cytoplasm. Both osuev1b mutant and OsUEV1B-RNA interference (RNAi) lines displayed serious symptoms of toxicity due to Pi overaccumulation. Some Pi starvation inducible and phosphate transporter genes were upregulated in osuev1b mutant and OsUEV1B-RNAi plants in association with enhanced Pi acquisition, and representative Pi starvation responses, including stimulation of acid phosphatase activity and root hair growth, were also activated in the presence of sufficient Pi. A yeast two-hybrid screen revealed an interaction between OsUEV1B and OsVDAC1, which was confirmed by bimolecular fluorescence complementation and firefly split-luciferase complementation assays. OsVDAC1 encoded a voltage-dependent anion channel protein localized in the mitochondria, and OsUbc13 was shown to interact with OsVDAC1 via yeast two-hybrid and bimolecular fluorescence complementation assays. Under sufficient Pi conditions, similar to osuev1b, a mutation in OsVDAC1 resulted in significantly greater Pi concentrations in the roots and second leaves, improved acid phosphatase activity, and enhanced expression of the Pi starvation inducible and phosphate transporter genes compared with wild-type DongJin, whereas overexpression of OsVDAC1 had the opposite effects. OsUEV1B or OsVDAC1 knockout reduced the mitochondrial membrane potential and adenosine triphosphate levels. Moreover, overexpression of OsVDAC1 in osuev1b partially restored its high Pi concentration to a level between those of osuev1b and DongJin. Our results indicate that OsUEV1B is required for rice phosphate homeostasis.

AM fungi increase uptake of Cd and BDE-209 and activities of dismutase and catalase in amaranth (Amaranthus hypochondriacus L.) in two contaminants spiked soil.

Soil co-contaminated with cadmium (Cd) and decabromodiphenyl ether (BDE-209) is a widespread environmental problem, especially in electronic waste contaminated surroundings. Accumulation of Cd and BDE-209 in crops has possibly harmful effects on local human health. In order to assess the potential of arbuscular mycorrhizal (AM) fungi and amaranth (Amaranthus hypochondriacus L.) in remediation of soil co-contaminated with Cd and BDE-209, pot trials were performed to investigate interactive effects of AM fungi, Cd and BDE-209 on growth of amaranth, uptake of Cd and BDE-209, distribution of chemical forms of Cd and activities of antioxidant enzymes in shoots and dissipation of BDE-209 in soil. The present results showed that shoot biomass of non-mycorrhizal plants was significantly inhibited by increasing of Cd addition (5-15 mg kg-1), but were only slightly declined with BDE-209 addition (5 mg kg-1). The interaction of Cd and BDE-209 reduced the proportions of ethanol- and d-H2O-extractable Cd in shoots, consequently alleviated Cd toxicity to plants and enhanced root uptake of Cd and BDE-209. Inoculation of AM fungi resulted in significantly greater shoot biomass as well as higher concentrations of Cd and BDE-209 compared with non-mycorrhizal treatment. Moreover, AM fungi played a beneficial role in relieving oxidative stress on amaranth by increasing the activities of dismutase (SOD) and catalase (CAT) in shoots and significantly improved the dissipation of BDE-209 in soil. The present study suggested that combination of AM fungi and amaranth may be a potential option for remediation of Cd and BDE-209 co-contaminated soils.

Microgravity Affects the Level of Matrix Polysaccharide 1,3:1,4-ß-Glucans in Cell Walls of Rice Shoots by Increasing the Expression Level of a Gene Involved in Their Breakdown.

The plant cell wall provides each cell with structural support and mechanical strength, and thus, it plays an important role in supporting the plant body against the gravitational force. We investigated the effects of microgravity on the composition of cell wall polysaccharides and on the expression levels of genes involved in cell wall metabolism using rice shoots cultivated under artificial 1 g and microgravity conditions on the International Space Station. The bulk amount of the cell wall obtained from microgravity-grown shoots was comparable with that from 1 g-grown shoots. However, the analysis of sugar constituents of matrix polysaccharides showed that microgravity specifically reduced the amount of glucose (Glc)-containing polysaccharides such as 1,3:1,4-ß-glucans, in shoot cell walls. The expression level of a gene for endo-1,3:1,4-ß-glucanase, which hydrolyzes 1,3:1,4-ß-glucans, largely increased under microgravity conditions. However, the expression levels of genes involved in the biosynthesis of 1,3:1,4-ß-glucans were almost the same under both gravity conditions. On the contrary, microgravity scarcely affected the level and the metabolism of arabinoxylans. These results suggest that a microgravity environment promotes the breakdown of 1,3:1,4-ß-glucans, which, in turn, causes the reduced level of these polysaccharides in growing rice shoots. Changes in 1,3:1,4-ß-glucan level may be involved in the modification of mechanical properties of cell walls under microgravity conditions in space.

Maize roots and shoots show distinct profiles of oxidative stress and antioxidant defense under heavy metal toxicity.

Heavy metal accumulation in agricultural land causes crop production losses worldwide. Metal homeostasis within cells is tightly regulated. However, homeostasis breakdown leads to accumulation of reactive oxygen species (ROS). Overall plant fitness under stressful environment is determined by coordination between roots and shoots. But little is known about organ specific responses to heavy metals, whether it depends on the metal category (redox or non-redox reactive) and if these responses are associated with heavy metal accumulation in each organ or there are driven by other signals. Maize seedlings were subjected to sub-lethal concentrations of four metals (Zn, Ni, Cd and Cu) individually, and were quantified for growth, ABA level, and redox alterations in roots, mature leaves (L1,2) and young leaves (L3,4) at 14 and 21 days after sowing (DAS). The treatments caused significant increase in endogenous metal levels in all organs but to different degrees, where roots showed the highest levels. Biomass was significantly reduced under heavy metal stress. Although old leaves accumulated less heavy metal content than root, the reduction in their biomass (FW) was more pronounced. Metal exposure triggered ABA accumulation and stomatal closure mainly in older leaves, which consequently reduced photosynthesis. Heavy metals induced oxidative stress in the maize organs, but to different degrees. Tocopherols, polyphenols and flavonoids increased specifically in the shoot under Zn, Ni and Cu, while under Cd treatment they played a minor role. Under Cu and Cd stress, superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR) activities were induced in the roots, however ascorbate peroxidase (APX) activity was only increased in the older leaves. Overall, it can be concluded that root and shoot organs specific responses to heavy metal toxicity are not only associated with heavy metal accumulation and they are specialized at the level of antioxidants to cope with.

Overexpression of Peroxidase Gene GsPRX9 Confers Salt Tolerance in Soybean.

Peroxidases play prominent roles in antioxidant responses and stress tolerance in plants; however, their functions in soybean tolerance to salt stress remain unclear. Here, we investigated the role of a peroxidase gene from the wild soybean (Glycine soja), GsPRX9, in soybean tolerance to salt stress. GsPRX9 gene expression was induced by salt treatment in the roots of both salt-tolerant and -sensitive soybean varieties, and its relative expression level in the roots of salt-tolerant soybean varieties showed a significantly higher increase than in salt-sensitive varieties after NaCl treatment, suggesting its possible role in soybean response to salt stress. GsPRX9-overexpressing yeast (strains of INVSc1 and G19) grew better than the control under salt and H2O2 stress, and GsPRX9-overexpressing soybean composite plants showed higher shoot fresh weight and leaf relative water content than control plants after NaCl treatment. Moreover, the GsPRX9-overexpressing soybean hairy roots had higher root fresh weight, primary root length, activities of peroxidase and superoxide dismutase, and glutathione level, but lower H2O2 content than those in control roots under salt stress. These findings suggest that the overexpression of the GsPRX9 gene enhanced the salt tolerance and antioxidant response in soybean. This study would provide new insights into the role of peroxidase in plant tolerance to salt stress.

The response of a model C3/CAM intermediate semi-halophyte Mesembryanthemum crystallinum L. to elevated cadmium concentrations.

Many areas exhibiting increased concentrations of soluble salts are simultaneously polluted with heavy metals (HM), and halophytes with extended tolerance to heavy metal toxicity seem to represent a promising tool for their phytoremediation. In this study, the response of the soil-grown C3-CAM (Crassulacean acid metabolism) intermediate halophyte Mesembryanthemum crystallinum (common ice plant) to increased concentrations of Cd (0.01-1 mM) was investigated. None of the tested Cd treatments affected growth parameters or tissue water content of either C3 or CAM-performing plants. Chlorophyll a fluorescence confirmed high tolerance of the photosynthetic apparatus of both metabolic states towards Cd. Plants performing both photosynthesis types accumulated significant Cd amounts only under the highest (1 mM) treatment, and the metal was primarily deposited in the roots, which are features typical of an excluding strategy. Upon the application of 1 mM Cd solution CAM-performing plants, due to the NaCl pre-treatment applied for CAM induction, were exposed to significantly higher amounts of bioavailable Cd in comparison with those of C3-performing plants. As a result, roots of CAM plants accumulated over 4-fold higher Cd amounts when compared with C3 plants. In our opinion, enhanced Cd-accumulating potential observed in CAM-performing plants was the effect of osmotic stress episode and resulting modifications e.g. in the detoxifying capacity of the antioxidative system. Increased antioxidative potential of NaCl pre-treated plants was pronounced with significantly higher activity of CuZnSOD (copper-zinc superoxide dismutase), not achievable in C3 plants subjected to high Cd concentrations. Moreover, the applied Cd doses induced SOD activity in a compartment-dependent manner only in C3 plants. We confirmed that none of the applied Cd concentrations initiated the metabolic shift from C3 to CAM.

Sulfur mediated improved thiol metabolism, antioxidant enzymes system and reduced chromium accumulation in oilseed rape (Brassica napus L.) shoots.

Chromium (Cr) pollution is at a worrying level in a region of oilseed rape production in China. Sulfur (S) is an indispensable element for plants that has been confirmed to play an important role in regulating plant response to heavy metal stress. The present study was conducted to examine the role of S in alleviating Cr toxicity in oilseed rape. Cr stress strongly induced oxidative stress and inhibited plant growth. Application of S significantly enhanced the tolerance of oilseed rape exposed to Cr stress by activating several detoxification mechanisms including the ascorbate-glutathione (AsA-GSH) enzyme defense system and GSH production. The Cr and phytochelatins (PC) contents in the root under S treatment were markedly higher than those under Cr stress. The transcript abundances of the heavy metal transporters HMA2 and HMA4 were lower under S treatment than under Cr treatment. Most Cr was restricted to roots, and the translocation factor (TF) of Cr was markedly decreased in oilseed rape. In conclusion, our study revealed that S application is advantageous to oilseed rape defense against Cr toxicity and inhibits Cr translocation from roots to shoots.

CRISPR/Cas9-Mediated Mutagenesis of Carotenoid Cleavage Dioxygenase 8 (CCD8) in Tobacco Affects Shoot and Root Architecture.

Strigolactones (SLs) are a class of phytohormones that regulate plant architecture. Carotenoid cleavage dioxygenase (CCD) genes are involved in the biosynthesis of SLs and are identified and characterized in many plants. However, the function of CCD genes in tobacco remains poorly understood. In this study, two closely related genes NtCCD8A and NtCCD8B were cloned from tobacco (Nicotiana tabacum L.). The two NtCCD8 genes are orthologues of the tomato (Solanum lycopersicum) carotenoid cleavage dioxygenase 8 (SlCCD8) gene. NtCCD8A and NtCCD8B were primarily expressed in tobacco roots, but low expression levels of these genes were detected in all plant tissues, and their transcript levels significantly increased in response to phosphate limitation. NtCCD8A and NtCCD8B mutations were introduced into tobacco using the CRISPR/Cas9 system and transgenic tobacco lines for both ntccd8 mutant alleles were identified. The ntccd8a and ntccd8b mutant alleles were inactivated by a deletion of three nucleotides and insertion of one nucleotide, respectively, both of which led to the production of premature stop codons. The ntccd8 mutants had increased shoot branching, reduced plant height, increased number of leaves and nodes, and reduced total plant biomass compared to wild-type plants; however, the root-to-shoot ratio was unchanged. In addition, mutant lines had shorter primary roots and more of lateral roots than wild type. These results suggest that NtCCD8 genes are important for changes in tobacco plant architecture.

Changes in phenolic acid composition and associated enzyme activity in shoot and kernel fractions of brown rice during germination.

Phenolic acid composition and activities of two associated enzymes such as PAL (phenylalanine ammonia-lyase) and CW-PRX (cell wall peroxidase) in brown rice (BR) were examined during a germination for 4 days. Shoot and kernel fractions of the germinated brown rice were separated, and soluble extracts and insoluble residues of the fractions were analyzed. In the shoot fraction, the PAL activity and soluble phenolic acid content reached to its maximum on the second day of atmospheric germination, and decreased thereafter. In contrast, the amount of insoluble phenolic acids and CW-PRX activity continuously increased during the germination for 4 days. Comparing the shoot fractions, the kernel fraction exhibited lower activities of PAL and CW-PRX, but showed an increase in total phenolic acid content during germination. Germination raised the antioxidant activity of brown rice, especially in the shoot fraction which contained more phenolic acids than the kernel fraction.

Expression of OsWNK9 in Arabidopsis conferred tolerance to salt and drought stress.

With No Lysine (WNK) kinase belongs to ser/thr protein kinase group in which conserved catalytic lysine (K) residue of subdomain II is shifted to subdomain I. In this study, we cloned full-length coding region of WNK9 from Oryza sativa (OsWNK9) and performed in silico studies to confirm the presence of all kinase signature regulatory elements. The transcript analysis revealed that OsWNK9 was strongly down regulated under salinity, drought and ABA stress in shoots. Constitutive expression of OsWNK9 in Arabidopsis thaliana imparted increased tolerance to salt, drought, and ABA stress. Transgenic lines showed healthy phenotypes such as green leaves, achieved higher fresh weight and longer roots under salt, drought and ABA stress as compared to wild-type (WT). Transgenic plants showed better seed germination, higher chlorophyll retention and less water loss under salt and drought stress compared to WT. Promoter/gene expression studies revealed that OsWNK9 were expressed throughout plant tissues with higher expression in roots. Subcellular localization studies of OsWNK9 showed their presence in the nucleus. The transcript analysis of abiotic stress marker genes and ABA dependent genes showed they were highly expressed in transgenic lines compared to WT in response to salt and drought stress. The endogenous ABA level under salt and drought stress in transgenic lines was higher than WT. The results indicated that OsWNK9 may regulate salt and drought response in ABA dependent manner.

LPCAT1 controls phosphate homeostasis in a zinc-dependent manner.

All living organisms require a variety of essential elements for their basic biological functions. While the homeostasis of nutrients is highly intertwined, the molecular and genetic mechanisms of these dependencies remain poorly understood. Here, we report a discovery of a molecular pathway that controls phosphate (Pi) accumulation in plants under Zn deficiency. Using genome-wide association studies, we first identified allelic variation of the Lyso-PhosphatidylCholine (PC) AcylTransferase 1 (LPCAT1) gene as the key determinant of shoot Pi accumulation under Zn deficiency. We then show that regulatory variation at the LPCAT1 locus contributes significantly to this natural variation and we further demonstrate that the regulation of LPCAT1 expression involves bZIP23 TF, for which we identified a new binding site sequence. Finally, we show that in Zn deficient conditions loss of function of LPCAT1 increases the phospholipid Lyso-PhosphatidylCholine/PhosphatidylCholine ratio, the expression of the Pi transporter PHT1;1, and that this leads to shoot Pi accumulation.

DNA METHYLTRANSFERASE1-mediated shoot regeneration is regulated by cytokinin-induced cell cycle in Arabidopsis.

DNA methylation plays a critical role in diverse biological processes of plants. Arabidopsis DNA METHYLTRANSFERASE1 (MET1) represses shoot regeneration by inhibiting WUSCHEL (WUS) expression, which is essential for shoot initiation. However, the upstream signals regulating MET1 expression during this process are unclear. We analyzed the signals regulating MET1 expression using a number of established strategies, such as genetic analysis, confocal microscopy, quantitative real-time PCR and chromatin immunoprecipitation. MET1 expression patterns underwent dynamic changes with the initiation of WUS during shoot regeneration. The cell cycle regulator E2FA was characterized as an upstream factor directly promoting MET1 expression. Moreover, cytokinin promoted MET1 expression partially by enhancing CYCD3 expression. Our findings reveal that MET1-mediated shoot regeneration is regulated by the cytokinin-induced cell cycle, and provide new insights into the regulation of DNA methylation in shoot regeneration.

Bioinformatics Analysis of Phylogeny and Transcription of TAA/YUC Auxin Biosynthetic Genes.

Auxin is a main plant growth hormone crucial in a multitude of developmental processes in plants. Auxin biosynthesis via the tryptophan aminotransferase of arabidopsis (TAA)/YUCCA (YUC) route involving tryptophan aminotransferases and YUC flavin-dependent monooxygenases that produce the auxin indole-3-acetic acid (IAA) from tryptophan is currently the most researched auxin biosynthetic pathway. Previous data showed that, in maize and arabidopsis, TAA/YUC-dependent auxin biosynthesis can be detected in endoplasmic reticulum (ER) microsomal fractions, and a subset of auxin biosynthetic proteins are localized to the ER, mainly due to transmembrane domains (TMD). The phylogeny presented here for TAA/TAR (tryptophan aminotransferase related) and YUC proteins analyses phylogenetic groups as well as transmembrane domains for ER-membrane localisation. In addition, RNAseq datasets are analysed for transcript abundance of YUC and TAA/TAR proteins in Arabidopsis thaliana. We show that ER membrane localisation for TAA/YUC proteins involved in auxin biosynthesis is already present early on in the evolution of mosses and club mosses. ER membrane anchored YUC proteins can mainly be found in roots, while cytosolic proteins are more abundant in the shoot. The distribution between the different phylogenetic classes in root and shoot may well originate from gene duplications, and the phylogenetic groups detected also overlap with the biological function.

Chilling-responsive DEMETER-LIKE DNA demethylase mediates in poplar bud break.

Annual dormancy-growth cycle is a developmental and physiological process essential for the survival of deciduous trees in temperate and boreal forests. Seasonal control of shoot growth in woody perennials requires specific genetic programmes responding to environmental signals. The environmental-controlled mechanisms that regulate the shift between winter dormancy and the growth-promoting genetic programmes are still unknown. Here, we show that dynamics in genomic DNA methylation levels are involved in the regulation of dormancy-growth cycle in poplar. The reactivation of growth in the apical shoot during bud break process in spring is preceded by a progressive reduction of genomic DNA methylation in apex tissue. The induction in apex tissue of a chilling-dependent poplar DEMETER-LIKE 10 (PtaDML10) DNA demethylase precedes shoot growth reactivation. Transgenic poplars showing downregulation of PtaDML8/10 caused delayed bud break. Genome-wide transcriptome and methylome analysis and data mining revealed that the gene targets of DEMETER-LIKE-dependent DNA demethylation are genetically associated with bud break. These data point to a chilling-dependent DEMETER-like DNA demethylase mechanisms being involved in the shift from winter dormancy to a condition that precedes shoot apical vegetative growth in poplar.

Effect of Exogenous Nitro Oxide on Chilling Tolerance, Polyamine, Proline, and γ-Aminobutyric Acid in Bamboo Shoots (Phyllostachys praecox f. prevernalis).

The effects of exogenous nitro oxide (NO) on chilling resistance and the metabolism of polyamine, proline, and γ-aminobutyric acid of bamboo shoots were investigated. Bamboo shoots were dipped in 0.07 mM sodium nitroprusside (SNP) and stored at 1 °C for 56 days. During the storage, the development of chilling injury of SNP treated bamboo shoots was inhibited with decreased accumulation of malonaldehyde and electrical leakage. At the end of storage, the chilling injury incidence of treated bamboo shoots decreased by 37.9% while their malonaldehyde content and electrical leakage were 8.8% and 18.6% lower than that of the control, respectively. Interestingly, the endogenous NO, polyamines, γ-aminobutyric acid, and proline contents of treated bamboo shoot also significantly increased. Consistently, the metabolisms of these nitrogenous compounds were stimulated in treated bamboo shoots, according to their higher (20.2%-49.8%) related enzyme activities, including nitric oxide synthase, arginine decarboxylase, ornithine decarboxylase, glutamate decarboxylase, orn-δ-aminotransferase, and Δ1-pyrroline-5-carboxylate synthetase. The results indicated that the SNP treatment enhanced chilling tolerance of bamboo shoots, which might associate with the activated metabolism of polyamines, γ-aminobutyric acid, and proline. SNP treatment might be an alternative technology to avoid chill injury during cold storage of bamboo shoots.

Potential of different AM fungi (native from As-contaminated and uncontaminated soils) for supporting Leucaena leucocephala growth in As-contaminated soil.

Arbuscular mycorrhizal (AM) fungi inoculation is considered a potential biotechnological tool for an eco-friendly remediation of hazardous contaminants. However, the mechanisms explaining how AM fungi attenuate the phytotoxicity of metal(oid)s, in particular arsenic (As), are still not fully understood. The influence of As on plant growth and the antioxidant system was studied in Leucaena leucocephala plants inoculated with different isolates of AM fungi and exposed to increasing concentrations of As (0, 35, and 75 mg dm-3) in a Typic Quartzipsamment soil. The study was conducted under greenhouse conditions using isolates of AM fungi selected from uncontaminated soils (Acaulospora morrowiae, Rhizophagus clarus, Gigaspora albida; and a mixed inoculum derived from combining these isolates, named AMF Mix) as well as a mix of three isolates from an As-contaminated soil (A. morrowiae, R. clarus, and Paraglomus occultum). After 21 weeks, the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were determined in the shoots in addition to measuring plant height and mineral contents. In general, AM fungi have shown multiple beneficial effects on L. leucocephala growth. Although the activity of most of the stress-related enzymes increased in plants associated with AM fungi, the percentage increase caused by adding As to the soil was even greater for non-mycorrhizal plants when compared to AM-fungi inoculated ones, which highlights the phytoprotective effect provided by the AM symbiosis. The highest P/As ratio observed in AM-fungi plants, compared to non-mycorrhizal ones, can be considered a good indicator that the AM fungi alter the pattern of As(V) uptake from As-contaminated soil. Our results underline the role of AM fungi in increasing the tolerance of L. leucocephala to As stress and emphasize the potential of the symbiosis L. leucocephala-R. clarus for As-phytostabilization at moderately As-contaminated soils.

Variable Levels of Glutathione S-Transferases Are Responsible for the Differential Tolerance to Metolachlor between Maize (Zea mays) Shoots and Roots.

Glutathione S-transferases (GSTs) play important roles in herbicide tolerance. However, studies on GST function in herbicide tolerance among plant tissues are still lacking. To explore the mechanism of metolachlor tolerance difference between maize shoots and roots, the effects of metolachlor on growth, GST activity, and the expression of the entire GST gene family were investigated. It was found that this differential tolerance to metolachlor was correlated with contrasting GST activity between the two tissues and can be eliminated by a GST inhibitor. An in vitro metolachlor-glutathione conjugation assay confirmed that the transformation of metolachlor is 2-fold faster in roots than in shoots. The expression analysis of the GST gene family revealed that most GST genes are expressed much higher in roots than shoots, both in control and in metolachlor-treated plants. Taken together, higher level expression of most GST genes, leading to higher GST activity and faster herbicide transformation, appears to be responsible for the higher tolerance to metolachlor of maize roots than shoots.

The role of gibberellins in improving the resistance of tebuconazole-coated maize seeds to chilling stress by microencapsulation.

Chilling stress during germination often causes severe injury. In the present study, maize seed germination and shoot growth under chilling stress were negatively correlated with the dose of tebuconazole in an exponential manner as predicted by the model Y = A + B × e(-x/k). Microencapsulation was an effective means of eliminating potential phytotoxic risk. The gibberellins (GAs) contents were higher after microencapsulation treatment than after conventional treatment when the dose of tebuconazole was higher than 0.12 g AI (active ingredient) kg-1 seed. Further analysis indicated that microencapsulation can stimulate ent-kaurene oxidase (KO) activity to some extent, whereas GA 3-oxidase (GA3ox) and GA 2-oxidase (GA2ox) activities remained similar to those in the control. Genes encoding GA metabolic enzymes exhibited different expression patterns. Transcript levels of ZmKO1 increased in the microcapsule treatments compared to the control. Even when incorporated into microcapsules, tebuconazole led to the upregulation of ZmGA3ox1 at doses of less than 0.12 g AI kg-1 seed and to the upregulation of ZmGA3ox2 when the dose was higher than 0.12 g AI kg-1 seed. With increasing doses of microencapsulated tebuconazole, the transcript levels of ZmGA2ox4, ZmGA2ox5 and ZmGA2ox6 exhibited upward trends, whereas the transcript levels of ZmGA2ox7 exhibited a downward trend.