Trp-574-Leu mutation and metabolic resistance by cytochrome P450 gene conferred high resistance to ALS-inhibiting herbicides in Descurainia sophia.

Descurainia sophia (flixweed) is a troublesome weed in winter wheat fields in North China. Resistant D. sophia populations with different acetolactate synthetase (ALS) mutations have been reported in recent years. In addition, metabolic resistance to ALS-inhibiting herbicides has also been identified. In this study, we collected and purified two resistant D. sophia populations (R1 and R2), which were collected from winter wheat fields where tribenuron-methyl provided no control of D. sophia at 30 g a.i. ha-1. Whole plant bioassay and ALS activity assay results showed the R1 and R2 populations had evolved high-level resistance to tribenuron-methyl and florasulam and cross-resistance to imazethapyr and pyrithiobac­sodium. The two ALS genes were cloned from the leaves of R1 and R2 populations, ALS1 (2004 bp) and ALS2 (1998 bp). A mutation of Trp 574 to Leu in ALS1 was present in both R1 and R2. ALS1 and ALS2 were cloned from R1 and R2 populations respectively and transferred into Arabidopsis thaliana. Homozygous T3 transgenic seedlings with ALS1 of R1 or R2 were resistant to ALS-inhibiting herbicides and the resistant levels were the same. Transgenic seedlings with ALS2 from R1 or R2 were susceptible to ALS-inhibiting herbicides. Treatment with cytochrome P450 inhibitor malathion decreased the resistant levels to tribenuron-methyl in R1 and R2. RNA-Seq was used to identify target cytochrome P450 genes possibly involved in resistance to ALS-inhibiting herbicides. There were five up-regulated differentially expressed cytochrome P450 genes: CYP72A15, CYP83B1, CYP81D8, CYP72A13 and CYP71A12. Among of them, CYP72A15 had the highest expression level in R1 and R2 populations. The R1 and R2 populations of D. sophia have evolved resistance to ALS-inhibiting herbicides due to Trp 574 Leu mutation in ALS1 and possibly other mechanisms. The resistant function of CYP72A15 needs further research.

Rocket (Eruca vesicaria (L.) Cav.) vs. Copper: The Dose Makes the Poison?

The effects of copper addition, from various adsorbents, on the accumulation ability and glucosinolate content of cultivated rocket were studied. Different adsorbents (zeolite NaX, egg shells, substrate, fly ash) were treated with copper(II) solution with an adsorption efficiency of 98.36, 96.67, 51.82 and 39.13%, respectively. The lowest copper content and the highest total glucosinolate content (44.37 µg/g DW and 4269.31 µg/g DW, respectively) were detected in the rocket grown in the substrate with the addition of a substrate spiked with copper(II) ions. Rocket grown in the fly ash-substrate mixture showed an increase in copper content (84.98 µg/g DW) and the lowest total glucosinolate content (2545.71 µg/g DW). On the other hand, when using the egg shells-substrate mixture, the rocket copper content increased (113.34 µg/g DW) along with the total GSLs content (3780.03 µg/g DW), indicating the influence of an adsorbent type in addition to the copper uptake. The highest copper content of 498.56 µg/g DW was detected in the rocket watered with copper(II) solution with a notable decrease in the glucosinolate content, i.e., 2699.29 µg/g DW. According to these results rocket can be considered as a copper accumulator plant.

Effects of Dielectric Barrier Ambient Air Plasma on Two Brassicaceae Seeds: Arabidopsis thaliana and Camelina sativa.

We investigated low-temperature plasma effects on two Brassicaceae seeds (A. thaliana and C. sativa) using dielectric barrier discharge in air. Comparisons of plasma treatments on seeds showed distinct responses on germination rate and speed. Optimal treatment time giving optimal germination is 15 min for A. thaliana with 85% increase compared to control after 48 h of germination and 1 min for C. sativa with 75% increase compared to control after 32 h of germination. Such germination increases are associated with morphological changes shown by SEM of seed surface. For better understanding at the biochemical level, seed surfaces were analyzed using gas chromatography-mass spectrometry which underlined changes of lipidic composition. For both treated seeds, there is a decrease of saturated (palmitic and stearic) fatty acids while treated C. sativa showed a decrease of unsaturated (oleic and linoleic) acids and treated A. thaliana an increase of unsaturated ones. Such lipid changes, specifically a decrease of hydrophobic saturated fatty acids, are coherent with the other analyses (SEM, water uptake and contact angle). Moreover, an increase in A. thaliana of unsaturated acids (very reactive) probably neutralizes plasma RONS effects thus needing longer plasma exposure time (15 min) to reach optimal germination. For C. sativa, 1 min is enough because unsaturated linoleic acid becomes lower in treated C. sativa (1.2 × 107) compared to treated A. thaliana (3.7 × 107).

Selenium Alleviates the Adverse Effect of Drought in Oilseed Crops Camelina (Camelina sativa L.) and Canola (Brassica napus L.).

Drought poses a serious threat to oilseed crops by lowering yield and crop failures under prolonged spells. A multi-year field investigation was conducted to enhance the drought tolerance in four genotypes of Camelina and canola by selenium (Se) application. The principal aim of the research was to optimize the crop yield by eliciting the physio-biochemical attributes by alleviating the adverse effects of drought stress. Both crops were cultivated under control (normal irrigation) and drought stress (skipping irrigation at stages i.e., vegetative and reproductive) conditions. Four different treatments of Se viz., seed priming with Se (75 µM), foliar application of Se (7.06 µM), foliar application of Se + Seed priming with Se (7.06 µM and 75 µM, respectively) and control (without Se), were implemented at the vegetative and reproductive stages of both crops. Sodium selenite (Na2SeO3), an inorganic compound was used as Se sources for both seed priming and foliar application. Data regarding physiochemical, antioxidants, and yield components were recorded as response variables at crop maturity. Results indicated that WP, OP, TP, proline, TSS, TFAA, TPr, TS, total chlorophyll contents, osmoprotectant (GB, anthocyanin, TPC, and flavonoids), antioxidants (APX, SOD, POD, and CAT), and yield components (number of branches per plant, thousand seed weight, seed, and biological yields were significantly improved by foliar Se + priming Se in both crops under drought stress. Moreover, this treatment was also helpful in boosting yield attributes under irrigated (non-stress) conditions. Camelina genotypes responded better to Se application as seed priming and foliar spray than canola for both years. It has concluded that Se application (either foliar or priming) can potentially alleviate adverse effects of drought stress in camelina and canola by eliciting various physio-biochemicals attributes under drought stress. Furthermore, Se application was also helpful for crop health under irrigated condition.

Accumulation of potentially toxic elements by plants of North Caucasian Alyssum species and their molecular phylogenetic analysis.

Accumulations of potentially toxic metals were investigated in soils and five North Caucasian Alyssum species from metalliferous areas and non-metalliferous areas in Karachay-Cherkessia, Kabardino-Balkaria, Dagestan and the Krasnodar region. Analyses of field samples showed that chemical features of the soils significantly affected the concentrations of Ni, Co, Zn, but had less effect on Cu and Pb concentrations in the shoots of Alyssum. Variations in the degree of accumulating ability were found in the studied species, including hyperaccumulation of Ni in Alyssum murale (up to 12,100 mg kg-1), and significant accumulation of Zn in A. gehamense (up to 1700 mg kg-1). A comparative molecular genetic analysis of two A. murale populations, both Ni-hyperaccumulating population from Karachay-Cherkessia and non-hyperaccumulating population from Dagestan, indicated considerable genetic difference between them. This result supports the hypothesis that the selection of metal hyperaccumulator species with enhanced phytoremediation efficiency should be considered at the population level.

Biochemical basis of differential selenium tolerance in arugula (Eruca sativa Mill.) and lettuce (Lactuca sativa L.).

Selenium (Se) biofortification in crops provides a valuable strategy to enhance human Se intake. However, crops vary greatly with their capacity in tolerating and metabolizing/accumulating Se, and the basis underlying such variations remains to be fully understood. Here, we compared the effects of Se and its analog S treatments on plant growth and biochemical responses between a Se accumulator (arugula) and a non-accumulator (lettuce). Arugula exhibited an increased biomass production in comparison with untreated controls at a higher selenate concentration than lettuce (20 µM vs. 10 µM Na2SeO4), showing better tolerance to Se. Arugula accumulated 3-folds more Se and S than lettuce plants under the same treatments. However, the Se/S assimilation as assessed by ATP sulfurylase and O-acetylserine (thiol)lyase activities was comparable between arugula and lettuce plants. Approximately 4-fold higher levels of Se in proteins under the same doses of Se treatments were observed in arugula than in lettuce, indicating that Se accumulators have better tolerance to selenoamino acids in proteins. Noticeably, arugula showed 6-fold higher ascorbate peroxidase activity and produced over 5-fold more glutathione and non-protein thiols than lettuce plants, which suggest critical roles of antioxidants in Se tolerance. Taken together, our results show that the elevated Se tolerance of arugula compared to lettuce is most likely due to an efficient antioxidant defense system. This study provides further insights into our understanding of the difference in tolerating and metabolizing/accumulating Se between Se accumulators and non-accumulators.

Potential use of Schumannianthus dichotomus waste: the phytotoxic activity of the waste and its identified compounds.

The phytotoxic potential of the leaves and twigs of Schumannianthus dichotomus, discarded in the mat-making industry against four test plants (lettuce (Lactuca sativa L.), rapeseed (Brassica napus L.), foxtail fescue (Vulpia myuros (L.) C.C. Gmel.) and timothy (Phleum pratense L.)) was investigated and found strong phytotoxic activity. An assay-guided fractionation of S. dichotomus extarcts against cress (Lepidium sativum L.) through a series of column chromatography steps yielded two compounds, 8-(5-oxo-2,5-dihydrofuran-2-yl) octanoic acid (ODFO) and (E)-6-hydroxy-2,6-dimethylocta-2,7-dienoic acid (8-carboxylinalool). ODFO and 8-carboxylinalool showed strong phytotoxic activity against cress and timothy. The concentrations required for 50% growth inhibition (I50 value) of the seedlings of cress and timothy were 111.94-128.01 and 36.30-91.75 µM, respectively, for ODFO, but the values were much higher at 315.98-379.13 and 107.92-148.41 µM, respectively, for 8-carboxylinalool, indicating the stronger phytotoxic activity of ODFO. This study is the first to isolate ODFO and 8-carboxylinalool from S. dichotomus and their phytotoxic potential while ODFO is firstly encountered from any natural source. The growth inhibitory activity of the identified compounds may explain their role in the phytotoxic activity of S. dichotomus, which suggests the possible use of its leaves and twigs or its active constituents as natural bioherbicides.

Investigation of resistant level to tribenuron-methyl, diversity and regional difference of the resistant mutations on acetolactate synthase (ALS) isozymes in Descurainia sophia L. from China.

Descurainia sophia L. is one of the most notorious broadleaf weeds in winter wheat fields of China. In this study, 95 out of 163 (58.3%) D. sophia populations which were collected from provinces of Hebei, Shandong, Henan, Shanxi, Shaanxi and Jiangsu, have evolved resistance to tribenuron-methyl. The als1 and als2 were cloned in all test D. sophia populations, while als3 and als4 were identified only in some of the populations. Resistant mutations of Pro-197-Ser/Thr/Leu/His/Ala/Arg, Asp-376-Glu and Trp-574-Leu were identified in tribenuron-methyl-resistant (TR) D. sophia plants, while the Pro-197-Arg was first identified in D. sophia in this study. These resistant mutations displayed no preference between ALS1 and ALS2. However, Pro-197-Ser/Thr and Trp-574-Leu were identified in all ALS isozymes, while the other mutations were not. In addition, some resistant mutations displayed regional differences, the frequency of Pro-197-Ser in Shandong and Trp-574-Leu in Shanxi province is much higher than that in other provinces.

Deciphering lead tolerance mechanisms in a population of the plant species Biscutella auriculata L. from a mining area: Accumulation strategies and antioxidant defenses.

The uptake and distribution of Pb and the mechanisms involved in the metal tolerance have been investigated in a mine population of Biscutella auriculata. Seedlings were exposed to 125 µM Pb(NO3)2 for 15 days under semihydroponic conditions. The results showed an increase in the size of Pb-treated seedlings and symptoms of toxicity were not observed. ICP-OES analyses showed that Pb accumulation was restricted to root tissue. Imaging of Pb accumulation by dithizone histochemistry revealed the presence of the metal in vacuoles and cell wall in root cells. The accumulation of Pb in vacuoles could be stimulated by an increase in phytochelatin PC2 content. Pb did not promote oxidative damage and this is probably due the increase of antioxidative defenses. In the leaves, Pb produced a significant increase in superoxide dismutase activity, while in roots an increase in catalase and components of the Foyer- Halliwell-Asada cycle were observed. The results indicated that Biscutella auriculata has a high capacity to tolerate Pb and this is mainly due to a very efficient mechanism to sequester the metal in roots and a capacity to avoid oxidative stress. This species could therefore be very useful for phytostabilization and repopulation of areas contaminated with Pb.

Characterization of mechanisms involved in tolerance and accumulation of Cd in Biscutella auriculata L.

Biscutella auriculata L. is one of the rare species that is able to grow in a very contaminated mining area in Villamayor de Calatrava (Ciudad Real, Spain). In an effort to understand the mechanisms involved in the tolerance of this plant to high metal concentrations, we grew B. auriculata in the presence of 125 µM Cd(NO3)2 for 15 days and analysed different parameters associated with plant growth, nitric oxide and reactive oxygen species metabolism, metal uptake and translocation, photosynthesis rate and biothiol (glutathione and phytochelatins) content. Treatment with Cd led to growth inhibition in both the leaves and the roots, as well as a reduction of photosynthetic parameters, transpiration and stomatal conductance. The metal was mainly accumulated in the roots and in the vascular tissue, although most Cd was detected in areas surrounding their epidermal cells, while in the leaves the metal accumulated mainly in spongy mesophyll, stomata and trichrome. Based on the Cd bioaccumulation (5.93) and translocation (0.15) factors, this species denoted enrichment of the metal in the roots and its low translocation to the upper tissues. Biothiol analysis showed a Cd-dependent increase of reduced glutathione (GSH) as well as the phytochelatins (PC2 and PC3) in both roots and leaves. Cd-promoted oxidative damage occurred mainly in the leaves due to disturbances in enzymatic and nonenzymatic antioxidants, while the roots did not show significant damage as a result of induction of antioxidant defences. It can be concluded that B. auriculata is a new Cd-tolerant plant with an ability to activate efficient metal-sequestering mechanisms in the root surface and leaves and to induce PCs, as well as antioxidative defences in roots.

Endosperm prevents toxic amounts of Zn from accumulating in the seed embryo - an adaptation to metalliferous sites in metal-tolerant Biscutella laevigata.

Seed germination represents the first crucial stage in the life cycle of a plant, and the seed must contain all necessary transition elements for the development and successful establishment of the seedling. Problematically, seed development and germination are often hampered by elevated metal(loid) concentrations in industrially polluted soils, making their revegetation a challenging task. Biscutella laevigata L. (Brassicaceae) is a rare perennial pseudometallophyte that can tolerate high concentrations of trace metal elements. Yet, the strategies of this and other plant species to ensure reproductive success at metalliferous sites are poorly understood. Here we characterized several parameters of germination and used synchrotron X-ray fluorescence microscopy to investigate the spatial distribution and concentration of elements within B. laevigata seeds from two metallicolous and two non-metallicolous populations. We find that average germination time was shorter and the seed weight was lower in the metallicolous compared to the non-metallicolous populations. By allowing for at least two generations within one growth season, relatively fast germination at metalliferous sites accelerates microevolutionary processes and likely enhances the potential of metallicolous accessions to adapt to environmental stress. We also identified different strategies of elemental accumulation within seed tissues between populations. Particularly interesting patterns were observed for zinc, which was found in 6-fold higher concentrations in the endosperm of metallicolous compared to non-metallicolous populations. This indicates that the endosperm protects the seed embryo from accumulating toxic concentrations of metal(loid)s, which likely improves reproductive success. Hence, we conclude that elemental uptake regulation by the seed endosperm is associated with enhanced metal tolerance and adaptation to metalliferous environments in B. laevigata.

Evolution of phenotypic plasticity: Genetic differentiation and additive genetic variation for induced plant defence in wild arugula Eruca sativa.

Phenotypic plasticity is the primary mechanism of organismal resilience to abiotic and biotic stress, and genetic differentiation in plasticity can evolve if stresses differ among populations. Inducible defence is a common form of adaptive phenotypic plasticity, and long-standing theory predicts that its evolution is shaped by costs of the defensive traits, costs of plasticity and a trade-off in allocation to constitutive versus induced traits. We used a common garden to study the evolution of defence in two native populations of wild arugula Eruca sativa (Brassicaceae) from contrasting desert and Mediterranean habitats that differ in attack by caterpillars and aphids. We report genetic differentiation and additive genetic variance for phenology, growth and three defensive traits (toxic glucosinolates, anti-nutritive protease inhibitors and physical trichome barriers) as well their inducibility in response to the plant hormone jasmonic acid. The two populations were strongly differentiated for plasticity in nearly all traits. There was little evidence for costs of defence or plasticity, but constitutive and induced traits showed a consistent additive genetic trade-off within each population for the three defensive traits. We conclude that these populations have evolutionarily diverged in inducible defence and retain ample potential for the future evolution of phenotypic plasticity in defence.

Evidence that tolerance of Eutrema salsugineum to low phosphate conditions is hard-wired by constitutive metabolic and root-associated adaptations.

MAIN CONCLUSION: The extremophyte Eutrema salsugineum (Yukon ecotype) has adapted to an environment low in available phosphate through metabolic and root-associated traits that enables it to efficiently retrieve, use, and recycle phosphorus. Efficient phosphate (Pi) use by plants would increase crop productivity under Pi-limiting conditions and reduce our reliance on Pi applied as fertilizer. An ecotype of Eutrema salsugineum originating from the Yukon, Canada, shows no evidence of decreased relative growth rate or biomass under low Pi conditions and, as such, offers a promising model for identifying mechanisms to improve Pi use by crops. We evaluated traits associated with efficient Pi use by Eutrema (Yukon ecotype) seedlings and 4-week-old plants, including acquisition, remobilization, and the operation of metabolic bypasses. Relative to Arabidopsis, Eutrema was slower to remobilize phosphorus (P) from senescing leaves, primary and lateral roots showed a lower capacity for rhizosphere acidification, and root acid phosphatase activity was more broadly distributed and not Pi responsive. Both species produced long root hairs on low Pi media, whereas Arabidopsis root hairs were well endowed with phosphatase activity. This capacity was largely absent in Eutrema. In contrast to Arabidopsis, maximal in vitro rates of pyrophosphate-dependent phosphofructokinase and phosphoenolpyruvate carboxylase activities were not responsive to low Pi conditions suggesting that Eutrema has a constitutive and likely preferential capacity to use glycolytic bypass enzymes. Rhizosphere acidification, exudation of acid phosphatases, and rapid remobilization of leaf P are unlikely strategies used by Eutrema for coping with low Pi. Rather, equipping an entire root system for Pi acquisition and utilizing a metabolic strategy suited to deficient Pi conditions offer better explanations for how Eutrema has adapted to thrive on alkaline, highly saline soil that is naturally low in available Pi.

Analysis of Arabidopsis thaliana HKT1 and Eutrema salsugineum/botschantzevii HKT1;2 Promoters in Response to Salt Stress in Athkt1:1 Mutant.

Soil salinity imposes a serious threat to the productivity of agricultural crops. Among several other transporters, high-affinity K+ transporter (HKT)'s play an important role in reducing the phytotoxicity of Na+. Expression of Eutrema salsugineum (a halophyte) HKT1;2 is induced upon salt exposure. To elucidate the role of its promoter, we compared the sequences of HKT1;2 promoters from E. salsugineum (1822 bp) and E. botschantzevii (1811 bp) with Arabidopsis thaliana HKT1;1 (846 bp) promoter. In silico analysis predicted several cis-acting regulatory elements (GT-1 elements, core motifs of DRE/CRT, MYC/MYB-recognition sites and ACGT elements). Activities of the three promoters were analyzed by measuring HKT1;1 and/or HKT1;2 transcript level in the Athkt1;1 mutant plants. NaCl tolerance of the transgenics was also assessed. Our results depicted that expressing either AtHKT1;1 or EsHKT1;2 coding regions under the control of AtHKT1;1 promoter, almost reversed the hypersensitivity of the mutant for salt, on contrarily, when AtHKT1;1 coding sequence expressed under either Es or EbHKT1;2 promoters did not. Changes in shoot Na+/K+ concentrations under salt exposure is significantly consistent with the complementation ability of the mutant. The transcript concentration for genes under the control of either of Eutrema promoters, at control level was very less. This may suggest that either an important upstream response motif is missed or that A. thaliana misses a transcriptional regulator that is essential for salt-inducible HKT1 expression in Eutrema.

Resistance of Rapistrum rugosum to tribenuron and imazamox due to Trp574 or Pro197 substitution in the acetolactate synthase.

Ten putative resistant and two susceptible Rapistrum rugosum populations originating from Greece were studied for resistance to acetolactate synthase (ALS)-inhibiting herbicides, using dose-response assays, sequencing of als gene and in vitro ALS activity assays. The dose-response assays showed that one (P1) out of ten putative resistant populations was cross-resistant to tribenuron and imazamox, while another population (P4) was resistant to tribenuron only. All populations were susceptible to MCPA at the recommended rate. Gene sequencing of als revealed that the P4 population had a point mutation at Pro197 by His providing resistance to tribenuron, whereas the P1 had a Trp574 by Leu point mutation conferring cross-resistance to tribenuron and imazamox. The in vitro activity of the ALS enzyme indicated I50 values (tribenuron concentration required for 50% reduction of the ALS activity) ranging from 66.68 to 137.01 µM, whereas the respective value for the S populations ranged from 0.29 to 0.54 µM. These results strongly support that two R. rugosum populations evolved resistance to ALS-inhibiting herbicides due different point mutations in the als gene.

Salicylic acid and jasmonic acid restrains nickel toxicity by ameliorating antioxidant defense system in shoots of metallicolous and non-metallicolous Alyssum inflatum Náyr. Populations.

The presence of heavy metals in the soils is undoubtedly one of the prime abiotic stresses in the world. There are a considerable amount of plant yield losses because of heavy metal stress. The goal of this study was to assess the morphological, physiological and biochemical changes in Alyssum inflatum Nyár. populations upon exposure to different levels of nickel (Ni) (0, 100, 200, 400) µM, salicylic acid (SA) (0, 50, 200) µM and jasmonic acid (JA) (0, 5, 10) µM. Results showed that there were no considerable interpopulation differences, including the shoot Ni concentrations. Reversing the effects of Ni, SA and JA decreased due to Ni accumulation in both populations. By increasing the levels of Ni stress, the fresh weight (FW) of shoot decreased, whereas the application of SA + JA elevated the FW of the shoot in NM plants. Also, SA + JA mitigated Ni oxidative effects by reducing H2O2 concentration in both populations. The results revealed that the exposure of both M and NM plants to high Ni concentration increased superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) activities compared to control in both populations. Conversely, APX activity was inhibited in NM plants. Furthermore, SA and JA treatments reversed the detrimental effects of Ni on carotenoid content and reduced the content of proline in plants exposed to Ni stress. All the above suggests that SA and JA confer tolerance to Ni stress in two population of A. inflatum via several mechanisms.

Analysis of OJIP Chlorophyll Fluorescence Kinetics and QA Reoxidation Kinetics by Direct Fast Imaging.

Chlorophyll fluorescence kinetic analysis has become an important tool in basic and applied research on plant physiology and agronomy. While early systems recorded the integrated kinetics of a selected spot or plant, later systems enabled imaging of at least the slower parts of the kinetics (20-ms time resolution). For faster events, such as the rise from the basic dark-adapted fluorescence yield to the maximum (OJIP transient), or the fluorescence yield decrease during reoxidation of plastoquinone A after a saturating flash, integrative systems are used because of limiting speed of the available imaging systems. In our new macroscopic and microscopic systems, the OJIP or plastonique A reoxidation fluorescence transients are directly imaged using an ultrafast camera. The advantage of such systems compared to nonimaging measurements is the analysis of heterogeneity of measured parameters, for example between the photosynthetic tissue near the veins and the tissue further away from the veins. Further, in contrast to the pump-and-probe measurement, direct imaging allows for measuring the transition of the plant from the dark-acclimated to a light-acclimated state via a quenching analysis protocol in which every supersaturating flash is coupled to a measurement of the fast fluorescence rise. We show that pump-and-probe measurement of OJIP is prone to artifacts, which are eliminated with the direct measurement. The examples of applications shown here, zinc deficiency and cadmium toxicity, demonstrate that this novel imaging platform can be used for detection and analysis of a range of alterations of the electron flow around PSII.

Mucor sp.-An endophyte of Brassicaceae capable of surviving in toxic metal-rich sites.

The aim of this study was to assess the biodiversity of endophytic fungi from Arabidopsis arenosa growing on a post mining waste dump and to evaluate their role in plant adaptation to metal toxicity. Severeal of the fungi were beneficial for the plant. Among them, a fungus belonging to the Mucor genus, was found to interact with a broad range of plants, including Brassicaceae metallophytes. Mucor sp. was shown to be highly tolerant to elevated levels of Zn, Cd, and Pb and to accelerate plant-host growth under either toxic-metal stress or control conditions. When inoculated with Mucor sp., A. arenosa under toxic-metal stress acquired more N and showed significantly down-regulated catalase activity, which suggests suppression of toxic-metal-induced oxidative stress. We used the model plant-A. thaliana to evaluate the dynamics of plant-tissue colonization by the fungus as monitored with qPCR and to analyze the host's transcriptome response during early stages of the interaction. The results revealed the induction of a plant-defense and stress-related response on the 5th day of co-culture, which was in accord with the decrease of fungal abundance in shoots on the 6th day of interaction. Presented results demonstrate the importance of endophytic fungi in plant toxic-metal tolerance.

Cross-resistance to diflufenican and picolinafen and its inheritance in oriental mustard (Sisymbrium orientale L.).

BACKGROUND: An oriental mustard population (P40) was identified as resistant to diflufenican by screening at the field rate. As diflufenican and picolinafen both target phytoene desaturase (PDS), cross-resistance to picolinafen was suspected. The mechanism of resistance and its inheritance to diflufenican and picolinafen were investigated. RESULTS: At the lethal dose (LD50 ) level, population P40 was 237-fold more resistant to diflufenican and seven-fold more resistant to picolinafen compared to two susceptible populations. Population P40 also had a significantly higher resistance to diflufenican (237-fold) than a previously described P3 population (143-fold). In addition to the Leu-498-Val mutation in PDS identified in all individuals of the P3 and P40 populations, a Glu-425-Asp mutation was also found in P40. Neither mutation was detected in any individuals of the susceptible population. As the segregation of phenotype and genotype of the F2 individuals fitted the model for a single dominant allele, resistance to both diflufenican and picolinafen is likely encoded on the nuclear genome and is dominant. CONCLUSION: Resistance to diflufenican and picolinafen in the P40 population is likely conferred by Leu-498-Val and Glu-425-Asp mutations in the PDS gene. Inheritance of resistance to these herbicides is managed by a single dominant gene. © 2018 Society of Chemical Industry.

Combined effects of NaCl and Cd2+ stress on the photosynthetic apparatus of Thellungiella salsuginea.

Plants show complex responses to abiotic stress while, the effect of the stress combinations can be different to those seen when each stress is applied individually. Here, we report on the effects of salt and/or cadmium on photosynthetic apparatus of Thellungiella salsuginea. Our results showed a considerable reduction of plant growth with some symptoms of toxicity, especially with cadmium treatment. The structural integrity of both photosystems (PSI and PSII) was mostly maintained under salt stress. Cadmium induced a considerable decrease of both PSI and PSII quantum yields and the electron transport rate ETR(I) and ETR(II) paralleled by an increase of non-photochemical quenching (NPQ). In addition, cadmium alone affects the rate of primary photochemistry by an increase of fluorescence at O-J phase and also the photo-electrochemical quenching at J-I phase. A positive L-band appeared with (Cd) treatment as an indicator of lower PSII connectivity, and a positive K-band reflecting the imbalance in number of electrons at donor and acceptor side. In continuity to our previous studies which showed that NaCl supply reduced Cd2+ uptake and limited its accumulation in shoot of divers halophyte species, here as a consequence, we demonstrated the NaCl-induced enhancement effect of Cd2+ toxicity on the PSII activity by maintaining the photosynthetic electron transport chain as evidenced by the differences in ψO, φEo, ABS/RC and TR0/RC and by improvement of performance index PI(ABS), especially after short time of treatment. A significant decrease of LHCII, D1 and CP47 amounts was detected under (Cd) treatment. However, NaCl supply alleviates the Cd2+ effect on protein abundance including LHCII and PSII core complex (D1 and CP47).