Selected reactive oxygen species and antioxidant enzymes in common bean after <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">phaseolicola</Emphasis> and <Emphasis Type="Italic">Botrytis cinerea</Emphasis> infection

Phaseolus vulgaris cv. Korona plants were inoculated with the bacteria Pseudomonas syringae pv. phaseolicola (Psp), necrotrophic fungus Botrytis cinerea (Bc) or with both pathogens sequentially. The aim of the experiment was to determine how plants cope with multiple infection with pathogens having different attack strategy. Possible suppression of the non-specific infection with the necrotrophic fungus Bc by earlier Psp inoculation was examined. Concentration of reactive oxygen species (ROS), such as superoxide anion (O₂ ^?) and H₂O₂ and activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) were determined 6, 12, 24 and 48 h after inoculation. The measurements were done for ROS cytosolic fraction and enzymatic cytosolic or apoplastic fraction. Infection with Psp caused significant increase in ROS levels since the beginning of experiment. Activity of the apoplastic enzymes also increased remarkably at the beginning of experiment in contrast to the cytosolic ones. Cytosolic SOD and guaiacol peroxidase (GPOD) activities achieved the maximum values 48 h after treatment. Additional forms of the examined enzymes after specific Psp infection were identified; however, they were not present after single Bc inoculation. Subsequent Bc infection resulted only in changes of H₂O₂ and SOD that occurred to be especially important during plant–pathogen interaction. Cultivar Korona of common bean is considered to be resistant to Psp and mobilises its system upon infection with these bacteria. We put forward a hypothesis that the extent of defence reaction was so great that subsequent infection did not trigger significant additional response.     

Selenium Improves Physiological Parameters and Alleviates Oxidative Stress in Shoots of Lead-Exposed <Emphasis Type="Italic">Vicia faba</Emphasis> L. <Emphasis Type="Italic">minor</Emphasis> Plants Grown Under Phosphorus-Deficient Conditions

The goal of the study was to investigate the effects of exogenous selenium (Se) on the tolerance of faba bean plants to lead (Pb) stress under P-deficient conditions. The bean plants were grown for 2 weeks on Hoagland solution supplied with Pb (0, 50 μM) and Se (0, 1.5, or 6 μM), separately or simultaneously. It was shown that Pb did not affect shoot growth but caused major damage in the leaves, which was accompanied by Pb accumulation in these tissues. The exposure of the shoots to Pb led to significant changes in the biochemical parameters: the MDA content, glutathione peroxidase (GSH-Px), guaiacol peroxidase (GPOX), and catalase (CAT) activity increased. Furthermore, Pb intensified O ₂ ^?? and H₂O₂ production. Both the Se concentrations used increased the chlorophyll b, chlorophyll a+b, and carotenoid content in the faba bean plants. Selenite also generally enhanced CAT, GPOX, and GSH-Px activities and the T-SH level. Our results imply that the degree of disturbances caused by Pb could be partially ameliorated by Se supplementation. Selenite at a lower dose alleviated Pb toxicity by decreased H₂O₂ and O ₂ ^?? production and decreased the GSH-Px, GPOX, and CAT activities. The beneficial effect of the higher selenite concentration could be related to reduction of lipid peroxidation in the shoots of the Pb-treated plants. However, the effect of Se on the Pb-stressed plants greatly depended on the selenite dose in the nutrient solution.     

Elevated CO<Subscript>2</Subscript> influences host plant defense response in chickpea against <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

Global atmospheric concentration of CO₂ is likely to increase from 350 to 750 ppm over the next 100 years. The present studies were undertaken to understand the effects of elevated CO₂ on enzymatic activity and secondary metabolites in chickpea in relation to expression of resistance to pod borer, Helicoverpa armigera. Fifteen-day-old chickpea plants [ICCL 86111—resistant and JG 11—commercial cultivar] grown in the greenhouse were transferred to open-top chambers (OTC) and kept under 350, 550 and 750 ppm of CO₂. Twenty neonates of H. armigera were released on each plant at 7 days after shifting the pots to the OTCs. Un-infested plants were maintained as controls. After 7 days of infestation, the activities of defensive enzymes [peroxidase (POD), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL)] and amounts of total phenols and condensed tannins increased with an increase in CO₂ concentration in chickpea. The nitrogen balance index was greater in plants kept at 350 ppm CO₂ than in plants kept under ambient conditions. The H. armigera-infested plants had higher H₂O₂ content; amounts of oxalic and malic acids were greater at 750 ppm CO₂ than at 350 ppm CO₂. Plant damage was greater at 350 ppm than at 550 and 750 ppm CO₂. This information will be useful for understanding effects of increased levels of CO₂ on expression of resistance to insect pests to develop strategies to mitigate the effects of climate change.     

Activities of photosystem II and antioxidant enzymes in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L<Emphasis Type="Italic">.</Emphasis>) cultivars exposed to chilling temperatures

This study was carried out to determine the effect of chilling on both cold-acclimated and non-acclimated chickpea (Cicer arietinum L.) cultivars (Gökçe and Can?tez 87). Chickpea seedlings grown in soil culture for 12 days were subjected to chilling temperatures (2 and 4°C for 12 days) after maintaining in cold-acclimation (10°C, 7 days) or non-acclimation (25°C, 7 days) periods. The lowest values of growth parameters were obtained with cold-acclimated plants, whereas non-acclimated plants exhibited the lowest water content values, especially at 2°C. There was no effect of cold-acclimation period on chlorophyll fluorescence parameters. Plants subjected to chilling temperatures after cold-acclimation were more tolerant with respect to chlorophyll fluorescence parameters, and Gökçe had better photosystem II (PSII) photochemical activity. In the chilling treatments, total chlorophyll (a + b) content reduced, especially at 2°C, while anthocyanin and flavonoid contents increased to a greater extent in Gökçe and carotenoid content of the cultivars did not change. Malondialdehyde (MDA) content was higher for Can?tez 87, mostly at 2°C, while proline accumulation was greater for Gökçe. The cold-acclimation period led to a remarkable increase in antioxidant enzyme activities of both cultivars. The superoxide dismutase (SOD) activity was much higher in Gökçe for both chilling temperatures and the ascorbate peroxidase (APX) activity increased only in the cold-acclimated 4°C treatments. Similarly, with APX activity, the glutathione reductase (GR) and peroxidase (POD) activities of cultivars were higher in cold-acclimated plants at both the chilling temperatures, mostly in Gökçe. The results of this study indicate that cold-acclimation increased the cultivars ability to withstand the chilling temperatures. The lower MDA content and higher antioxidant and photochemical activities in Gökçe indicated an enhanced chilling tolerance capacity of this cultivar to protect the plant from oxidative damage.     

Characterization of a field-grown transgenic pineapple clone containing the genes <Emphasis Type="Italic">chitinase</Emphasis>, <Emphasis Type="Italic">AP24</Emphasis>, and <Emphasis Type="Italic">bar</Emphasis>

We previously introduced the bar gene, along with chitinase and AP24 genes, into the pineapple genome. The present report focuses on the evaluation of the first vegetative generation of a transgenic clone containing these genes. Three materials were compared: macropropagated controls (non-transformed), micropropagated controls (non-transformed), and micropropagated transformed plants. From each group, 50% of the plants were sprayed with FINALE^® 3 mo after the experiment initiation. The characterization was performed after 1 yr of field growth. FINALE^® killed all non-transgenic plants. Plants that survived the herbicide application showed 2n?=?50 chromosomes in their roots after 1 yr in the field. Micropropagated transformed plants sprayed with FINALE^® did not show phenotype differences from micropropagated transformed plants not sprayed with the herbicide. Between the micropropagated transformed plants sprayed with FINALE^® and the micropropagated control plants, the following differences were observed: modifications in levels of cell wall-linked, free and total phenolics, and total proteins. Moreover, changes of the fruit mass without crown were also recorded. Between the micropropagated transformed plants sprayed with FINALE^® and the macropropagated control plants, levels of chlorophyll b, total chlorophyll pigments, and proteins were different. Furthermore, activities of phenylalanine ammonia-lyase, superoxide dismutase, and glutamine synthetase were dissimilar. The plant height and diameter, and the crown height were also different. Until now, we have evaluated transformed pineapple plants during hardening and field growth. Although some unexpected variations were recorded, we believe they are not relevant enough to justify rejection of transgenesis as an important tool for pineapple genetic improvement.     

<Emphasis Type="Italic">Piriformospora indica</Emphasis>-mediated salinity tolerance in <Emphasis Type="Italic">Aloe vera</Emphasis> plantlets

Piriformospora indica, a root endophytic fungus, has been reported to promote growth of many plants under normal condition and allow the plants to survive under stress conditions. However, its impact on an important medicinal plant Aloe vera L. has not been well studied. Therefore, this study was undertaken to investigate the effect of P. indica on salinity stress tolerance of A. vera plant. P. indica inoculated and non-inoculated A. vera plantlets were subjected to four levels of salinity treatment- 0, 100, 200 and 300 mM NaCl. The salinity stress decreased the ability of the fungus to colonize roots of A. vera but the interaction of A. vera with P. indica resulted in an overall increase in plant biomass and greater shoot and root length as well as number of shoots and roots. The photosynthetic pigment (Chl a, Chl b and total Chl) and gel content were significantly higher for the fungus inoculated A. vera plantlets, at respective salinity concentrations. Furthermore, the inoculated plantlets had higher phenol, flavonoid, flavonol, aloin contents and radical scavenging activity at all salinity concentrations. The higher phenolic and flavonoid content may help the plants ameliorate oxidative stress resulting from high salinity.     

Physiological and cellular responses to fluoride stress in tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis>) leaves

Tea (Camellia sinensis (L.) O. Kuntze) hyper-accumulates fluoride (F), mainly in the leaves. To understand how tea copes with the stress caused by F, we tracked photosynthesis, antioxidant defense, and cell ultrastructure under different F concentrations (0–50 mg L^?1). High F (≥5 mg L^?1) caused decreases in photosynthetic and chlorophyll fluorescence parameters. Activated oxygen metabolism was altered by F, as manifested in increasing lipid peroxidation, electrolyte leakage (EL), and accumulation of H₂O₂. The activities of ascorbate peroxidase (APX, EC 1.11.1.1) and catalase (CAT, EC 1.11.1.6) increased at 0–5 mg L^?1 F, but sharply decreased less than 10–50 mg L^?1 F. The activity of manganese superoxide dismutase (Mn-SOD, EC 1.15.1.1) decreased with increasing F concentration. Expression of genes encoding antioxidant enzymes were in accordance with their measured activities. The results suggest that the antioxidant enzymes in the tea plant can eliminate reactive oxygen species (ROS) at <5 mg L^?1 F, but not at 20–50 mg L^?1 F. High F increased the number of epidermal hairs on tea leaves and decreased the stomatal aperture, reducing water loss. The leaf cellular structure appeared normal under 1–50 mg L^?1 F, although starch grains in chloroplast increased with increasing F. Proline and betaine play important roles in osmotic regulation in tea plant tolerating F stress. ROS scavenging and greater number of epidermal hairs are likely parts of the tea plant F-tolerance mechanism.     

Over-Expression of <Emphasis Type="Italic">PmHSP17.9</Emphasis> in Transgenic <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Confers Thermotolerance

Small heat shock proteins (sHSPs) have been shown to be involved in stress tolerance. However, their functions in Prunus mume under heat treatment are poorly characterized. To improve our understanding of sHSPs, we cloned a sHSP gene, PmHSP17.9, from P. mume. Sequence alignment and phylogenetic analysis indicated that PmHSP17.9 was a member of plant cytosolic class III sHSPs. Besides heat stress, PmHSP17.9 was also upregulated by salt, dehydration, oxidative stresses and ABA treatment. Leaves of transgenic Arabidopsis thaliana that ectopically express PmHSP17.9 accumulated less O₂ ^? and H₂O₂ compared with wild type (WT) after 42 °C treatment for 6 h. Over-expression of PmHSP17.9 in transgenic Arabidopsis enhanced seedling thermotolerance by decreased relative electrolyte leakage and MDA content under heat stress treatment when compared to WT plants. In addition, the induced expression of HSP101, HSFA2, and delta 1-pyrroline-5-carboxylate synthase (P5CS) under heat stress was more pronounced in transgenic plants than in WT plants. These results support the positive role of PmHSP17.9 in response to heat stress treatment.     

Physiological and biochemical responses of <Emphasis Type="Italic">Camellia sinensis</Emphasis> to stress associated with <Emphasis Type="Italic">Empoasca vitis</Emphasis> feeding

The tea green leafhopper, Empoasca vitis, is the most serious pest in plantations of tea, Camellia sinensis. Beyond physical damage to the leaves, tea yields may be affected if feeding stress causes physiological and biochemical changes in the tea plant, which affected the quality and flavor of the tea. Yet the effect of feeding stress, induced by E. vitis, is largely unknown. We measured the injury index and the physiological and biochemical responses of C. sinensis to stress by E. vitis feeding in a series of laboratory trials. Using 2-year-old C. sinensis plants, we tested the effects of leafhopper feeding at different densities—0, 5, 10, and 20 leafhoppers—and different durations of exposure—1, 4, 7, and 10 days—on potential changes in chlorophyll, tea polyphenols, nutrient content, activities of protective enzymes (peroxidase, POD; superoxide dismutase, SOD; and catalase, CAT), and the lipid peroxidation (MDA). We found that the injury indices for tea leaves increased continuously as the density of E. vitis increased in the same day, and simultaneously, as the time of leafhoppers damage increased, the injury indices for tea leaves also increased. Our results also indicated that feeding by E. vitis caused a considerable decline in chlorophyll a, chlorophyll b, total chlorophyll in tea leaves and soluble carbohydrate content, and an increase in tea polyphenols. Soluble protein content showed a direct increasing relationship with the increasing leafhopper density and the duration of exposure. Throughout the period of E. vitis exposure, there was highly significant difference in the activities of protective enzymes and MDA content. Additionally, POD, SOD, and CAT activities in tea leaves were elevated significantly with the increase of leafhopper density. Lipid peroxidation (MDA) content also increased after the exposure to leafhopper feeding. Overall, our results indicate that although C. sinensis displays a certain level of tolerance to E. vitis feeding stress, higher density of leafhoppers, and longer exposure duration, can cause severe damage to tea leaves and also a decline in plant defense of tea, so as to affect the tea quality.     

Characterizing the growth of <Emphasis Type="Italic">Sarcoptes scabiei</Emphasis> infrapopulations

The plant stress hypothesis posits that a herbivore’s reproductive success increases when it feeds on stressed plants, while the plant vigor hypothesis predicts that a herbivore preferentially feeds on more vigorous plants. We examined these opposing hypotheses by growing spider mites (Tetranychus urticae) on the leaves of stressed and healthy (vigorous) cucumber plants. Host plants were grown under controlled conditions at low, moderate, and high concentrations of NaCl (to induce salinity stress), at low, moderate, and high fertilizer concentrations (to support growth), and without these additions (control). The effects of these treatments were evaluated by measuring fresh and dry plant biomass, carotenoid and chlorophyll content, antioxidant enzyme activity, and concentrations of PO₄^3?, K⁺, and Na⁺ in plant tissues. The addition of low concentrations of fertilizer increased dry mass, protein, and carotenoid content relative to controls, suggesting a beneficial effect on plants. The highest NaCl treatment (2560 mg L^?1) resulted in increased Na⁺ and protein content relative to control plants, as well as reduced PO₄^3?, K⁺, and chlorophyll levels and reduced catalase and ascorbate peroxidase enzyme activity levels. Analysis of life table data of T. urticae mites raised on leaves from the aforementioned plant groups showed the intrinsic rate of increase (r) for mites was 0.167 day^?1 in control specimens, 0.125 day^?1 for mites reared on plants treated with a moderate concentration of fertilizer (10 mL L^?1), and was highest (0.241 day^?1) on plants grown under moderate salinity conditions (1920 mg L^?1 NaCl). Reproductive success of T. urticae did not differ on plants watered with a moderate concentration of NaCl or a high concentration of fertilizer. The moderately-stressed plants formed a favorable environment for the development and reproduction of spider mites, supporting the plant stress hypothesis.     

Management of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae) in strawberry fields with <Emphasis Type="Italic">Neoseiulus californicus</Emphasis> (Acari: Phytoseiidae) and acaricides

The purpose of this study was to evaluate the performance of Neoseiulus californicus (McGregor) for the control of Tetranychus urticae Koch in commercial strawberry fields, under greenhouse conditions, in association or not with the use of acaricides. The N. californicus strain used in this study was tolerant or resistant to several pesticides. Three experiments were carried out in the State of São Paulo, Brazil. For the first experiment, the initial infestation of T. urticae was 87.1 active stages per leaflet. Two applications of propargite were made on the first and the 14th day of the experiment. Approximately 2 h after each propargite application, N. californicus was released at a rate of 3.0 and 1.9 adult mites per plant, respectively, for each application. The population of T. urticae decreased from 87.1 to 2.8 mites per leaflet in the first three weeks. After this period, the population of T. urticae was maintained at low levels (≤1.5 mites/leaflet) until the end of the experiment (10th week). Propargite and dimethoate sprayed on the strawberry field did not affect significantly the population of this predaceous mite. For the second experiment, the infestation of T. urticae was 29.1 mites per leaflet, when the acaricide chorfenapyr was applied on the strawberry field. The release of N. californicus (2 mites per plant) was made 2 weeks after spraying the acaricide. The population of T. urticae was maintained at low levels (≤2.8 mites/leaflet) for 8 weeks (evaluation period). The T. urticae infestations in plots with N. californicus were significantly lower than in non-release plots, for the experiments 1 and 2. In the third experiment, the initial infestation of T. urticae was 40.5 mites per leaflet (55.5 active stages/leaflet on release plants; 25.5 active stages/leaflet on non-release plants). Three releases of N. californicus (average rate of 3.0 adult mites/plant), without any acaricide application, were not sufficient to reduce significantly the T. urticae population in release plots (release plants + non-release plants) in 6 weeks from the first release, however, the spider mite population decreased from 55.5 to 7.8 active stages per leaflet on release plants, during this period. Interplant dispersal of N. californicus was low in this strawberry field with high infestation of T. urticae. The studies indicate the viability of the use of this strain of N. californicus for the control of T. urticae in strawberry fields under greenhouse conditions, especially in association with selective acaricides.     

CAT and MDH improve the germination and alleviate the oxidative stress of cryopreserved <Emphasis Type="Italic">Paeonia</Emphasis> and <Emphasis Type="Italic">Magnolia</Emphasis> pollen

Researches have reported that reactive oxygen species (ROS)-induced oxidative stress plays an important role in cell cryodamage during cryopreservation. In the current study, pollen from Magnolia denudata and Paeonia lactiflora ‘Zi Feng Chao Yang’ was cryopreserved and incubated with exogenous catalase (CAT) and malate dehydrogenase (MDH) immediately after thawing. The effect of CAT and MDH on the germination of cryopreserved pollen was measured. Based on that, the ROS level, lipid peroxidation and antioxidants activities in fresh pollen, cryopreserved pollen added with or without CAT or MDH were determined to investigate their relationship with oxidative stress. Pollen from Magnolia and Paeonia showed a significant loss of germination, but marked increase of ROS and malondialdehyde (MDA) production after cryostorage. Antioxidant profiles in them were also enhanced. CAT and MDH addition increased the post-LN pollen germination of Magnolia and Paeonia significantly. Their germination rate achieved the highest with 100 IU ml^?1 MDH and 400 IU ml^?1 CAT application, respectively. Compared to their untreated controls, ROS and MDA accumulation reduced significantly in cryopreserved Magnolia pollen treated with 100 IU ml^?1 MDH, while superoxide dismutase (SOD) activity improved markedly. In the case of Paeonia, significantly lower level of ROS and MDA, but higher activity of CAT and SOD were observed in cryopreserved pollen treated with 400 IU ml^?1 CAT. In conclusion, pollen deterioration after cryopreservation is associated with ROS-induced oxidative stress. Exogenous CAT and MDH can reduce the oxidative damage through the activity stimulation of antioxidant enzymes, and play a protective role in the pollen during cryopreservation.     

Oxidative enzymes activity during abiotic and biotic stresses in <Emphasis Type="Italic">Zea mays</Emphasis> leaves and roots exposed to Cu,methyl jasmonate and <Emphasis Type="Italic">Trigonotylus caelestialium</Emphasis>

The activities of antioxidative enzymes, i.e. superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and guaiacol peroxidase (GPX), in the leaves and roots of Zea mays L. plants exposed to abiotic (methyl jasmonate, MJ, or/and copper, Cu) and biotic (Trigonotylus caelestialium) factors were examined. The contribution of MJ as a signal molecule in the defense mechanism against abiotic and biotic stresses was studied. All plants were cultivated hydroponically and divided into three groups: not treated by abiotic factors (control), treated by MJ only (MJ) and by MJ and Cu (MJ + Cu) and in each group half of the plants were exposed to T. caelestialium attack. The enzymatic activities of SOD, CAT, APX, and GPX in the leaves were higher in the insect-treated than non-insect-treated control plants, but lower in both MJ + Cu- or MJ- and insect-treated plants. In the roots, the enzyme activities were elevated in all insect-treated plants with the highest rise in MJ + Cu, in comparison with the MJ-treated plants. The results showed that MJ and MJ + Cu were efficient in reducing the activity of the antioxidative enzymes in the leaves under the insect influence by elevating enzyme activity in the roots.     

Physiological responses and antioxidant enzyme changes in <Emphasis Type="Italic">Sulla coronaria</Emphasis> inoculated by cadmium resistant bacteria

Plant growth promoting bacteria (PGPB) may help to reduce the toxicity of heavy metals on plants growing in polluted soils. In this work, Sulla coronaria inoculated with four Cd resistant bacteria (two Pseudomonas spp. and two Rhizobium sullae) were cultivated in hydroponic conditions treated by Cd; long time treatment 50 µM CdCl₂ for 30 days and short time treatment; 100 µM CdCl₂ for 7 days. Results showed that inoculation with Cd resistant PGPB enhanced plant biomass, thus shoot and root dry weights of control plants were enhanced by 148 and 35% respectively after 7 days. Co-inoculation of plants treated with 50 and 100 µM Cd increased plant biomasses as compared to Cd-treated and uninoculated plants. Cadmium treatment induced lipid peroxidation in plant tissues measured through MDA content in short 7 days 100 µM treatment. Antioxidant enzyme studies showed that inoculation of control plants enhanced APX, SOD and CAT activities after 30 days in shoots and SOD, APX, SOD, GPOX in roots. Application of 50 µM CdCl₂ stimulated all enzymes in shoots and decreased SOD and CAT activities in roots. Moreover, 100 µM of CdCl₂ increased SOD, APX, CAT and GPOX activities in shoots and increased significantly CAT activity in roots. Metal accumulation depended on Cd concentration, plant organ and time of treatment. Furthermore, the inoculation enhanced Cd uptake in roots by 20% in all treatments. The cultivation of this symbiosis in Cd contaminated soil or in heavy metal hydroponically treated medium, showed that inoculation improved plant biomass and increased Cd uptake especially in roots. Therefore, the present study established that co-inoculation of S. coronaria by a specific consortium of heavy metal resistant PGPB formed a symbiotic system useful for soil phytostabilization.     

Physiological alterations due to field salinity stress in melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.)

This study was aimed to assess physiological responses of melon (Cucumis melo L.) cultivars to salinity stress under field conditions. Seventeen melon cultivars including 16 widely distributed native and one exotic (‘Galia’) were subjected to 2-year (2014–2015) field salinity stress. Leaf relative water content (RWC), membrane stability index (MSI), pigments [chlorophyll a, b, total chlorophyll (TChl), carotenoid (Car) and their ratios], malondialdehyde (MDA), H₂O₂ content, proline content (Pro), total soluble sugar content (TSC), salinity tolerance and susceptibility indices as well as yield were evaluated. The results of combined analysis of variance showed significant genotypic variation for all the traits and significant effect of salinity stress on all the traits with the exception of Chla/Chlb and TChl/Car ratios. Overall, field salinity stress caused an increase in leaf MDA, H₂O₂, Chla, Chlb, TChl, Car, Pro and TSC and caused a reduction in leaf MSI and RWC as well as yield. The results of correlation coefficients showed that accumulation of osmolytes (proline and TSC) led to an increase in RWC and a decrease in MDA contents. In addition, the results of multiple regression analysis showed that leaf MDA, TSC, MSI and Chla contents were the most important predictors of yield justifying 72% total variation of yield under saline conditions. These results may highlight a dynamic interplay among biomarkers for lipid peroxidation (MDA), sugar osmolytes (TSC) and photosynthetic pigment (Chla) to maintain cell viability and cell wall integrity under salinity stress conditions in melon.     

Exogenous 5-aminolevulinic acid alleviates the detrimental effects of UV-B stress on lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L) seedlings

One of the abiotic stress factors affecting plant metabolism is ultraviolet-B (UV-B) radiation. 5-Aminolevulinic acid (ALA), a key precursor of porphyrin biosynthesis, promotes plant growth and crop yields. To investigate the alleviating effects of exogenous ALA on the damages caused by UV-B exposure, two different concentrations [10 ppm (ALA1) and 25 ppm (ALA2)] of ALA were applied to lettuce seedlings for 24 h and then they were exposed to 3.3 W m^?2 UV-B. Results showed that UV-B treatment significantly decreased chlorophyll a and b (Chl a and b) concentration, enhanced the activity of antioxidant enzymes, total phenolic concentration, soluble sugar contents, expression of phenylalanine ammonia lyase (PAL) and γ-tocopherol methyltransferase (γ-TMT) genes, the concentration of malondialdehyde (MDA), hydrogen peroxide (H₂O₂), and the rate of superoxide radical (\({\text{O}}_{2}^{ - }\)) generation in the lettuce seedlings when compared to the control. Pre-treatment with exogenous ALA significantly enhanced UV-B stress tolerance in lettuce seedlings by decreasing the reactive oxygen species. On the other hand, ALA application caused more increases in the PAL and γ-TMT gene expression, antioxidant enzymes activities, Chl a and b concentration, total phenolic content, antioxidant capacity and the concentrations of soluble sugars. Obtained results indicated that UV-B radiation exerts an adverse effect on lettuce seedlings, and some of the negative effects of UV-B radiation can be alleviated by exogenous ALA.     

Accumulation and tolerance characteristics of lead in <Emphasis Type="Italic">Althaea rosea</Emphasis> Cav. and <Emphasis Type="Italic">Malva crispa</Emphasis> L.

Two ornamental plants of Althaea rosea Cav. and Malva crispa L. were exposed to various concentrations of lead (Pb) (0, 50, 100, 200 and 500 mg·kg^?1) for 70 days to evaluate the accumulating potential and the tolerance characteristics. The results showed that both plant species grown normally under Pb stress, and A. rosea had a higher tolerance than M. crispa, while M. crispa had a higher ability in Pb accumulation than A. rosea. Besides, lower Pb concentration (50 mg·kg^?1) stimulated the shoot biomass in both plant species. Pb accumulation in plants was consistent with the increase of Pb levels, and the main accumulation sites were the roots and the older leaves. In addition, the photosynthetic pigments content and chlorophyll fluorescence parameters were influenced by Pb stress. In such case, both of the plants could improve the activities of antioxidant enzymes of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX), and the contents of the total soluble sugar and soluble protein, which reached the highest value at Pb 100 mg·kg^?1, as well as the accumulation of the total thiols (T-SH) and non-protein thiols (NP-SH) to adapt to Pb stress. Thus, it provides the theoretical basis and possibility for ornamental plants of A. rosea and M. crispa in phytoremediation of Pb contaminated areas.     

Growth and nodulation in <Emphasis Type="Italic">Alnus sieboldiana</Emphasis> in response to <Emphasis Type="Italic">Frankia</Emphasis> inoculation and nitrogen treatments

Key message

We investigated a Frankia – Alnus sieboldiana symbiosis, including the minimum inoculum dose for constant nodulation, the period of time to nodulation after inoculation, and the effects of N on nodulation.

Abstract

Frankia is a nitrogen-fixing actinomycete that forms root nodules in some dicotyledonous plants, which are called actinorhizal. We studied nodule formation in Alnus sieboldiana, an actinorhizal plant, after inoculation with a Frankia isolate to establish techniques for Frankia inoculation and the cultivation of inoculated plants. Root nodules formed on seedlings of A. sieboldiana by 2 weeks after inoculation, and N₂ fixation measured by acetylene reduction activity started 3 weeks after inoculation. Nodulation was observed after inoculation with a Frankia isolate at 0.001 μL packed cell volume (pcv). The number of nodules formed on the seedlings inoculated with Frankia at more than 0.05 μL pcv was not significantly different. Nodule development and N₂ fixation were reduced when inoculated seedlings were treated weekly with 15 mM NH₄NO₃-N. In contrast, treatment with 3.75 or 0.9375 mM NH₄NO₃-N did not inhibit nodule development or N₂ fixation of inoculated seedlings by 15 weeks of N treatment.

     

Correction to: Shoot regeneration process and optimization of <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in <Emphasis Type="Italic">Sinningia speciosa</Emphasis>

The objective of this research was to induce mitotic chromosome doubling in Anemone sylvestris L. The mitosis inhibitor oryzalin was directly added to the induction medium at 1, 2, 5, 10 and 15 μM for 8, 10 or 12 weeks of cultivation. Three tetraploid plants (2n?=?4x?=?32), 0.8% (polyploidization efficiency), were obtained from diploid plants (2n?=?2x?=?16) in three treatments (1 μM for 10 weeks, 5 μM for 8 weeks and 8 μM for 10 weeks). Ploidy level was confirmed by flow cytometry. Morphological characteristics (e.g. flower diameter, total plant height, leaf area) and chlorophyll content differences between diploid and tetraploid A. sylvestris were observed together with polyphenol content and antioxidant activity. The inter primer binding sites markers were used for evaluation of polymorphism. New genotypes with different morphological and biological characteristics were obtained through somatic polyploidization. The tetraploid plants were stronger, more vigorous and had an early flowering, which is essential for its use as an ornamental plant. The iPBS analysis showed unique amplicons that can be used for the purposes of molecular identification of tetraploid plants of A. sylvestris in the future. The results demonstrate the first report of in vitro induction of tetraploids of A. sylvestris.