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.     

Responses of He-Ne laser on agronomic traits and the crosstalk between UVR8 signaling and phytochrome B signaling pathway in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> subjected to supplementary ultraviolet-B (UV-B) stress

UV-B acclimation effects and UV-B damage repair induced by a 632.8-nm He-Ne laser were investigated in Arabidopsis thaliana plants in response to supplementary UV-B stress. There was an increasing trend in growth parameters in the combination-treated plants with He-Ne laser and UV-B light compared to those stressed with enhanced UV-B light alone during different developmental stages of plants. The photosynthetic efficiency (Pn) and survival rates of seedlings were significantly higher in the combination treatments than UV-B stress alone. The expression of UVR8, phytochrome B (PhyB), and their mediated signal responsive genes such as COP1, HY5, and CHS were also significantly upregulated in plants with the laser irradiation compared with other groups without the laser. Levels of flavonol accumulation in leaves and capsule yield of He-Ne laser-treated plants were increased. The phyB-9 mutants were more sensitive to enhanced UV-B stress and had no obvious improvements in plant phenotypic development and physiological damage caused by enhanced UV-B stress after He-Ne laser irradiation. Our results suggested that UVR8 and its mediated signaling pathway via interaction with COP1 can be induced by He-Ne laser, and these processes were dependent on cytoplasmic PhyB levels in plant cells, which might be one of the most important mechanisms of He-Ne laser on UV-B protection and UV-B damage repair. These current data have also elucidated that the biostimulatory effects of He-Ne laser on Arabidopsis thaliana plants would happen not only during the early growth stage but also during the entire late developmental stage.     

Kinetin Regulates UV-B-Induced Damage to Growth,Photosystem II Photochemistry,and Nitrogen Metabolism in Tomato Seedlings

Cytokinins are a class of plant growth regulators that regulate several developmental processes in plants, and recently their role in counteracting the deleterious effects of abiotic stresses has been noted. The impacts of kinetin (10 µM, KN; an artificial cytokinin) on growth, photosystem II photochemistry, and nitrogen metabolism in tomato seedlings exposed to two levels (UV-B₁, ambient+?1.2 kJ m^?2 day^?1, and UV-B₂, ambient+?2.4 kJ m^?2 day^?1) of enhanced UV-B radiation were analyzed under open field condition. The growth, pigment contents, carbonic anhydrase activity, photosynthetic O₂ yield, and values of chlorophyll a fluorescence parameters: F _v/F ₀, F _v/F _m or φP₀, ψ ₀, φE ₀, and PI_ABS declined, whereas the values of energy flux parameters (ABS/RC, TR₀/RC, ET₀/RC, and DI₀/RC) of PS II, efficiency of water splitting complex (F ₀/F _v), and respiratory rate of O₂ uptake increased under UV-B stress. Likewise, UV-B exposure at both doses significantly inhibited the activity of enzymes involved in nitrogen metabolism: nitrate reductase, nitrite reductase, glutamine synthetase, and glutamate synthase. In contrast, an enhancing effect on glutamate dehydrogenase activity was observed under UV-B stress. Exogenous KN resulted in a significant attenuation in UV-B-induced negative effects on growth, pigments, photosynthesis, and nitrogen metabolism. The study concludes that exogenous KN improved the growth performance of tomato seedlings by attenuating the damaging effects of UV-B radiation on photochemistry of PS II and nitrogen metabolism, and the alleviating effect against the low dose (UV-B₁) of UV-B was more pronounced.     

Effects of medium-wave ultraviolet radiation on levels and spectrum of polyamines in leaves and roots of wild-growing plants

The work continues serial studies on short-term effects of medium-wave ultraviolet radiation (UV-B) at 12.5 kJ/m² on plants. Special attention is paid to the rapid response of the antioxidant system. Free and conjugated forms of putrescine polyamines (putrescine, spermine, and spermidine), as well as those of cadaverine, are recognized to be constituents of the antioxidant system. These compounds were analyzed in plants 24 h after UV-B irradiation. Thellungiella salsuginea (Pallas) O.E.Schulz, Salvia officinalis L, Plantago major L., and Geum urbanum L. grown in aquatic culture under phytotron conditions were examined. The results support the hypothesis that putrescine plays the chief role in the plant defense response against medium-wave ultraviolet irradiation. Three of four plants manifested an increase in the content of this polyamine in leaves. It is the change that determines the enhanced total level of free polyamines. We failed to reveal a general tendency in dynamics of levels of conjugated forms of spermine, spermidine, and cadaverine; only conjugates of putrescine demonstrated a distinct increase. This study allows a conclusion that contributions of particular polyamines to the protective response primarily depend on the species to which the investigated plant belongs. It is likely that conjugated polyamines can be reserved as a pool necessary for rapid recovery of free polyamine levels.     

<Emphasis Type="Italic">Piriformospora indica</Emphasis> Alleviates Salinity by Boosting Redox Poise and Antioxidative Potential of Tomato

More than 20% of irrigated land has been influenced by salt stress, decreasing crop production. In this research, we investigated the effect of different levels of salinity (0, 50, 100 and 150 mM NaCl) and the efficiency of Piriformospora indica on growth, biochemical traits, antioxidative defense system in tomato (Solanum lycopersicum L.). NaCl stress reduced chlorophyll content, height and biomass of plants. Higher level of salinity (150 mM) declined the plant height by 22.65%, total dry weight by 56.44% and total chlorophyll by 44.34%, however, P. indica inoculation raised plant height by 43.47%, dry weight by 69.23% and total chlorophyll content by 48.09%. Salinity stress increased H₂O₂, malondialdehyde (MDA), superoxide anion and 1,1-diphenyl-2-picrylhydrazyl (DPPH) level in leaves and roots tomato seedlings. However, P. indica inoculation reduced H₂O₂, MDA and superoxide anion and enhanced DPPH compared to non-inoculated plants at all NaCl levels. The total phenol and flavonoids increased with NaCl treatment. On the other hand, the total phenolic and flavonoid increased more in P. indica inoculated plants compared to non-inoculated ones. Moreover, inoculation of P. indica implicated noteworthy improvement of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR), and glutathione reductase (GR) activity in tomato upon salinity. Notably, colonization with P. indica significantly improved the content of reduced ascorbic acid (AsA), glutathione (GSH) and redox ratio in the tomato plants under salinity resulting in reduced redox state. Our findings confirmed that salinity had negative effect on tomato seedling; however, P. indica inoculation increased tolerance to salinity by improving the content of phenolic compounds, non-enzymatic antioxidants, and increasing the activity of antioxidant enzymes.     

Salt tolerance conferred by expression of a global regulator IrrE from <Emphasis Type="Italic">Deinococcus radiodurans</Emphasis> in oilseed rape

As salinity is a major threat to sustainable agriculture worldwide, cultivation of salt-tolerant crops becomes increasingly important. IrrE acts as a global regulator and a general switch for stress resistance in Deinococcus radiodurans. In this study, to determine whether the irrE gene can improve the salt tolerance of Brassica napus, we introduced the irrE gene into B. napus by the Agrobacterium tumefaciens-mediated transformation method. Forty-two independent transgenic plants were regenerated. Polymerase chain reaction (PCR) analyses confirmed that the irrE gene had integrated into the plant genome. Northern as well as Western blot analyses revealed that the transgene was expressed at various levels in transgenic plants. Analysis for the T₁ progenies derived from four independent transformants showed that irrE had enhanced the salt tolerance of T₁ in the presence of 350 mM NaCl. Furthermore, under salt stress, transgenic plants accumulated more compatible solutes (proline) and a lower level of malondialdehyde (MDA), and they had higher activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). However, agronomic traits were not affected by irrE gene overexpression in the transgenic B. napus plants. This study indicates that the irrE gene can improve the salt tolerance of B. napus and represents a promising candidate for the development of crops with enhanced salt tolerance by genetic engineering.     

Molecular cloning and functional characterization of a Cu/Zn superoxide dismutase gene (<Emphasis Type="Italic">CsCSD1</Emphasis>) from <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Superoxide dismutase (SOD) proteins, which are widely present in the plant kingdom, play vital roles in response to abiotic stress. However, the functions of cucumber SOD genes in response to environmental stresses remain poorly understood. In this study, a SOD gene CsCSD1 was identified and functionally characterized from cucumber (Cucumis sativus). The CsCSD1 protein was successfully expressed in E. coli, and its overexpression significantly improved the tolerance of host E. coli cells to salinity stress. Besides, overexpression of CsCSD1 enhanced salinity tolerance during germination and seedling development in transgenic Arabidopsis plants. Further analyses showed that the SOD and CAT (catalase) activities of transgenic plants were significantly higher than those of wild-type (WT) plants under normal growth conditions as well as under NaCl treatment. In addition, the expression of stress-response genes RD22, RD29B and LEA4-5 was significantly elevated in transgenic plants. Our results demonstrate that the CsCSD1 gene functions in defense against salinity stress and may be important for molecular breeding of salt-tolerant plants.     

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 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.     

Combined effect of ethylene- and salicylic acid-signaling insensitive mutation on Arabidopsis response to low temperature

The roles of ethylene (ET) or salicylic acid (SA) in plant response to low temperature (LT, 5 °C) have been implicated. However, the combined effect of ET- and SA-signaling on plant growth and metabolism under LT remains to be evaluated. In this study, we comparatively analyzed the response of Arabidopsis ethylene insensitive (ein) 2-1 (an ET insensitive mutant), nonexprressor of pathogenesis relative (npr)1-1 (an SA insensitive mutant) and double mutant ein2-1/npr1-1 plants to LT. The results show that a LT of 5 °C induced plant growth retardation to a less degree in ein2-1, an intermediate degree in npr1-1, but a much larger in ein2-1/npr1-1 compared to the wild-type (WT) plants. The LT susceptibility of the ein2-1/npr1-1 plants was correlated to a lower net photosynthetic rate and proline content, and a higher content of H₂O₂ and malondialdehyde and electrolyte leakage relative to the WT plants. Lower activities of superoxide dismutase, peroxidase, and catalase, as well as a lower glutathione content and a ratio of its reduced form to its oxidized form were also observed in the double mutant plants as compared with the WT plants. However, at normal conditions (23 °C), all the tested physiological and biochemical parameters were comparable between the ein2-1/npr1-1 and WT plants, and plant growth was even better in the double mutant than in the WT plants. On the contrary, most of the above-mentioned parameters were advantageous in the ein2-1 and npr1-1 plants over the WT plants under the LT conditions. These data suggest that a parallel function or physiological redundancy of nonexpressor of pathogenesis relative 1 and ethylene insensitive 2 existed in the Arabidopsis plant response to the LT. On the other hand, an interaction between ET- and SA-signaling occurred during this process.     

Plant species-specificity and effects of bioinoculants and fertilization on plant performance for nickel phytomining

Aims

To investigate the effects of fertilization and bacterial inoculation on the growth, health and Ni phytoextraction capacity of three Ni-hyperaccumulators, Odontarrhena bracteata, O. inflata and O. serpyllifolia.

Methods

Plants were grown for three months in serpentine soil fertilized with inorganic NPK or amended with cow manure and inoculated with five rhizobacterial strains (previously isolated from O. serpyllifolia). Shoot and root dry weight (DW) yields, Ni accumulation and removal, nutritive status and stress indicators were determined.

Results

Plants grown in manure-amended soil showed significantly higher DW yields, improved nutritive status and higher total Ni phytoextracted. Some bacterial inoculants enhanced Ni removal due to the stimulation in growth and/or increase in shoot Ni concentration but this depended on the plant species, soil type and inoculant. Pseudoarthrobacter oxydans strain SBA82 enhanced shoot DW yield of all three Odontarrhena spp. in at least one soil type. Paenarthrobacter sp. strain LA44 and Stenotrophomonas sp. strain MA98 promoted growth of O. serpyllifolia and O. bracteata. Inoculated plants showing growth promotion presented lower activities of antioxidative enzymes, and concentrations of malondialdehyde (MDA) and H₂O₂, indicating a protective effect of these inoculants on the plants.

Conclusion

Rhizobacterial inoculants applied in combination with manure can improve plant growth and health, and Ni phytoextraction, in some hyperaccumulating Odontarrhena spp.

     

Overexpression of a peroxiredoxin gene from <Emphasis Type="Italic">Tamarix hispida</Emphasis>, <Emphasis Type="Italic">ThPrx1</Emphasis>, confers tolerance to oxidative stress in yeast and Arabidopsis

Peroxiredoxins (Prxs) are ubiquitous thiol-specific antioxidant enzymes that are critically involved in cell defense and protect cells from oxidative damage. In this study, a putative Type II Prx (ThPrx1) was identified and characterized from Tamarix hispida. The expression of ThPrx1 is highly induced in response to hydrogen peroxide (H₂O₂) and methyl viologen (MV) stresses. When expressed ectopically, ThPrx1 showed enhanced tolerance against oxidative stress in yeast and Arabidopsis. In addition, transgenic Arabidopsis plants overexpressing ThPrx1 displayed improved seedling survival rates and increased root growth and fresh weight gain under H₂O₂ and MV treatments. Moreover, transgenic Arabidopsis plants showed decreased accumulation of H₂O₂, superoxide (O₂^??) and malondialdehyde (MDA), increased superoxide dismutase (SOD) activity compared to wild-type (WT) plants under oxidative stress. Moreover, transgenic plants maintained higher photosynthesis efficiency and lower electrolyte leakage rates than that of WT plants under stress conditions. These results clearly indicated that ThPrx1 plays an important role in cellular redox homeostasis under stress conditions, leading to the maintenance of membrane integrity and increased tolerance to oxidative stress.     

Physiological and biochemical changes in tomato cultivar PT-3 with dual inoculation of mycorrhiza and PGPR against root-knot nematode

Arbuscular mycorrhizal fungi (AMF) and plant growth promoting rhizobacteria (PGPR) have potential to control soil-borne diseases including plant-parasitic nematodes. First, the effects of dual inoculation of mycorrhiza (Rhizophagus irregularis) and two stains of pseudomonads (Pseudomonas jessenii strain R62 and Pseudomonas synxantha strain R81) on tomato (Solanum lycopersicum cv. PT-3) growth were tested. Further, the physiological and biochemical changes caused by these beneficial organisms during infection by the root-knot nematode Meloidogyne incognita were studied. The experiment was conducted under glass house conditions and carried out up to one month after nematode inoculation. Plants treated with dual or individual inoculation of AMF and PGPR showed significantly enhanced plant growth and reduced nematode infection. In addition, they exhibited potent activity of phenolics (28 %) and defensive enzymes i.e. peroxidase (PO; 1.26 fold), polyphenyloxidase (PPO; 1.35 fold) and superoxide dismutase (SOD; 1.09 fold) while a significant reduction in malondialdehyde (MDA; 1.63 fold) and hydrogen peroxide (H₂O₂; 1.30 fold) content was recorded when compared to the nematode-infected plants. These findings indicate the feasibility of AMF and PGPR individually or in combinations as potential biocontrol agents for the management of root-knot nematodes.     

Interaction of gibberellic acid and 24-epibrassinolide in the regulation of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> seedling scotomorphogenesis

The effects of GA₃, 24-epibrassinolide (EBL), and their combination on morphogenesis of Arabidopsis thaliana (L.) Heynh seven-day-old seedlings were studied. Four plant lines were analyzed: wild type Ler and ga4-1 mutant, belonging to the Landsberg erecta ecotype and wild type Col and det2 mutant, both of the Columbia ecotype. In ga4-1 and det2, GA_4/1-and brassinosteroid-deficient mutants, the highest hypocotyl growth response to the lack of hormones was noted. The cotyledon shape and size were dependent on EBL, and the root length was both GA₃-and EBL-regulated, indicating organ specificities in the responses to these hormones. Simultaneous treatment of dark-grown plants with GA₃ and EBL exerted an additive stimulatory effect on the root growth of det2, reduced the inhibitory effect of EBL on hypocotyl elongation of ga4-1, and enhanced the effect of EBL on hypocotyl and cotyledon elongation of det2.     

UV-B stress-induced ABA production in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> mutants defective in ethylene signal transduction pathway

In this study, we examined the influence of UV-B radiation (280–320 nm) on ABA accumulation in 14-day-old Arabidopsis thaliana (L.) Heynh plants of wild type (WT), ethylene receptor mutant (etr1-1), and mutant with a constitutively active ethylene signal transduction pathway (ctr1-1). ABA content in nonirradiated WT plants was twice higher than in each mutant. UV-B irradiation caused dose-dependent ABA accumulation in WT plants. In the etr1-1 mutant, the amount of accumulated ABA was significantly less. In the ctr1-1 mutant, ABA content didn’t increase after UV-B irradiation. These data suggest that start of stress-induced ABA formation requires the adjustable ethylene signal pathway. In the ctr1-1 mutant, a constitutively active (nonadjustable) ethylene signal pathway blocks stress-induced ABA accumulation.     

Overexpression of <Emphasis Type="Italic">IrlVHA</Emphasis>-<Emphasis Type="Italic">c</Emphasis>, A Vacuolar-Type H<Superscript>+</Superscript>-ATPase c Subunit Gene from <Emphasis Type="Italic">Iris lactea</Emphasis>, Enhances Salt Tolerance in Tobacco

Vacuolar-type H⁺-ATPase (V-ATPase), a multi-subunit endomembrane proton pump, plays an important role in plant growth and response to environmental stresses. In the present study, transgenic tobacco that overexpressed the V-ATPase c subunit gene from Iris lactea (IrlVHA-c) was used to determine the function of IrlVHA-c. Quantitative PCR analysis showed that IrlVHA-c expression was induced by salt stress in I. lactea roots and leaves. Subcellular localization of green fluorescent protein (GFP) as marker combined with FM4-64 staining showed that the IrlVHA-c-GFP was localized to the endosomal compartment in tobacco cells. Compared with the wild-type, the IrlVHA-c transgenic tobacco plants exhibited greater seed germination rates, root length, fresh weight, and higher relative water content (RWC) of leaves under salt stress. Furthermore, the IrlVHA-c transgenic tobacco leaves have lower stomatal densities and larger stomatal apertures than wild-type. Under salt stress, superoxide dismutase (SOD) activity in the transgenic tobacco was significantly enhanced. Moreover, the level of malondialdehyde (MDA) in the transgenic tobacco was significantly lower than that in wild-type plants under salt stress. Taken together, these results suggested that the IrlVHA-c plays an important role in salt tolerance in transgenic tobacco by influencing stomatal movement and physiological changes.