The objective of this study was to relate the activation of enzymatic antioxidant system to the production of reactive oxygen species induced by salt stress. Rice (Oryza sativa L.) genotypes BRS Bojuru and BRS Pampa, tolerant and sensitive to salinity, respectively, were subjected to 150 mM NaCl for 0, 6, 24, 48, and 72 h. A significant increase of superoxide anion and H₂O₂ and a decrease in malondialdehyde (MDA) content were observed in the tolerant genotype, whereas in the sensitive genotype, there was no change in superoxide anion content, reduced H₂O₂ content, and increased MDA content. The superoxide dismutase (SOD) activity increased significantly in both genotypes, and increases in amounts of transcript were observed for OsSOD3Cu/Zn and OsSODA1-Mn in the tolerant genotype and for OsSOD4-Cu/Zn, OsSOD3-Cu/Zn, OsSODCc1-Cu/Zn, OsSOD-Fe, and OsSODA1-Mn in the sensitive genotype. The activities of catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were not significantly and consistently changed, but OsCATA, OsAPX2 and OsGR1 were induced in both genotypes. OsCATB transcription was increased in the tolerant genotype and OsCATC and OsAPX3 in the sensitive genotype under salinity. It is concluded that OsAPX3, OsGR2, OsGR3, and OsSOD3-Cu/Zn genes are the most suitable to distinguish tolerant from sensitive genotypes under salt stress.

Sulphur (S) is incorporated into diverse primary and secondary metabolites that play important roles in proper growth and development of plants. In cereals, a fraction of the nitrogen (N) accumulated in developing grains is guaranteed by amino acid remobilization from vegetative tissues, a contribution that becomes critical when soil nutrients are deficient. Glutamine synthetase (GS) and amino acid transporters (AAT) are key components involved in N assimilation and recycling. The aim of the present study was to evaluate the effect of S availability on the expressions of HvGS and several selected HvAAT genes in barley plants and on the phloem exudation rate of amino acids. To this end, two independent experiments were designed to impose low S availability conditions to barley plants. Low S availability caused a decrease in the phloem exudation rate of amino acids as well as in the gene expression of all the HvGS genes and five of the six HvAAT genes analyzed. The strong correlation found between the phloem amino acid exudation rate and HvGS1-1, HvGS1-2, HvAAP7, and HvProT1 gene expression may indicate the participation of these genes in the regulation of amino acid remobilization through the phloem.

The aim of this study was to identify single nucleotide polymorphism (SNP) markers genetically linked to root elongation rate (RER) in sugar beet (Beta vulgaris L.). A population of 244 F₃ individuals, obtained from the cross between lines L01 (a low RER) and L18 (a high RER), was phenotyped by measuring RER of 11-d-old seedlings grown in a hydroponic culture. Two DNA bulks of 50 F₃ individuals with extreme phenotypes were used for bulk segregant analysis by restriction-associated DNA sequencing. A total of 20 376 SNPs were identified. Single nucleotide polymorphisms were filtered to reduce the number of the false positive and mapped on candidate chromosomal regions of the B. vulgaris reference genome. One of the total of SNPs selected, SNP10139, was strongly linked to RER (P < 0.01). The pattern of association between the SNP10139 genotype and RER was also evaluated on a breeding line panel comprising 40 low and 40 high RER individuals with different allele frequencies between groups (P < 0.01). The SNP10139 sequence was mapped on the B. vulgaris peptide transporter (PTR) gene, a carrier that influences root elongation in Arabidopsis thaliana. Our results suggest that SNP10139 influence RER in sugar beet, and sequence information can be used in marker-assisted selection programs.

Drought is a severe environmental constraint to plant productivity. Being a multidimensional stress, it triggers a wide variety of plant responses ranging from physiological, biochemical to molecular levels. One of the inevitable consequences of drought stress is an increase in reactive oxygen species (ROS) production in different cellular compartments, namely the chloroplasts and mitochondria. This enhanced ROS production is, however, kept under tight control by a versatile and cooperative antioxidant system that modulates intracellular ROS content and sets the redoxstatus of the cell. Furthermore, ROS production under stresses functions as an alarm signal that triggers defence or acclimation. Specific signal transduction pathways involve, e.g., H₂O₂ as a secondary messenger. ROS signalling under drought is linked to abscisic acid (ABA) and Ca²⁺ fluxes. At molecular levels, several drought-responsive genes, transcription factors, aquaporins, late embryogenesis abundant proteins, heat shock proteins, and dehydrins have been identified. This review discusses recent understanding on molecular responses and protective mechanisms of drought stress.

In a previous study, we have identified and characterized gene from wheat (Triticum aestivum L.) encoding F-box protein and named it TaFBA. In this paper, transgenic tobacco (Nicotiana tabacum L.) plants overexpressing TaFBA1 displayed accelerated growth early, but the rate slowed gradually at later stages of growth, and the mature transgenic plants were even shorter in stature and flowered later than did the wild type (WT). Treatment with gibberellin (GA) conferred an accelerated growth rate to the transgenic tobacco plants at later stages, similar to that of WT, whereas growth was inhibited more seriously in WT than in transgenic tobacco when plants were treated with a GA biosynthesis inhibitor. The content of GA in transgenic tobacco plants was higher at early developmental stages, but it was lower at later growth stages than in WT. Some GA biosynthesis genes were down regulated, which was accompanied with elevated expression of a GA catabolism gene. Thus, our results suggest that TaFBA1 is possibly involved in the regulation of plant growth and development, and that it may be related to the production, metabolism, and proper function of GA.

The plastidic thioredoxin F-type (TrxF) protein plays an important role in plant saccharide metabolism. In this study, a gene encoding the TrxF protein, named SlTrxF, was isolated from tomato. The coding region of SlTrxF was cloned into a binary vector under the control of 35S promoter and then transformed into Arabidopsis thaliana. The transgenic Arabidopsis plants exhibited increased starch accumulation compared to the wild-type (WT). Real-time quantitative PCR analysis showed that constitutive expression of SlTrxF up-regulated the expression of ADP-glucose pyrophosphorylase (AGPase) small subunit (AtAGPase-S1 and AtAGPase-S2), AGPase large subunit (AtAGPase-L1 and AtAGPase-L2) and soluble starch synthase (AtSSS I, AtSSS II, AtSSS III and AtSSS IV) genes involved in starch biosynthesis in the transgenic Arabidopsis plants. Meanwhile, enzymatic analyses showed that the major enzymes (AGPase and SSS) involved in the starch biosynthesis exhibited higher activities in the transgenic plants compared to WT. These results suggest that SlTrxF may improve starch content of Arabidopsis by regulating the expression of the related genes and increasing the activities of the major enzymes involved in starch biosynthesis.

Seasonal growth is characteristic for many tree species including almond. Varying conditions during the season are responsible for growth cessation, bud set, dormancy entry, cold hardening, and bud burst. Here, we report the characterization of an almond homologue of the Arabidopsis GIGANTEA (AtGI) gene (designated as PdGI, GenBank accession No. KJ502316). We propose a role for this gene in the transition to dormancy and cold acclimation. The complementary DNA (cDNA) sequence of PdGI was 4 322 bp long and contained an open reading frame of 3 512 bp. The deduced amino acid sequence of PdGI shared 76 % identity with AtGI. The expression of PdGI at ambient day/night temperatures of 22/20 ºC was differentially regulated under a 16-h or 12-h photoperiod, increasing during the day and decreasing after dusk. However, this diurnal regulation was disrupted when plants were transferred to cold (12 ºC) conditions. In addition, we have assessed the expression of PdGI and putative almond homologues of the downstream target genes CONSTANS (PdCO-like) and FLOWERING LOCUS T (PdFT-like) in flower buds and shoots from adult trees during the bud break period in autumn and early winter. Our results show a clear increase in transcript abundance towards anthesis, suggesting a role of these genes in flower development.

This study investigated the role of the protein kinase MEK1/2 in the regulation of ascorbate and glutathione metabolism by nitric oxide in Agropyron cristatum leaves. The results showed that NO donor, sodium nitroprusside (SNP), induced the increased activities of ascorbate peroxidase, glutathione reductase, monodehydroascorbate reductase, dehydroascorbate reductase, L-galactono-1,4-lactone dehydrogenase, and γ-glutamylcysteine synthetase (γ-ECS), the transcription of respective genes, and the content of reduced ascorbate, reduced glutathione, total ascorbate, and total glutathione. All the above increases, except the activity of γ-ECS, were suppressed by pre-treatment with MEK1/2 inhibitors 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one and 1,4-diamino-2,3-dicyano-1,4-bis(o-aminophenylmercapto) butadiene. The results of Western blot showed that SNP induced the increase in the phosphorylation of MEK1/2. Our results suggested that SNP induced the phosphorylation of MEK1/2, which, in turn, up-regulated the ascorbate and glutathione metabolism in A. cristatum leaves.

Glutathione transferases (GSTs) mainly catalyze the nucleophilic addition of glutathione to a large variety of hydrophobic molecules participating to the vacuole compartmentalization of many toxic compounds. In this work, the putative tolerance of transgenic tobacco plants over-expressing CsGSTU genes towards the chloroacetanilide herbicide alachlor was investigated. Our results show that the treatment with 0.0075 mg cm^-3 of alachlor strongly affects the growth of both wild type and transformed tobacco seedlings with the sole exception of the transgenic lines overexpressing CsGSTU2 isoform that are barely influenced by herbicide treatment. In order to correlate the in planta studies with enzyme properties, recombinant CsGSTs were in vitro expressed and tested for GST activity using alachlor as substrate. The recombinant GSTU2 enzyme was twice more active than GSTU1 in conjugating alachlor to GSH thus indicating that CsGSTU2 might play a crucial role in the plant defense against the herbicide. Moreover, as a consequence of the infiltration with a bacterial suspension of the P. syringae pv. tabaci, transgenic tobacco plants but not wild type plants bestowed the capability to limit toxic metabolite diffusion through plant tissues as indicated by the absence of chlorotic halos formation. Consequently, the transgenic tobacco plants described in the present study might be utilized for phytoremediation of residual xenobiotics in the environment and might represent a model for engineering plants that resist to pathogen attack.

The response of higher plants to ionising radiation depends on factors related to both radiation properties and plant features including species, cultivar, age, and structural complexity of the target organ. Adult plants of dwarf tomato were irradiated with different doses of X-rays to investigate possible variations in leaf morpho-anatomical traits, photosynthetic efficiency, and genomic DNA. In order to assess if and how responses depend on leaf developmental stage, we analysed two types of leaves; nearly mature leaves (L1) and actively developing leaves (L2), whose lamina size corresponded to 70 and 25 %, respectively, of the lamina size of the fully expanded leaves. The results show that the X-rays prevented full lamina expansion of the L2 leaves at all doses and induced early death of tissue of plants irradiated with doses higher than 20 Gy. Most anatomical modifications were not clearly dose-dependent and the radiation-induced increase in phenolic compounds was irrespective of dose. At high doses of X-rays (50 and 100 Gy), photochemical efficiency decreased significantly in both leaf types, whereas total chlorophyll content significantly decreased only in the L2 leaves. The random amplification of polymorphic DNA data show that the X-rays induced mutagenic effects in the L2 leaves even at low doses despite the absence of severe phenotypic alterations. Genetic structure found in the population of samples corroborates the results of anatomical and eco-physiological analyses: the 20 Gy dose seems to mark the threshold dose above which genetic alterations, structural anomalies, and perturbations in the photosynthetic apparatus become significant, especially in the actively expanding leaves.

Phytocyanins (PCs) are ancient blue copper-binding proteins in plants that bind to single type I copper atoms and function as electron transporters. PCs play an important role in plant development and stress resistance. Many PCs are considered to be chimeric arabinogalactan proteins (AGPs). Previously, 38, 62, and 84 PC genes were identified in Arabidopsis thaliana, Oryza sativa, and Brassica rapa, respectively. In this study, we identified 30 putative PC genes in the orchid Phalaenopsis equestris through comprehensive bioinformatics analysis. Based on phylogeny and motif constitution, the P. equestris phytocyanins (PePCs) were divided into five subclasses: 10 early nodulin-like proteins, 10 uclacyanin-like proteins, five stellacyanin-like proteins, four plantacyanin-like proteins, and one unknown protein. Structural and glycosylation predictions suggested that 16 PePCs were glycosylphosphatidylinositol-anchored proteins localized to the plasma membrane, 22 PePCs contain N-glycosylation sites, and 14 are chimeric AGPs. Phylogenetic analysis indicated that each subfamily was derived from a common ancestor before the divergence of monocot and dicot lineages and that the expansion of the PC subfamilies occurred after the divergence of orchids and Arabidopsis. The number of exons in PC genes was conserved. Expression analysis in four tissues revealed that nine PC genes were highly expressed in flowers, stems, and roots, suggesting that these genes play important roles in growth and development in P. equestris. The results of this study lay the foundation for further analysis of the functions of this gene family in plants.

The interaction between phosphorus (P) and other media components alters root development and masks the plant response and thus limits the ability to correctly identify P-deficiency response (pdr) mutants. This study aims to assess changes in root development caused by different composition of growth media normally used in Arabidopsis research and to study their effects on pdr-mutant screening. Primary root growth of four genotypes was analyzed in media differing in P concentrations: half-strength Murashige and Skoog (1/2 MS) and Somerville and Ogren (SO). The effects of nitrogen source and Fe on root growth were investigated in each medium separately and in a mixture. We found that the primary root length of all genotypes grown on 1/2 MS was reduced in comparison with plants grown on SO medium. The mutant pdr9 was the most sensitive in 1/2 MS, This mutant was also hypersensitive to Fe that intensified its sensitivity to ammonium. Ammonium increased the root inhibition caused by Fe also in wild-type plants. In conclusion, on the basis of our study we recommend to use SO medium, which ensures an efficient selection to screen for pdr mutants through root growth. Moreover, nitrogen sources in the media other than nitrate should be taken carefully.

Selenium (Se) is an essential trace element for humans and animals. A hydroponic experiment was performed to study the effects of sulphur (S) on Se uptake, translocation, and assimilation in wheat (Triticum aestivum L.) seedlings. Sulphur starvation had a positive effect on selenate uptake and the form of Se supplied greatly influenced Se speciation in plants. Compared with the control plants, Se uptake by the S-starved plants was enhanced by 4.81-fold in the selenate treatment, and selenate was readily transported from roots to shoots. By contrast, S starvation had no significant effect on selenite uptake, and selenite taken up by roots was rapidly converted to organic forms and tended to accumulate in roots. X-ray absorption near edge spectroscopy (XANES) analysis showed that organic forms of selenium, including selenocystine, Se-methyl-selenocysteine (MeSeCys), and selenomethionine-Se-oxide, were dominant in the plants exposed to selenite and accounted for approximately 90 % of the total Se. Whereas selenate remained as the dominant species in the roots and shoots exposed to selenate, with little selenate converted to selenite and MeSeCys. Besides, sulphur starvation increased the proportion of inorganic Se species in the selenate-supplied plants, but had no significant effects on Se speciation in plants exposed to selenite. The present study provides important knowledge to understand the associated mechanism of Se uptake and metabolism in plants.

The cucumber genes CsPDR8/CsABCG36 and CsPDR12/CsABCG40 encode two similar pleiotropic drug resistance proteins (ABCG_PDRs) belonging to the large ABC family of multispecific ATP-dependent transporters. We have already shown that the amount of root CsPDR8/CsABCG36 and CsPDR12/CsABCG40 transcripts is markedly elevated by phytohormones related to the plant response to environmental constraints, suggesting the involvement of both genes in hormone-mediated reactions to stresses. To further characterize the function and regulation of CsPDR8/CsABCG36 and CsPDR12/CsABCG40, we determined the subcellular localization of the predicted CsPDR8/CsABCG36 and CsPDR12/CsABCG40 proteins in cucumber and performed a transcriptional analysis of genes encoding these proteins under different abiotic stresses (heavy metals, salinity, osmotic stress, and oxidative stress) and redox perturbations. In addition, the activities of antioxidative enzymes as well as the content of hydrogen peroxide and superoxide were measured in cucumber roots to monitor the redox perturbations under all experimental conditions. Western blot analysis of membrane fractions prepared from cucumber roots with specific antibodies raised against the peptides corresponding to sequences unique to CsPDR8/CsABCG36 and CsPDR12/CsABCG40 revealed that both proteins localize to the plasma membrane. The transcript abundance and the plasma membrane protein content closely correlated with the stress severity and the hydrogen peroxide content but not with the superoxide anion content. Based on the results obtained so far, we may conclude that CsPDR8/CsABCG36 and CsPDR12/CsABCG40 are up-regulated under multiple stress conditions and redox perturbations and that the H₂O₂ and stress-related phytohormones can act as signaling molecules affecting the expression of both cucumber genes.

In the present study, three Arabidopsis thaliana pop2 mutant lines with different T-DNA insertions in a gene coding γ-aminobutyric acid transaminase (GABA-TA) were screened for seed germination percentage, stress-induced oxidative damage, and GABA content and metabolism under various abiotic stresses including high temperature (42 °C), low temperature (4 °C), salinity (NaCl), and osmotic stress (mannitol). All mutant lines showed a decreased germination under all the stress treatments with a significant reduction in the pop2-1 and pop2-3 mutant lines. Content of GABA and MDA increased significantly in all pop2 mutants and wild type (WT) seedlings in response to all the treatments. However, content of GABA and MDA was lower in all pop2 mutants comparing to the WT under the same treatments. GABA increased already after 30 min and increased significantly after 2 h at 42 °C especially in the pop2-3 and WT seedlings. In response to the cold treatment, GABA content increased up to 4-fold compared to the control in all pop2 mutants and WT seedlings. In response to the NaCl treatment, GABA accumulated slightly in the WT and all pop2 mutants. On the contrary, GABA content increased significantly in the pop2, pop2-1, and pop2-3 mutants and WT under all mannitol treatments.

As result of a close evolutionary relationship between Triticeae B, G, and S genomes, the exchange of genetic material between them is possible and may be beneficial for broadening the genetic diversity of cultivated bread wheat. However, the extent to which regulatory networks are conserved remains poorly researched. Here, the structural organization and transcriptional activity of the B, S, and G genome copies of a gene encoding flavonoid biosynthesis enzyme chalcone-flavanone isomerase (CHI) were explored using introgression lines which differ from the wild type by carrying a non-bread wheat Chi-1 gene. Chi-S1, Chi-G1, and Chi-B1 all mapped to a comparable region of chromosomes 5S, 5G, and 5B, respectively. Nucleotide sequences of Aegilops speltoides Chi-S1 and Triticum timopheevii Chi-G1 were determined and compared with T. aestivum Chi-B1 sequences. The enzymes encoded by these three genes shared the same predicted tertiary structure and active sites. However, the replacement of Chi-B1 by Chi-S1 or Chi-G1 in a wheat background resulted in a significant decrease in the global amount of the Chi-1 transcript present in the seedling shoot indicating divergence in regulation of expression of the orthologous Chi-1 genes among Triticeae ssp.

One of the largest families of transcriptional regulators contains WRKY proteins. They play important roles in plant defense responses. In this study, a novel WRKY gene, CsWRKY2, was isolated from the tea [Camellia sinensis (L.) O. Kuntze] plant. The full-length cDNA of CsWRKY2 was 2 050 bp in length and encoded a 522-amino acid peptide chain containing two typical WRKY domains and two zinc finger motifs, suggesting that CsWRKY2 was member of the WRKY group I family. A subcellular localization assay shows that CsWRKY2 was localized to the nucleus. Real time qPCR analysis shows that CsWRKY2 expression was higher in leaves than in other organs and was induced by cold (4 °C), drought stress, and exogenous abscisic acid (ABA). Additionally, ABA content was enhanced after the cold or drought stress and the effects were relieved by an ABA biosynthesis inhibitor. Furthermore, the expression of CsWRKY2 was up-regulated by exogenous ABA under the cold and drought stresses and down-regulated by an ABA biosynthesis inhibitor. Our findings indicate that CsWRKY2 played an important role in plant defense responses to the cold and drought stresses by participating in the ABA signaling pathway, downstream to ABA.

Nitric oxide (NO) and 5-aminolevulinic acid (5ALA) play fundamental roles in plant responses to environmental stresses, but their cross-talk in antioxidant defense in cold-stressed Elymus nutans Griseb. have not been investigated. We herein report that 5ALA and NO donor, sodium nitroprusside (SNP), alleviated cold stress-induced plant growth inhibition and lipid peroxidation in roots of two E. nutans ecotypes (Damxung, DX and Zhengdao, ZD). However, application of an NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (PTIO) differentially blocked these protective effects indicating that an inhibition of NO accumulation reduced 5ALA-enhanced cold resistance. Application of exogenous 5ALA or NO markedly up-regulated activities of superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase, enhanced reduced glutathione accumulation and reduced glutathione to oxidized glutathione ratio, activated plasma membrane (PM) H⁺-ATPase, and reduced Na⁺/K⁺ ratio in roots of the two E. nutans ecotypes. Moreover, in the presence of 5ALA, nitric oxide synthase (NOS) activity and NO release in cold-resistant DX were higher than those in cold-sensitive ZD. Conversely, both NO treatment and inhibition of endogenous NO accumulation by PTIO or NOS inhibitor N^ω-nitro-L-arginine did not induce 5ALA production. These results suggest that NO might be acting as a downstream signal to mediate 5ALA-induced cold resistance by activating antioxidant defense and PM H⁺-ATPase and maintaining Na⁺ and K⁺ homeostasis.

Screening salt-sensitive mutants is a powerful method to identify genes associated with salt tolerance. We used forward genetic screening with sodium azide-mutated rice (Oryza sativa L. cv. Tainung 67) to identify mutants showing hypersensitivity to salt stress. A new mutant line, named salt hypersensitive 1 (shs1) and exhibiting a severe salt-sensitivity when grown under a high NaCl concentration, was identified; the salt hypersensitivity was caused by duplicate recessive epistasis with mutations likely in two different loci. The shs1 salt sensitive phenotypes included a decreased seed germination rate, reduced shoot height and root length, severe and quick wilting, and overaccumulation of sodium ions in shoots as compared with wild-type plants. In addition, shs1 showed a decreased photosynthetic efficiency and enhanced hydrogen peroxide (H₂O₂) production under the salt stress. An increased superoxide dismutase activity and decreased catalase activity were responsible for the hyperaccumulation of H₂O₂ in shs1. The hypersensitivity of shs1 to the salt stress might be caused by an impaired antioxidant machinery and cellular Na⁺ homeostasis.

Histone deacetylases (HDACs) catalyze histone deacetylation and play an important role in suppression of gene transcription in multiple cellular processes. HDACs are widely distributed in eukaryotes, however, detailed characterrization of HDACs in woody plants is not available. In this study, the sequences of reduced potassium dependency 3/histone deacetylase 1 (RPD3/HDA1) family proteins in black cottonwood (Populus trichocarpa Torr. & A. Gray) were characterized and their expression patterns in response to cold and salt stresses were determined. The RPD3/HDA1 proteins had conserved HDAC domains and can be divided into three classes based on sequence similarity and phylogenetic analysis. The transcripts of the HDAC genes were detected in different amounts in leaves, stems, and roots. The expressions of HDAC genes HDA902, HDA903, HDA904, HDA909, and HDA912 were up-regulated in a cold stress. Interestingly, in a salt stress, most of the HDAC genes were down-regulated. These results indicate that the poplar HDAC genes were regulated by the cold and salt stresses, and the members of the RPD3/HDA1 family play a role in stress responses.

The amount and composition of phytohormones, sugars, and some other leaf characteristics depending on a crop load were evaluated in apple (Malus domestica Borkh. cv. Ligol grafted on P 60 rootstock) trees in order to prevent biennial bearing. The crop load was adjusted to 12 (control, unthinned), 8, 4, and 0 (non-fruiting) inflorescences (or fruits) per cm² of trunk cross-sectional area (TCSA). Inflorescences were removed in May before flowering. Phytohormones were analyzed in axillary buds and leaves in September. Results show that, in contrast to the unthinned trees, thinning to 4 fruits cm^-2(TCSA) resulted in a significant decrease of yield per tree, but a significant increase of fruit mass, return bloom, and leaf area. The heavy crop load resulted in suppressed bloom in the following year. Composition and content of phytohormones was changed considerably. Moreover, thinning resulted in an increased hexose accumulation. Such data suggest that flowering inhibition depended on the phytohormones that were exported to buds and on sugar-hormone signalling cross-talk.

Alternative respiration pathway (AP) is an important pathway which can be induced by environment stresses in plants. In the present study, we show a new mechanism involving the AP in nitrogen deprivation-induced tolerance of Poa annua callus to salt stress. The AP capacity markedly increased under a 600 mM NaCl treatment or nitrogen deprivation pretreatment and reached a maximum under the nitrogen deprivation pretreatment combined with the NaCl treatment (-N+NaCl). Malondialdehyde (MDA) and H₂O₂ content and Na⁺/K⁺ ratio significantly increased under the 600 mM NaCl treatment but less under the-N+NaCl treatment. Moreover, both the nitrogen deprivation and the NaCl stress stimulated the plasma membrane (PM) H⁺-ATPase activity and increased pyruvate content. The maximal stimulating effect was found under the-N+NaCl treatment. When the AP capacity was reduced by salicylhydroxamic acid (SHAM, an inhibitor of AP), content of MDA and H₂O₂ and Na⁺/K⁺ ratio dramatically increased, whereas PM H⁺-ATPase activity decreased. Moreover, exogenous application of pyruvate produced a similar effect as the nitrogen deprivation pretreatment. The effects of SHAM on the Poa annua callus were counteracted by catalase (a H₂O₂ scavenger) and diphenylene iodonium (a plasma membrane NADPH oxidase inhibitor). Taken together, our results suggest that the nitrogen deprivation enhanced the capacity of AP by increasing pyruvate content, which in turn prevented the Poa annua callus from salt-induced oxidative damages and Na⁺ over-uptake.

Jasmonate ZIM domain (JAZ) proteins are key regulators of the jasmonic acid (JA) signaling pathway. Repressors of JAZ remain bound to the myelocytomatosis 2 (MYC2) or MYC3/MYC4 transcription factors in the absence of JA and negatively regulate transcription of the JA responsive genes. In the presence of JA, JAZ proteins interact with coronative insensitive 1 (COI1), the recognition molecule of E3 ubiquitin ligase SCF^COI1 (COI1 stabilized by Skp, cullin, F-box containing complex), get ubiquitinated, and subsequently degraded by the 26S proteasome. However, there is a dearth of knowledge about this gene family in monocot cereals, specifically its role in finger millet is unknown till date. Here we present the isolation and characterization of a novel JAZ family repressor gene from nonsequenced Eleusine coracana (EcJAZ) utilizing available genome information of Oryza, Sorghum, and Setaria. The EcJAZ sequence showed the presence of a conserved ZIM domain, the Jas motif, and N-terminal motif 7 like other Group1 TIFY sequence containing proteins. We observed coronatine (an analog of JA-Ile) dependent and time dependent degradation of recombinant EcJAZ that thereby fulfilled the basic characteristic of the JAZ proteins. We found a proteasome inhibitor N-(phenylmethoxy) carbonyl-L-leucyl-N-[(1R)-1-formyl-3-methylbutyl]-L-leucinamide) (MG132) mediated degradation inhibition of EcJAZ that supported its 26S proteasome mediated degradation. Our study shows the nuclear localization of GFP-EcJAZ by Agrobacterium mediated transient transformation of onion scale epidermal cells. In Eleusine leaves, transcription of EcJAZ increased 4.2-fold by salt stress and 5.5-fold by coronatine application; thus ascertained its inducibility by the abiotic stress as well as by bioactive JA-Ile. Taken together, all these results contribute to our understanding of the JA signaling pathway in Eleusine coracana.

Carrot (Daucus carota L.) chromosomes are small and uniform in shape and length. Here, mitotic chromosomes were subjected to multicolour fluorescence in situ hybridization (mFISH) with probes derived from conserved plant repetitive DNA (18-25S and 5S rDNA, telomeres), a carrot-specific centromeric repeat (Cent-Dc), carrot-specific repetitive elements (DCREs), and miniature inverted-repeat transposable elements (MITEs). A set of major chromosomal landmarks comprising rDNA and telomeric and centromeric sequences in combination with chromosomal measurements enabled discrimination of carrot chromosomes. In addition, reproducible and unique FISH patterns generated by three carrot genome-specific repeats (DCRE22, DCRE16, and DCRE9) and two transposon families (DcSto and Krak) in combination with telomeric and centromeric reference probes allowed identification of chromosome pairs and construction of detailed carrot karyotypes. Hybridization patterns for DCREs were observed as pericentromeric and interstitial dotted tracks (DCRE22), signals in pericentromeric regions (DCRE16), or scattered signals (DCRE9) along chromosomes similar to those observed for both MITE families.

PopW, a harpin protein identified from Ralstonia solanacearum, has multiple beneficial effects in plants, promoting plant growth and development, increasing crop yield, and inducing resistance to pathogens. Tobacco plants transformed with popW, the PopW-encoding gene, exhibited a promoted growth rate and enhanced resistance to Tobacco mosaic virus (TMV). Here, it is documented that the transgenic tobacco plants overexpressing popW exhibited a higher resistance to R. solanacearum YN10 infection compared with that of the wild-type plants. In the popW-expressing tobacco lines, an enhanced H2O2 accumulation and hypersensitive reaction (HR) were activated in the inoculated site. In addition, the resistance was accompanied with increased transcripts in numbers of genes related to defense (including HR), reactive oxygen species (ROS) scavenging, and salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) production. These results suggest that popW acted as positive regulator in tobacco resistance against R. solanacearum via modulation of SA-, JA-, and ET-mediated signaling pathways. We report for the first time that the expression of a harpin-encoding gene in vivo improved plant resistance to R. solanacearum.

Members of the Brassicaceae family disperse their seeds through a mechanism commonly referred to as fruit dehiscence or pod shatter. Pod shatter is influenced by variations in valve margin structure and by the molecular control pathways related to valve development. Anatomical patterns of the dehiscence zone from Brassica napus L., Brassica rapa L., Brassica carinata L., and Sinapis alba L., representing fruit types differing in pod shatter resistance, were compared using histological staining. The pod shatter-susceptible plant B. napus showed an increased lignin deposition at the vascular bundle of the replum as well as an increased separation of cell layers. In pod shatter-resistant plants S. alba, B. rapa, and B. carinata, we observed two layers of lignified valve margin cells. From these four species, we isolated and identified homologs of SHATTERPROOF (SHP1, SHP2), INDEHISCENT (IND), ALCATRAZ (ALC), FRUITFULL (FUL), AGAMOUS (AG), NAC SECONDARY WALL THICKENING PROMOTING FACTOR1 (NST1), and SEEDSTICK (STK) genes involved in fruit development and pod shatter in Arabidopsis. Transcriptional analysis of these eight genes was performed by real-time polymerase chain reaction and the results demonstrate that differences in the expression patterns of the eight genes may be associated with dehiscence variation within these four species.

Globally, drought is the main factor that reduces common bean yield. For this reason, breeding alternatives, such as molecular marker-assisted selection, that focus on various functional genes directly involved in the response to water stress, such as those encoding late embryogenesis abundant (LEA), early response to dehydration (ERD), and dehydrin proteins, have been implemented. The aim of this study was to identify differentially expressed genes of Phaseolus vulgaris in drought-tolerant cultivars Pinto Saltillo (PS) and Pinto Villa (PV), and drought-susceptible cultivars Bayo Madero (BM) and Canario 60 (C60) in vegetative and reproductive stages. Relative water content (RWC) in leaf tissue was measured. Twenty-eight P. vulgaris genes obtained from GenBank and from a subtractive suppressive library from the PS cultivar were analysed, and their expression profiles were examined by reverse transcription polymerase chain reaction (RT-PCR). Then, cDNA arrays were developed and hybridised to confirm expression which was finally validated by quantitative PCR (qPCR). The usefulness of the identified genes as selection criteria for the tolerance of different genotypes to drought was examined using cDNA arrays. Expression of 21 genes was induced by drought. The cDNA arrays confirmed that expression of 19 of these genes increased in the vegetative stage upon exposure to the drought, and a higher expression was observed in the reproductive stage compared with vegetative stage V4. Only five genes induced by the drought were found to have a lower expression in the susceptible cultivars compared with the tolerant ones. During recovery after the drought in the reproductive stage, 13 of the 21 induced genes remained transcriptionally active including LEA3 and dehydrin. The RWC during the drought in vegetative stage V4 decreased by about 55 % in all cultivars, but at the onset of flowering, it increased to 80 % in PV and PS. In contrast, in the susceptible cultivars, it remained at 55 %. Using qPCR validation, expression induction was confirmed in the drought-tolerant cultivars. Polyubiquitin2, LEA3, LEA4, and dehydrin were useful genes for selecting drought-tolerant genotypes under field conditions.

The first rate-limiting enzyme of the mevalonate pathway during isoprenoid biosynthesis is 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR). In this study, the expression pattern of the MdHMGR2 gene in Malus domestica suggests that MdHMGR2 was expressed in a tissue-specific manner and was significantly induced by ethephon (ETH), indoleacetic acid (IAA), methyl jasmonate (MeJA), and salicylic acid (SA). The MdHMGR2 promoter was isolated, sequenced, and analyzed through bioinformatics tools, and the results suggest the presence of various putative cis-acting elements responsive to different hormones. Activity of β-glucuronidase (GUS) driven by the full length MdHMGR2 promoter and its 5'deletion fragments was detected in transgenic Arabidopsis thaliana. A strong GUS activity was observed in seedlings, roots, newly growing true leaves, anthers, and stigmas in transgenic Arabidopsis containing the full MdHMGR2 promoter. The results indicate that a region from -1050 to -827 was crucial for promoter activity. In addition, the MdHMGR2 promoter was induced in response to ETH, IAA, MeJA, and SA. The analysis suggests that an ethylene-responsive element in the region from -1050 to -1005 was required for the ethylene inducibility.

To study the effects of application of salicylic acid (SA) on the antioxidant system and photosystem II (PS II) in wheat seedlings we used two different experiments. The first method was carried out by immersing roots in Hoagland's nutrient solution containing 0, 0.25, or 2.5 mM SA, and the second method was performed by spraying two-week-old seedlings with the same SA concentrations. After 24 h, chlorophyll fluorescence, thylakoid membrane proteins, antioxidant enzyme activities, and reactive oxygen species were measured. The low concentration of SA caused a significant increase in the antioxidant enzyme activities. However, the treatment with 2.5 mM SA resulted in an increase in the non-photochemical quenching coefficient and a decrease in the antioxidant enzyme activities, the quantum yield of PS II photochemistry, and the photochemical quenching, especially in the first method of application. All these results indicate that the effects of SA on PS II and the antioxidative defense system were dependent on the concentration used and the method of application.

Very little is known about lipid transfer proteins from flax (Linum usitatissimum L.). In the present work, three genes encoding a lipid transfer protein (LTP) were isolated from flax, two of which encoded Type-1 and one Type-2 LTPs with molecular masses of about 9 and 7 kDa, respectively. The analysis of deduced amino acid sequence reveals that only Type 2 of the L. usitatissimum leaf specific LTP (LuLTP_Ls) had an N terminal signal peptide consisting of 23 amino acids. The phylogenetic analyses of LuLTP_Ls suggest their closest relatedness with respective proteins from Dimocarpus longan and Vitis vinifera. The gene expression analysis shows that LTP Type 1 genes, which include LuLTP_Ls1 and LuLTP_Ls3, were progressively expressed during leaf development, whereas LuLTP_Ls4 (Type 2) was expressed only at initial and terminal senescence stages of cotyledons. The results suggest that both types of LuLTP_Ls were differentially yet significantly expressed in cotyledons implicating their function in transport and scavenging lipidic skeletons for the benefit of other developing parts of the plant.