The nuclear factor Y (NF-Y) is one of the largest transcription factor families in plants consisting of NF-YA, NF-YB, and NF-YC subunits. It could play important roles in various processes such as flowering time, seed development, and response to drought. In this study, 6 NF-YA, 13 NF-YB, and 8 NF-YC proteins were identified and characterized in Prunus mume. Analyses of a conserved domain indicated that the PmNF-Y subunits shared an elevated degree of homology with the corresponding Arabidopsis NF-Y ones. Phylogenetic analysis showed that each NF-Y subunit family from Prunus mume and Arabidopsis could be divided into 4 or 2 clades based on their full-length proteins. The gene expression patterns of all 27 PmNF-Y genes were examined under abscisic acid (ABA), osmotic, salt, and H₂O₂ treatments using real-time quantitative PCR analyses. PmNF-YA1/2/4/5/6, PmNF-YB3/4/8/10/11/13, and PmNF-YC1/2/4/5/6/8 were found to be up-regulated under the ABA and osmotic treatments. PmNF-YA1/2/3/4/5/6, PmNF-YB1/3/8/10/11/13, and PmNF-YC1/2/5/6/8 were obviously induced by the H₂O₂. In addition, only PmNF-YA2 and PmNF-YB3 expressions were enhanced under the salt stress. These findings could provide an entry point to investigating the roles of PmNF-Y genes during abiotic stress responses.

Root epidermis and apoplastic barriers (endodermis and exodermis) are the critical root structures involved in setting up plant-soil interface by regulating free apoplastic movement of solutes within root tissues. Probing root apoplast permeability with "apoplastic tracers" presents one of scarce tools available for detection of "apoplastic leakage" sites and evaluation of their role in overall root uptake of water, nutrients, or pollutants. Although the tracers are used for many decades, there is still not an ideal apoplastic tracer and flawless procedure with straightforward interpretation. In this article, we present our experience with the most frequently used tracers representing various types of chemicals with different characteristics. We examine their behaviour, characteristics, and limitations. Here, we show that results gained with an apoplastic tracer assay technique are reliable but depend on many parameters-chemical properties of a selected tracer, plant species, cell wall properties, exposure time, or sample processing.

Eragrostis curvula (Schrad) Nees (weeping lovegrass) represents important cultivated forage in semiarid regions, and the most useful cultivars are tetraploid and reproduce by pseudogamous diplosporous apomixis. We previously produced a series of genetically related E. curvula lines that provide a suitable system for the identification of gene(s) involved in diplosporous apomixis and ploidy, including a natural apomictic tetraploid (T), a diploid sexual line (D), and a tetraploid sexual plant (C). A collection of expressed sequence tags (ESTs) was generated from cDNA libraries obtained from panicles of the D, T, and C, and leaves of the T. The present study aimed to analyze the repetitive content of these four cDNA libraries and further identify and characterize transposable element (TE)-related ESTs. Repetitive sequences were identified through the interface RepeatMasker (RM) using the database Repbase Update and further classification of TEs was performed manually from the RM output. The different contribution of ESTs with identity to TEs among libraries was further evaluated, and such differences were validated through RT-qPCR. We found that the percentage of repetitive content in the leaf cDNA library was almost double than in inflorescence libraries, with retrotransposons contributing mostly in all libraries. The expression of TE-related ESTs was compared in cDNA samples extracted from D, T, and C leaves or inflorescences revealing that seven mRNAs containing MuDR-like DNA transposons, Gypsy-like, and Copia-like retrotransposons were differentially represented according to tissue, reproductive mode, or ploidy. The euploid series of Eragrostis curvula is a useful model to the study of epigenomic changes produced after changes in ploidy. The present work constitutes the first detailed report on repetitive sequences of Eragrostis curvula at the transcriptome level.

In vitro orchid micropropagation is efficient biotechnological strategy for conservation and commercial plantlet production. However, micropropagated plantlets generally need to adapt to survive severe changes in humidity, irradiance, and growing medium that accompany the transfer to ex vitro conditions. Such adaptive cellular changes would give insights into the phenotypic plasticity of the model plant Cattleya xanthina (L.) Van den Berg. Therefore, we aimed to evaluate structural changes in the leaves of C. xanthina cultivated in vitro and acclimatized ex vitro using qualitative and quantitative analyses. During acclimatization, we observed a higher accumulation of dry mass, a greater convexity of the outer surface of epidermal cells, an increased deposition of epicuticular waxes, a greater elongation of mesophyll parenchymatic cells, and finally, the presence of chloroplasts with organized thylakoids and well-developed grana. Stomatal density was not changed. Furthermore, a gradual acclimatization allows this species the best adaptation to a new environment.

Calcium ion (Ca²⁺) is essential secondary messenger in plant signaling networks. In this study, the effect of Ca²⁺ on oxidative damage caused by a high irradiance (HI) was investigated in the leaves of two cultivars of tall fescue (Arid3 and Houndog5). Pretreatment of the tall fescue leaves with a CaCl₂ solution significantly increased Ca²⁺ content and intrinsic HI tolerance due to a decreased ion leakage and content of malondialdehyde, hydrogen peroxide, and superoxide radicals. Moreover, the activities of superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase increased in both the cultivars in the presence of Ca²⁺ under the HI stress. In contrast, treatments with a Ca²⁺ chelator ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) or a plasma membrane Ca²⁺ channel blocker LaCl₃ reversed these effects. On the other hand, a pronounced increase in nitric oxide synthase-like activity and NO release by exogenous Ca²⁺ treatment was observed in the tolerant Arid3 plants after exposure to the HI, whereas only a small increase was observed in more sensitive Houndog5. Moreover, the inhibition of NO production by 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide or N ^ω-nitro-L-arginine blocked the protective effect of exogenous Ca²⁺, whereas the inhibition of Ca²⁺ by EGTA or LaCl₃ had no influence on the protective effect of NO. The results indicate that NO might be involved in the Ca²⁺-induced activities of antioxidant enzymes further protecting against HI-induced oxidative damage. This protective mechanism was found to be more efficient in Arid3 than in Houndog5.

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.

We tested the capability of plants to utilize a mixture of amino acids and peptides as the exclusive source of nitrogen. The aim of this study was to find out how such a nutrition affected plantlet growth, photosynthetic performance, and N assimilation metabolism in tobacco (Nicotiana tabacum L., cv. Petit Havana SR1) grown in vitro. Plantlets grown in a casein hydrolysate-supplemented (CA) medium were compared to plantlets grown in a complete Murashige-Skoog (MS) medium, plantlets grown in an ammonium-deficient medium (N1), or plantlets grown in a nitrate-reduced medium (N2). In addition, the plantlets were grown in the presence or absence of 1.5 % (m/v) saccharose as an additional carbon source. Casein hydrolysate, similarly as inorganic N limitation, reduced generally plantlet growth, whereas no significant effects were observed on photosynthetic parameters evaluated by chlorophyll a fluorescence. Although addition of saccharose stimulated the plantlet growth particularly in the MS, it showed a rather negative influence both on the growth and on the photochemical efficiency of photosystem II in the plantlets grown in the CA and N1. The activities of enzymes involved in N assimilation, such as nitrate reductase (NR) and glutamine synthetase (GS), were lower in the plantlets grown in the CA, N1, and N2, both in leaves and in roots. On the other hand, glutamate synthase and glutamate dehydrogenase were employed by the plantlets grown in the CA. The presence of saccharose in the growth medium stimulated mainly NR and GS activities in the MS grown plantlets, whereas enzyme activities of the plantlets grown on the N1, N2, and CA were not significantly influenced. We proved that the tobacco plantlets can utilize casein hydrolysate as the sole source of N particularly during their photoautotrophic cultivation. Contrary to positive effects of photomixotrophic nutrition for the MS grown plantlets, exogenous sugar seemed to diminish the ability of the casein hydrolysate-supplemented plantlets to utilize efficiently the additional C source.

To investigate whether exogenous sucrose can protect cucumber from water stress, cucumber (Cucumis sativus L.) seedlings were pretreated with 90 mM sucrose or 90 mM mannitol for 1 d and then were dehydrated with 10 % (m/v) polyethylene glycol (PEG) 6000 for further 1 d. Dehydration inhibited plant growth and decreased osmotic potential and relative water content (RWC) in leaves. The pretreatment with 90 mM sucrose further reduced the osmotic potential but increased the RWC and alleviated the growth inhibition. Compared with the PEG treatment alone, the combination of sucrose + PEG increased the activities of superoxide dismutase, guaiacol peroxidase, glutathione reductase, dehydroascorbate reductase, monodehydroascorbate reductase, ascorbate peroxidase, and glutathione peroxidase, and elevated the content of endogenous sucrose, glucose, and fructose together with the activities of soluble acid invertase and neutral invertase. This was in accordance with the enhanced transcription of genes encoding copper/zinc superoxide dismutase, guaiacol peroxidase, and glutathione reductase. Furthermore, the sucrose pretreatment decreased the content of malondialdehyde and hydrogen peroxide and increased the content of ascorbate, reduced glutathione, and proline under the dehydration. Taken together, the pretreatment with 90 mM sucrose, but much less with mannitol, induced antioxidants, proline, and soluble sugars and thus reduced dehydration-caused damage to the cucumber seedlings.

Barley (Hordeum vulgare L.), an important food and fodder crop, is potentially tolerant to salinity. To identify quantitative trait loci (QTLs) controlling salt tolerance, the population of 162 recombinant inbred lines (RILs) derived from F8 generation of Arigashar (an extremely salt tolerant Iranian six-rowed barley landrace) crossed with Igri (a salt semi-sensitive two-rowed cultivar) were evaluated. The growth of shoots, roots, and coleoptiles, and root numbers are four important growth characteristics severely affected by salt stress at seedling growth stages. A linkage map was constructed using 106 AFLP and SSR markers spanning six barley chromosomes including 2(2H), 3(3H), 4(4H), 7(5H), 6(6H), and 1(7H). Out of totally 26 detected QTLs, 17 QTLs were found effective for salt tolerance at 250 and 350 mM NaCl which localized on chromosomes 2H, 3H, 4H, 6H, 7H, and linkage group L1, whereas considering equivalent overlapped QTLs with a pleiotropic effect led to detection of totally 9 distinctive QTLs (QClgH2.1b, QSdgH2.1b, QSlgH2.1c, QNrgH2.1b, QTwgH2.2c, QSdg3Hb, QSlg4Hb1, QClg4Hb, and QSlg6Hc2) effective for salinity tolerance. 2(2H), 4(4H), and 6(6H) were major chromosomes harboring QTLs which effectively controlled salt tolerance in the Igri×Arigashar population. An interesting QTL, QTwg4Hc, was localized on chromosome 4H in the XE41-M61 marker distance that controls several traits including shoot and coleoptile lengths and shoot fresh mass under salt stress. A dense marker cluster around a resistance gene could offer a starting point for positional cloning.

Protein synthesis is a ubiquitous and essential process in all organisms, including plants. It is primarily regulated at translation initiation stage which is mediated through a number of translation initiation factors (eIFs). It is now becoming more apparent that in addition to synthesis of proteins, eIFs also regulate various aspects of plant development and their interaction with environment. Translation initiation factors, such as eIF3, eIF4A, eIF4E, eIF4G, and eIF5A affect different processes during vegetative and reproductive growth like embryogenesis, xylogenesis, flowering, sporogenesis, pollen germination, etc. On the contrary, eIF1A, eIF2, eIF4, and eIF5A are associated with interaction of plants with different abiotic stresses, such as high temperature, salinity, oxidative stress, etc. Similarly, eIF4E and eIF4G have roles in interaction with many viruses. Therefore, the translation initiation factors are important candidates for improving plant performance and adaptation. A large number of genes encoding eIFs can functionally be validated and utilized through genetic engineering approaches for better adaptability and performance of plants by inhibiting/minimizing or increasing expression of desired eIF(s).

The effects of a short (30 min) heat shock (HS) on plants subsequently grown under a salinity stress (SS, 200 mM NaCl) for 10 d were investigated in barley (Hordeum vulgare L.) cv. Tokak 157/37. The maximum temperature for HS allowing plant survival was 45 °C. The root length was significantly decreased by SS, whereas HS alone did not affect root growth. Interestingly, HS stimulated root elongation under SS. An osmotic adjustment was promoted in leaves by SS. On the contrary, HS increased the osmotic potential in leaves in the absence of SS, and partly counteracted the effect of SS in the HS+SS treatment. Cu/Zn-SOD, HvAPX, HvCAT2, HSP17, HSP18, and HSP90 were transcribed in leaves of HS-treated plants, but not in control plants. The HSP70 was constitutively transcribed in both the SS and control plants, but after HS, a shorter amplicon was also observed. The genes coding antioxidants, Cu/Zn-SOD, HvCAT2 and HvAPX, were differentially influenced by SS or HS+SS in the roots and leaves. In the roots, the mRNA content of BAS1, HvDRF1, HvMT2, and HvNHX1 increased after the HS treatment. In a recovery experiment in which plants were grown to maturity after HS and HS+SS stress exposure, the plant height increased and the time to maturity was reduced in comparison with SS. Our results show that HS could stimulate plant growth and reduce some of the negative effects of SS, and that it affected the transcription of several stress-related genes.

Cytosine methylation (C meth) is a ubiquitous regulator of gene transcription. The maize gene ZmGRP3 is a notable marker for root initiation, but how its root-specific transcription is regulated is not understood. Here, bisulfite sequencing and a C meth sensitive Southern blot assay were used to show that the transcription of ZmGRP3 was promoted by a reduction in the extent of C meth both in its promoter and in the vicinity of its translation start site. The result was validated by revealing the effect of 5-aza-2-deoxycytidine on the C meth status in various seedling organs. An analysis of methylation patterns indicates that the C meth of cytosine-guanine dinucleotides was the most important determinant of transcription, however, the C meth of CHG or CHH trinucleotides had little influence.

Two barley (Hordeum vulgare L.) cultivars, Czech spring cv. Amulet and Syrian landrace Tadmor, were subject to different salinity treatments: 1) the NaCl concentration was gradually increased from 0 (the control) to either 100 (a moderate salt stress) or 300 mM NaCl (a high salt stress), 2) the NaCl concentration was increased directly either from 0 to 300 mM NaCl or from 100 to 300 mM NaCl, and 3) a recovery when all variants were transferred back to control conditions and cultivated for seven additional days before sampling. The following parameters were determined: water saturation deficit (WSD), osmotic potential (ψ_s), leaf proline content, maximum quantum yield of photosystem (PS) II photochemistry (measured as variable to maximum chlorophyll a fluorescence ratio, F_v/F_m), and relative accumulation of dehydrins (DHN). Both quantitative and qualitative differences in dehydrins were found between NaCl-treated Amulet and Tadmor. A principal component analysis (PCA) of all experiment data revealed a differential ability of Amulet and Tadmor to recover after the 300 mM NaCl treatments indicating better salt tolerance in Tadmor. Correlation analyses have shown statistically significant correlations between WSD, ψ_s, proline, and DHN.

Lipase class 3 is part of the triacylglycerol lipase family involved in lipid degradation, esterification, and transesterification processes in plants. In this study, a lipase class 3 gene and promoter from oil palm (Elaeis guineensis Jacq.) were isolated and characterized by Northern blot, Southern blot, oil palm genome sequence, and transient expression GUS assay. The full-length lipase class 3 (FLL1) deduced polypeptide encoded 483 amino acids and was identical to that deduced from lipase (EgLip1) cDNA (GI: 409994625). It contained the lipase consensus sequence, GxSxG motif, and a putative catalytic triad and had a 3-dimensional protein model similar to that of a lipase from Giberella zeae with a 50 % identity. The Northern blot and reverse transcription polymerase chain reaction (RT-PCR) show that FLL1 was predominantly expressed in the mesocarp and the expression increased as fruits reached maturity. A lower expression was detected in germinated seedlings and especially in roots. The expression of FLL1 was also enhanced in the mesocarp of cold treated fruits. A high oil accumulation in the mesocarp during fruit development makes this tissue a suitable target for a genetic modification, hence the isolation of the FLL1 promoter. The transient expression of the β-glucuronidase (GUS) gene driven by the FLL1 promoter detected the GUS expression in mesocarp slices, especially in vascular bundles. This suggests the potential role of using the promoter as tool to direct the expression of a transgene to the mesocarp of transgenic oil palm.

The growth of the Cd-hyperaccumulator Solanum nigrum L. and its physiological responses to a short-term (7 d) Cd stress and to exogenous methyl jasmonate (MeJA) were investigated. Compared with the leaves of S. nigrum, the roots were more liable to Cd and showed a significantly decreased dry mass and increased malondialdehyde content. Cd accumulation in the shoots and roots of S. nigrum were proportional to the Cd concentration in the hydroponic solution. The application of a low concentration of MeJA (0.01 μM) significantly reduced the translocation/accumulation of Cd in both the shoots and roots compared with a 40 mg dm^-3 Cd treatment only. Moreover, 40 mg dm^-3 Cd significantly decreased the activity of leaf superoxide dismutase, but 0.01 μM MeJA restored it. MeJA also enhanced the activity of catalase in the leaves but showed no significant effect on peroxidase activity. The content of both endogenous jasmonic acid (JA) and MeJA in the leaves of S. nigrum increased with the increase of exogenous MeJA concentration.

The GhMYB9 encodes a R2R3 MYB transcription factor in the upland cotton (Gossypium hirsutum L.) genome. Our studies show that GhMYB9 predominantly expressed in flowers and fibers. To gain a better understanding of its regulatory mechanism, we isolated the 5'-flanking region of GhMYB9 which was 1 487 bp in length. The cis-acting element prediction shows that this region contained the basic structure of the core promoter elements (TATA-box, CAAT-box) and the transcription start site (TSS). Other motifs, such as defense and stress responsiveness (TC-rich repeats), anaerobic induction (ARE), and MYB binding sites involved in drought-inducibility (MBS), were also found. Histochemical assay shows that the GhMYB9 promoter governed β-glucuronidase (GUS) expression mainly in seeds, fibers, and flowers of transgenic cotton. Also, the activity of the promoter was induced by auxin in fibers of transgenic cotton. This is consistent with its transcript abundance in different tissues. A further deletion analysis confirms that a promoter region from -1 231 to -860 was required for auxin response. Our findings provide a useful reference for the understanding of the transcriptional regulation mechanism of the GhMYB9 gene.

Actinidia kolomikta (Rupr. & Maxim.) Maxim. leaves showed dramatic colour changes during plant growth phases, and we studied structure and optical properties of variegated leaves. Leaf surface cells were smooth, and there were no surface appendages (wax or trichomes) observed in variegated leaves. Palisade tissue cells in white and pink leaves were looser and contained relatively fewer chloroplasts. White leaves contained many intercellular spaces between the epidermal and mesophyll cells or within the palisade cell layer. Variegated leaves had three distinct radiation reflection patterns: a bright white area, a spotted pattern, and a polygonal pattern. Reflectance at 450-1100 nm from the adaxial surface of white leaves was greater than that of green leaves, but anthocyanin accumulation in pink leaves decreased the reflectance at 500-600 nm. When variegated leaves turned green, the reflectance at 500-600 nm increased. On abaxial surfaces, the reflectance of variegated leaves was similar to green leaves at 450-700 nm. In conclusion, reflection patterns and the formation of variegated leaves of A. kolomikta were significantly correlated with the leaf anatomy. The white and pink colours of leaves were a result of an internal reflection between air spaces and cells in the leaves, chlorophyll deficiency in palisade tissue, and anthocyanin accumulation. Variegated leaves turned green when the chlorophyll content in palisade tissue increased.

A photoperiod-sensitive soybean [Glycine max (L.) Merr] cv. ZhongDou 24 (ZD24) exhibiting delayed flowering when grown under long-days (LD, a 16-h photoperiod) was used to identify the genetic control of flowering delay. A differential expression profiling technique enabled identification of a gene fragment that was up-regulated under LD. This fragment was homologous to a gene encoding methionine synthase (MS) in soybean and was named GmMS. The RNA content confirmed that GmMS was expressed in roots, stems, and leaves of soybean grown under LD. The highest expression was in stems. Full length GmMS, encoding 763 amino acids, was transferred into tobacco plants. The ectopic expression of GmMS in tobacco resulted in delayed flowering. Other effects included stunting, an increased MS activity and methionine content, a higher content of alcohol-soluble proteins and of chlorophylls, and a lower content of anthocyanins.

Extracellular ATP (eATP) has been considered as signalling compound to mediate several physiological processes. Here we show that eATP played a role in alleviating the salicylic acid (SA)-induced inhibition of cell viability and respiration in tobacco (Nicotiana tabacum L. cv. Bright Yellow-2) suspension cells. Exogenous SA at higher concentrations (0.05, 0.1, 0.3, 0.5, or 0.7 mM) caused a significant reduction in respiratory O₂ uptake or cell viability. The addition of exogenous ATP alleviated the SA-induced reductions in cell viability and respiration, but the effect was dependent on the concentrations of either exogenous SA or ATP. Further study shows that the alleviative effects of exogenous ATP were abolished by the addition of GdCl₃ (an inhibitor of Ca²⁺ channels) or EGTA (a Ca²⁺ chelator). These results suggest that this role of extracellular ATP could be related to a Ca²⁺ signalling pathway.

In this study, proteome and activities of glutathione (GSH)-related enzymes were investigated in detached leaves of Arabidopsis thaliana treated with ochratoxin A (OTA) alone or supplemented with buthionine sulfoximine (BSO, a specific inhibitor of the first step in GSH biosynthesis). A comparative proteomic study using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization-time of flight tandem mass spectrometry (MALDI-TOF/TOF MS/MS) identified 12 differentially expressed proteins mainly involved in GSH metabolism, energy metabolism, sugar metabolism, and photosynthesis. The treatment with OTA significantly enhanced the activities of glutathione-S-transferase (GST) and glutathione reductase (GR) through up-regulating the corresponding genes (GSTF7, GR1), an the diminishing effect of BSO on them counteracted the results. However, both OTA and BSO decreased the activity of ascorbate peroxidase (APX), and OTA also decreased the monodehydroascorbate reductase (MDHAR) and glutathione peroxidase (GPX) activities. Briefly, the OTA-induced phytotoxicity to the A. thaliana detached leaves was increased slightly by addition of BSO, and the fluctuation in GSH synthesis, GSH metabolism and disorder of cellular metabolism happened.

Coleus forskohlii (syn. Plectranthus barbatus) is a widely used medicinal plant and its main bioactive constituents are diterpenes forskolin and isoforskolin. The present study aimed to construct a cDNA library to identify expressed sequence tags related to terpene biosynthesis in C. forskohlii. We constructed a high quality normalized full-length cDNA library which reached the requirements (abundance, integrity, and library content) for isolating full-length genes. A total of 4 224 cDNA clones were sequenced and 2 394 unigenes were assembled with an average unigene size of 753 bp. A total of 2 100 (87.7 %) unigenes were functionally classified using gene ontologies, and 1 716 (71.7 %) unigenes were assigned to establish pathway associations in KEGG mapings. Notably, 64 unigenes putatively participated in the biosynthesis of secondary metabolites, in which 17 unigenes were identified that might be involved in the biosynthesis of the terpenoid backbone and monoterpenes, diterpenes, and triterpenes.

The effects of copper oxide nanoparticles (CuONPs) on germinating seedlings of green pea (Pisum sativum L.) were studied. The seedlings were grown in a half-strength Murashige and Skoog semisolid medium containing 0, 50, 100, 200, 400, and 500 mg dm^-3 CuONPs for 14 d under controlled growth chamber conditions. Exposures to 100, 200, 400, and 500 mg dm^-3 CuONPs significantly reduced plant growth (shoot and root lengths) and increased reactive oxygen species (ROS) generation and lipid peroxidation. Gene expression study using real-time polymerase chain reaction showed no significant change in the expression of genes coding CuZn-superoxide dismutase (CuZnSOD), catalase (CAT), and ascorbate peroxidase (APX) in shoots. However in roots, a significant increase in the expression of the CuZnSOD gene was observed under the exposures to 100, 200, 400, and 500 mg dm^-3 CuONPs, in the expression of the CAT gene under 100 and 200 mg dm^-3 CuONPs, and in the expression of APX under 200 and 400 mg dm^-3 CuONPs.

Tospoviruses are devastating plant viruses causing severe economic losses in a diverse range of crops worldwide. Here, we describe the development and evaluation of an RNA interference (RNAi) broad-spectrum virus resistance strategy based on a unique and short hairpin-RNA-generating construct (pNhpRNA). This construct was designed from a region of the nucleocapsid gene (N) of Tomato spotted wilt virus (TSWV) that showed a high sequence identity to the corresponding region in the related species Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV). To test the effectiveness of the pNhpRNA construct, we developed a silencing reporter assay based on three fusion proteins in which the complete viral N gene sequence from each of the three tospoviruses was fused in frame to the green fluorescent protein (GFP) sequence. Co-agroinoculation of these constructs with pNhpRNA into leaves of Nicotiana benthamiana resulted in a strong silencing phenotype determined by GFP decay and suppression of the three N genes at the RNA and protein levels. To test the potential of the pNhpRNA construct to generate virus-resistant plants, we infiltrated the whole shoots of N. benthamiana with pNhpRNA. When these infiltrated plants were mechanically inoculated with the mentioned viruses 100, 70, and 60 % resistance phenotypes to TSWV, GRSV, and TCSV, respectively, were observed. The induction of a broad tospovirus resistance with a simple construct and a minimized off-target effect are the main contributions of pNhpRNA.

In shaded wheat (Triticum aestivum L.) leaves, the suppression of blue radiation (BR) triggers senescence. This phenomenon is correlated to an increase in oxidative stress symptoms and a decrease of catalase (CAT) activity, among other traits. Previous data suggest that the radiation signal transduction pathway may involve changes in Ca²⁺ and H₂O₂ homeostasis. For better a understanding of the interaction among the spectral composition of radiation, Ca²⁺ availability, and the antioxidant metabolism in the regulation of shade-induced senescence, detached wheat leaves were placed in a growth chamber and exposed to either blue (B, high BR transmittance) and/or green (G, very low BR transmittance) Lee® filters in the absence or presence of 0.8 mM verapamil (a Ca²⁺ channels blocker), 4.0 mM EGTA (a Ca²⁺chelator), or 8.0 mM 3-amino-1,2,4-triazole (a CAT inhibitor). At defined time points, the leaf samples were analyzed for changes in chlorophyll content, specific activities of CAT, ascorbate peroxidase (APX), and guaiacol peroxidase (POX), CAT isozymes, and gene expression of CAT1, CAT2, and two senescence markers (TaSAG1 and TaSAG3). BR transmittance decreased the chlorophyll degradation rate and SAG genes expression either in leaves continuously exposed under the B filter, as well as in leaves previously exposed under the G filter. The effect of BR was associated with the maintenance of a high CAT (but not APX and POX) activity, and it was suppressed either in the presence of 3-AT or when Ca²⁺ availability was decreased. BR altered the CAT activity both at the transcriptional and at the posttranscriptional level. Nevertheless, different responses of CAT isozymes and CAT genes expression profiles to specific treatment combinations indicate that they differed in their regulatory pathways.

A segregating F4 population from the cross between drought sensitive (Yecora Rojo) and drought tolerant (Pavon 76) genotypes was made to identify molecular markers linked to a wheat (Triticum aestivum L.) abscisic acid (ABA) content at two water regimes. The parents and 150 F₄ lines were evaluated phenotypically for drought tolerance using two irrigation treatments [0.25 and 0.75 m³(H₂O) m^-2(soil)]. Forty different target region amplification polymorphism (TRAP) primer combinations, 98 different sequence-related amplified polymorphism (SRAP) primer combinations, and 400 simple sequence repeat (SSR) primers were tested for polymorphism among the parental genotypes and the F₄ lines. Seven loci in the F₄ lines treated with the drought stress were identified. Single quantitative trait loci (QTLs) were located on chromosomes 1B, 2A, 3A, 5D, and 7B and each of them explained from 15 to 31 % of phenotypic variance with a LOD value of 7.2 to 15.7. Five QTLs were located on chromosome 4A and six QTLs on chromosome 5A. In control (well-watered) F₄ lines, two QTLs were mapped on chromosome 3B and one QTL on each chromosome 5B and 5D. Statistically the most significant groups of QTLs for the ABA content were identified in the regions of chromosomes 3B, 4A, and 5A mostly near to Barc164, Wmc96, and Trap9 markers. Therefore, these markers linked to QTLs for the drought-induced ABA content can be further used in breeding for drought tolerance in wheat.

Drought stress presents a considerable threat to the global crop production. As a dominant source of vegetarian diet, cereals and grain-legumes remain crucial to meeting the growing dietary demands worldwide. Therefore, breeding cultivars of these staple crops with enhanced drought tolerance stands to be one of the most sustainable solutions to enhance food production in changing climate. Given the context, a more focused survey of environment-defined germplasm sets is imperative to comprehend such adaptive traits. In parallel, uncovering the genetic architecture and the molecular networks that collectively contribute towards drought tolerance is urgently required through rationally combining large-scale genomics, proteomics, and metabolomics data. Also, attention needs to be directed to reasonably quantify the epistatic as well as environmental influences, thereby warranting deployment of analyses like metaquantitative trait loci (QTL) that encompass multiple environments and diverse genetic backgrounds. Further, innovative techniques like genomic selection (GS) and genome wide association study (GWAS) would help to capture the quantitative variation underlying drought tolerance. Equally importantly, integration of physiological traits-based techniques with ever-evolving 'omics' technologies and the new-generation phenotyping platforms will be of immense importance in advancing our existing knowledge about the genetically-complex and poorly-understood phenomena, such as plant drought response, and a deeper understanding would likely to provide a great impetus to the progress of crop breeding for drought tolerance.

The effects of proline and/or glycine betaine (GB) application on growth, photosynthetic pigments, H₂O₂ content, and activities of antioxidant enzymes in rice (Oryza sativa L. cv. KDML105) under salt stress were investigated. The H₂O₂ content and the activities of superoxide dismutase (SOD), glutathione reductase (GR), and ascorbate peroxidase (APX) but not catalase (CAT) increased under salinity. Under 160 mM NaCl, the CAT activity was maintained on the pre-stress level in the presence of proline, whereas in the presence of GB, the GR activity increased more than without GB application. A co-application of 30 mM proline and 1 mM GB did not reduce the increase in H₂O₂ caused by the NaCl stress more than applying each of the osmoprotectants and no synergistic effect on the antioxidant enzymes was observed. However, the application of both the osmoprotectants was the most effective in alleviating degradation of photosynthetic pigments.

Strawberry (Fragaria × ananassa) contains anthocyanins which are important secondary metabolites and key contributors to the antioxidant capacity and nutritional value of the fruit. Anthocyanin biosynthetic genes have been identified. However, the detailed mechanism responsible for anthocyanin accumulation and regulation of biosynthetic genes during strawberry fruit ripening remain unclear. In the present study, we examined the effect of a Fragaria × ananassa myeloblastosis 1 homolog, FaMYB1, on anthocyanin accumulation in the strawberry fruit receptacle. Expression analysis shows that FaMYB1 transcripts increased in response to irradiance but not to abscisic acid treatments. Down-regulation of FaMYB1 was achieved in planta using Agrobacterium-mediated RNA interference (RNAi). As a result, FaMYB1-RNAi fruits exhibited a significant increase in anthocyanin content. Conversely, overexpression of FaMYB1 resulted in a decrease in anthocyanin content. Overexpression of FaMYB1 also significantly reduced expression of genes encoding anthocyanidin synthase and flavonoid glycosyltransferase, whereas down-regulation of FaMYB1 resulted in a significant decrease in the amount of transcripts of leucoanthocyanidin reductase. These data suggest that FaMYB1 might negatively control anthocyanin biosynthesis in the strawberry fruit at the branching-point of anthocyanin/proanthocyanidin biosynthesis.

Litchi (Litchi chinensis Sonn.) is an important tropical and subtropical evergreen woody fruit tree, and it has been shown that nitric oxide (NO) could promote litchi flowering. NO responsive genes of litchi (cv. Nuomici) primordia were identified through a suppression subtractive hybridization (SSH) library screen. We obtained 1 563 expressed sequences tags (ESTs) that were enriched in the NO treated inflorescence primordia. We then used a reverse Northern analysis to identify 728 true NO responsive ESTs, the sequences of which have been further analyzed. They represent 70 litchi unique genes that could be classified into 9 categories: 14 % of them were involved in transport facilitation, 7 % in transcription regulation, 9 % in stress response, 7 % in sugar metabolism, 9 % in secondary metabolism, 10 % in intracellular signalling, and 44 % in other metabolism, whereas 11 % were genes with unknown functions, and 7 % were genes with no hit found. Next, we performed a real-time quantitative polymerase chain reaction (RT-qPCR) to determine the expression of selected candidate genes during a time-course of NO treatment and of normal floral tissue development.

Apple (Malus × domestica Borkh.) is a perennial woody plant that undergoes a period of dormancy (in cv. Jonathan between late September and mid-December) to survive freezing temperatures of winter. DORMANCY-ASSOCIATED MADS-BOX (DAM) genes play important roles in the regulation of growth cessation and terminal bud formation in peach. To understand the role of DAM orthologs in apple, we isolated and characterized four DAM-like genes (designated as MdDAMa, MdDAMb, MdDAMc, and MdDAMd) and monitored their expression in apical buds throughout the season by real-time quantitative polymerase chain reaction analyses. The transcription of MdDAMa peaked in October and that of MdDAMc was elevated from August to October, whereas MdDAMb and MdDAMd were practically undetectable. The tandemly arranged genes MdDAMa/MdDAMb and MdDAMc/MdDAMd were localized to chromosomes 16 and 8, respectively. Based on these observations, we infer that MdDAMa and MdDAMc acted in a dominant fashion on each locus and were correlated with the period of endodormancy.