Histone modification is a ubiquitous regulator of gene transcription. Arabidopsis CUP-SHAPED COTYLEDON (CUC) genes serve as a marker for shoot apical meristem initiation, but how they are regulated during shoot regeneration from in vitro culture, it is not yet understood. Here, the histone modification status of CUC1, CUC2, and CUC3 was analysed using a combination of chromatin immunoprecipitation (ChIP) and real time quantitative PCR. The activation of CUC1 and CUC2 was associated with an increased level of histone H3K4 trimethylation and/or H3K9 acetylation, as well as a reduced level of H3K9 demethylation in various parts of their promoter and coding sequences. Histone modification is suggested to play an important role in regulating CUC1 and CUC2 expression during shoot regeneration.

Pathogenesis-related (PR) proteins play key roles in plant disease resistance. Here, we isolated and characterized pathogenesis-related PR2 gene encoding β-1,3-glucanase from Brassica juncea and named it BjPR2 (GenBank accession number DQ359125). The amino acid sequence of BjPR2 showed ~99 % similarity with β-1,3-glucanase of Brassica rapa, B. napus, and B. oleracea. BjPR2 transcription was rapidly increased after Alternaria brassicae infection, salicylic acid application, and wounding, but the induction was delayed in response to jasmonic acid. To investigate the transcriptional regulation of BjPR2 gene, its promoter was isolated. In silico analysis of BjPR2 promoter showed cis-regulatory elements upstream of TATA and CAAT boxes responsive to defense, hormones, wounding, and plant developmental stage. Homozygous Arabidopsis thaliana lines were developed with plasmid construct having β-glucuronidase (GUS) reporter gene driven by BjPR2 promoter. The analysis of GUS protein in Arabidopsis lines showed that BjPR2 promoter drived distinct pattern of pathogen inducible expression after fungal infection (Alternaria brassicae, Erysiphe orontii), phytohormones, and wounding. It also showed age dependent and organ specific expressions. BjPR2 promoter drove strong GUS activity in Arabidopsis seedlings and showed organ specific expression at the later growth stages (lateral organ junctions, leaf serrate, base of siliques, and receptacle). Due to stress-inducible and tissue specific nature, the BjPR2 promoter can serve as a potential candidate in genetic engineering.

Aluminum (Al) is the main limiting factor for crop production in acidic soils. Efflux of organic acids is one of the mechanisms that determine Al-tolerance, and an Al-activated citrate transporter (multidrug and toxic compound extrusion) MATE1 gene is involved in different species. The contribution of the rye MATE1 gene (ScMATE1) depends on the rye (Secale cereale L.) cultivars and the crosses analyzed; there is no information about different rye species. The cDNA sequences, phylogenetic relationships, Al-tolerance, citrate exudation, and expression of the ScMATE1 gene were analyzed in several cultivars and wild species/subspecies of the Secale genus. Genotypes highly tolerant to Al were found within this genus. For the first time, sequences of the cDNA of the ScMATE1 gene were isolated and characterized in wild ryes. At least two copies of this gene were found likely to be related to Al-tolerance. The sequence comparison of 13 exons of ScMATE1 revealed variability between species, but also inter- and intra-cultivars. Variations were found in the Al-induced expression of ScMATE1 gene, as well as its contribution to Al-tolerance. The pattern of citrate exudation was inducible in most of the species/subspecies studied and constitutive in few. The phylogenetic analysis indicated that ScMATE1 is orthologue of two genes (HvMATE1 and TaMATE1) involved in the Al stress response in barley and wheat, respectively, but not orthologue of SbMATE, implicated in Al-tolerance in sorghum. ScMATE1 is involved in the response to Al stress in ryes, but its contribution to Al-tolerance is complex, and like in other species, there are tolerant and sensitive alleles in the different cultivars and species studied.

Zinc nutrition of tea (Camelia sinensis) have a great impact on acclimation to dehydration stress and on improvement of stress recovery. Dehydration stress induced by withholding water for 7 d decreased relative water content, dry mass of leaf, and content of antioxidants like ascorbate and glutathione and increased H₂O₂ content and lipid peroxidation and changed activities of superoxide dismutase, catalase, peroxidase, and glutathione reductase. A pre-treatment with 0.1 and 0.5 mM ZnSO₄ minimized these effects and increased Zn uptake. Further, foliar spray with 0.1 and 0.5 mM ZnSO₄ before rehydration lowered H₂O₂ generation, increased content of antioxidants and activities of antioxidative enzymes, and decreased lipid peroxidation. The present findings suggest that zinc regulates water stress responses and recovery after rehydration in tea.

WHIRLY transcription factors play critical roles in responses to biotic and abiotic stresses, but their other biological functions remain unclear. In this study, SlWHY2, a member of the WHIRLY family, was isolated from Solanum lycopersicum. The role of SlWHY2 was studied using transgenic tobacco plants. Real-time quantitative polymerase chain reaction analysis showed that SlWHY2 expression was induced by polyethylene glycol, NaCl, salicylic acid, hydrogen peroxide, and bacterial pathogens. SlWHY2 overexpression in tobacco caused enhanced tolerance to drought stress, as indicated by lower accumulation of malondialdehyde and relative electrolyte leakage, as well as higher relative water content and activities of superoxide dismutase and ascorbate peroxidase. Moreover, higher expression of cytochrome oxidase 1 (NtCOX1) and open reading frame 1 (NtORF1) were observed under drought in the transgenic lines. This suggested that overexpression of SlWHY2 enhanced tolerance to drought stress by regulating the transcription of mitochondrial genes and stabilizing mitochondrial function. Transgenic tobacco also displayed greater resistance to Pseudomonas solanacearum infection as evidenced by lower reactive oxygen species content and higher expression of defence-related genes. Overall, these findings suggest that SlWHY2 acts as a positive regulator in response to biotic and abiotic stresses.

The presence of solar radiation is one of the most important environmental factors, which is required for the optimal growth and development of plants. The absence of it (e.g. in the night period or artificially prolonged darkness) can alter the light-dependent signalling and regulation pathways and may induce new defence responses. Antioxidant enzymes as components of the plant defence system play a crucial role in the detoxification of reactive oxygen species (ROS) induced by several stressors; however, their regulation can be different in the light or in the dark. In this review we summarize the current knowledge about the physiological and molecular aspects of dark-modulated key antioxidant enzymes (superoxide dismutase, catalase, and ascorbate peroxidase) in different plant species and discuss their roles in different developmental processes (seedling growth and development or senescence) and in responses to environmental stresses (cold, chilling, heat, and biotic stress). Moreover, the hormonal regulation of respective gene transcription and the changes in activity of various isoenzymes at subcellular level are also summarized. Based on this knowledge, modification of these antioxidant enzymes may be used to increase the yield and stress tolerance of cultivated plants in the changing environment.

Glycine-rich RNA-binding proteins (GR-RBPs) are involved in RNA processing and also some of them are output signals of the circadian clock. In tomato, one GR-RBP gene family (LeGRP1) is composed by three highly homologous genes (LeGRP1a-c); each one rendering three transcriptional products: the un-spliced pre-RNA (preLegrp1a-c), the mature mRNA (mLegrp1a-c) and the alternatively spliced mRNA (asLegrp1a-c). To get insight into their regulation and impact on RNA metabolism in fruits, Solanum lycopersicum cv. Micro-Tom was transformed with preLeGRP1a fused to the polygalacturonase promoter, which drives expression to fruits from the mature green stage. Our results demonstrated a complex positive regulation of LeGRPs, in which LeGRP1a overexpression led to the induction of the others LeGRP1 members. Even though the LeGRP1 transcription and the content of three LeGRPs proteins were affected, the overall LeGRP protein circadian rhythm profile was similar in transgenic and wild type (WT) fruits. However, when the fruits were kept at a chilling temperature after harvest, total protein content was significantly higher in transgenic than in WT fruits, and the content of some free amino acids was modified. The results obtained suggest a probable role of LeGRP1s: structural rearrangements and/or stabilization of mRNA to allow efficient processing of fruits under cold conditions.

Uridine diphosphate glucose dehydrogenase (UGDH) plays an important role in biosynthesis of hemicellulose by catalyzing oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in biosynthesis of the plant cell wall. In this study, a UGDH ortholog referred to as LgUGDH was isolated from Larix gmelinii using PCR and rapid amplification of cDNA ends techniques. Real-time PCR shows that the LgUGDH gene was expressed primarily in larch stems in addition to its roots and leaves, and Southern blot analysis indicates that UGDH is encoded by two paralogous genes in L. gmelinii. Overexpression of LgUGDH increased the content of soluble sugars and hemicelluloses and enhanced vegetative growth and cold tolerance in transgenic Arabidopsis thaliana. These results reveal that L. gmelinii UGDH participates in sucrose/polysaccharide metabolism and cell wall biosynthesis and may be a good candidate gene for enhancing plant growth, cold tolerance, and hemicellulose content.

Synthetic pathogen-inducible promoters (SPIP) hold a great promise to meet the demands for a desired temporal and spatial regulation of transgenes. Four pathogen-inducible cis-elements (F-box, S-box, Gst1-box, and W-box) and the minimal cauliflower mosaic virus 35S (CaMV 35S) promoter (-46 to +8 TATA box) were used to design SPIP. Eight SPIP were synthesized and named FSGW, FSWG, GWFS, GWSF, SFGW, SFWG, WGFS, and WGSF according to the order of cis-element dimers. They were used to replace the CaMV 35S promoter in the plasmid pBI121 to control expression of the β-glucuronidase (gus) gene. The transcriptional properties of each SPIP were evaluated in homozygous T₃ lines of transgenic Arabidopsis thaliana by histochemical staining gus expression and real time quantitative PCR. FSGW and FSWG had a very low basal level and a poor inducibility. The other six SPIP showed different levels of background and inducibility. Using Ralstonia solanacearum, the spores of Phytophthora capsici, and salicylic acid as inducing factors, GWSF showed the advantages of a low basal expression, rapid response, and efficient transcriptional activity in the rosette leaves of five-week-old plants. The results indicate that the permutation and combination of the cis-elements had important effects on transcriptional activities of SPIP. Synthetic pathogen-inducible promoters like GWSF are valuable because it can potentially be further improved to apply to plant genetic engineering for disease resistance.

The proposed work describes a protocol for high-frequency in vitro regeneration through nodal segments and shoot tips in Decalepsis arayalpathra, a critically endangered medicinal liana of the Western Ghats. Nodal segments were more responsive than shoot tips in terms of shoot proliferation. Murashige and Skoog's (MS) basal medium supplemented with 5.0 μM 6-benzyladenine (BA) was optimum for shoot initiation through both the explants. Among different combinations of plant growth regulators and growth additive screened, MS medium added with 5.0 μM BA + 0.5 μM indole-3-acetic acid + 20.0 μM adenine sulphate effectuated the highest response: 11.8 shoots per nodal segment and 5.5 shoots per shoot tip with mean shoot length of 9.2 and 4.8 cm, respectively. Half-strength MS medium with 2.5 μM α-naphthalene acetic acid was optimum for in vitro root induction. The plantlets with the well developed shoot and root were acclimatized in Soilrite™ with 92 % survival rate in the field conditions. During acclimatization, chlorophyll content, net photosynthetic rate, stomatal conductance, and transpiration rate were gradually changed in dependence of formation of new leaves. Further, the changes in activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) as well as activity of carbonic anhydrase were also observed: a continuous rise in SOD activity, but a rise and fall in the activities of CAT, APX, and GR were also noticed. Maximum fresh mass (3.1 g plant^-1), dry mass (0.35 g plant^-1) of roots and 2-hydroxy-4-methoxybenzaldehyde content of 9.22 μg cm^-3(root extract) were recorded after 8 weeks of acclimatization.

The effects of silver nanoparticles (AgNPs), silver ions (Ag⁺), and polyvinylpyrrolidone (PVP) on mitosis and expression of a gene encoding cyclin-dependent kinase 2 (cdc2) in onion roots were compared. Three concentrations (5, 10, and 15 mg dm^-3) were employed in combination with three incubation times (3, 6, and 9 h). PVP enhanced mitotic index and cdc2 expression. Both silver forms decreased mitotic index and cdc2 expression. Genotoxicity of both silver forms were indicated by three major distinguishable classes of chromosome aberrations: spindle disturbances, clastogenic aberrations, and chromosome stickiness. Concerning Ag⁺ treatments, significant enhancements in occurrence of any chromosome aberration type was associated with significant decrease in mitotic index. On the other hand, disturbed spindle in AgNPs treatments was observed even in absence of significant reduction in mitotic index suggesting that AgNPs inhibit cellular events occurring during mitosis to proceed normally rather than starting of cell division.

Osmotic stress causes a series of morphological, physiological, biochemical, and molecular changes that alters plant growth, development, and productivity around the globe. Phytohormones, nutrients, and transcription factors may induce adaptive responses to osmotic stress in plants. We evaluated the effect of osmotic stress induced by 18.5 % polyethylene glycol (PEG) or 100 mM NaCl on growth, content of abscisic acid (ABA), chlorophyll (Chl), sodium, and potassium, and the expression of multifunctional NAC transcription factors in rice cultivars (the salt-tolerant Cotaxtla and salt-sensitive Tres Ríos). The PEG and NaCl decreased shoot height and increased ABA content in both cultivars, and reduced root length in cv. Tres Ríos. The PEG increased Chl content in cv. Cotaxtla leaves. NaCl reduced shoot K⁺ content in cv. Tres Ríos and increased shoot and root Na⁺ content in both cultivars, thus resulting in a decreased K⁺/Na⁺ ratio. Of the 57 NAC genes evaluated, two of them were repressed (Os10g42130 and Os07g04560) and two other induced (Os02g34970 and OsNAC10) in cv. Cotaxtla in response to PEG, whereas three of them were repressed (Os10g42130, Os07g04560 and Os08g10080), and six induced (Os02g56600, Os02g34970, Os11g08210, Os05g34830, OsNAC6, and OsNAC10) in response to NaCl. In the cv. Tres Ríos, we found two genes repressed (Os10g42130 and Os07g04560), and five induced (Os08g33910, Os03g60080, Os06g51070, OsNAC6, and OsNAC10) in response to PEG, while only two genes were repressed (Os10g42130 and Os07g04560) but 13 induced (Os03g21060, Os08g39110, Os03g60080, Os01g15640, Os06g51070, Os09g33490, Os04g40130, Os12g29330, Os02g36880, Os11g08210, Os05g34830, OsNAC6, and OsNAC10) by NaCl. Osmotic stress affected more severely cv. Tres Ríos than cv. Cotaxtla plants. These different responses might be regulated by ABA and NAC transcription factors.

Brassinosteroids (BRs) are plant hormones that were isolated for the first time in the 1970s. This group currently includes more than 70 compounds that differ in their structure and physiological activity. BRs are present in plants in a free form or in the form of conjugates. BRs are known as plant growth regulators, but they also play a role in the plant response to environmental stresses. In the case of plants that are exposed to low/high temperature, exogenous BRs can counteract growth inhibition and reduce biomass losses as well as increase plant survival. BRs show a multidirectional activity in regulating the metabolism of plants exposed to extreme temperatures. The following BRs actions can be distinguished: changes in membrane physicochemical properties, regulation of the expression of selected genes (including stress-responsive genes), as well as indirect effects on metabolism through other hormones or signalling molecules (such as hydrogen peroxide). This review summarizes the current knowledge about the effects of BRs on the physiological and biochemical processes that occur in plants during exposure to low or high temperatures.

Moth orchids (Phalaenopsis) are among the top-traded blooming potted plants in the world. To explore mitochondrial DNA (mtDNA) markers for species identification, we located simple sequence repeats in the mtDNA of Phalaenopsis aphrodite subsp. formosana and then pre-screened them for polymorphic markers by their comparison with corresponding mtDNA regions of P. equestris. The combination of 13 selected markers located in intergenic spacers could unambiguously distinguish 15 endemic moth orchids. Five most variable markers with polymorphic information content (PIC) ≥ 0.7 could be combined to classify 18 of 19 endemic moth orchids including parental strains most commonly used in breeding programs. The sequences of four selected mtDNA regions were highly variable, and one region (MT2) could be used to completely distinguish 19 endemic moth orchids. Though mitochondrial introns were highly conserved among moth orchids, evolutionary hotspots, such as variable simple sequence repeats and minisatellite repeats, were identified as useful markers. Furthermore, a marker technology was applied to reveal the maternal inheritance mode of mtDNA in the moth orchids. Moreover, phylogenetic analysis indicates that the mtDNA was nonmonophyletic below the Phalaenopsis genus. In summary, we have revealed a set of mtDNA markers that could be used for identification and phylogenetic study of Phalaenopsis orchids.

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.

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.

Boron (B) is an essential plant micronutrient. Two major B-transport proteins have been recently identified and partially characterized: BOR1, a high-affinity B efflux transporter involved in xylem loading, and NIP5;1, a plasma-membrane boric-acid channel involved in B uptake. To date, studies of these B transporters have investigated their expression individually (mainly as mRNA), and only in response to variation in B availability (mostly B deficiency); the influence of other factors, such as plant resource status, has not been studied. To address this, we grew geranium (Pelargonium × hortorum cv. Maverick White) plants under ambient or elevated CO₂ concentration, different sub-saturating irradiances, and different B availability. For comparison we also grew three other species (Arabidopsis thaliana, Azolla caroliniana, and Hordeum vulgare) under broad range of B supply. Relative accumulation of BOR1 and NIP5;1 proteins were measured using protein-specific antibodies and Western blotting or ELISA. Elevated CO₂ significantly increased content of NIP5;1, while increases in irradiance increased BOR1 content, but decreased NIP5;1 content. Across species, content of both transporters often decreased with increasing B availability, but sometimes remained unchanged or even increased, depending on CO₂, irradiance, species, or transporter. Content of BOR1 and NIP5;1 was correlated with root proteins, B content, and sugar content (for high CO₂ only), as well as B uptake, but CO₂ and irradiance often affected these relationships. Thus, relative accumulation of BOR1 and NIP5;1 is influenced not only by B content, as expected, but by other environmental factors as well.

This review reports the physiological and metabolic changes in plants during development under elevated atmospheric carbon dioxide concentration and/or limited-nitrogen supply in order to establish their effects on leaf senescence induction. Elevated CO₂ concentration and nitrogen supply modify gene expression, protein content and composition, various aspects of photosynthesis, sugar metabolism, nitrogen metabolism, and redox state in plants. Elevated CO₂ usually causes sugar accumulation and decreased nitrogen content in plant leaves, leading to imbalanced C/N ratio in mature leaves, which is one of the main factors behind premature senescence in leaves. Elevated CO₂ and low nitrogen decrease activities of some antioxidant enzymes and thus increase H₂O₂ production. These changes lead to oxidative stress that results in the degradation of photosynthetic pigments and eventually induce senescence. However, this accelerated leaf senescence under conditions of elevated CO₂ and limited nitrogen can mobilize nutrients to growing organs and thus ensure their functionality.

Root cortical aerenchyma (RCA) is suggested to reduce metabolic cost for root growth, but it might lower water uptake by plants. The objective of this work was to evaluate the effects of drought and phosphorus on the RCA development along the root axis and to elucidate its role in water stress tolerance of two maize genotypes. Plants of drought-tolerant DKB390 and drought-sensitive BRS1010 genotypes were grown in Vermiculite at field capacity of 100, 75, 50, and 25 % and supplied with 0.1, 0.4, and 0.8 mM phosphorus. Growth parameters, RCA, and plant P content were evaluated for all plants. Higher RCA development was observed in DKB390 than in BRS1010. Drought reduced the percentage of RCA in the root-hair zone of both genotypes but increased its development in the root maturation zone. Phosphorus limitation enhanced RCA development only in the DKB390. Under drought stress, DKB390 showed resilient growth whereas growth was inhibited in BRS1010. Higher root P content was related to its higher supply. Therefore, RCA formation was induced either by drought or by phosphorus limitation, while no interaction was evident. The RCA development varied along the root axis in order to balance water and phosphorus uptake and the drought response was genotype dependent.

Heat stress, one of the major abiotic stresses in wheat, affects chlorophyll fluorescence and chlorophyll content and thereby photosynthesis. To identify quantitative trait loci (QTLs) associated with these traits under terminal heat stress, 251 recombinant inbred lines (RILs) derived from a cross HD 2808/HUW510 were phenotyped. Using composite interval mapping, 40 QTLs were identified; 17 were related to conditions after timely sowing and 23 to heat stress after late sowing. The various parameters of chlorophyll fluorescence were associated with 23 QTLs, which were located on chromosomes 1A, 2A, 3A, and 2D and explained 3.67 to 18.04 % of phenotypic variation, whereas chlorophyll content was associated with 17 QTLs on chromosomes 2A, 2B, 2D, 5B, and 7A explaining 3.49 to 31.36 % of phenotypic variation. Most of the identified QTLs were clustered on chromosome 2D followed by 2A and 1A. The QTL Qchc.iiwbr-2A for chlorophyll content linked with marker gwm372 was stable over conditions and explained 3.81 to 18.05 % of phenotypic variation. In addition, 7 epistatic QTL pairs were also detected which explained 1.67 to 11.0 % of phenotypic variance. These identified genomic regions can be used in marker assisted breeding after validation for heat tolerance in wheat.

Vetiver grass [Vetiveria zizanioides (L.) Nash] displays comprehensive abiotic stress tolerance closely related to fine maintenance of plant water relation mediated by plasma membrane intrinsic proteins (PIPs). Two open reading frame sequences of PIPs (867 and 873 bp) were cloned from vetiver grass and named as VzPIP1;1 and VzPIP2;1, respectively. Expression of green fluorescent protein revealed only subcellular localization of VzPIP2;1 in the plasma membrane. Agrobacterium tumefaciens mediated transgenic (VzPIP2;1) soybean plants had a higher water content in above-ground parts under sufficient water supply through enhancing transpiration as compared to the non-transgenic plants but displayed a more severe drought injury because of a lower photosynthesis and a higher transpiration rate. However, A. rhizogenes mediated transgenic soybean plants kept a higher water content in above-ground parts by improving root water transport and kept a more effective photosynthesis under normal and drought conditions.

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.

In Arabidopsis, EXPORTIN1A (HIT2/XPO1A) and EXPORTIN1B (XPO1B) mediate the translocation of nuclear export sequence (NES)-bearing proteins from nucleus to cytoplasm. However, a mutation in HIT2/XPO1A but not in XPO1B induces sensitivity to high irradiance (HI). Arabidopsis thaliana heat stress elements A4a and A5 (AtHsfA4a and AtHsfA5) are involved in plant responses to HI and possess NESs; therefore, their nucleo-cytoplasmic partitioning was analyzed. In wild-type and xpo1b mutant cells, AtHsfA4a normally remained in the cytoplasm but became concentrated in the nucleus following exposure to HI, whereas AtHsfA5 was constitutively distributed in both cytoplasm and nucleus. However, in hit2/xpo1a mutant, AtHsfA4a and AtHsfA5 were always confined to the nucleus, regardless of the irradiance. Although AtHsfA4a can enhance the ability of plants to scavenge H₂O₂, and AtHsfA5 is a repressor of AtHsfA4a, athsfa5 but not athsfa4a mutant plants exhibited HI sensitivity. Additionally, athsfa4a plants expressing AtHsfA4aΔNES were sensitive to HI, but athsfa5 plants expressing AtHsfA5ΔNES were not. Meanwhile, hit2/athsfa4a double mutant was more tolerant to HI than hit2. These results indicate that both AtHsfA4a and AtHsfA5 were HIT2/XPO1A-specific substrates. Long-term accumulation of AtHsfA4a contributed to the hit2 HI-sensitive phenotype independent of the scavenging ability of H₂O₂, and the presence of AtHsfA5 could mitigate this adverse effect.

Auxins are one of the main regulators of in vitro plant growth and development. However, the mechanisms, by which auxins, such as 1-naphthaleneacetic acid (NAA), affect in vitro root and leaf anatomy and photosystem function, remain unclear. Accordingly, the aim of the present study was to analyze the effect of different NAA concentrations on the anatomy and photosynthetic performance of in vitro-propagated Aechmea blanchetiana and to determine whether such a treatment affects micropropagated plants after acclimatization. In vitro-established A. blanchetiana plants were transferred to culture media that contained 0, 2, 4, or 6 μM NAA, and after 50 d, they were transplanted into plastic seedling trays with a commercial substrate and cultivated for 60 d in a greenhouse. The plants were evaluated after a 50-d in vitro NAA exposure (growth traits, chlorophyll α fluorescence, and root and leaf anatomy) and after 60 d of acclimatization in the greenhouse (root and leaf growth). Changes induced by NAA in root anatomy might improve uptake of minerals and sugars from the medium, thereby increasing the in vitro growth. In the leaves, the lowest chlorenchyma thickness and sclerenchyma area were observed in plants grown without NAA, and NAA exposure also improved photosystem II activity. The highest ex vitro growth rate was observed for plants that were propagated with 4 μM NAA. Therefore, the use of NAA during in vitro propagation can improve the anatomical and physiological quality of A. blanchetiana plants, as well as to improve ex vitro transfer.

Brassinosteroids (Brs) have drawn wide attention due to their protective role against toxicity of heavy metals in plants. To better understand the role of Br in arsenic (As) and cadmium (Cd) uptake by rice plants, a hydroponic experiment was conducted to investigate the combined effect of 24-epibrassinolide (Br24) or 28-homobrassinolide (Br28) and iron plaque (IP) on As and Cd uptake and accumulation in rice seedlings. Six-week-old seedlings were sprayed with 0.2 or 0.02 μM Br24 or Br28 and grown in nutrient solution for 3 d, and then 20 or 60 mg Fe²⁺ dm^-3 (Fe20 and Fe60) was used to induce root IP formation for 3 d. These seedlings with or without Br and with or without IP were exposed to solution containing 0.5 mg dm^-3 As^III or Cd for 9 d. The results showed that rice growth decreased when Br24 were applied, but it increased when combination of Br24 and IP was applied. Fe concentrations in dithionite-citratebicarbonate (DCB) extracts were increased after 0.2 or 0.02 μM Br24 application in the absence of IP, but decreased by Br24 in the presence of IP. In the absence of IP, As and Cd content in leaves was significantly reduced by 0.02 μM Br24 and 0.2 μM Br28, respectively. The As content in leaves was also reduced by the combination of 0.02 and 0.2 μM Br28 and IP, and the Cd content in leaves was reduced by the combined effect of 0.2 μM Br24 and IP. These results indicate that Br24 and Br28 could impede As and Cd accumulation, and the interactions between Br and IP may have a potential in restricting the transport of As and Cd into rice shoots.

The effects of copper and zinc salts on transgenic canola plants expressing rice transcription factor (TF) OsMYB4 were investigated. Transgenic plants (TPs), which showed a high OsMyb4 expression in response to either Cu or to Zn excess, were used for the current study. In leaves of TPs, the content of Cu was equal and the content of Zn was significantly higher than in non-transformed plants (NTPs). The TPs grown on an extremely high concentration of heavy metals (HMs; 150 μМ CuSO₄ or 5 000 μМ ZnSO₄) were able to survive for more than 15 d, while NTPs died after 7 - 9 d of incubation. This indicates that expression of OsMyb4 in canola plants improved their HM tolerance. The TPs tolerance to HMs was confirmed by a higher shoot biomass than that in NTPs. Excess of HMs caused oxidative stress (indicated by increase in malondialdehyde content) especially in leaves of NTPs. This data suggests a protective role of the OsMyb4 TF in oxidative stress. The HMs caused a lower decrease in activities of superoxide dismutase and guaiacol peroxidase in TPs than in NTPs. Higher tolerance of TPs to HMs was also suggested by a considerable increase in the content of low-molecular phenolic compounds, including flavonoids and anthocyanins, as well as proline (a potential antioxidant and chaperone). These data suggest that OsMYB4 may play a role as a positive regulator of phenylpropanoid pathway and proline synthesis. The created canola OsMyb4 TPs may be useful for future applications in phytoremediation of HM-polluted soils.

Identifying important regulative elements and pathways related to dwarfism in cotton is a major challenge in cotton breeding. Isobaric tags for relative and absolute quantitation-based proteomics of stem terminal buds from upland cotton (Gossypium hirsutum L.) dwarf line LA-1 and high near-isogenic line LH-1 was performed. Moreover, transcriptional expression of differentially-expressed proteins (DEPs) belonging to phytohormone signal transduction, hormone biosynthesis, and ubiquitin system were analysed using quantitative real-time polymerase chain reaction. A total of 4 849 proteins were identified from LA-1 and LH-1, 697 of which showed differential accumulations. Most of the DEPs have catalytic, binding, and transporter activity and are involved in metabolism- and protein processing-related pathways. In particular, seven DEPs, including two gibberellin (GA) receptors, three cytokinin (CK) receptors, CKoxidase, and CK-N-glucosyltransferase were up-regulated in LA-1, and GA20-oxidase was down-regulated in LH-1. Our results suggest that the DELLA-independent GAsignalling pathway is the primary cause of dwarfism in LA-1 and indicate that CKresponse element 1-2, GA-insensitive dwarf, CKoxidase, and GA3-β-dioxygenase are potential indicators of dwarf cotton. The profiling of DEPs may offer a valuable resource for cotton breeding.

Multiple allele-inherited male sterility has been widely used by breeders of Brassica rapa L. ssp. pekinensis, but the molecular mechanisms of male sterility are not yet clear. In this study, we isolated the full-length cDNA of a new gene (not included in the Brassica database). This gene, comprising 1 054 bp, encodes a 39.99 kDa protein with a Gly-Asp- Ser-Leu (GDSL)-lipase domain that is a member of the lipolytic protein GDSL family. The sequence of candidate gene is the most similar to extracellular lipase 6 (EXL6) of Arabidopsis and was therefore designated BrEXL6 and submitted to NCBI (accession No. JX131630.1). Reverse transcription semi-quantitative PCR and Western blot analysis showed that BrEXL6 and its encoded protein were significantly more expressed in fertile buds than in sterile buds. Quantitative PCR and in situ hybridization showed that BrEXL6 was highly expressed in the anthers of fertile buds, especially anthers at the pollen-development stages, but only weakly expressed in other tissues and floral organs of fertile plants and whole sterile plants. These results suggest that BrEXL6 is a pollen development-related gene. The results of this study provide clues for understanding the mechanisms underlying multiple allele-inherited male sterility.

Papaya (Carica papaya L.) is susceptible to viral diseases caused by Papaya mosaic virus (PapMV) and Papaya ringspot virus (PRSV), which limit fruit production and affect economic yield. The symptoms produced by both the viruses are similar in early stages of infection and include vein and leaf chlorosis, which develop into mosaic at later stages of infection when leaf lamina can get reduced in size and distorted with a shoe-string aspect. Digital image analyses, such as fractal dimension (FD) and lacunarity (λ) were used here to examine papaya tissue after single and mixed infections of PRSV and PapMV. Morphological changes, such as hypoplasia, hyperplasia, and neoplasia are described in tissues with viral infections. Furthermore, we quantified these changes and suggest three ranges of tissue damage according to their λ values in rank 1 (0.01 to 0.39), rank 2 (0.4 to 0.79), and rank 3 (0.8 to 1). Our analyses suggest that in synergism and antagonism there is a competition of both viruses to occupy the same mesophyll cells, as indicated by their intermediate values of lacunarity.