Harpin, an elicitor molecule of bacterial origin induces hypersensitive response (HR) in non-host plants. In an attempt to induce male sterility, harpin was tagged with a signal peptide and expressed downstream to tapetum-specific TA29 promoter resulting in extracellular secretion, subsequent degeneration of tapetum and development of male sterility in tobacco. Putative transgenics were analyzed by PCR amplification of transgene, semiquantitative RT-PCR analysis from total RNA extracts from anther tissue with transgene specific probe, Western blotting using polyclonal antibody raised against harpin, by transmission and scanning electron microscopy, and by confocal microscopy of anthers and pollen at various stages of development. Varying degrees of male sterility (30-100 %) was observed with plants showing complete and partial male sterility as well as several morphological variations were seen especially in leaves and flowers. Further, some of the transgenics showed un-induced of HR-like local lesions in the vegetative tissues. Harpin_Pss got deposited on the pollen grains upon tapetal degeneration resulting in significant alterations in the morphology of pollen cell wall. However, megagametogenesis was not affected in complete and partial male sterile plants and female gametes were completely fertile. The complete male sterility was attributed to premature tapetal cell death due to sufficient extracellular harpin_Pss accumulation whereas insufficient protein content might be the reason for partial male sterility. These findings indicate the possible use of cytotoxic harpin_Pss for the development of male sterile plants.

Turnip (Brassica rapa L.) plants were grown in Perlite with low (< 2.5 μM) or adequate (25 μM) boron supply under well-watered and drought conditions for 12 weeks. Dry mass of leaves and roots was reduced under drought by about 61 and 56 % in plants supplied adequately with B, while up to 84 and 74 % under B starvation. Drought reduced B content by about 70 and 82 % for B-sufficient and B-deficient plants, respectively. According to the chlorophyll fluorescence parameters, the photosynthesis processes conserved their normal activities under low B supply in well-watered plants, while a serious damage to photosystem 2 occurred under drought stress. Stomatal limitation was the most important cause for a 17 % lower net photosynthetic rate (P_N) of drought stressed B-sufficient plants. In B-deficient plants, however, both stomatal and non-stomatal limitations were involved in 53 % reduction of P_N. Low B supply reduced strongly leaf water potential.

Limited information is available on the role of brassinosteroids (BRs) in response of plants to nutrient deficiency. To understand the functions of BRs in response to iron deficiency, we investigated the effect of 24-epibrassinolide (EBR) on activities of ferric-chelate reductase (FCR), H⁺-ATPase, Ca²⁺-ATPase, nitrate reductase (NR), antioxidant enzymes, Fe and other minerals content and distribution, chlorophylls, soluble protein, free proline, reactive oxygen species, and malondialdehyde in peanut (Arachis hypogea L.) plants subjected to Fe deficiency (10^-5 M Fe(III)-EDTA) with foliar application of EBR (0, 10^-8, 5.0×10^-8, 10^-7, 5.0×10^-7, and10^-6 M). Results show that EBR increased Fe translocation from roots to shoots and increased Fe content in cell organelles. Activities of antioxidant enzymes increased and so the ability of resistance to oxidative stress was enhanced. As result of enhancement of H⁺-ATPase and Ca²⁺-ATPase activities, the inhibition of Fe, Ca, Mg, and Zn uptake and distribution was ameliorated. Chlorophyll, soluble protein, and free proline content also increased and consequently, chlorosis induced by Fe deficiency was alleviated. The results demonstrate that EBR had a positive role in regulating peanut growth and development under Fe deficiency and an optimal concentration appeared to be 10^-7 M.

Phenylalanine ammonia lyase (PAL) is a specific branch point enzyme of primary and secondary metabolism. The Citrus reticulata Blanco PAL gene was cloned and designated as CrPAL1. The cDNA sequence of CrPAL1 was 2 166 bp, encoding 721 amino acid residues. Sequence alignment indicates that CrPAL1 shared a high identity with PAL genes found in other plants. Both the dominant and catalytic active sites of CrPAL1 were similar to PAL proteins observed in Petroselinum crispum. Phylogenetic tree analysis indicates that CrPAL1 was more closely related to PALs in Citrus clementina × C. reticulata and Poncirus trifoliata than to those from other plants. Subcellular localization reveals that CrPAL1-green fluorescent protein fusion protein was specifically localized in the plasma membrane. Activity of PAL as well as CrPAL1 expression increased under Fe deficiency. A similar result was noted for total phenolic content. The root exudates of C. reticulata strongly promoted reutilization of apoplastic Fe in roots. Furthermore, Fe was more desorbed from the cell wall under Fe deficiency than in sufficient Fe supply.

Physiological changes associated with senescence of flowers and abscission of floral parts in Oncidesa (formerly Oncidium) cv. Gower Ramsey are caused by a plant hormone ethylene which is produced by pollinia cap dislodgment during postharvest handling and transportation. The ethylene receptor gene OgERS1 of Oncidesa has been previously cloned and characterized. To analyze promoter activity of OgERS1, transgenic Arabidopsis thaliana plants were generated to express the ß-glucuronidase (GUS) reporter gene under the control of 5'-upstream sequence of OgERS1 from Oncidesa. The expression pattern of the OgERS1 promoter at the cellular level was investigated by analysis of GUS activity. This promoter can activate gene expression in both actively dividing young tissues and abscission-related aging tissues. Expression of GUS was detected in the shoot meristem uniquely in 10 to 30 d-old-plants and was found in flower buds, axillary buds, flower stems, and abscission layers during later development. In 2- to 3-week-old transgenic Arabidopsis, exogenous ethylene, glucose, lactose, and maltose enhanced promoter activity implying that crosstalk between sugar and an ethylene receptor may exist. However, indole-3-acetic acid, benzylaminopurine, abscisic acid, heat, wounding, salinity, drought, and flooding slightly suppressed promoter activity. These results demonstrate that the promoter of OgERS1 was developmentally and environmentally regulated, and imply a potential for application of this bi-functional promoter to increase branching or enhanced dwarfing.

MicroRNAs (miRNAs) are small non-coding RNAs that play crucial regulatory roles in diverse developmental processes via cleavage or translational inhibition of their target mRNAs. Although a growing number of miRNAs and their targets have been predicted and discovered via experimentation in many plants, little is known about conserved miRNAs and their target genes in Pinus densata. In the present study, the conserved miRNAs, miR171 and miR482, from Pinus densata were characterized. Analysis of miR171 and miR482 reveal that these miRNAs were highly conserved in other plant species. In addition, the precursors of miR171 and miR482 were validated by real time-PCR and sequencing. Using real-time quantitative PCR, miR171 and miR482 as well as their corresponding targets were found to be differentially expressed in needles, stems, and roots of Pinus densata. Furthermore two target genes, one GRAS family transcription factor protein gene and one nucleotide-binding site leucine-rich repeat (NBS-LRR) resistance protein gene, were experimentally verified to be the targets of pde-miR171 and pde-miR482, respectively, using RNA ligase-mediated 5'-rapid amplification of cDNA ends (RLM-RACE).

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.

Secretion of organic acids (OAs) by roots has been suggested to be an important mechanism of Al resistance in many species. In Stylosanthes, the participation of OAs in the mechanism of Al resistance is poorly understood. We aimed to study the production and secretion of OAs by two Brazilian Stylosanthes species with different Al resistance. Stylosanthes capitata and S. guianensis were treated with Al at different concentrations in 0.5 mM CaCl₂ (pH 4.0), and then root elongation, Al and OA content, OA secretion into the external solution, and the activity of citrate synthase (CS) were measured. Al-induced secretion of citric acid was also evaluated in the presence of protein synthesis and anion channel inhibitors. S. guianensis accumulated lower amounts of Al in its roots and displayed less inhibition of root elongation compared to S. capitata. Citric and malic acids were the most abundant OAs in the roots, and their content decreased with the Al treatment, except for citric acid in S. guianensis. Citric acid was the only OA secreted into the nutrient solution by the Al-treated plants of both species, but more by S. guianensis. Citrase synthase activity decreased in S. capitata but increased in S. guianensis with the Al treatment, and it may have a crucial role in the maintenance of citric acid content in the roots of S. guianensis. The use of anion channel and protein synthesis inhibitors reveal that anion channels were likely involved in the secretion of citric acid, and channel protein transcription was up-regulated by exposure to Al in Stylosanthes.

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.

Plants are subjected to several abiotic stresses that adversely affect growth, metabolism and yield. The dynamic research in plant genetics complemented by genome sequencing has opened up avenues to address multiple problems caused by abiotic stresses. Though many drought-induced genes have been phytoengineered in a wide range of plants, the drought signal transduction pathways, and the alteration of plant sensing and signaling systems to adverse environments still remain an intriguing subject for comprehensive investigation. To impart enhanced drought tolerance in plants, a thorough perception of physiological, biochemical and gene regulatory networks is essential. Recent functional genomics tools have facilitated the progress in our understanding of stress signaling and of the linked molecular regulatory networks. This has revealed several stress-inducible genes and various transcription and signaling factors that regulate the drought stress-inducible systems. Translational genomics of these drought specific genes using model plants have provided encouraging outcomes, but the in-depth knowledge of the specific roles of various metabolites in plant stress tolerance will lead to evolvement of strategies for the phytoengineering of drought tolerance in plants in future.

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.

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.

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.

Dicer proteins belong to the RNase III family of proteins, which are key components in small RNA biogenesis. In Solanum lycopersicum, seven Dicer-like (DCL) genes have been identified and have been named SlDCL. In this study, we cloned the full-length sequence of the SlDCL genes including untranslated regions using RNA ligase-mediated rapid amplification of cDNA ends. Our analysis indicates that 7 SlDCLs were located on 5 tomato chromosomes (6, 7, 8, 10, and 11). The gene structure of the SlDCLs covered long genomic regions and contained more than 20 exons. Phylogenetic analysis divided the seven SlDCL members into four subgroups. In general, all seven SlDCLs were expressed in all organs but more in flowers and fruits than in the other parts. Moreover, the expressions of some genes changed slightly after treatment with ethylene or 1-methylcyclopropene suggesting their likely roles in plant responses to ethylene. Our findings provide essential information on SlDCL genes in tomato and will aid in the functional classification of DCL families in plants.

The life cycle of flowering plants is partially defined by environmental cues like day length and temperature. In the model plant Arabidopsis thaliana and temperate cereals, such as barley (Hordeum vulgare) and wheat (Triticum spp.), differences in life cycle control have been associated with a natural variation in FLOWERING LOCUS C (FLC) and VERNALIZATION 1-3 (VRN1-3). In sugar beet (Beta vulgaris L.), variation in vernalization requirement and life cycle is determined by a major gene at the B locus. This gene has recently been identified as a pseudo-response regulator (PRR) gene BOLTING TIME CONTROL 1 (BTC1). A second gene in beet with homology to BTC1 and ARABIDOPSIS PSEUDO RESPONSE REGULATOR 7 (APRR7) in Arabidopsis was identified and termed Beta vulgaris PSEUDO RESPONSE REGULATOR 7 (BvPRR7). We functionally characterized BvPRR7 by transgenic analysis in Arabidopsis and expression profiling during development in beet. We show that BvPRR7 was diurnally regulated and responded to cold. Constitutive expression of BvPRR7 distorted diurnal rhythms and caused late flowering in Arabidopsis suggesting a conserved function of BvPRR7 in clock regulation. Conceivably, the retention of a functional role of BvPRR7 in clock regulation may have facilitated the evolution of a distinct role as major floral regulator of the second PRR7 homolog in beet, BTC1.

Carbon monoxide (CO) is considered as a new emerging cell signal molecule which is involved in plant growth, development, and acquisition of stress tolerance. In recent years, hydrogen sulfide (H₂S) has been found to have similar functions, but crosstalk between CO and H₂S in the acquisition of heat tolerance in plants is not clear. In this study, pretreatment of tobacco (Nicotiana tabaccum L.) cells cultured in a suspension with a CO donor hematin significantly increased survival percentage of cells under a heat stress and regrowth ability after the heat stress, alleviated a decrease in cell vitality, and accumulation of malondialdehyde. In addition, treatment with hematin enhanced the activity of L-cysteine desulfhydrase, a key enzyme in H₂S biosynthesis, which in turn induced accumulation of endogenous H₂S in tobacco cells. Interestingly, hematin-induced heat tolerance was enhanced by addition of NaHS, a H₂S donor, but weakened by specific inhibitors of H₂S biosynthesis DL-propargylglycine or its scavenger hypotaurine. Furthermore, pretreatment with hemoglobin (a CO scavenger) and zinc protoporphyrin IX (a CO specific synthetic inhibitor) had no significant effect on NaHS-induced heat tolerance of tobacco cells. These results suggest that CO pretreatment could improve the heat tolerance of tobacco suspension cultured cells, and H₂S might exert its signal role downstream to CO-induced heat tolerance.

Water use efficiency (WUE) is an worth attempting trait to discover the genomic regions governing it, especially in view of the diminishing water resources for the crop plants in general and rice in particular. In order to address this, the present investigation was aimed at identification of genomic regions governing WUE employing a recombinant inbred line population derived from a cross between INRC10192, a high WUE landrace, and IR64, a high yielding cultivar. A total of 36 quantitative trait loci (QTLs) were detected under control as well as drought conditions on chromosomes 1, 2, 4, 8, 9, 10, and 11. Among all, the QTLs with the marker intervals RM486-RM6703, RM6703-RM11484, RM404-RM447, RM24879-RM171, and RM229-RM332 on chromosomes 1, 8, 10, and 11 were found to govern the water use efficiency related traits such as carbon isotope discrimination, specific leaf area, leaf width, and relative water content. Nine major QTL intervals were targeted for candidate gene identification using gene ontology (GO) and transcriptome-based analyses. Overrepresented GO terms in the targeted QTLs were found to be associated with the genes/pathways controlling stomatal regulatory mechanism, stress responsive genes or transcription factors, and saccharide biosynthesis pathways under stress situation. Hence, these genes or genomic regions are potential candidates for development of high WUE rice cultivars.

The role of peroxisomes in the oxidative injury induced by the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) in leaves of pea (Pisum sativum L.) plants was studied. Applications of (2,4-D) on leaves or to root substrate increased the superoxide radical production in leaf peroxisomes. Foliar application also increased H₂O₂ contents in leaf peroxisomes. Reactive oxygen species (ROS) overproduction was accompanied by oxidative stress, as shown by the changes in lipid peroxidation, protein carbonyls, total and protein thiols, and by the up-regulation of the activities of superoxide dismutase, ascorbate peroxidase, glutathione reductase, catalase, glucose 6-phosphate dehydrogenase and NADP⁺-dependent isocitrate dehydrogenase. Foliar or root 2,4-D applications also induced senescence symptoms in pea leaf peroxisomes, as shown by the decrease of protein content and glycolate oxidase and hydroxypyruvate reductase activities, and by the increase of endopeptidase, xanthine oxidase, isocitrate lyase and acyl-CoA oxidase activities as well as of 3-ketoacyl-CoA thiolase and thiol-protease protein contents. 2,4-D did not induce proliferation of pea leaf peroxisomes but induced senescence-like morphological changes in these organelles. Results suggest that peroxisomes might contribute to 2,4-D toxicity in pea leaves by overproducing cell-damaging ROS and by participating actively in 2,4-D-induced leaf senescence.

Two genes encoding CAX2-like proteins were isolated from a Zn/Cd hyperaccumulating ecotype (HE) and nonhyperaccumulating ecotype (NHE) of Sedum alfredii Hance, and they were named SaCAX2h and SaCAX2n, respectively. Both SaCAX2h:eGFP and SaCAX2n:eGFP proteins were localized to the vacuolar membrane of tobacco epidermal cells and yeast mutants. Heterologous expression of SaCAX2h or SaCAX2n in the Δzrc1 yeast mutant increased Cd content in yeast cells. Yeast complementary assay also revealed that both the transporters could suppress Ca and Mn hypersensitivity and enhance Ca and Mn accumulation in a K667 yeast mutant. The expression patterns of the two genes were different under the Cd treatment. Transcription of SaCAX2h was down-regulated in roots and up-regulated in shoots whereas transcription of SaCAX2n was down-regulated in shoots after the exposition to Cd. Furthermore, over-expression of SaCAX2h enhanced metal accumulation in the tobacco plants. The Cd content increased by 17-19 % in shoots and 31-36 % in roots; the Ca content of the transgenic plants increased by 31-32 % in shoots, and the Mn content increased by 60-79 % in shoots and 22-29 % in roots. These results indicate that SaCAX2h was responsible for Ca and Mn sequestration into vacuoles, and over-expression of SaCAX2h enhanced Cd accumulation in the transgenic tobacco.

Several studies have been performed to elucidate the role of brassinosteroids (BRs) in plant growth and development. However, information on the role of BR signaling in nutrient uptake is limited. This study explores the relationship between BRs and ammonium transporter 1 (AMT1) expression in Arabidopsis roots. We found that BR treatment reduced the expression of AMT1 genes and that a BR receptor BRI1 mutant bri1-5 reversed its BR-repressed expression. Furthermore, the BR signaling transcription factor, BES1, regulates AMT1 expression in roots. NH₄ ⁺-mediated repression of AMT1;1, AMT1;2, and AMT1;3 was suppressed in a gain-of-function BES1 mutant (bes1-D). This mutant was more sensitive to methyl-ammonium and contained a higher ammonium content compared to wild-type plants. However, BES1 failed to bind E-box elements present in the promoter region of the AMT1 genes. Furthermore, NH₄ ⁺-mediated glutamine synthetase (GS) and glutamine oxoglutarate aminotransferase (GOGAT) gene expressions were partially inhibited, and GS activity was slightly lower in the bes1-D mutant relative to that observed in wild-type En2 roots. NH₄ ⁺-mediated AMT1 suppressions are known to be caused by N-metabolites rather than NH₄ ⁺ itself, and glutamine application inhibited AMT1 expression in both En2 and bes1-D indicating that BES1 activation inhibited NH₄ ⁺-mediated GS/GOGAT induction, which might in turn inhibit AMT1 repression. In conclusion, the present study demonstrates that BR regulated nitrogen uptake and assimilation via the BR signaling pathway.

Ttranscription factors WRKY play vital roles in response to biotic and abiotic stresses, and previous studies have predominantly focused on model plants and fairly limited research has been performed with tomato. In the present study, a novel pathogen-induced WRKY gene named SpWRKY6 was isolated from the late blight resistant tomato (Solanum pimpinellifolium) cultivar L3708 using in silico cloning and reverse transcription polymerase chain reaction (RT-PCR) methods. Multiple sequence alignment with other plant WRKYs indicates that SpWRKY6 contains two WRKY domains and belongs to group I WRKY transcription factors. Furthermore, some cis-acting elements associated with responses to environmental stresses were observed in the promoter region of this gene. Gene expression patterns were determined by analyzing microarray data of SpWRKY6 in tomato and of an orthologous gene from Arabidopsis thaliana using the Genevestigator tool. The results reveal a very strong biotic and abiotic stress responsive behaviour of this gene. Moreover, bioinformatics results were confirmed by real time quantitative polymerase chain reaction and show that SpWRKY6 expression was rapidly induced after infection with Phytophthora infestans and Botrytis cinerea, respectively. Expression of SpWRKY6 was up-regulated by application of various phytohormones including salicylic acid, methyl jasmonate, and abscisic acid. Likewise, the SpWRKY6 expression was induced by NaCl, drought, heat, cold, and HgCl₂ treatments.

The effects of exogenous salicylic acid (SA), sodium nitropusside (SNP, a nitric oxide donor), or their combination on dwarf polish wheat (Triticum polonicum L.) seedlings under UV-B stress were studied. The UV-B stress significantly decreased plant height, shoot dry mass, pigment content, net photosynthetic rate, intercellular CO₂ concentration, stomatal conductance, transpiration rate, and variable to maximum chlorophyll fluorescence ratio (F_v/F_m) in all plants, but less in the presence of SA, SNP, and their combination. On the other hand, there were considerable increases in malondialdehyde (MDA), proline, O₂ ^*-, and H₂O₂ content under the UV-B stress. When SA, SNP, and their combination were applied, content of MDA, proline, H₂O₂, and O₂ ^*- were less increased. Moreover, there were considerable increases in activities of superoxide dismutase, peroxidase, ascorbate peroxidase, and glutathione reductase under the UV-B stress and more in the presence of SA, SNP, and their combination. Therefore, it is considered that SA, SNP, and especially their combination could alleviate UV-B stress in dwarf polish wheat.

To investigate the antioxidative response of glutathione metabolism in Urtica dioica L. to a cadmium induced oxidative stress, activities of glutathione reductase (GR), glutathione-S-transferase (GST), and glutathione peroxidase (GSH-Px), content of reduced (GSH) and oxidized (GSSG) glutathione, lipid peroxidation (LPO), and also accumulation of Fe, Zn, Mn, Cu besides Cd were determined in the roots, stems, and leaves of plants exposed to 0 (control), 0.045, and 0.09 mM CdCl₂ for 58 h. Whereas the Cd content continuously increased in all organs, the Fe, Zn, Mn, and Cu content decreased in dependence on the applied Cd concentration and incubation time. The Cd treatment resulted in increased GR and GST activities in all organs, however, GSH-Px activity was dependent on Cd concentration and plant organ. The GSH/GSSG ratio maintained above the control level in the stems at both Cd concentrations. The LPO was generally close to the control values in the roots and stems but it increased in the leaves especially at 0.09 mM Cd.

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.

Ascorbate (AsA) is an important antioxidant that can scavenge reactive oxygen species to protect plant cells against oxidative stress. Guanosine 5'-diphosphate (GDP)-L-galactose phosphorylase (GGP) is a key enzyme in the AsA biosynthetic pathway. To investigate the functions of GGP in AsA synthesis and oxidative stress tolerance in tomato, antisense lines with a reduced expression of SlGGP were obtained. Photobleaching after treatment of leaf disks with methyl viologen was more severe in transgenic lines compared to wild type (WT) plants. Moreover, compared with the WT plants, the transgenic plants showed a higher content of hydrogen peroxide, superoxide anion, malondialdehyde, as well as ion leakage, but a lower content of AsA and chlorophylls, ascorbate peroxidase activity, net photosynthetic rate, and maximal photochemical efficiency of photosystem II. Results of real-time quantitative polymerase chain reaction show that suppression of the SlGGP gene in the transgenic plants reduced their oxidative stress tolerance.

Leaf anatomy and irradiance-dependent leaf transmittance changes serving as irradiance acclimation mechanisms in leaves were studied in two ecologically contrasting Tradescantia species, a shade plant T. fluminensis Vell. and a sun plant T. sillamontana Matuda, grown at different irradiances. A dramatic increase in leaf thickness (2 to 4-fold) under a high growth irradiance (800 μmol m^-2 s^-1) compared with a low growth irradiance (60 μmol m^-2 s^-1), achieved mainly by expansion of the epidermis, was recorded in both species. The effect took place on the background of modest changes in mesophyll thickness (1.8-fold in T. fluminensis and 1.15-fold in T. sillamontana) and chloroplast size (0.8-fold in T. fluminensis and an insignificant change in T. sillamontana). Mesophyll structure and growth irradiance response did not seem to facilitate significantly light-dependent chloroplast (avoidance) movement in these species. Nevertheless, an exceptionally large (2 to 4-fold) irradiance-induced increase in light transmittance attributable to chloroplast avoidance movement was revealed. This increase by far exceeded that in other higher plants according to available literature. The magnitude of the irradiance-dependent transmittance changes positively correlated both with the rate of photosystem II recovery and with the extent of xanthophyll deepoxidation in the leaves. This was opposite to a negative correlation observed between the same parameters in different plant species. We hypothesize that, at the evolutionary timescale, chloroplast avoidance movement might adjust independently from other photoprotective mechanisms, e.g., non-photochemical quenching, whereas, on the ontogenetic timescale, adjustment of these mechanisms inevitably follows the same trend.

In northern China, freezing injury is observed frequently in the rare species Magnolia wufengensis but not in the more common species Magnolia denudata. To investigate the role of the phytohormone abscisic acid (ABA) on frost tolerance in these two species, exogenous ABA was applied to the seedlings and then physiological and biochemical responses were measured during cold acclimation. Shoot growth cessation was stimulated by ABA in M. wufengensis but not in M. denudata. Abscisic acid inhibited shoot growth in M. wufengensis but not in M. denudata. Treatment with ABA stimulated leaf senescence in both species, and this effect was greater in M. denudata. For both species, ABA-treated plants exhibited bud dormancy sooner and had an increased tolerance to freezing, decreased water content and increased accumulation of proline, glucose, and fructose in shoots. These effects were generally greater for M. denudata. Freezing tolerance was significantly correlated with content of water, proline, glucose, and fructose for both species, but freezing tolerance was significantly correlated with raffinose content only in M. wufengensis. We conclude that exogenous ABA could increase cold acclimation and improve cold hardiness of both Magnolia species, although M. denudata was more responsive to ABA than M. wufengensis, which might result from a greater dehydration and accumulation of proline and certain soluble sugars.

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.

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.

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.