Festulolium cultivars are widely utilized in Lithuania because they are persistent under abiotic stresses and are high yielding. However, changing climate challenges the existing Festulolium cultivars to adapt to new growing conditions and still maintain the yield. In this study, we aimed at evaluating the yield stability of two Festulolium cultivars in field trials under fluctuating Lithuanian conditions. The mean total dry matter yield (DMY) of both Festulolium cultivars fluctuated greatly between the years and ANOVA analysis showed a significant effect of environment on total DMY as well as DMY of each cut, but the genotype × environment interaction was not significant. There was a high difference between the total DMY of 1^st year and 2^nd year of use of plots in each year of observation. The highest DMYs were harvested in the years 2015 and 2016. Dry matter yield of the 1^st cut was the largest component of the total DMY for most of the years. The plants overwintered the first winter after sowing very well over the whole study period, resulting in excellent spring growth. The winter survival scores of 2^nd year of use of plots were lower than 1^st year of use and strongly correlated with the 1^st cut DMY of 2^nd year of use (r = 0.81). Spring growth of plants at 2^nd year of use was poorer, the correlation between winter survival and spring growth of 2^nd year of use was 0.62. The scores of regrowth after the cuts of 1^st and 2^nd years of use were very similar for most of the experimental years and moderately correlated with the sum of DMYs after cuts (r = 0.55 and r = 0.5, respectively).

Fusarium head blight (FHB) represents a damaging disease of cereal spikes caused by mycotoxin-producing Fusarium fungi revealing adverse effects on grain quality and yield. Chinese spring wheat (Triticum aestivum L.) cv. Sumai 3 represents a major source of FHB resistance thanks to Fhb1 QTL on 3BS chromosome conferring resistance to FHB and encoding a lectin-like protein. The aim of our study lied in a comparison of proteome response to Fusarium culmorum inoculation and mycotoxin deoxynivalenol (DON) application at 10 d after inoculation (dai) in spikes of resistant wheat cultivar Sumai 3 and susceptible wheat cultivar SW Kadrilj. Proteome analysis revealed profound impact of Fusarium inoculation and mycotoxin application on plant energy metabolism. Fusarium inoculation decreased photosynthesis and ATP biosynthesis and increased the level of stress-protective proteins (chaperones such as Hsc70, lectins). Genotype related differences observed at 10 dai indicated an active acclimation in Sumai 3 as indicated by increased content of some enzymes involved in phenolics biosynthesis (phenylalanine ammonia lyase PAL, BAHD acyltransferase), jasmonate biosynthesis (lipoxygenase LOX), and oligosaccharide biosynthesis (sucrose synthase SuSy, UDP-glucose uridylyltransferase) which are an important part of glycoproteins such as lectins and other pathogen-responsive biomolecules. The study thus provides data on plant acclimation to Fusarium infection which may underlie superior resistance of Sumai 3 cultivar.

Superoxide dismutase (SOD) is a crucial reactive oxygen species (ROS) scavenger, which converts superoxide radical to H₂O₂, so it is thought to enhance abiotic stress tolerance by reducing ROS and thus avoiding oxidative damage. In this study, we isolated a salt- and oxidative stress-responsive copper-zinc (Cu/Zn) SOD encoding gene TaSOD5 from wheat. The ectopic overexpression of TaSOD5 in Arabidopsis thaliana increased total SOD and Cu/Zn SOD activities and enhanced tolerance to salt stress. Arabidopsis ectopically expressing TaSOD5 possessed a superior resistance to oxidative stress stimulated by exogenous H₂O₂. Ectopic overexpression of TaSOD5 elevated the activities of both ROS scavengers and an O₂^.- producer - NADPH oxidase. These findings show that Cu/Zn SOD enhanced salt tolerance via regulating the machinery of redox homeostasis rather than improving SOD activity alone.

Zinc deficiency is a worldwide problem for crops including potato (Solanum tuberosum L.), the fourth most important crop worldwide. The zinc/iron-regulated transporter-like protein (ZIP) transporter family is thought to play key roles in Zn uptake and transport. However, little is known about the potato ZIP family. In this study, 12 genes encoding members of the ZIP family were identified in the potato genome. The 12 StZIP genes were predicted to encode proteins of 220 - 407 amino acids harboring 5 - 9 putative transmembrane domains (TMDs), and 11 of these proteins had a variable region rich in histidine residues between TMDIII and TMDIV. A phylogenetic analysis divided the StZIPs into four groups on the basis of gene structure and conserved motifs. Furthermore, the StZIP expression profiles were determined under Zn-deficiency in both high and low Zn-content genotypes. Four differentially expressed genes, StZIP6, -9, -11, and -12, were identified in tubers of the two genotypes under Zn-deficiency, and StZIP11 and StZIP12 may have a more prominent function in Zn uptake and accumulation in potato tubers owing to their higher expressions. Thus, the results provide useful information for further studying the functions of StZIP genes.

Tamarix ramosissima is a deciduous shrub that resides in arid and semi-arid regions. Although of ecological and medicinal values, some Tamarix species are considered invasive as they have dominated the riparian zones of dryland in some parts of the world. Here, the complete chloroplast (cp) genome of T. ramosissima was sequenced and analyzed, showing a size of 156 150 bp and a GC content of 36.5 %. The plastome displayed a typical quadripartite structure, consisting of a pair of inverted repeat (IR) regions of 26 554 bp, separated by a large single copy (LSC) region of 84 795 bp, and a small single copy (SSC) region of 18 247 bp. The cp genome encoded 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. A total of 32 repeat sequences and 64 simple sequence repeat (SSR) were identified in the plastome, and an obvious A/T bias was observed in the majority of the SSRs detected. By comparing the T. ramosissima cp genome with those of the other four Tamaricaceae species, a number of divergence hotspots were identified among these plastomes. Together with SSRs and long repeats identified, these divergence hotspots could be developed as potential molecular markers facilitating species discrimination and evolutionary studies. Using plastome sequences, we re-investigated the phylogenetic relationship among 19 species, and T. ramosissima was found to be a sister of Tamarix chinensis. Taken together, our study provides valuable genomic resources to deepen the understanding of plant photosynthetic mechanism and phylogenomics.

The importance of Na⁺/H⁺ antiporters in salt tolerance in plants has been demonstrated in many studies, but much less is known about their protective role during drought stress. To study their possible contribution to water deficit tolerance, two closely related soybean Na⁺/H⁺ antiporters belonging to the intracellular NHX exchanger protein family, GmNHX3 and GmNHX1, were evaluated in transgenic Arabidopsis thaliana. A. thaliana plants ectopically expressing GmNHX3 or GmNHX1 displayed a more drought-tolerant phenotype compared to wild-type plants, which was accompanied by an increase in relative water content and chlorophyll content during stress conditions. Both GmHNX1 and GmHNX3 transgenic lines accumulated higher amounts of Na⁺ and K⁺ cations, showed increased antioxidant enzyme activities and less membrane damage due to lipid peroxidation under water deficit, as compared to non-transformed plants. Furthermore, plants expressing GmNHX3 showed an increased sensitivity to abscisic acid as deduced from stomatal closure and seed germination inhibition studies. Finally, a significant up-regulation of abiotic stress-related genes was observed in both transgenic lines compared to wild-type plants in response to abscisic acid and mannitol treatments. These results demonstrate that GmNHX3 and GmNHX1 antiporters confer protection during drought stress in A. thaliana and hence are potential genetic targets to improve drought tolerance in soybean and other crops.

Cellulose is the most abundant polysaccharide produced by plants. In the form of rigid microfibrils surrounding the cells, cellulose constitutes the load-bearing cell wall element that controls cell growth and shape. Cellulose microfibrils are laid down outside the cell by the multimeric plasma membrane-inserted cellulose synthase complexes (CSCs), which move along underlying cortical microtubules (CMTs). In plants, CSCs are shaped as rosettes with six lobes symmetrically arranged in a hexagonal structure. In Arabidopsis, the CSC is composed of at least three functionally non-redundant cellulose synthase (CESA) glycosyltransferases in both primary and secondary cell walls. The number, organization, and interactions of CESA proteins within the CSC have been debated for many years on the basis of numerous lines of evidence provided by electron microscopy, biochemical and genetic approaches, spectroscopic techniques, as well as computational modeling. The Arabidopsis thaliana model was extremely useful in elucidating the molecular composition of CSC and enabled to elucidate the specialized functions of distinct AtCESA isoforms. Several additional, non-CESA proteins involved in cellulose synthesis and its regulation were also identified in Arabidopsis. This review outlines the latest findings on CSC organization, trafficking, and plant-specific proteins directly associated with the complex and interconnecting CESAs with CMTs.

The myeloblastosis (MYB) transcription factors are essential for plant stress responses. They can enhance plant tolerance to abiotic stresses (e.g., drought, salinity, and cold) via improved physiological and biochemical responses including the accumulation of metabolites. In this study, we constructed a Panax ginseng MYB4 (PgMYB4) gene expression vector and established the stable transgenic Arabidopsis thaliana lines to study the effects of this gene on plant stress tolerance. The germination rate and seedling taproot length were greater for the PgMYB4-overexpressing plants than for the wild-type plants. Accordingly, the overexpression of PgMYB4 in Arabidopsis enhanced seedling tolerance to drought, salt, and cold conditions. Under drought stress, the relative chlorophyll content decreased less, the proline content increased more, and the water loss rate decreased more in the transgenic plants than in the wild type. The expressions of stress-related genes responsive to dehydration 19A, responsive to dehydration 22, responsive to desiccation 29A, cold-regulated 15A, cold-regulated 47, and pyrroline-5-carboxylate synthase 1 were significantly upregulated in the transgenic Arabidopsis plants. Under high salt stress, the kinesin 1 (KIN1) expression was significantly upregulated in the transgenic plants. In response to the low temperature stress, the dehydration-responsive element binding protein 2A and KIN1 expressions increased dramatically in the transgenic Arabidopsis plants. Thus, PgMYB4 positively regulated the stress tolerance gene networks, which promoted the expression of anti-stress effector genes. This gene may be useful for ginseng breeding programs aiming to develop new cultivars with enhanced stress tolerance.

The seasonal course of xylem sap parameters (electrical conductivity EC, potassium concentration [K⁺], and pH) of three conifers (Pinus cembra, Picea abies, and Larix decidua) growing at the alpine timberline was monitored. We also looked into possible effects of [K⁺] and pH on the difference in hydraulic conductivity (Δk_s). In all studied species, EC, [K⁺], and pH varied considerably over the year, with pH ranging between 7.3 (February) and 5.8 (June) and [K⁺] changing between 0.4 (January) and 2.5 mM (June). The Δk_s was overall low with positive values during winter (up to +20 %) and negative values in summer (-15 % in August). Samples perfused with alkaline solutions showed higher Δk_s. Xylem sap parameters in all conifers under study were surprisingly variable over the year thus indicating either effects upon seasonal changes in environmental factors or active adjustments, or both. Although Δk_s values over the year were minor, observed induction of Δk_s by high pH might indicate a role for hydraulic adjustment in harsh winter periods.

An efficient in vitro regeneration protocol through somatic embryogenesis was established from stem transverse thin cell layers (tTCLs) of Dendrobium aqueum Lindley, an imperiled orchid. This study outlines the induction and successive maturation stages of D. aqueum somatic embryos (SEs). The tTCLs (~ 0.5 mm thick) cultured on halfstrength Murashige and Skoog (MS) medium containing cytokinins and auxins, either individually or in combination, produced embryogenic callus (EC). Treatment with 0.5 mg dm^-3 zeatin induced EC in 41.42 % of tTCLs. As many as 42.66 globular SEs per tTCL were formed in the presence of 1.5 mg dm^-3N⁶-(2-isopentyl) adenine (2iP) but only on 10.33 % of explants. The combined treatment of 2iP (1.5 mg dm^-3) and 0.5 mg dm^-3 6-benzyladenine resulted in 34 globular SEs on 14.7 % of tTCLs whereas the combination of 2iP and 1.0 mg dm^-3 indole-3-butyric acid (IBA) induced 7.4 globular SEs on 52.33 % of tTCLs. Supplementation of activated charcoal, amino acids, and antioxidants alleviated browning at all the concentrations tested, but the EC response declined. The addition of 0.5 mg dm^-3 polyvinylpyrrolidone to 1.5 mg dm^-3 2iP and 1.0 mg dm^-3 IBA produced 24 SEs on 19.89 % of tTCLs suggesting that the EC and SEs can be effectively induced by individual cytokinins whereas the synergistic treatments with other compounds can only enhance the induction of EC. Histological observations of EC showed the formation of globular SEs from sub-epidermal regions. Successive developmental stages of globular SEs and the intermediate stage of protocorm like bodies until the formation of plantlets were observed. The plantlets obtained through SEs showed no morphological variations, and inter simple sequence repeat profiles also confirmed the genetic fidelity of in vitro-derived progeny with high monomorphism (97.78 %). In conclusion, the use of stem tTCLs is an effective method to produce SEs through indirect somatic embryogenesis in D. aqueum.

Responses of rice to submergence have been extensively studied, but the molecular network behind the tolerance to this stress is still incomplete. Transcription factors (TFs) are important players in gene transcription regulation during stresses. Here, we analyzed expression of WRKY genes and morphological and anatomical changes in different rice cultivars under submergence. When they were submerged for 48 h, changes in root number, fresh and dry masses, and aerenchyma development were observed. Although accumulations of WRKY transcripts were observed in both shoots and roots, root tissues showed higher accumulation with a peak already after 6 h under submergence. Especially transcriptions of OsWRKY11 and OsWRKY56 were high, more than 100-fold in comparison with controls. The WRKY promoter analysis showed that some cis-regulatory elements could be characterized as stress-responsive elements and linked to oxygen depletion. In the promoter of OsWRKY62, two cis-regulatory elements were found: ARE and GC-motif. These elements are known to be involved in oxygen deficiency responses. In addition, the W-box cis-regulatory element, the target of WRKY transcription factors, was found in OsWRKY11, OsWRKY56, and OsWRKY62, suggesting a feedback control acting on the upregulation of WRKY transcription factors. Genes involved in the submergence stress and resulting aerenchyma development had a W-box in their promoter regions, which also suggested regulation by WRKYs. Overall, the results support the role of WRKY transcription factors in rice submergence tolerance and unveil their action in other tolerance mechanisms.

The GRAS transcription factors encoding proteins ranging from 400 to 700 residues are recognized by their conserved C terminus. Here, a set of 23 CmGRAS genes was identified from a scan of the chrysanthemum (Chrysanthemum morifolium) transcriptome. A phylogenetic analysis implied that nine of these genes could be assigned orthologs to the GRAS gene family, and that four of them formed two pairs of paralogs. A phylogenetic analysis of the GRAS protein family based on the chrysanthemum and recent study of eight representative species of angiosperms showed that most of the CmGRAS genes belonged to a recognized sub-group. CmGRAS4 and CmGRAS10 were strongly transcribed in flowers and roots, respectively. The DELLA subfamily transcript abundance of the CmGRAS19 and CmGRAS20 was high in the reproductive tissues and they were responsive to phytohormones and stresses. Establishment of the orthology relationships between the known representative GRAS genes and CmGRAS, and transcriptional profiles of CmGRASs after phytohormone treatments or stresses will facilitate subsequent functional analyses in the GRAS gene family.

Tomato (Solanum lycopersicum) plants have four fructokinase genes, SlFRK1-4. The SlFRK4 is expressed only in pollen whereas the other three are expressed in all plant parts. While SlFRK2 and SlFRK3 are involved in vascular tissue development and affects the shape, size, and cell-wall width of xylem vessels and xylem fibers, the role of SlFRK1 has not been studied previously. The current work investigates the expression of SlFRK1 using transgenic tomato plants expressing the β-glucuronidase reporter gene under the SlFRK1 promoter, as well as the role of SlFRK1 using transgenic plants with antisense suppression of SlFRK1. The SlFRK1 promoter is expressed primarily in vascular tissues and specific suppression of SlFRK1 reduces water transport in stems, but has no other anatomical or phenotypic effects. Combined suppression of SlFRK1 and SlFRK2 severely inhibited plant growth and an anatomical analysis revealed a reduction in secondary xylem area and distorted phloem fibers characterized by thin cell walls and reduced lignification. The results suggest that SlFRK1 is involved in vascular tissue development and hydraulic conductivity in tomato plants and that SlFRK1 is important for normal phloem fiber development, together with SlFRK2.

Among the 400 soybean (Glycine max) landraces, we selected 3 tolerant (KAS150-9, KAS160-15, and KAS170-9) and 3 susceptible lines (KAS160-14, KAS160-20, and KAS201-6-1) by the survival percentage and injury scores. Susceptible lines showed decrease in chlorophyll content and increase in glucose and malondialdehyde (MDA) contents under waterlogging stress, while tolerant lines did not change significantly. For AFLP analysis, 8 EcoRI (+3) and 8 MseI (+3) primers used in 32 primer combinations generated a total of 2 566 bands with a mean of 80 bands per primer combination, of which 1 117 (43.5 %) were clearly polymorphic between the tolerant and susceptible lines. A genetic similarity coefficient, based on cluster analysis using an unweighted pair grouping method of average (UPGMA), was 0.79 for the tolerant group, while the susceptible landraces were genetically less related, with a genetic similarity coefficient of 0.17. The 10 reproducible polymorphic PCR products present in the 3 tolerant or susceptible lines were sequenced and converted into sequence tagged site (STS) markers. These STS primer sets were designated GmWT01-GmWT06 and GmWS01-GmWS04. Two STS primer sets, GmWT06 and GmWS02, generated a single monomorphic PCR product identical in size to the original AFLP fragments. For the broad application of these STS markers in marker-assisted selection (MAS) for soybean genotypes tolerant to waterlogging stress, two developed STS markers are being evaluated with putative waterlogging tolerant mutant lines induced by γ-radiation in soybean mutation breeding programs.

Downy mildew caused by Hyaloperonospora parasitica is a serious fungal disease in non-heading Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). Pathogenesis-related 5 (PR-5) genes play an important role in plant resistance to disease invasion. In this study, a gene encoding pathogenesis-related 5-like (PR-5L) protein, named BcPR-5L, was successfully cloned from non-heading Chinese cabbage. The cDNA sequence of BcPR-5L is 747 bp in length. It encoded a protein of molecular mass of 25.78 kDa, an isoelectric point of 4.42, and containing 248 amino acids. Multiple sequence alignment indicated that BcPR-5L protein was highly homologous to other PR-5L proteins identified in 13 different species, with the highest homology to Brassica rapa. We analyzed the subcellular localization of BcPR- 5L protein by using onion epidermal cells and found that it is localized in the membrane. Real time quantitative PCR analyses revealed that the expression of BcPR-5L gene was significantly upregulated after H. parasitica infection, and the expression in the resistant cultivar was higher than that in the susceptible cultivar. In summary, our data suggest that BcPR-5L gene may play an important role in the resistance of non-heading Chinese cabbage to H. parasitica infection.

In plants, class III T2 RNases involves two groups of structurally similar proteins, but with different biological functions: S-RNases and non-S-RNases. S-RNases have been involved in self-incompatibility whereas non-S-RNases have been implicated in stress responses. Here we report a novel class III RNase termed NaPi/S_x-RNase, which works both in self-incompatibility and in response to phosphate deficiency. The NaPi/S_x-RNase gene was identified in roots of Nicotiana alata grown in the absence of inorganic phosphate. Phylogenetic analysis showed that NaPi/S_x-RNase was included within the class III RNase T2 group. The NaPi/S_x-RNase was expressed in styles and its temporal expression increased in parallel to stylar development, with a slight decrease after anthesis. Progeny analysis showed that NaPi/S_x-RNase and S₁₀₇-RNase, a functional allele of the self-incompatibility system, segregated in a 1:1 ratio. The progeny segregation of a semicompatible cross, in which NaPi/S_x-RNase was shared by the two parents, exhibited a pattern consistent with a functional S-RNase allele. Considering genetic segregation, primary structure, and physiological role, the NaPi/S_x-RNase may be either an S-RNase with diversified functions or a non-S-RNase linked to the S-locus. To our knowledge, this is the first evidence for a specific function of the S-locus other than the self-incompatibility reaction. These results support the hypothesis that the self-incompatibility and stress responses may have evolved from a common origin.

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

Elymus nutans is an important alpine perennial forage of the Pooideae subfamily, that can survive subzero temperatures. To understand the molecular mechanisms underlying cold tolerance in E. nutans, we performed the transcriptional analysis by RNA-Seq in two genotypes, the tolerant Damxung (DX) and the sensitive Gannan (GN), under cold stress. The new E. nutans transcriptomes comprised 200 520/200 836 and 181 331/211 973 transcripts in leaves/crowns of DX and GN, respectively. More cold-stress-related genes were identified in leaves than in crowns of both genotypes throughout the whole cold stress. The most prominent functional category in leaves of both genotypes at 3 h of stress was transcriptional regulation. Brassinosteroid and jasmonic acid mediated signalling pathways play central roles in regulating downstream protective responses in DX after 24 h of cold stress. Prolonged cold stress caused more severe transcriptome responses in crowns and leaves of DX compared to GN. The most significant transcriptomic changes in both genotypes were associated with the response to abiotic stresses and the oxidation-reduction processes, implying reprogramming of the cellular metabolism as an adaptation to cold stress. This study reveals mechanisms of genotype- and organ-specific cold stress response in E. nutans and thus provides a basis for future breeding strategies aimed at improving the tolerance of cold-sensitive plants.

Gibberellins (GAs) are a large family of tetracyclic diterpenoids, controlling important aspects of growth and development throughout the plant life cycle. To explore the possibility that gibberellin A₃ (GA₃) signalling induces epigenetic alteration(s), we carried out a field experiment study using Nicotiana tabacum as a model system. The GA₃ application on leaves resulted in increased plant-height, foliage density, leaf cell area, and trichome density. The plants exposed to GA₃ also exhibited: 1) increased chromatin de-condensation, 2) reduced global DNA methylation, 3) reduced DNA methyltransferases (NtDNMTs) activities accompanied by decreased amounts of NtMET1 and NtCMT3 transcripts, and 4) partial restoration of phenotype and expression of epigenetically silenced reporter transgene. Based on these observations, we propose that GA₃ application induces complex epigenetic re-programming, which may lead to distinct developmental phenotypes. These results could provide an important insight for future studies on epigenetic mechanism(s) in other important crops.

The miR171 is a conserved microRNA (miRNA) family and has been shown to participate in plant growth and development. However, the precise function of miR171 in Pinus densata remains largely unclear. Mature miR171 sequence comparison reveals high similarity between Arabidopsis thaliana and P. densata and the pre-miR171 could fold into a characteristic stem-loop hairpin structure. Genes encoding GRAS (GAI-RGA-SCR) family transcription factors and actin binding protein were identified as targets of pde-miR171 using a modified RNA ligase mediated 5' rapid amplification of cDNA ends (RLM-RACE). Furthermore, the interaction between pde-miR171 and Arabidopsis SCL6 (SCARECROW-LIKE6) was further validated through transient co-expression of both genes in Nicotiana benthamiana leaves. Next, results of real-time quantitative PCR demonstrated that the expression of pde-miR171 was significantly up-regulated in miR171-overexpressing plants than in wild-type plants, which was inversely correlated with the expression of Arabidopsis SCL6 genes. In addition, overexpression of pde-miR171 in Arabidopsis induced larger leaves and earlier flowering under long-day conditions compared with the wild type. The findings presented here suggest that miR171 derived from a P. densata precursor together with its target gene SCL6 may play important roles in the regulation of primary root growth, leaf shape, and flowering time in plants.

Gibberellin 2-oxidases (GA2oxs) irreversibly convert bioactive gibberellins (GAs) and their immediate precursors into inactive GAs via 2-β hydroxylation and so regulate gibberellin content in plants. However, to the best of our knowledge, little has been known about the GA2oxs and its function in cool season turfgrass Poa pratensis. In this study, rapid amplification of cDNA end (RACE) was employed to isolate PpGA2ox from P. pratensis. The open reading frame of PpGA2ox was 1 047 bp in length, corresponding to 348 amino acids. PpGA2ox was localized in both nucleus and cytoplasm. The expression of PpGA2ox could be up-regulated by 10 μM gibberellic acid, 5 μM methyl jasmonate, or 10 μM indole-3-acetic acid. In addition, its native promoter could drive GUS expression in both leaf apex and shoot apical region. Moreover, overexpression of PpGA2ox in Arabidopsis led to GA-deficiency leading to dwarf phenotype, delayed flowering time, and increased chlorophyll content. Our study suggests that PpGA2ox could be a candidate gene for breeding new cultivars of P. pratensis.

Salinity is one of the major abiotic constraints to agriculture. The physiological and molecular mechanisms of salt tolerance have been studied in plants for many years. The regulation of osmosis and ion homeostasis is crucial. A lot of important components involved in plant responses to salt stress have been identified. Among them, ion transporters and channels take an essential role in ion homeostasis, mainly for Na⁺, Cl^-, and K⁺. Until now, many cation antiporters important for salt tolerance in plants have been characterized. Among them, the monovalent cation/proton antiporters (CPA) family is one of the most important families, including sodium proton exchangers (NHXs), K⁺-efflux antiporters (KEAs), and cation/H⁺ exchangers (CHXs). Here, the current knowledge of the plant CPA family in responses to salt stress was reviewed. The regulation mechanisms were also included and discussed.

The tubular ray flower (turf) mutant of sunflower is characterized by a switch of ray flowers from zygomorphic to near-actinomorphic disc flowers. In sunflower, floral symmetry of ray and disc flowers is specified by the activity of members of a CYCLOIDEA (CYC) gene family. The turf mutant is generated by the insertion of a CACTA-like transposable element (TE), named Transposable element of turf1 (Tetu1), in the coding sequence of the HaCYC2c gene. The TEinsertion changes the reading frame of turf-HaCYC2c for the encoded protein and leads to a premature stop codon. Tetu1 is a non-autonomous version of a CACTA TEcarrying the minimum sequences necessary for transposition in the presence of autonomous elements in the sunflower genome. In the previous analysis, performed in more than 11 000 plants homozygous for the turf-HaCYC2c allele, the absence of chimerism and the segregation rate of derived-progenies from reverted phenotypes suggest that Tetu1 transpositions are restricted to a time shortly before and/or during meiosis. Here, we report the analysis of F₅ and F₆ progenies, derived from an F₄ progeny of the cross turf × Chrysanthemoides2, where plants with a chimeric inflorescence were detected. Tetu1 showed active excision in all progenies taken into consideration and named High Frequency of Tetu1 Transposition (HFTT). Within a total of 449 plants, Tetu1 excision generated a 13.81 % of non-chimeric revertants but also a 5.12 % of plants with somatic sectors of variable size in the outmost whorl of the inflorescence. These unexpected results suggest variations in tissue specificity and time of TEexcision. The excision of Tetu1 was confirmed by DNA molecular screening of non-chimeric and chimeric revertants and transcription analysis of the HaCYC2c gene. In HFTT progenies, sequence analyses excluded significant DNA changes with respect to the original Tetu1 transposon as well as to the adjacent 5'- and 3'-TE regions. Genetic and epigenetic regulatory mechanisms were proposed to explain the time and frequency of Tetu1 transposition in HFTT progenies.

Wild rice genotypes are rich in genetic diversity. This has potential to improve agronomic rice by allele mining for superior traits. Late embryogenesis abundant (LEA) proteins are often associated with desiccation tolerance and stress signalling. In the present study, a group 3 LEA gene, Wsi18 from the wild rice Oryza nivara was expressed under its own inducible promoter element in stress susceptible cultivated indica rice (cv. IR20). The resulting transgenic plants cultivated in a greenhouse showed enhanced tolerance to soil water deficit. Transgenic plants had higher grain yield, plant survival rate, and shoot relative water content compared to wild type (WT) IR20. Cell membrane stability index, proline and soluble sugar content were also greater in transgenic than WT plants under water stress. These results demonstrate the potential for improving SWS tolerance in agronomically important rice cultivar by incorporating Wsi18 gene from a wild rice O. nivara.

Leek is an economically important vegetable. In model plants, the CONSTANS (CO) and CONSTANS-like (COL) genes play central roles in plant flowering modulation. However, none of leek CO homolog has been identified, because of limited gene resources obtained in this crop. Here, we reported the transcriptome analysis of leek, along with the identification of putative leek CONSTANS-like (COL) (ApCOL) genes. A total of 189 713 non-redundant transcripts were de novo assembled by using about 128.9 million clean sequence reads, of which, 48 621 were achieved for functional annotation. Thereafter, the search for putative ApCOL genes against the assembled transcripts was performed, and 17 genes were identified. The 17 putative ApCOL proteins, together with 16 function-known COL proteins published for other species, were subjected to phylogenetic analysis and divided into four groups. Some putative ApCOL members showed high sequence similarity with published COL proteins involved in flowering regulation. Expression analysis revealed that, among the 17 putative ApCOL genes, eight, two, and three genes showed higher expression in leaves, cauloids, and roots, respectively. The discovery of putative ApCOL genes and the characterization of their expression patterns will provide a basis for future clarification of their functions in leek growth and development.

DNA methylation is one of the epigenetic mechanisms regulating gene expression in plants in response to environmental conditions. In this study, analysis of methylation patterns was carried out in order to assess the effect of water stress in two contrasting wheat genotypes using methylation-sensitive amplified polymorphism (MSAP). The results revealed that demethylation was higher in drought-tolerant genotype (C306) as compared to drought-sensitive genotype (HUW468) after experiencing drought stress. Comparisons of different MSAP patterns showed a high percentage of polymorphic bands between tolerant and susceptible wheat genotypes (from 74.79 % at anthesis to 88.89 % at tillering). Furthermore, differential DNA methylation in roots and leaves also revealed tissue-specific methylation of genomic DNA. Interestingly, 54 developmental stage-specific bands and 23 bands that were found contrasting between these two wheat genotypes were detected. Furthermore, a few sites with stable DNA methylation differences were identified between drought-tolerant and drought-sensitive cultivars, thus providing genotype-specific epigenetic markers. These results not only provide data on differences in DNA methylation changes but also contribute to dissection of molecular mechanisms of drought response and tolerance in wheat.

Medicinal plants are a rich source of secondary metabolites, extensively used in traditional health care systems. High altitude biodiversity encompasses the diversified and valuable medicinal plant species. The extreme environmental conditions of high altitude region viz. fluctuating temperatures, high UV radiation, salinity, low oxygen concentration, and high wind velocity limits the plant growth and distribution. Yet, how medicinal plants respond to these extreme conditions is not sufficiently understood. Therefore, addressing plant acclimation to different stresses presents an opportunity to unravel adaptive mechanism of medicinal plants along altitude gradient. This article reviews the recently published research that highlights the major role of proteins in plant adaptation to extreme environmental conditions. In the last few decades, climate change has made a profound impact on high altitude plants. Stress conditions alter cellular homeostasis of plants. With the advent of proteomics, it has become evident that stresses induce changes in proteome by synthesis/expression of novel stress responsive proteins. These proteins constitute a highly organized, complex network that leads to changes in the molecular, biochemical, physiological, and morphological responses of plants. Herein, we comprehensively discuss the proteomics of medicinal plants and its role in adaptation along altitude gradient. This review aims to provide impetus to current research in medicinal plants ranging from developmental to stress biology and to generate basis for genetic engineers and plant breeders to produce next-generation medicinal plants.

Sulfur dioxide is a widespread air pollutant and it also acts as a signaling molecule in various processes in mammals. However, the role of SO₂ in programmed cell death (PCD) in plants is unclear. Here we studied the role of SO₂ in gibberellin (GA)-treated wheat aleurone layers. The results showed that 100 μM SO₂ donor (NaHSO₃/Na₂SO₃) could effectively delay PCD and inhibit the coalescence of small protein storage vacuoles (PSVs) in aleurone cells treated with GA. Also, SO₂ could reduce the accumulation of hydrogen peroxide and superoxide anion in GA-treated aleurone layers. In this process, SO₂ could sustain higher activities of catalase, guaiacol peroxidase, ascorbate peroxidase, and superoxide dismutase and lower activities of lipoxygenase and polyphenol oxidase by comparing with GA alone. In addition, an induction of endogenous H₂S and NO was observed in SO₂-treated aleurone layers. The application of NO scavenger cPTIO could accelerate PCD in SO₂ or H₂S treated aleurone cells, suggesting that NO alleviated PCD by acting downstream of SO₂ and H₂S. In conclusion, these results imply that SO₂ could delay PCD in GA-treated wheat aleurone layers by enhancing cellular antioxidative capacity and H₂S/NO signals act downstream of SO₂.

Heterotrimeric guanine-nucleotide-binding proteins (G-proteins) play important roles in signal transduction and regulate responses to various stresses. Although G-protein signaling pathways have been extensively identified and characterized in model plants, there is little knowledge in non-model and especially in woody plants. Mulberry is an economically and ecologically important perennial tree, which is adaptable to many environmental stresses. In this study, we identified and cloned six G-protein genes including one Gα, one Gβ, two Gγ, one RGS (regulator of G-protein signaling protein) and one RACK1 (receptor for activated C kinase 1) involved in G-protein signaling. Sequence and phylogenetic analysis revealed that Morus G-proteins are evolutionarily conserved compared with those of other plants. A real-time quantitative reverse transcription polymerase chain reaction analysis showed that Morus G-protein signaling genes were ubiquitously but differentially expressed in various tissues. The expression of all of these genes was affected by abiotic stresses and signal molecules, which indicated that Morus G-protein signaling may be involved in environmental stress and defense responses.

A large-scale expression profiling study was performed to investigate candidate genes associated with the two quantitative trait loci (QTLs) for aluminum (Al) tolerance (Alt1 and Alt2). They have been identified in rye and localized on chromosomes 6R and 3R, respectively. Materials employed were hexaploid wheat (cv. Chinese Spring), and two wheat-rye addition lines (3R-AL and 6R-AL). Seedlings were treated with and without Al for 24 h to examine genes up-regulated or down-regulated by Al. Measurements of root growth at different Al concentrations showed the Al tolerance was higher in 3R-AL than in 6R-AL and wheat. Initial transcriptomic results revealed that more genes changed expression (>10 fold) in the wheat and in the 6R-AL line (moderately tolerant) than in the 3R-AL line (highly tolerant). A method was developed to determine whether candidate genes are involved in Al tolerance or in responses to Al toxicity. Real-time qPCRs were carried out in a subset of six genes with known function in near isogenic rye lines 389 (Al-sensitive) and 390 (Al tolerant). All six genes were up-regulated by Al in line 389 but not in line 390, indicating that they were involved in Al stress response but not in Al tolerance mechanisms. Subsequent analysis of Arabidopsis lines with knockout mutations in homologues of these six genes showed an Al sensitivity similar to the wild-type, providing more evidence towards their participation in the response to stress rather than to Al tolerance. Once the stress response genes were ruled out, the focus was turned to the identification of tolerance genes by studying transcripts up-regulated and down-regulated in the tolerant 3R line with respect to wheat and 6R line. Finally, a list of candidate genes that could be conferring increased tolerance was obtained.