Tomato spotted wilt virus (TSWV) has significant economic impact on horticulture worldwide. One of the five proteins encoded by TSWV genome is the multifunctional NSs protein, which is a viral suppressor of RNA silencing (VSR) besides functioning as the effector of Tsw resistance gene in resistant pepper cultivars. In this study we demonstrate the in vivo self-interaction of NSs protein using bimolecular fluorescence complementation and yeast two-hybrid assays, and propose that a highly charged alpha helix located at the second half of the protein is required for self-interaction. Furthermore, we confirmed that self-interaction is not required for its effector function on pepper. Moreover, self-interaction is dispensable for gene silencing suppressor activity, although it enhances the suppression efficiency.

Garlic mustard (Alliaria petiolata) is an herbaceous biennial plant native to Europe. In Ukraine, in addition to becoming a serious invader, garlic mustard can serve as a host to several viruses, which may affect agricultural crops. In view of this, the purpose of the study was to identify the virome of garlic mustard growing in Ukraine. Plant samples collected in Kyiv regions were tested for the presence of cucumber mosaic virus (CMV), turnip mosaic virus (TuMV), turnip yellow mosaic virus (TYMV), watermelon mosaic virus II (WMV), and turnip crinkle virus (TCV) by serological and/or molecular methods. According to the results found in the present study, symptomatic A. petiolata obtained in 2021 were infected with CMV (60%), TuMV (20%), or co-infected with CMV + TuMV (20%). TYMV, WMV II, and TCV were not detected in any of the collected samples. The cDNA fragments encoded the coat protein (CP) gene of CMV and TuMV were sequenced and named as CMV-Ap and TuMV-Ap, respectively. In phylogenetic analysis, the CMV-Ap closely resembled the German isolate MW582807 (Sarracenia sp.), with 99.8% nucleotide identity and belongs to subgroup II of CMV. In the phylogenetic tree, TuMV-Ap clustered with isolates AP017803, AP017764, AP017791, and JQ073722, and represented the highest identity (98.6%) to Iranian isolate IRNTRa9 (AP017803) from Rapistrum rugosum and Turkish isolate TUR49 (AP017872) from Raphanus raphanistrum. The sequences of CMV-Ap and TuMV-Ap were deposited in the GenBank under Accession Numbers MZ540213 and OM799323, respectively. The results obtained in the study indicate the important role of infected garlic mustard as alternative host and natural reservoir of CMV and TuMV from which these economically important viruses can spread to other wild and cultivated plants. This is the first molecular evidence of TuMV infection in A. petiolata from Ukraine.

Leaves of Lotus japonicus infected by powdery mildew accumulate isoflavonoids, especially vestitol and sativan as protective compounds, whereas their flavonol-glycosides or phenylpropanoic acids content were rather decreased. For a better understanding of the induction of phytoalexin production in L. japonicus, the influence of important signalling molecules, namely methyl jasmonate and salicylic acid, was tested by leaf application, up to 15 d. No effects after spraying the leaves with 0.5 mM salicylic acid were observed regarding the plant isoflavonoid content, whereas methyl jasmonate showed moderate effect on vestitol accumulation. Chitosan application led to a strong increase in vestitol content, sativan and vestitol derivative also increased. In a similar way to the effect of powdery mildew infection, the content of ferulic acid and kaempferol glycosides decreased, except for one compound, identified as kaempferol-6-deoxyhexose, that is the result of acetylation of kaempferol glycosides. The application of chitosan also led to an approximately 2.5- to 3.5-fold increase in hydrogen peroxide content, indicating the involvement of H₂O₂ in the chitosan signalling pathway. The activity of key enzyme of the phenolic biosynthesis, phenylalanine amonia lyase (PAL), was stimulated as well as the expression of 3 PAL-isogenes (LjPAL4, LjPAL6, and LjPAL9), whereas LjPAL8 decreased. Moreover, the expression of two key enzymes of vestitol and sativan biosythesis: pterocarpan reductase and isoflavone synthase were also strongly induced. The data suggest that chitosan acts as an elicitor of the fungal attack in Lotus japonicus that do not lead to a general stimulation of the metabolism of phenols, but rather to a specific induction of isoflavonoid-phytoalexine production, especially for production of vestitol.

It is common knowledge that R2R3-MYB transcription factors play significant roles in plant biological and physiological processes, especially in the phenylpropanoid metabolism pathway. The cultivated strawberry (Fragaria × ananassa Duch.) is an octoploid (2n = 8x = 56) species from the Rosaceae family and it is also an important fruit crop species. However, the function of R2R3-MYB genes in strawberry remains largely unknown. Here, based on the genome of the cultivated strawberry cv. Reikou, 66 FanMYB genes were found and systematically analyzed. RNA-seq analysis revealed that some FanMYBs exhibited tissue-specific expressions and were methyl jasmonate (MeJA)-responsive. Phylogenetic relationships and protein-protein interaction analysis suggested that 13 FanMYBs were likely associated with phenylpropanoid metabolism. Out of these genes, FanMYB22, FanMYB36, FanMYB47, FanMYB49, and FanMYB63 were post-transcriptionally regulated by miR858 according to the degradome data analysis, suggesting the conservation and complex regulation network in F. × ananassa. Current findings provide a useful resource for future research on the function of FanMYBs and the regulatory mechanism of the phenylpropanoid pathway in strawberry.

Bacterial leaf blight (BLB) is a common disease that affects rice development and yield. The effects of major nutrients, especially nitrogen, on rice BLB susceptibility have been considered when devising rational fertilization strategies. However, the defense mechanism of rice against BLB under phosphate (Pi)-deficient conditions remains uncertain. Jasmonic acid (JA) is a phytohormone produced by rice plants to respond to abiotic and biotic stresses. Here, the involvement of the JA pathway in rice response to Xanthomonas oryzae pv. oryzae (Xoo) under low Pi was investigated in two contrasting rice cultivars G299 and G22. Expressions of JA-related genes under low Pi and Pi-related genes under JA treatment were assessed. The resistant capacity of G299 and G22 against Xoo infection was also investigated. In the JA-sensitive and Pi-sensitive cv. G299, JA-related genes were highly expressed under low Pi, and low Pi-responsive genes were strongly upregulated under JA treatment. Neither JA nor Pi pathways were activated in the JA-tolerant and low Pi-tolerant cv. G22. Low Pi strongly enhanced rice resistance to Xoo in cv. G299. Our study demonstrated that Pi deficiency confers rice resistance to Xoo. The JA pathway modulates the response to low Pi, depending on the cultivar. Pi-response genes are involved in Pi stress and may participate in the regulation of overall plant growth under various abiotic stresses. These findings provide new insights into the interaction between phosphate deficiency and the JA pathway and the subsequent effect on plant disease resistance.

The “Centre for Experimental Plant Biology”, a joint project of the Institute of Experimental Botany of the Czech Academy of Sciences and CEITEC (represented by Mendel and Masaryk Universities), focused on elucidating  the mechanisms of plant responses to abiotic and biotic stresses and their combinations at the cellular level, in intact plants during vegetative and reproductive stages, and fruit development. The consortium demonstrated the importance of shared research facilities, complementary approaches, and knowledge exchange, addressing demanding questions  in plant biology. The consortium made breakthrough in plant-pathogen interactions, including identification of  exocyst-syntaxin cooperation in non-host resistance. The results confirmed the fundamental role of phytohormones in stress responses, including negative correlation of leaf bioactive gibberellins with drought stress, and the role of cytokinins in ROS homeostasis, sulphur metabolism, and heat stress responses, including volatile emission. Molecular analyses revealed expansin-mediated cell wall remodelling, brassinosteroid-mediated regulation of root growth through PIN2, the role of ALBA and LARP6C proteins in pollen development under abiotic stress, and heat stress impact on fertilization rate, embryo and seed development. Gene Set Enrichment and RNA-Seq analyses allowed to identify crucial genes involved in the apple scab resistance network. The main results obtained during the five-year project are summarised here.

Populus euphratica belongs to Salicaceae family and grows in extreme desert environments. At present, the identification of the AP2/ERF gene family of transcription factors in Salicaceae is rare, and the role of the AP2/ERF gene family in P. euphratica under salt stress and exogenous hormones has not been reported. In this study, 197, 210, 231, 192, and 147 AP2/ERF genes were identified in P. euphratica, Populus trichocarpa, Populus deltoides, Salix sinopurpurea, and Arabidopsis thaliana, respectively. The 197 AP2/ERF gene family members of P. euphratica were divided into five subfamilies, namely, AP2 (35), RAV (5), ERF (96), DREB (65), and Soloist (1), by sequence alignment and phylogenetic analysis. In addition, these genes were scattered across 19 chromosomes. The detection of 10 motifs in the P. euphratica AP2/ERF gene family revealed that motif-8 and motif-9 only appeared in the ERF subfamily and DREB subfamily, respectively. Transcriptome data showed that PeAP2/ERF genes had different expression patterns under salt stress, abscisic acid (ABA) and gibberellic acid (GA₃) treatments, suggesting that the genes PeERF002, PeERF037, PeERF082, PeERF090, and PeAP2-14 may play important roles under salt stress and exogenous hormone treatments. This study provides a reference for the functional study of the PeAP2/ERF gene, and it also lays a foundation for the breeding strategy to improve the salt tolerance of P. euphratica.

Emmenopterys henryi Oliv., a monotypic genus in family Rubiaceae with high scientific and application value, is currently endangered. Adaptability of E. henryi to environment is important for its cultivation, management, and application. In this paper, the effect of abscisic acid (ABA) on the photosynthesis of E. henryi under salt stress was discussed. Results showed that 0.3 % (m/v) NaCl led to reduction of net photosynthetic rate (P_N), instantaneous carboxylation efficiency (ICE), and Rubisco activity but the increase of intercellular CO₂ concentration (c_i), which were remarkable relieved by ABA pretreatment. NaCl also induced reduction of the maximal efficiency of PS II photochemistry (F_v/F_m), the photochemical quenching (qP), the non-photochemical quenching (NPQ), the actual PS II efficiency [Y(II)], the regulated-energy dissipation [Y(NPQ)], whereas the increase of the unregulated energy dissipation [Y(NO)]. ABA application remarkably mitigated the depression of F_v/F_m, qP, Y(II) and the increase of Y(NO), but further promoted NPQ and Y(NPQ) in E. henryi due to high salinity. In addition, decrease of chlorophyll content, accumulation of MDA, and ion leakage in E. henryi due to high salinity were significantly alleviated by ABA pretreatment. Fluridone, an inhibitor of endogenous ABA synthesis, partially or completely reversed ABA mediated effects. These results suggested that ABA might improve the photosynthesis of E. henryi under salt stress via allocating more energy to photochemical reactions, or/and mitigating the decrease of Rubisco activity, or/and protecting photosynthetic apparatus by reinforcement of heat dissipation and alleviating oxidative damage.

Lycium barbarum Thunb. and Lycium ruthenicum Murray (wolfberries) have been utilized as traditional medicinal and nutritional plants in China for centuries. Much research has been focused on their high quality, yet the molecular mechanisms underlying morphological differences remain unclear. In this study, a comparative analysis of morphological and cytological characteristics indicated that significant differences existed. Meanwhile, transcriptomic analyses of the flower and fruit were performed at different developmental stages, and a total of 54 795 differentially expressed genes (DEGs) were screened. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that these DEGs were significantly enriched in substance metabolism, catalytic activity, single organism process, starch and sucrose metabolism, carotenoid biosynthesis, amino sugar and nucleotide sugar metabolism, phenylpropanoid biosynthesis, and other pathways. Based on these significantly enriched pathways, the ratio between nonsynonymous and synonymous substitution rates (Ka/Ks), and numerous studies related to flower and fruit development, we preliminarily screened eight transcription factor families related to flower and fruit development and counted the number of potential transcription factor genes. These candidate genes could provide a basis for future functional verification, helping to further research on the molecular mechanism of morphological differences in the two Lycium species.

Two ginseng species Panax vietnamensis and Panax stipuleanatus are precious medicinal plants restricted in several Vietnam provinces. They are very limited and endangered due to degraded habitats and over-harvesting. To preserve these two species, we used eight nuclear microsatellite markers to investigate genetic variability from the nine populations with 246 individuals for these two ginseng species. Our findings showed a moderate genetic heterozygosity in two species, P. vietnamensis (H_E = 0.386) and P. stipuleanatus (H_E = 0.342). Deficiency of heterozygosity was observed in all the studied populations of P. vietnamensis and three populations of P. stipuleanatus. Some populations had high allelic richness for both species. Private alleles were determined in all the studied populations of P. vietnamensis and two P. stipuleanatus populations. Genetic differentiation was low in two ginseng species. However, habitat loss, over-utilization and over-harvesting can be the main causes of reduced genetic heterozygosity. Neighbor-joining tree and discriminant analysis of principal components detected three major genetic groups. Finally, based on our findings, we propose in situ conservation of populations with high expected heterozygosity, allelic richness, and private alleles. Seed collection should be performed for ex-situ conservation as genetic pools in the future.

The artichoke (Cynara cardunculus subsp. scolymus) is an intriguing source of indigestible sugar polymers such as inulin-type fructans. Artichoke represents an important component of a traditional Mediterranean diet and its edible parts are a good source of many high added value compounds such as inulin, a polymer showing relevant prebiotic properties. Compared to the cultivated varieties, the wild cardoon (C. cardunculus var. sylvestris) growing naturally under harsh conditions and well-adapted to many marginal areas, could have a good potential for use in sustainable production in stressed lands. Here, we evaluated by enzymatic assay, the amount of inulin-type fructans both in artichoke and wild cardoon in the two different organs, heads and rhizomes. The expression pattern of the genes encoding the two key enzymes sucrose:sucrose 1-fructosyltransferase and fructan 1-fructosyltransferase, involved in fructan biosynthesis, have been also evaluated. Our results showed that the amount of inulin-type fructans was higher in the wild cardoon than in the artichoke heads, together with a higher expression of the two key genes involved in the fructan biosynthetic pathway. A conspicuous content of inulin-type fructans was found also in the rhizome, supporting the significant role of these compounds in the storage and in protection from cold and/or winter stresses.

Dendrobium williamsonii and Dendrobium cariniferum (Orchidaceae) are endangered perennial herbs, and they are very similar in morphology. Chloroplast genome sequencing technology provides a powerful tool for molecular analysis to get more infomation for phylogenetic analysis and identification of Dendrobium species. In this study, the complete chloroplast genomes of Dendrobium williamsonii and Dendrobium cariniferum were assembled and characterized using Illumina NovaSeq 6000. The genome sizes are 159 695 and 159 479 bp, including pairs of inverted repeats (27 055 and 27 024 bp) each separated by small single-copy regions (18 451 and 18 488 bp) and large single-copy regions (87 134 and 86 943 bp). The chloroplast genome overall GC content was 37.11% and 37.13%, respectively. Each chloroplast genome encoded the same number (147) of genes, including 88 protein-coding genes, 51 tRNA genes, and 8 rRNA genes. Comparative analysis of chloroplast genomes revealed the high degree of divergence included accD-psaL and ycf4 -cemA. The phylogenetic tree showed the two Dendrobium species formed only one small clade. A pair of primers that could effectively identify the two Dendrobium species were also screened. This study will provide theoretical basis for species identification, genetic breeding, and evolution of Dendrobium.

The seasonal patterns of changes in the content of lipophilic antioxidants β-carotene (β-Car), zeaxanthin (Zx), α-tocopherol (α-Toc), plastoquinone (PQ)/plastoquinol (PQH₂) were studied in the assimilating shoots of evergreen shrub Ephedra monosperma J.G. Gmel ex C.A. Mey under natural conditions of Central Yakutia. The shortening of the photoperiod and the seasonal decrease in temperature induced a 1.4-fold increase in α-Toc content. The fall in the average daily temperature from 0.1 to -8.1°C in October led to a decrease in the content of β-Car as a result of the accumulation of rhodoxanthin (Rhd). In this period a sharp increase in the content of Zx retained overnight was also detected. In winter, elevated content of Zx and α-Toc persisted. During September, the content of PQH₂ increased by 2.5 times and PQ by 1.4 times (compared to July). The beginning of exposure to freezing average daily temperatures from -3 to -5°C led to the depletion of the total PQ pool by 18%. However, the content of PQ_tot in the winter months was 1.5 times higher than at the end of July. The results revealed different timing and temperature ranges of variation for individual antioxidants during the development of frost resistance in ephedra.

The present work aimed to evaluate the effect of heat stress on common bean (Phaseolus vulgaris L.) genotypes during the reproductive phase as a function of the inoculation of plants with Bacillus subtilis. The treatments were established by inoculating two strains of B. subtilis (AP-3 and AP-12) and a control. The plants were subjected to heat stress when they reached the reproductive stage, with an increase in temperature to 28/33°C. The duration of the stress period was ten days. Flowering, biochemical, and gene expression evaluations were performed. There was the interaction of B. subtilis AP-3 with the bean cultivar IAC-Imperador, reducing flower abortion, promoting the formation of new flower buds, and increasing the content of proline and guaiacol peroxidase activity in plant tissues. However, there was a reduction of transcription of genes encoding the 1-carboxylic acid-1aminocyclopropane oxidase and ethylene response factors and an increase of the Δ¹-pyrroline-5-carboxylate synthetase1 gene. These results suggest that B. subtilis may modulate some metabolic pathways in response to high-temperature stress during the reproductive phase of the common bean. This also confirms that Bacillus strains represent a useful option to moderate abiotic stresses.

Late embryogenesis abundant (LEA) proteins are important for promoting the growth and stress tolerance of plants. They are widely involved in plant growth regulation and responses to hormones and environmental factors. However, knowledge of the functions of the LEA gene in ginseng species remains limited. In this study, a Panax ginseng LEA gene (PgLEA) expression vector was constructed, and stable transgenic Arabidopsis lines were established. The PgLEA protein was classified in the LEA-2 subgroup. Reverse-transcription quantitative PCR analysis showed that the expression of PgLEA increased under 300 mM NaCl or 10 % (m/v) polyethylene glycol treatments. Under salt and osmotic stresses, overexpression of PgLEA in transgenic Arabidopsis plants improved germination rate, root length, and survival rate compared to wild-type plants. In response to drought or salt stress, transgenic plants increased proline accumulation, decreased malonaldehyde content and ion leakage. Furthermore, the transgenic plants exhibited significantly increased activity of superoxide dismutase, peroxidase, and catalase, and reduced accumulation of hydrogen peroxide and superoxide. Moreover, overexpression of PgLEA affected the expression of genes related to salt/drought stress. Taken together, PgLEA is a positive regulator of drought and salinity stress, and positively functioned in pleiotropic effects through regulating osmotic balance, reactive oxygen species scavenging and inducing transcription of stress-related genes. PgLEA may enable ginseng plants to adapt to adverse environments. The data presented herein imply that PgLEA may be useful for breeding new stress-tolerant ginseng cultivars.

Long-term low temperatures restrict the regrowth of winter wheat (Triticum aestivum L.), thus decreasing agricultural output. Non-enzymatic expansins, which are related to plant growth, have been reported to respond to drought, salinity, and low-temperature stress. We obtained an expansin 3 gene, TaEXPA9. It is located in winter wheat cv. Dongnong with high cold hardiness. We analyzed the expression patterns of TaEXPA9-A/B/D in this cultivar and conducted a subcellular localization analysis of TaEXPA9-A/B/D in the onion epidermis. Transgenic Arabidopsis thaliana line with EXPA9-A/B/D overexpression was obtained to examine the effects of the orthologous genes of these expansins on plant growth and low-temperature stress resistance. The results showed that EXPA9-A/B/D expression significantly increased at 4 °C, it was higher in the roots than in shoots, and EXPA9-A/B/D was localized in the cell wall. The roots were well-developed in the transgenic A. thaliana, and the growth-related markers and setting rate were better than in the wild-type. Recovery was stronger in the transgenic plants after freezing stress. At low-temperature stress, the antioxidant enzyme activities and content of osmoregulatory substances in the TaEXPA9-A/B/D-overexpressing A. thaliana plants were significantly higher than in the wild-type plants, and the degree of membrane lipid peroxidation was lower. In summary, TaEXPA9 orthologous genes participate in the low-temperature stress response, and they might be of great importance in molecular breeding.

Tomato yellow leaf curl virus (TYLCV), a whitefly-transmitted single-stranded DNA (ssDNA) virus, causes the most important viral disease of tomato worldwide. TYLCV-mediated disease is mainly controlled via extensive insecticide sprays aiming at the whitefly vector. RNA-based vaccination was proven to be a non-transgenic approach leading to efficient plant virus control. In this work, double-stranded RNA (dsRNA) molecules deriving from sequences of the C4 and V2 genes of TYLCV-Mild were produced in vitro and topically applied onto tomato plants along with the virus (via agroinfiltration). DsC4 and dsV2 application reduced disease incidence to 23 and 46 %, respectively, while TYLCV positive control reached 64 %. Bioinformatics analysis of the virus-specific small interfering RNAs (vsiRNAs) from TYLCV-infected tomato revealed 'hot' and 'cold' spots in the TYLCV-Mild genome. Interestingly, the viral C-strand had twofold siRNA reads when compared to that of the V-strand. Overall, vsiRNAs of negative and positive polarity were almost equal (53.5 vs. 46.6 %); vsiRNAs of negative polarity prevailed at the V-strand. Stem-loop RT-PCR validated the presence of six vsiRNAs (hot or cold spots) in TYLCV-Mild-infected and dsRNA-treated tomato. The exogenously applied dsRNA was found to rapidly move systemically in tomato and was detected for 54 days post treatment (dpt). The applied dsRNA molecules were successfully processed by the Dicer-like proteins (DCLs) in tomato since small interfering RNAs (siRNAs) deriving from the dsRNA were detected for at least 54 dpt. This consists the first report of dsRNA-based vaccination applied against a monopartite geminivirus.

Thioredoxins (Trx) are small multifunctional redox proteins that contain thioredoxin conserved domain and active site WCXXC. The Trx family has an important role in multiple processes, including electron transport, seed germination, redox regulation, biotic and abiotic stresses resistance, etc. Although Trx genes have been extensively characterized in some plants, they have not been reported in peanut until now. The identification of AhTrx genes provides potential candidate genes for studying their effects and regulatory mechanisms in peanut (Arachis hypogaea L.) growth and development, especially under aluminium (Al) stress. It is also helpful to further analyze the Al resistance pathway in plants. Seventy AhTrx genes were identified using a genome-wide search method and conservative domain analysis. Then the basic physicochemical properties, phylogenetic relationship, gene structure, chromosomal localization, and promoter prediction were studied by the bioinformatic methods. Furthermore, the expressions of AhTrx genes under different Al treatment times in two peanut cultivars were tested using a real-time quantitative polymerase chain reaction. Seventy AhTrx genes were identified and characterized. Phylogenetic tree analysis showed that all AhTrx members could be classified into 9 groups with different conserved domains. Motif 1 was found to exist in every sequence, with an active site. Furthermore, the gene structures showed that the AhTrx family was complicated and changeable during evolution. The chromosomal localization indicated that the distribution and density of the Trx family on 20 peanut chromosomes were uneven. Predictive promoter analysis indicated that AhTrx proteins might play a role in phytohormones synthesis and stress response. Finally, the expression patterns of the AhTrx genes showed that every gene was differently expressed under Al treatment in different peanut cultivars, some were obvious, others had no significant difference, some were at a high level, while others were at a low level. This study systematically identifies the Trx gene family in peanut, providing some candidates for further study on its effects and regulatory mechanism under Al stress in peanut.

The present study aimed to determine if the needles of male and female European yew (Taxus baccata L.) trees differ in their content of basic compounds (proteins, amino acids, and carbohydrates), and whether the observed differences result only from the gender factor or if they are also associated with the needles' age and season. The study was conducted on male and female European yew specimens, collected from the Botanical Garden of the Jagiellonian University in Krakow during three seasons. Male specimens had significantly higher content of insoluble carbohydrates compared to the females ones. In the first year of the needles life, the dry mass and content of soluble carbohydrates increased significantly. In the third year of the needles' life, the content of amino acids increased significantly and the content of soluble proteins decreased. The highest differences between the genders in the individual months were observed in the soluble carbohydrates and amino acids amounts and the least in the soluble proteins. The growth of the new needles seems to be at least partly sustained by carbohydrates remobilization from the older needles. In conclusion, male and female yews differ in their metabolism. These gender differences may vary with the needles' age. For this reason, the needle samples of different age should not be mixed, and the sampling time should be chosen carefully. For one-year-old needles, a potentially interesting gender marker may be an increased content of free amino acids in June and July, and high content of soluble carbohydrates in January, while for two-year-old needles, an increased content of free amino acids in male individuals from September. March seems to be an interesting month, as it shows significant differences between the genders in terms of all biochemical features studied in this research.

To reveal the regulation mechanism of terpenoid biosynthesis in the leaves of Eucalyptus camaldulensis, the content of volatiles in eucalyptus leaves and the transcriptome databases of young and mature leaves were analyzed. The results showed that E. camaldulensis contains 92 and 89 kinds of volatile substances in the young and mature leaves, respectively. Among them, the content of 1,8-cineole, β-pinene, and other substances was significantly different in young and mature leaves. A total of 99 802 unigenes were obtained from the transcriptome database of young and mature leaves of E. camaldulensis and 18 441 genes displayed obviously differential expressions during both developmental stages. There were 6 982 up-regulated unigenes and 11 461 down-regulated unigenes in the young leaf stage compared to the mature leaf stage. The key genes for terpenoid biosynthesis, including limonene synthase-10, limonene synthase-11, myrcene synthase-1, α- pinene synthase-2, and 1,8-cineole synthase-2, were selected for further analysis to explore the mechanism of gene regulation and genetic transformation. The expressions of key genes were validated by RT-qPCR, and their expressions were consistent with RNA-seq data. WRKY, MYB, NAC, and bHLH transcription factors (TFs) displayed important regulatory effects on the above key genes. Thus, a regulatory network model of terpenoid biosynthesis was constructed using target genes and TFs during leaf development in E. camaldulensis. These results provide theoretical evidence for understanding the terpenoid biosynthesis in plants and reference for terpenoids utilization by genetic engineering methods in E. camaldulensis.

Grasslands play a significant role in livestock fodder production and thus, contribute to food security worldwide while providing numerous additional ecosystem services. However, how agroclimatic conditions and adverse weather events relevant for grasslands will change across the European grassland areas has not been examined to date. Using a single reference setup for soil and management over 476 European sites defined by climate stations, we show the probability of eight selected adverse weather events with the potential to significantly affect grassland productivity under climate change and how these events vary regionally across Europe. Changes in these eight key agroclimatic indicators create markedly specific spatial patterns. We found that by 2050, the exposure of the south and west European grasslands to heat and drought may double in comparison with today and that the area with frequent occurrences of heat and drought will expand northwards. However, across Ukraine, Belarus, and the Baltic countries to southern Finland and Sweden, the likelihood of these events is likely to decrease. While changing cultivars and management strategies are unavoidable, shifting grassland production to other regions to reduce the risk may not be possible as the risk of adverse events beyond the key grassland-growing areas increases even further. Moreover, we found marked changes in the overall thermal and water regimes across European regions. The effect of adverse weather events in the future could be different in other regions of the world compared to regions in Europe, emphasizing the importance of conducting similar analyses for other major grassland producing regions. To mitigate the impact of climate change, new ways of maintaining grassland productivity need to be developed. These methods include more efficient selection of species mixtures for specific regions, including increased use of legumes and forbs; incorporation of new genetic resources, including the development of hybrid cultivars, such as Festulolium hybrids; and incorporation of state-of-the-art technologies in breeding programs and new grazing management.

The family of pyrabactin resistance 1 (PYR1)/PYR1-like (PYL) regulatory components of ABA receptors (RCAR) play a vital role in the initial step of ABA signaling. To understand the expression mode of PYL genes in response to various abiotic stresses in Chinese cabbage (Brassica rapa ssp. pekinensis), the members of BrPYL gene family were first identified with the relevant bioinformatics software. And then, the relative expressions of identified BrPYLs after heat, cold, drought, and salt treatments for 0, 24, 48, and 72 h were determined via real-time quantitative PCR. Here, we identified 24 PYLs in the B. rapa genome. Based on the phylogenetic analysis, these BrPYL genes were divided into three classes and distributed on ten chromosomes in Chinese cabbage. Most of BrPYL genes in the same group have similar gene structures and intron numbers. There were seven genes (BrPYL5, BrPYL8, BrPYL22, BrPYL3, BrPYL18, BrPYL11, and BrPYL21) from Group A with two introns and one gene (BrPYL19) from Group D with one intron. Analysis of conserved motifs suggested that every group contained motif 2 containing the Polyketide_cyc2 domain. Subsequently, the prediction of cis-acting elements indicated that BrPYL genes had 5 stress-related elements and 5 hormone-related elements, among which the number of MYC (dehydration reaction) was the highest, suggesting that BrPYL genes could respond to hormones and abiotic stresses. Expression patterns under four abiotic stresses showed that the expressions of BrPYL4, BrPYL11, BrPYL21, and BrPYL23 responded to these stresses at different time points. To conclude, we identified the BrPYL genes and build the BrPYLs expression mode in response to various abiotic stresses. This study provides a theoretical basis for stress-resistance breeding of Chinese cabbage.

Midvein is an important structure of the upright leaf of rice, and its normal development is essential to the formation of a common plant type of rice (Oryza sativa L.). To reveal the effect of midvein deficiency on photosynthesis-related characteristics, leaf microstructure, and vein characteristics, the photosynthetic features between the midvein-deficient mutant dl-14 and wild-type Huanghuazhan plants were analyzed. The results indicated that the midvein area of the dl-4 mutant lacked large intercellular space and instead it was filled with mesophyll cells. Moreover, the vein density of the dl-14 mutant was significantly higher than that in cv. Huanghuazhan. Chlorophyll (Chl) a, Chl b, and carotenoid content were markedly elevated in dl-14. In terms of photosynthetic characteristics, we observed that under high irradiance and high CO₂ concentration, the net photosynthetic rate of dl-14 plants was significantly higher than that of Huanghuazhan plants, but its water use efficiency was significantly lower. In addition, several major photosynthetic parameters, including characteristics of chlorophyll fluorescence (the efficiency of excitation capture of open PS II center, photochemical quenching, effective quantum yield of PS II photochemistry, and electron transfer rate) were significantly higher in dl-14 plants compared to Huanghuazhan plants, but the nonphotochemical quenching of dl-14 mutant was significantly lower than that of Huanghuazhan. These findings indicate that the dl-14 mutant has higher vein density, stronger photon conversion ability, and weaker radiation dissipation ability. This study can provide theoretical support for breeders to use the midvein-deficient mutant.

Trehalose is a nonreducing disaccharide that is involved in the regulation of plant responses to a variety of environmental stresses. Trehalose 6-phosphate synthase (TPS) and trehalose 6-phosphate phosphatase (TPP) are two key enzymes in trehalose synthesis and they are widely distributed in higher plants. At present, TPS family genes have been systematically identified and analyzed in many plant species, but the TPP family genes have been rarely studied. In this study, ten TPS and six TPP genes in cannabis (Cannabis sativa L.) were identified at the genomic level. The phylogenetic tree of TPS and TPP family members in cannabis, Arabidopsis, and rice was constructed, and all the genes were divided into three subgroups: Class I, Class II, and Class III. The number of exons and motif types among Class I members was exactly the same, as were Class II members, but the gene structure and motif types of Class III members were slightly different. There were four pairs of CsTPSs and CsTPPs that had gene duplication, indicating that gene duplication events played an important role in the amplification of TPS and TPP families in cannabis. The results of expression analysis under abiotic stresses showed that 68.75 % of CsTPS and CsTPP genes were significantly induced by at least one abiotic stress. Among these genes, the expression of CsTPS1, CsTPS9, and CsTPPA was highest under at least one abiotic stress. These three genes may play a key role in abiotic stress responses. Most of the CsTPS and CsTPP genes that are closely located in the evolutionary tree have the same or similar functions. To our knowledge, this is the first paper that systematically reports the TPS and TPP gene families in cannabis.

Paspalum fasciculatum Willd. ex Flüggé grows in mining soils which are Cd- and Pb-contaminated where it exhibits tolerance to Pb and the ability to extract Pb from these soils. To elucidate tolerance mechanisms to Pb-stress, liquid chromatography with tandem mass spectrometry (LC-MS/MS) was used to quantify changes in the accumulation of proteins in leaves. We identified 323 proteins involved in primary metabolism and response to biotic or abiotic stresses. Although proteins involved in the processes of photosynthesis and saccharide and energy metabolism presented the greatest amount of down-regulated proteins, the plant was able to maintain photosynthetic functions and obtain energy to sustain the vital balance. P. fasciculatum based their tolerance on increased antioxidant defenses, improving the protection and repair of proteins and transduction signals to coordinate physiological response to Pb-stress. Our results provide important information to understand the tolerance mechanisms in P. fasciculatum and could be important in future molecular studies on the resistance and accumulation of Pb in plants.

Cauliflower is one of the most important popular vegetables in China. Cauliflower (Brassica oleracea L. var. botrytis) produces glucosinolates, which are secondary metabolites that have anti-cancer properties. The choice of suitable reference genes (RGs) for gene expression studies has a significant effect on experimental outcomes. In this study, we selected 15 candidates as RGs and analyzed them under heat, cold, drought, and salt stresses and methyl jasmonate (MeJA) treatment. The precision of the real-time quantitative polymerase chain reaction data was assessed using four methods (geNorm, NormFinder, BestKeeper, and ΔCt). The results revealed that EF1-β and ACT7; ACT1, EF1-β, and TUB6; TUA2 and EF1-β; HIS and ACT7; and ACT7 and ACT3 were the most stable combinations of genes for heat stress, cold stress, drought stress, salt stress, and MeJA treatment, respectively. The expression stability of the RGs fluctuated under different experimental conditions. Choosing the optimal RG for the specific experiment is therefore essential. This work provides relevant information for further gene expression studies on cauliflower and other closely related species.

Drought and salinity, which can alter the water balance, disrupt the ionic equilibrium, and create reactive oxygen species (ROS), are capable of destroying plant tissues. In this study, transcriptomics, proteomics, and metabolomics have been used to elucidate various abiotic stress responses. In transcriptional signaling pathways, abscisic acid (ABA) is one of the plant phytohormones that regulate the stress response. On the other hand, several regulons and factors of transcription contributed in the reaction to osmotic stresses, as well as in ABA-dependent/independent signaling pathways. However, the findings display that intricate molecular reaction of plants under stress conditions may be controlled by complicated regulative networks of gene expression and signal transduction, as well as by the interaction between them. From the point of view of proteomics, protein modifications in response to stress can be considered as a molecular tool to improve the resistance of plants to environmental stresses. These studies have provided new information about the significance of several gene and protein networks involved in the response of plants to salinity and drought, and the induction of tolerance. Moreover, identifying the crucial pathways which are involved in salinity and drought resistance can open doors for the establishment of commercial-resistant crop cultivars, and might be very useful in the next-generation crop breeding strategies to produce plants with salinity and drought-resistant traits.

An efficient plant regeneration protocol has been established for two commercial Populus hybrid clones, MC (Populus × euramericana) and UNAL (Populus × interamericana). The culture of internode segments on Murashige and Skoog (MS) medium with 0.5 μM α-naphthalene acetic acid (NAA) and 4 μM N⁶-benzyladenine for 7 weeks (2 weeks in absence of activated charcoal and 5 weeks in its presence) resulted in the highest frequency of shoot regeneration (100 % for MC and 82 % for UNAL). All regenerated shoots longer than 2 cm rooted on half-strength MS medium, independent of the addition of 0.1 μM NAA. Nevertheless, shoots developed better-formed roots in NAA-free medium, which had a positive effect on the acclimatization of plants. In order to know the cellular processes underlying in vitro shoot organogenesis, a histological study was made in UNAL internode-explants. Results revealed that in vitro culture caused swelling around the cut-off zones in all explants, but only those undergoing organogenesis formed proliferation centers under subepidermal cells, which led to formation of bud primordia. Moreover, in vivo tissues and explants with different in vitro response showed different immunolabelling patterns when they were treated with fluorescentmonoclonal antibodies directed to several pectin-polysaccharides of the cell wall. Results allow us to assign a predominant role of homogalacturonan with a low degree of methyl-esterification in the initiation of bud primordia, role of β-1,4-D-galactan side chains of rhamnogalacturonan-I in the cellular differentiation, role of α-1,5-L-arabinan side chains of rhamnogalacturonan-I and of homogalacturonan with a high degree of methyl-esterification in cell division and growth.