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International audience; Evergreen plants in boreal biomes undergo seasonal hardening and dehardening adjusting their photosynthetic capacity and photoprotection; acclimating to seasonal changes in temperature and irradiance. Leaf epidermal ultraviolet (UV)-screening by flavonols responds to solar radiation, perceived in part through increased ultraviolet-B (UV-B) radiation, and is a candidate trait to provide cross-photoprotection. At Hyytiälä Forestry Station, central Finland, we examined whether the accumulation of flavonols was higher in leaves of Vaccinium vitis-idaea L. growing above the snowpack compared with those below the snowpack. We found that leaves exposed to colder temperatures and higher solar radiation towards the top of hummocks suffered greater photoinhibition than those at the base of hummocks. Epidermal UV-screening was highest in upper-hummock leaves, particularly during winter when lower leaves were beneath the snowpack. There was also a negative relationship between indices of flavonols and anthocyanins across all leaves suggesting fine-tuning of flavonoid composition for screening vs. antioxidant activity in response to temperature and irradiance. However, the positive correlation between the maximum quantum yield of photosystem II photochemistry (Fv/Fm) and flavonol accumulation in upper hummock leaves during dehardening did not confer on them any greater cross-protection than would be expected from the general relationship of Fv/Fm with temperature and irradiance (throughout the hummocks). Irrespective of timing of snow-melt, photosynthesis fully recovered in all leaves, suggesting that V. vitis-idaea has the potential to exploit the continuing trend for longer growing seasons in central Finland without incurring significant impairment from reduced duration of snow cover.

It remains unclear how global climate change affects dioecious plants that may be especially vulnerable to climate drivers, because they often exhibit skewed sex ratios and eco-physiological specialization in certain microhabitats. In this study, female and male saplings of Populus cathayana were employed to explore sex-specific responses and the effects of sexual competition under elevated temperature (ET), elevated CO2 (EC) and combination of elevated temperature and CO2 (ETC). The results demonstrated that elevated temperature and CO2 interactively modulated sexual competition and responses of P. cathayana. Moreover, competition patterns affected the eco-physiological responses of P. cathayana to climate change treatments. Under both intra- and inter-sexual competition, biomass components, photosynthetic parameters and carbon-related metabolites of females were most strongly affected by ET, while males exhibited a higher photosynthesis and resource use efficiency, and a better biomass accumulation and carbon balance mechanism when compared to females when experiencing intra-sexual competition under EC. The competitive pressure of females on males in inter-sexual competition was intensified by ET, while it was alleviated by ETC. We conclude that climate change drivers and competition patterns differently regulate the sex-specific responses and competitive intensity of males and females, which may have a crucial effect on sex ratios, spatial sexual segregation, biomass production and carbon sequestration in dioecious species in the future.

Forest trees frequently interact with a diverse range of microorganisms including dark septate endophytes (DSEs) and fungal pathogens. Plant defense responses to either individual pathogens or endophytes have been widely studied, but very little is known on the effect of coinfection on host defenses. To study the impact of coinfection or tripartite interaction on plant growth and host defenses, Norway spruce (Picea abies (L.) Karst) seedlings were inoculated with a DSE Phialocephala sphaeroides or with a root pathogen Heterobasidion parviporum Niemela & Korhonen or coinfected with both fungi. The results showed that the DSE promoted the root growth of spruce seedlings. Control seedlings without any inoculum were subjected to sequencing and used as a baseline for identification of differentially expressed genes (DEGs). RNA-seq analysis of seedlings inoculated with P. sphaeroides, infected with H. parviporum or coinfected with both fungi resulted in a total of 5269 DEGs. The majority of DEGs were found in P. sphaeroides-inoculated seedlings. Lignin biosynthesis pathways were generally activated during fungal infections. The pattern was distinct with endophyte inoculation. The majority of the genes in the flavonoid biosynthesis pathway were generally suppressed during fungal infections. A specific transcriptional response to P. sphaeroides inoculation was the increased transcripts of genes involved in jasmonic acid biosynthesis, mitogen-activated protein kinases signaling pathway, plant hormone signal transduction and calcium-mediated signaling. This may have potentially contributed to promoting the root growth of seedlings. Although the coinfection suppressed the induction of numerous genes, no negative effect on the growth of the spruce seedlings occurred. We conclude that the subsequent H. parviporum infection triggered reprogramming of host metabolism. Conversely, the endophyte (P. sphaeroides), on the other hand, counteracted the negative effects of H. parviporum on the growth of the spruce seedlings.

We conducted genomic characterization based on SNP and SilicoDArT markers on the invasive Himalayan balsam (Impatiens glandulifera) plants originating from native and non-native regions of their distribution. When genetic relationships were explored by PCoA using SNP and SilicoDArT marker data, the first, second, and third principal coordinates explained altogether 37.4% and 31.0% of the variability, respectively. Samples from the UK, Canada, and Pakistan were grouped together, while Indian plants were clearly distinct based on SNP markers but relatively close to the UK-Canada-Pakistan group based on SilicoDArT markers. Constructed trees differentiated individuals into clusters resembling the PCoA patterns. The Bayesian BAPS analysis performed for the SNP data revealed that the individuals were distributed in seven clusters, representing samples from each of the four Finnish populations, India, Pakistan, and the combination of the UK and Canada. Similar clustering was visible in the UPGMA tree. The Indian cluster did not display any ancestral gene flow with the others, while the Pakistani cluster showed ancestral gene flow only with the combined UK and Canada cluster. Furthermore, the latter cluster displayed ancestral gene flow with the Finnish populations varying from 0% to 3.1%. The BAPS analyses conducted for the SilicoDArT data differ slightly: The individuals were distributed in nine clusters, and the Indian cluster exhibited ancestral gene flow with the mixed cluster including Canadian, Pakistani, and UK samples, and one Finnish sample. The AMOVA showed that 45% and 26% of variation was present among the I. glandulifera groups/populations and the rest within them based on SNP and SilicoDArT markers, respectively. The Bayesian BAPS analyses and the gene flow networks were the most informative tools for resolving relationships among native and introduced plants. It is notable that the small sample sizes for non-Finnish plant materials may affect the accuracy of the gene flow and other estimates.

Hop (Humulus lupulus L.): Traditional and Present Use, and Future Potential. Hop (Humulus lupulus) is best known for its use in beer brewing owing to its bittering flavor and floral aroma. Today, the brewing industry uses as much as 98% of the produced hop crop worldwide. However, there are many other uses, some of them known since prehistoric times. Hops, the cone–like female structures called strobili, are the most frequently used part of the hop plant, but other tissues are of interest as well. The present review compiles existing knowledge of the chemical and pharmacological properties, traditional and present uses and further use potential, genetic resources, and breeding attempts in H. lupulus, and discusses climate change challenges to hop production. It contains hundreds of phytochemicals, and some of the secondary metabolites have definite potential pharmacological and medicinal value, but further investigations are desirable. Hop substances are potential alternatives, e.g., in antimicrobial, cancer, metabolic syndrome, and hormone replacement therapy treatments, as well as insecticides, preservatives, and fragrances. There are presently a few hundred cultivated hop varieties, and new cultivars are being developed and tested. Future hop breeding efforts with different quality and adaptation targets can utilize existing genetic resources, such as wild populations and landraces present in many regions.

Species differ in their biological susceptibility to extinction, but the set of traits determining susceptibility varies across taxa. It is yet unclear which patterns are common to all taxa, and which are taxon-specific, with conse-quences to conservation practice. In this study we analysed the generality of trait-based prediction of extinction risk across terrestrial (including freshwater) vertebrates, invertebrates and plants at a global scale. For each group, we selected five representative taxa and within each group we explored whether risk can be related to any of 10 potential predictors. We then synthesized outcomes across taxa using a meta-analytic approach. High habitat specificity was a consistent predictor across vertebrates, invertebrates and plants, being a universal predictor of risk. Slow life-history traits - large relative offspring size, low fecundity, long generation length -, and narrow altitudinal range were also found to be good predictors across most taxa, but their universality needs to be supported with additional data. Poor dispersal ability was a common predictor of extinction risk among invertebrate and plant taxa, but not consistently among vertebrates. The remaining traits (body size, micro -habitat verticality, trophic level, and diet breadth) were useful to predict extinction risk but only at lower tax-onomical levels. Our study shows that despite the idiosyncrasies among taxa, universal susceptibility to extinction exists and several traits might influence extinction risk for most taxa. Informing conservation prior-itization at lower taxonomic scales should however include taxon-specific trait-based predictors of extinction risk.

Potyviruses move to neighboring cells in the form of virus particles or a coat protein (CP)-containing ribonucleoprotein complex. However, the precise roles of RNA-binding residues in potyviral CP in viral cell-to-cell movement remain to be elucidated. In this study, we predicted the three-dimensional model of tobacco vein banding mosaic virus (TVBMV)-encoded CP and found nine residues presumably located in the CP RNA-binding pocket. Substitutions of the two basic residues at positions 192 and 225 (R192 and K225) with either alanine, cysteine, or glutamic acid abolished TVBMV cell-to-cell and systemic movement in Nicotiana benthamiana plants. These substitutions also reduced the replication of the mutant viruses. Results from the electrophoretic mobility shift assay showed that the RNA-binding activity of mutant CPs derived from R192 or K225 substitutions was significantly lower than that of wild-type CP. Analysis of purified virus particles showed that mutant viruses with R192 or K225 substitutions formed RNA-free virus-like particles. Mutations of R192 and K225 did not change the CP plasmodesmata localization. The wild-type TVBMV CP could rescue the deficient cell-to-cell movement of mutant viruses. Moreover, deletion of any of the other seven residues also abolished TVBMV cell-to-cell movement and reduced the CP RNA-binding activity. The corresponding nine residues in watermelon mosaic virus CP were also found to play essential roles in virus cell-to-cell movement. In conclusion, residues R192 and K225 in the CP RNA-binding pocket are critical for viral RNA binding and affect both virus replication and cell-to-cell movement. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Chocolate spot (CS), caused by Botrytis fabae Sard., is an important threat to global faba bean production. Growing resistant faba bean cultivars is, therefore, paramount to preventing yield loss. To date, there have been no reported quantitative trait loci (QTLs) associated with CS resistance in faba bean. The objective of this study was to identify genomic regions associated with CS resistance using a recombinant inbred line (RIL) population derived from resistant accession ILB 938. A total of 165 RILs from the cross Mélodie/2 × ILB 938/2 were genotyped and evaluated for CS reactions under replicated controlled climate conditions. QTL analysis identified five loci contributing to CS resistance on faba bean chromosomes 1 and 6, accounting for 5.0–23.4% of the total phenotypic variance. The sequences of SNP markers linked to resistance QTLs on chromosome 1 that have the largest effects encode multiple classes of putative disease and/or defense-related genes. The results of this study not only provide insight into disease-resistance QTLs, but also can be used as potential targets for marker-assisted breeding in faba bean genetic improvement for CS resistance.Key messageQTL mapping identified key genomic regions associated with chocolate spot resistance on faba bean chromosomes 1 and 6, which may serve as novel genetic tools to develop disease-resistant faba bean cultivars.

Bird‐mediated dispersal is presumed to be important in the dissemination of many different types of organisms, but concrete evidence remains scarce. This is especially true for biota producing microscopic propagules. Tree‐dwelling birds, such as woodpeckers, would seem to represent ideal dispersal vectors for organisms growing on standing tree trunks such as epiphytic lichens and fungi. Here, we utilize bird natural history collections as a novel source of data for studying dispersal ecology of plants, fungi, and microorganisms. We screened freshly preserved specimens of three Finnish woodpecker species for microscopic propagules. Samples were taken from bird feet, and chest and tail feathers. Propagules were extracted using a sonication–centrifugation protocol, and the material obtained was studied using light microscopy. Diverse biological material was recovered from all specimens of all bird species, from all positions sampled. Most abundant categories of discovered biological material included bryophyte fragments, fungal spores, and vegetative propagules of lichens. Also, freshwater diatoms, bryophyte spores, algal cells, testate amebae, rotifers, nematodes, pollen, and insect scales were identified. The method developed here is applicable to living specimens as well, making it a versatile tool for further research. Our findings highlight the potential of bird‐mediated dispersal for diverse organisms and showcase the use of natural history collections in ecological research., Woodpeckers are ideal candidates for bird‐mediated dispersal for a diverse set of organisms, especially epiphytes. We report high loads of diverse biological propagules extracted from feathers and feet of woodpecker specimens from natural history collections. The study highlights the potential of birds as dispersal vectors of lichens, fungi, bryophytes, and more.

Faba bean is susceptible to several leaf diseases caused by pathogenic fungi. With the increasing importance of faba bean in northern European cropping systems, the importance of its leaf diseases is likely to increase. The aim of this study was to discriminate the diseases and to test agronomic strategies for limiting their spread in a five-year experiment. Chocolate spot disease caused by Botrytis spp. and leaf blotches caused by an Alternaria/Stemphylium complex dominated, the severity of rust was lower, and downy mildew occurred in only one year. The severity of the diseases depended on weather conditions and cultivar, but not on sowing rate. Application of fungicides significantly decreased the severity of chocolate spot, leaf blotch and rust, but not downy mildew, and in no case was the disease progress stopped. None of the cultivars showed strong resistance to disease. The Alternaria/Stemphylium leaf blotch is an emerging disease of faba bean in northern Europe. Work continues on the identification of the responsible species in the chocolate spot and leaf blotch complexes and the determination of their relative importance in causing disease.

Plasma membrane aquaporin AtPIP2;5 is permeable to CO2 and contributes to mesophyll conductance of CO2 in leaves., The roles of different plasma membrane aquaporins (PIPs) in leaf-level gas exchange of Arabidopsis thaliana were examined using knockout mutants. Since multiple Arabidopsis PIPs are implicated in CO2 transport across cell membranes, we focused on identifying the effects of the knockout mutations on photosynthesis, and whether they are mediated through the control of stomatal conductance of water vapour (gs), mesophyll conductance of CO2 (gm), or both. We grew Arabidopsis plants in low and high humidity environments and found that the contribution of PIPs to gs was larger under low air humidity when the evaporative demand was high, whereas any effect of a lack of PIP function was minimal under higher humidity. The pip2;4 knockout mutant had 44% higher gs than wild-type plants under low humidity, which in turn resulted in an increased net photosynthetic rate (Anet). We also observed a 23% increase in whole-plant transpiration (E) for this knockout mutant. The lack of functional plasma membrane aquaporin AtPIP2;5 did not affect gs or E, but resulted in homeostasis of gm despite changes in humidity, indicating a possible role in regulating CO2 membrane permeability. CO2 transport measurements in yeast expressing AtPIP2;5 confirmed that this aquaporin is indeed permeable to CO2.

Snowmelt in alpine ecosystems brings ample water, and together with above-freezing temperatures, initiates plant growth. In this scenario, rapid activation of photosynthesis is essential for a successful life-history strategy. But, strong solar radiation in late spring enhances the risk of photodamage, particularly before photosynthesis is fully functional. We compared the photoprotective strategy of five alpine forbs: one geophyte not particularly specialised in subnival life (Crocus albiflorus) and four wintergreens differing in their degree of adaptation to subnival life, from least to most specialised: Gentiana acaulis, Geum montanum, Homogyne alpina and Soldanella alpina. We used distance to the edge of snow patches as a proxy to study time-dependent changes after melting. We postulated that the photoprotective response of snowbed specialists would be stronger than of more-generalist alpine meadow species. F-v/F-m was relatively low across wintergreens and even lower in the geophyte C. albiflorus. This species also had the largest xanthophyll-cycle pool and lowest tocopherol and flavonoid glycoside contents. After snow melting, all the species progressively activated ETR, but particularly the intermediate snowbed species G. acaulis and G. montanum. The photoprotective responses after snowmelt were idiosyncratic: G. montanum rapidly accumulated xanthophyll-cycle pigments, tocopherol and flavonoid glycosides; while S. alpina showed the largest increase in plastochromanol-8 and chlorophyll contents and the greatest changes in optical properties. Climate warming scenarios might shift the snowmelt date and consequently alter the effectiveness of photoprotection mechanisms, potentially changing the fitness outcome of the different strategies adopted by alpine forbs.

How the root metabolic profiles and rhizosphere bacterial communities of dioecious plants respond to soil properties and sex identity is largely unknown. In this study, we analyzed root phenolic metabolomes and rhizosphere bacterial microbiomes of Populus euphratica females and males in two P. euphratica plantations with different soil properties to reveal the relative importance of soil and sex effects, and to decipher associations of certain phenolic compounds with specific bacterial taxa. We found that the relative abundances of bacterial OTUs and phenolic metabolites were closely linked to soil properties and sex identity. Soil is the main filter influencing the root phenolic metabolic profiles and rhizosphere bacterial communities of P. euphratica, while sexes and their interactions with soil properties are secondary factors. Differences in the diversity and evenness of phenolic metabolites were affected by plant sex, but not by soil properties. Conversely, the diversity and evenness of bacterial communities were affected by soil properties independent of plant sex. A multiple regression model indicated the presence of associations between root phenolic metabolites and specific soil bacteria taxa. Furthermore, all bacterial phyla and families correlated with at least one phenolic metabolite. Especially, both Nitrosomonadaceae and Cytophagaceae positively correlated with salicylic acid. Thus, our study provides new insights into the ecological mechanism that maintains rhizosphere bacterial communities in P. euphratica plantations in the desert area.

Publisher Copyright: © 2021 Finnish Mycological Society. All rights reserved. Fungi that cause powdery mildew on plants are plant pathogenic parasites (Erysiphales) and can significantly reduce the ornamental value of plants and cause significant yield losses among cultivated plants. In this study, 94 plant accessions infected with powdery mildew were observed in Kumpula Botanic Garden, Helsinki, Finland, in 2015. The taxonomic affiliation and species richness of powdery mildew fungi were investigated. Morphological studies by microscope distinguished only 14 fungal species, whereas further comparisons of internal transcribed spacer (ITS) sequences enabled the identification of 28 species. Hence, ITS sequencing improved the reliability of species determination, as compared with the use of morphological characteristics only. The vegetation in an area of six hectares supported a wide range of fungi that cause powdery mildew as well as hyperparasitic microbes, which may balance the impact of pathogens in host plants. The findings of this study emphasize the role of botanical gardens in protecting biological diversity in urban areas.

Resinicolous fungi constitute a heterogeneous assemblage of fungi that live on fresh and solidified plant resins. The genus Sarea includes, according to current knowledge, two species, S. resinae and S. difformis. In contrast to other resinicolous discomycetes, which are placed in genera also including non-resinicolous species, Sarea species only ever fruit on resin. The taxonomic classification of Sarea has proven to be difficult and currently the genus, provisionally and based only on morphological features, has been assigned to the Trapeliales (Lecanoromycetes). In contrast, molecular studies have noted a possible affinity to the Leotiomycetes. Here we review the taxonomic placement of Sarea using sequence data from seven phylogenetically informative DNA regions including ribosomal (ITS, nucSSU, mtSSU, nucLSU) and protein-coding (rpb1, rpb2, mcm7) regions. We combined available and new sequence data with sequences from major Pezizomycotina classes, especially Lecanoromycetes and Leotiomycetes, and assembled three different taxon samplings in order to place the genus Sarea within the Pezizomycotina. Based on our data, none of the applied phylogenetic approaches (Bayesian Inference, Maximum Likelihood and Maximum Parsimony) supported the placement of Sarea in the Trapeliales or any other order in the Lecanoromycetes. A placement of Sarea within the Leotiomycetes is similarly unsupported. Based on our data, Sarea forms an isolated and highly supported phylogenetic lineage within the " Leotiomyceta". From the results of our multilocus phylogenetic analyses we propose here a new class, order, and family, Sareomycetes, Sareales and Sareaceae in the Ascomycota to accommodate the genus Sarea. The genetic variability within the newly proposed class suggests that it is a larger group that requires further infrageneric classification.

Aim The diversity and composition of natural communities are rapidly changing due to anthropogenic disturbances. Magnitude of this compositional reorganization varies across the globe, but reasons behind the variation remain largely unknown. Disturbances induce temporal turnover by stimulating species colonizations, causing local extinctions, altering dominance structure, or all of these. We test which of these processes drive temporal community changes, and whether they are constrained by natural environmental gradients. Moreover, we assess to what degree identity shifts translate to changes in dominance structure. Location Finland. Time period Observations 1985–2006, disturbance history > 140 years. Major taxa studied Vascular plants. Methods We investigated temporal turnover of boreal forest understorey in response to disturbance, here forest management, along a soil fertility gradient. We disentangle the roles of species gains, losses and abundance changes in driving temporal turnover in response to and after disturbance by comparing turnover rates in different forest age categories along a fertility gradient. We quantify temporal turnover using richness‐based complement of Jaccard’s similarity index and proportional‐abundance based dissimilarity index. We also test whether disturbance history or fertility influence the relationship between identity shifts and dominance structure. Results We found that the impact of disturbance on temporal turnover depends on soil fertility. The greatest turnover occurred in the most fertile forests immediately after disturbance. There, species gains and losses strongly altered dominance structure leading to high turnover, whereas undisturbed old forests and nutrient‐poor habitats were characterized by stable dominant species even when the majority of species shifted their identity. Main conclusions Our results suggest that human impacts on temporal biodiversity change vary along environmental gradients. In boreal forests, the fertile habitats have a higher probability than nutrient‐poor sites of changing their composition in response to anthropogenic disturbances. Resource availability and disturbance history may thus influence consequences of temporal turnover for ecosystem functioning.

SummaryScots pine (Pinus sylvestris) is the most widespread coniferous tree in the boreal forests of Eurasia and has major economic and ecological importance. However, its large and repetitive genome presents a challenge for conducting genome-wide analyses such as association studies and genomic selection. We present a new 50K SNP genotyping array for Scots pine research, breeding programs, and other applications. To select the SNP set, we first genotyped 480 Scots pine samples on a 407 540 SNP screening array, and identified 47 712 high-quality SNPs for the final array (called ‘PiSy50k’). Here, we provide details of the design and testing, as well as allele frequency estimates from the discovery panel, functional annotation, tissue-specific expression patterns, and expression level information for the SNPs or corresponding genes, when available. We validated the performance of the PiSy50k array using samples from breeding populations from Finland and Scotland. Overall, 39 678 (83.2%) SNPs showed low error rates (mean = 0.92%). Relatedness estimates based on array genotypes were consistent with the expected pedigrees, and the amount of Mendelian error was negligible. In addition, array genotypes successfully discriminate Scots pine populations from different geographic origins. The PiSy50k array will be a valuable tool for future genetic studies and forestry applications.Significance statementScots pine is an evolutionary, economically and ecologically impressive coniferous species but its gigantic genome has limited studying e.g. the genetic basis of its functional trait variation. We have developed a genotyping array that facilitates Scots pine genetic research and linking its trait variation to genetic polymorphisms and gene expression levels across the genome.

Microsoroideae is the third largest of the six subfamilies of Polypodiaceae, containing over 180 species. These ferns are widely distributed in the tropical and subtropical regions of the Old World and Oceania. We documented the spore ornamentation and integrated these data into the latest phylogenetic hypotheses, including a sampling of 100 taxa representing each of 17 major lineages of microsoroid ferns. This enabled us to reconstruct the ancestral states of the spore morphology. The results show verrucate ornamentation as an ancestral state for Goniophlebieae and Lecanoptereae, globular for Microsoreae, and rugulate surface for Lepisoreae. In addition, spore ornamentation can be used to distinguish certain clades of the microsoroid ferns. Among all five tribes, Lecanoptereae show most diversity in spore surface ornamentation. Electronic supplementary material The online version of this article (10.1007/s10265-020-01238-4) contains supplementary material, which is available to authorized users.

Coat proteins (CPs) play critical roles in potyvirus cell-to-cell movement. However, the underlying mechanism controlling them remains unclear. Here, we show that substitutions of alanine, glutamic acid, or lysine for the conserved residue tryptophan at position 122 (W-122) in tobacco vein banding mosaic virus (TVBMV) CP abolished virus cell-to-cell movement in Nicotiana benthamiana plants. In agroinfiltrated N. benthamiana leaf patches, both the CP and RNA accumulation levels of three W-122 mutant viruses were significantly reduced compared with those of wild-type TVBMV, and CP accumulated to a low level similar to that of a replication-deficient mutant. The results of polyprotein transient expression experiments indicated that CP instability was responsible for the significantly low CP accumulation levels of the three W-122 mutant viruses. The substitution of W-122 did not affect CP plasmodesmata localization or virus particle formation; however, the substitution significantly reduced the number of virus particles. The wild-type TVBMV CP could complement the reduced replication and abolished cell-to-cell movement of the mutant viruses. When the codon for W-122 was mutated to that for a different aromatic residue, phenylalanine or tyrosine, the resultant mutant viruses moved systemically and accumulated up to 80% of the wild-type TVBMV level. Similar results were obtained for the corresponding amino acids of W-122 in the watermelon mosaic virus and potato virus Y CPs. Therefore, we conclude that the aromatic ring in W-122 in the core domain of the potyviral CP is critical for cell-to-cell movement through the effects on CP stability and viral replication.

Chloroplast-to-nucleus retrograde signaling is essential for cell function, acclimation to fluctuating environmental conditions, plant growth and development. The vast majority of chloroplast proteins are nuclear-encoded, and must be imported into the organelle after synthesis in the cytoplasm. This import is essential for the development of fully functional chloroplasts. On the other hand, functional chloroplasts act as sensors of environmental changes and can trigger acclimatory responses that influence nuclear gene expression. Signaling via mobile transcription factors (TFs) has been recently recognized as a way of communication between organelles and the nucleus. In this study, we performed a targeted reverse genetic screen to identify dual-localized TFs involved in chloroplast retrograde signaling during stress responses. We found that CHLOROPLAST IMPORT APPARATUS 2 (CIA2) has a functional plastid transit peptide, and can be located both in chloroplasts and the nucleus. Further, we found that CIA2, along with its homolog CIA2-like (CIL) are involved in the regulation of Arabidopsis responses to UV-AB, high light and heat shock. Finally, our results suggest that both CIA2 and CIL are crucial for chloroplast translation. Our results contribute to a deeper understanding of signaling events in the chloroplast-nucleus cross-talk.

In Arabidopsis, two leaf-type ferredoxin-NADP(+) oxidoreductase (LFNR) isoforms function in photosynthetic electron flow in reduction of NADP(+), while two root-type FNR (RFNR) isoforms catalyse reduction of ferredoxin in non-photosynthetic plastids. As the key to understanding, the function of RFNRs might lie in their spatial and temporal distribution in different plant tissues and cell types, we examined expression of RFNR1 and RFNR2 genes using beta-glucuronidase (GUS) reporter lines and investigated accumulation of distinct RFNR isoforms using a GFP approach and Western blotting upon various stresses. We show that while RFNR1 promoter is active in leaf veins, root tips and in the stele of roots, RFNR2 promoter activity is present in leaf tips and root stele, epidermis and cortex. RFNR1 protein accumulates as a soluble protein within the plastids of root stele cells, while RFNR2 is mainly present in the outer root layers. Ozone treatment of plants enhanced accumulation of RFNR1, whereas low temperature treatment specifically affected RFNR2 accumulation in roots. We further discuss the physiological roles of RFNR1 and RFNR2 based on characterization of rfnr1 and rfnr2 knock-out plants and show that although the function of these proteins is partly redundant, the RFNR proteins are essential for plant development and survival.

Laccases (EC 1.10.3.2), a group of multi-copper oxidases (MCOs), play multiple biological functions and widely exist in many species. Fungal laccases have been extensively studied for their industrial applications, however, there was no database specially focused on fungal laccases. To provide a comparative genomics platform for fungal laccases, we have developed a comparative genomics platform for laccases and MCOs (http://laccase.riceblast.snu.ac.kr/). Based on protein domain profiles of characterized sequences, 3,571 laccases were predicted from 690 genomes including 253 fungi. The number of putative laccases and their properties exhibited dynamic distribution across the taxonomy. A total of 505 laccases from 68 genomes were selected and subjected to phylogenetic analysis. As a result, four clades comprised of nine subclades were phylogenetically grouped by their putative functions and analyzed at the sequence level. Our work would provide a workbench for putative laccases mainly focused on the fungal kingdom as well as a new perspective in the identification and classification of putative laccases and MCOs.

Aims Populus deltoides and P. euramericana are widely used in China as major forestry species. At present, little is known about their responses to nitrogen (N) deficiency when grown in monocultures or mixed plantations. The aim of this investigation was to analyze the growth, and morphological and physiological responses of P. deltoides and P. euramericana to different N levels under competition conditions. Methods We employed two Populus species (P. deltoides and P. euramericana) to discover how N deficiency affects plant traits under different competition types (P. deltoides × P. deltoides, intraspecific competition; P. euramericana × P. euramericana, intraspecific competition; P. deltoides × P. euramericana, interspecific competition). Potted seedlings were exposed to two N levels (normal N, N deficiency), and nitrogen- and competition-driven differences in growth, morphology and physiology were examined. Important Findings Under normal N conditions, interspecific competition significantly decreased the total root weight, root mass fraction (RMF), root–shoot ratio (R/S) and carbon/nitrogen ratio (C/N), and increased the leaf dry weight, leaf mass fraction and total leaf area of P. euramericana compared with intraspecific competition. The same conditions significantly affected the growth and morphological variables of P. deltoides, except for the dry weight of fine roots, R/S, specific leaf area, RMF, total nitrogen content and C/N compared with intraspecific competition. In addition, chlorophyll a (Chla), total chlorophyll (Tchl), carotenoid contents (Caro) and the carbon isotope composition (δ 13C) of P. deltoides were significantly lower in interspecific competition than in intraspecific competition, but no difference was detected in P. euramericana. The effects of N deficiency on P. deltoides under intraspecific competition were stronger than under interspecific competition. In contrast, the effects of N deficiency on P. euramericana between intraspecific and interspecific competition were not significantly different. These results suggest that under normal N condition, P. deltoides is expected to gain an advantage in monocultures rather than in mixtures with P. euramericana. Under N deficiency, the growth performance of P. euramericana was more stable than that of P. deltoides under both cultivation modes.

At the early stage of primary succession, there are deficient nutrient resources as well as competition stress among neighboring plants. Our aims were to elucidate the flexibility of tree seedlings' stoichiometric relationships and their effects on soil microbial communities, and to determine the driving forces of species turnover during primary succession through the evaluation of carbon (C) : nitrogen (N) : phosphorus (P) stoichiometric relationships. We conducted an experiment testing N addition effect on two species from the early stage of primary succession, under intra- and interspecific competition conditions. Our results showed that higher values of delta N-15-NO(3)(-)and delta C-13 were observed inPopulus purdomiiindividuals than inSalix rehderianaafter N application, which indicated a more efficient N uptake and water-use efficiency inP. purdomiiplants. Furthermore, under N addition, the intraspecific competition ofP. purdomiipresented a higher urease activity, microbial biomass C (MBC), microbial N:P ratio (MBN:MBP), and phylogenetic diversity compared to the intraspecific competition ofS. rehderiana. The results showed thatP. purdomiiseedlings influenced soil properties in a way that led to a positive feedback on their performance with an increasing N availability. In contrast,S. rehderianaseedlings influenced soil properties in a way that caused a negative feedback on their performance with increasing N. Such events can promote species turnover fromSalixtoPopulusduring succession. Additionally, DNA sequencing of soil bacterial communities showed differences in the composition of microbial communities in response to N fertilization and different competition patterns. Altogether, our results showed that plant, soil, and microbial community responses to N fertilization in a subalpine glacier forefield differed among tree species and competition patterns. This study brings new insight into mechanisms that drive species replacement and biogeochemical cycling during primary succession.

Plasmodesmata are small channels that connect plant cells. While recent technological advances have facilitated analysis of the ultrastructure of these channels, there are limitations to efficiently addressing their presence over an entire cellular interface. Here, we highlight the value of serial block electron microscopy for this purpose. We developed a computational pipeline to study plasmodesmata distributions and detect the presence/absence of plasmodesmata clusters, or pit fields, at the phloem unloading interfaces of Arabidopsis (Arabidopsis thaliana) roots. Pit fields were visualized and quantified. As the wall environment of plasmodesmata is highly specialized, we also designed a tool to extract the thickness of the extracellular matrix at and outside of plasmodesmata positions. We detected and quantified clear wall thinning around plasmodesmata with differences between genotypes, including the recently published plm-2 sphingolipid mutant. Our tools open avenues for quantitative approaches in the analysis of symplastic trafficking.

Fruticose lichens of the genus Usnea Dill. ex Adans. (Parmeliaceae), generally known as beard lichens, are among the most iconic epiphytic lichens in modern forest ecosystems. Many of the c. 350 currently recognized species are widely distributed and have been used as bioindicators in air pollution studies. Here we demonstrate that usneoid lichens were present in the Palaeogene amber forests of Europe. Based on general morphology and annular cortical fragmentation, one fossil from Baltic amber can be assigned to the extant genus Usnea. The unique type of cortical cracking indirectly demonstrates the presence of a central cord that keeps the branch intact even when its cortex is split into vertebrae-like segments. This evolutionary innovation has remained unchanged since the Palaeogene, contributing to the considerable ecological flexibility that allows Usnea species to flourish in a wide variety of ecosystems and climate regimes. The fossil sets the minimum age for Usnea to 34 million years (late Eocene). While the other similar fossils from Baltic and Bitterfeld ambers cannot be definitely assigned to the same genus, they underline the diversity of pendant lichens in Palaeogene amber forests.

In many regions, drought is suspected to be a cause of Scots pine decline and mortality, but the underlying physiological mechanisms remain unclear. Because of their relationship to ecohydrological processes, δ18O values in tree rings are potentially useful for deciphering long-term physiological responses and tree adaptation to increasing drought. We therefore analyzed both needle- and stem-level isotope fractionations in mature trees exposed to varying water supply. In a first experiment, we investigated seasonal δ18O variations in soil and needle water of Scots pine in a dry inner Alpine valley in Switzerland, comparing drought-stressed trees with trees that were irrigated for more than 10 years. In a second experiment, we analyzed twentieth-century δ18O variations in tree rings of the same forest, including a group of trees that had recently died. We observed less 18O enrichment in needle water of drought-stressed compared with irrigated trees. We applied different isotope fractionation models to explain these results, including the Péclet and the two-pool correction, which considers the ratio of unenriched xylem water in the needles to total needle water. Based on anatomical measurements, we found this ratio to be unchanged in drought-stressed needles, although they were shorter. The observed lower 18O enrichment in needles of stressed trees was therefore likely caused by increased effective path length for water movement within the leaf lamina. In the tree-ring study, we observed lower δ18O values in tree rings of dead trees compared with survivors during several decades prior to their death. These lower values in declining trees are consistent with the lower needle water 18O enrichment observed for drought-stressed compared with irrigated trees, suggesting that this needle-level signal is reflected in the tree rings, although changes in rooting depth could also play a role. Our study demonstrates that long-term effects of drought are reflected in the tree-ring δ18O values, which helps to provide a better understanding of past tree physiological changes of Scots pine.

Tree stems are an overlooked source of volatile organic compounds (VOCs). Their contribution to ecosystem processes and total VOC fluxes is not well studied, and assessing it requires better understanding of stem emission dynamics and their driving processes. To gain more mechanistic insight into stem emission patterns, we measured monoterpene, methanol, and acetaldehyde emissions from the stems of mature Scots pines (Pinus sylvestris L.) in a boreal forest over three summers. We analysed the effects of temperature, soil water content, tree water status, transpiration, and growth on the VOC emissions, and used generalized linear models to test their relative importance in explaining the emissions. We show that Scots pine stems are considerable sources of monoterpenes, methanol, and acetaldehyde, and their emissions are strongly regulated by temperature. However, even small changes in water availability affected the emission potentials: increased soil water content increased the monoterpene emissions within a day, whereas acetaldehyde and methanol emissions responded within two to four days. This lag corresponded to their transport time in the xylem sap from the roots to the stem. Moreover, the emissions of monoterpenes, methanol, and acetaldehyde were influenced by the cambial growth rate of the stem with six- to ten-day lags. This article is protected by copyright. All rights reserved.

Photosynthetic rate is concurrently limited by stomatal limitations and nonstomatal limitations (NSLs). However, the controls on NSLs to photosynthesis and their coordination with stomatal control on different timescales remain poorly understood. According to a recent optimization hypothesis, NSLs depend on leaf osmotic or water status and are coordinated with stomatal control so as to maximize leaf photosynthesis. Drought and notching experiments were conducted on Pinus sylvestris, Picea abies, Betula Pendula and Populus tremula seedlings in glasshouse conditions to study the dependence of NSLs on leaf osmotic and water status, and their coordination with stomatal control, on timescales of minutes and weeks, to test the assumptions and predictions of the optimization hypothesis. Both NSLs and stomatal conductance followed power-law functions of leaf osmotic concentration and leaf water potential. Moreover, stomatal conductance was proportional to the square root of soil-to-leaf hydraulic conductance, as predicted by the optimization hypothesis. Though the detailed mechanisms underlying the dependence of NSLs on leaf osmotic or water status lie outside the scope of this study, our results support the hypothesis that NSLs and stomatal control are coordinated to maximize leaf photosynthesis and allow the effect of NSLs to be included in models of tree gas-exchange.

Background and Aims The stomatal conductance (gs) of most plant species decreases in response to elevated atmospheric CO2 concentration. This response could have a significant impact on plant water use in a future climate. However, the regulation of the CO2-induced stomatal closure response is not fully understood. Moreover, the potential genetic links between short-term (within minutes to hours) and long-term (within weeks to months) responses of gs to increased atmospheric CO2 have not been explored. Methods We used Arabidopsis thaliana recombinant inbred lines originating from accessions Col-0 (strong CO2 response) and C24 (weak CO2 response) to study short- and long-term controls of gs. Quantitative trait locus (QTL) mapping was used to identify loci controlling short- and long-term gs responses to elevated CO2, as well as other stomata-related traits. Key Results Short- and long-term stomatal responses to elevated CO2 were significantly correlated. Both short- and long-term responses were associated with a QTL at the end of chromosome 2. The location of this QTL was confirmed using near-isogenic lines and it was fine-mapped to a 410-kb region. The QTL did not correspond to any known gene involved in stomatal closure and had no effect on the responsiveness to abscisic acid. Additionally, we identified numerous other loci associated with stomatal regulation. Conclusions We identified and confirmed the effect of a strong QTL corresponding to a yet unknown regulator of stomatal closure in response to elevated CO2 concentration. The correlation between short- and long-term stomatal CO2 responses and the genetic link between these traits highlight the importance of understanding guard cell CO2 signalling to predict and manipulate plant water use in a world with increasing atmospheric CO2 concentration. This study demonstrates the power of using natural variation to unravel the genetic regulation of complex traits.

peerReviewed

European mountain grasslands are increasingly affected by land-use changes and climate, which have been suggested to exert important controls on grassland carbon (C) and nitrogen (N) pools. However, so far there has been no synthetic study on whether and how land-use changes and climate interactively affect the partitioning of these pools amongst the different grassland compartments. We analyzed the partitioning of C and N pools of 36 European mountain grasslands differing in land-use and climate with respect to above- and belowground phytomass, litter and topsoil (top 23 cm). We found that a reduction of management intensity and the abandonment of hay meadows and pastures increased above-ground phytomass, root mass and litter as well as their respective C and N pools, concurrently decreasing the fractional contribution of the topsoil to the total organic carbon pool. These changes were strongly driven by the cessation of cutting and grazing, a shift in plant functional groups and a related reduction in litter quality. Across all grasslands studied, variation in the impact of land management on the topsoil N pool and C/N-ratio were mainly explained by soil clay content combined with pH. Across the grasslands, below-ground phytomass as well as phytomass- and litter C concentrations were inversely related to the mean annual temperature; furthermore, C/N- ratios of phytomass and litter increased with decreasing mean annual precipitation. Within the topsoil compartment, C concentrations decreased from colder to warmer sites, and increased with increasing precipitation. Climate generally influenced effects of land use on C and N pools mainly through mean annual temperature and less through mean an- nual precipitation. We conclude that site-specific conditions need to be considered for understanding the effects of land use and of current and future climate changes on grassland C and N pools.

Publisher Copyright: © 2022 Northern forest soils are a major carbon (C) reservoir of global importance. To estimate how the C balance in these soils will change, the roles of tree roots and soil microbes in C balance should first be decoupled. This study determined how the activity of heterotrophs and tree roots together with root-associated microbes in the rhizosphere varies in coniferous forest soils in boreal, hemiboreal, and temperate climates along a latitudinal gradient using a trenching approach. We created experimental plots without living tree roots, measured soil respiration (CO2 efflux) from these and from unmanipulated plots using the chamber technique, and partitioned the efflux into root-rhizosphere (RR) and heterotrophic (RH) respiration. The share of RR in ecosystem gross primary production (GPP) decreased from north to south in the Scots pine (Pinus sylvestris L.) and the Norway spruce (Picea abies (L.) Karst.) forests, with the exception of a mixed site, where the share of RR in GPP varied strongly between the years. RR per ground area and per root biomass were mainly independent of climate within the gradient. RH per ground area increased from north to south with temperature, while RH per soil C did not change with temperature. Soil moisture did not significantly affect the respiration components in the northernmost site, whereas soil moisture was positively connected with RH and negatively with RR in other Scots pine sites and positively connected with RR in pure Norway spruce stands. The dynamic ecosystem model LPJ-GUESS was able to capture the seasonal dynamics of RH and RR at the sites, but overall accuracy varied markedly between the sites, as the model underestimated RH in the southern site and RR elsewhere. Our study provides knowledge about the nature of soil respiration components. The valuable insights can be used in more accurate land-ecosystem modelling of forest ecosystems.

SummaryWhile pathogenic and mutualistic microbes are ubiquitous across ecosystems and often co-occur within hosts, how they interact to determine patterns of disease in genetically diverse wild populations is unknown.To test whether microbial mutualists provide protection against pathogens, and whether this varies among host genotypes, we conducted a field experiment in three naturally-occurring epidemics of a fungal pathogen, Podosphaera plantaginis, infecting a host plant, Plantago lanceolata, in the Åland Islands, Finland. In each population, we collected epidemiological data on experimental plants from six allopatric populations that had been inoculated with a mixture of mutualistic arbuscular mycorrhizal fungi or a non-mycorrhizal control.Inoculation with arbuscular mycorrhizal fungi increased growth in plants from every population, but also increased host infection rate. Mycorrhizal effects on disease severity varied among host genotypes and strengthened over time during the epidemic. Host genotypes that were more susceptible to the pathogen received stronger protective effects from inoculation.Our results show that arbuscular mycorrhizal fungi introduce both benefits and risks to host plants, and shift patterns of infection in host populations under pathogen attack. Understanding how mutualists alter host susceptibility to disease will be important for predicting infection outcomes in ecological communities and in agriculture.Plain Language SummaryBeneficial, ‘mycorrhizal’ fungi in roots help plants grow and may protect them from diseases caused by pathogenic microbes. This study shows that arbuscular mycorrhizal fungi can influence patterns of plant disease during pathogen outbreaks in a natural landscape.

Summary Experimental evidence that nonstomatal limitations to photosynthesis (NSLs) correlate with leaf sugar and/or leaf water status suggests the possibility that stomata adjust to maximise photosynthesis through a trade-off between leaf CO2 supply and NSLs, potentially involving source-sink interactions. However, the mechanisms regulating NSLs and sink strength, as well as their implications for stomatal control, remain uncertain. We used an analytically solvable model to explore optimal stomatal control under alternative hypotheses for source and sink regulation. We assumed that either leaf sugar concentration or leaf water potential regulates NSLs, and that either phloem turgor pressure or phloem sugar concentration regulates sink phloem unloading. All hypotheses lead to realistic stomatal responses to light, CO2 and air humidity, including conservative behaviour for the intercellular-to-atmospheric CO2 concentration ratio. Sugar- and water-regulated NSLs are distinguished by the presence/absence of a stomatal closure response to changing sink strength. Turgor- and sugar-regulated phloem unloading are distinguished by the presence/absence of stomatal closure under drought and avoidance/occurrence of negative phloem turgor. Results from girdling and drought experiments on Pinus sylvestris, Betula pendula, Populus tremula and Picea abies saplings are consistent with optimal stomatal control under sugar-regulated NSLs and turgor-regulated unloading. Our analytical results provide a simple representation of stomatal responses to above- and below-ground environmental factors and sink activity. Experimental evidence that nonstomatal limitations to photosynthesis (NSLs) correlate with leaf sugar and/or leaf water status suggests the possibility that stomata adjust to maximise photosynthesis through a trade-off between leaf CO2 supply and NSLs, potentially involving source-sink interactions. However, the mechanisms regulating NSLs and sink strength, as well as their implications for stomatal control, remain uncertain. We used an analytically solvable model to explore optimal stomatal control under alternative hypotheses for source and sink regulation. We assumed that either leaf sugar concentration or leaf water potential regulates NSLs, and that either phloem turgor pressure or phloem sugar concentration regulates sink phloem unloading. All hypotheses led to realistic stomatal responses to light, CO2 and air humidity, including conservative behaviour for the intercellular-to-atmospheric CO2 concentration ratio. Sugar-regulated and water-regulated NSLs are distinguished by the presence/absence of a stomatal closure response to changing sink strength. Turgor-regulated and sugar-regulated phloem unloading are distinguished by the presence/absence of stomatal closure under drought and avoidance/occurrence of negative phloem turgor. Results from girdling and drought experiments on Pinus sylvestris, Betula pendula, Populus tremula and Picea abies saplings are consistent with optimal stomatal control under sugar-regulated NSLs and turgor-regulated unloading. Our analytical results provide a simple representation of stomatal responses to above-ground and below-ground environmental factors and sink activity.

The continuously increasing atmospheric carbon dioxide concentration ([CO2]) has substantial effects on plant growth, and on the composition and structure of forests. However, how plants respond to elevated [CO2] (e[CO2]) under intra- and interspecific competition has been largely overlooked. In this study, we employed Abies faxoniana Rehder & Wilson and Picea purpurea Mast. seedlings to explore the effects of e[CO2] (700 p.p.m.) and plant–plant competition on plant growth, physiological and morphological traits, and leaf ultrastructure. We found that e[CO2] stimulated plant growth, photosynthesis and nonstructural carbohydrates (NSC), affected morphological traits and leaf ultrastructure, and enhanced water- and nitrogen (N)- use efficiencies in A. faxoniana and P. purpurea. Under interspecific competition and e[CO2], P. purpurea showed a higher biomass accumulation, photosynthetic capacity and rate of ectomycorrhizal infection, and higher water- and N-use efficiencies compared with A. faxoniana. However, under intraspecific competition and e[CO2], the two conifers showed no differences in biomass accumulation, photosynthetic capacity, and water- and N-use efficiencies. In addition, under interspecific competition and e[CO2], A. faxoniana exhibited higher NSC levels in leaves as well as more frequent and greater starch granules, which may indicate carbohydrate limitation. Consequently, we concluded that under interspecific competition, P. purpurea possesses a positive growth and adjustment strategy (e.g. a higher photosynthetic capacity and rate of ectomycorrhizal infection, and higher water- and N-use efficiencies), while A. faxoniana likely suffers from carbohydrate limitation to cope with rising [CO2]. Our study highlights that plant–plant competition should be taken into consideration when assessing the impact of rising [CO2] on the plant growth and physiological performance.

Disease suppressive soils are important for managing soil-borne diseases that cannot be controlled with chemicals. One such disease is the potato common scab caused by Streptomyces species. Suppressiveness against common scab can develop spontaneously in fields where potato is grown for years without interruption, and this has been attributed to non-pathogenic Streptomyces strains. Streptomyces spp. have been used as inoculants in biological control, but their long-term effects have gained less attention. In our previous studies, a nonpathogenic Streptomyces strain (Str272) isolated from a potato common scab lesion suppressed common scab in field trials lasting over 5 years. In this study, bacterial communities in the tuberosphere i.e. in the soil adjacent to potato tubers, were analysed by next generation sequencing (NGS). The aim was to compare bacterial communities in untreated control plots to those in which seed tubers were treated with Str272 in one or several growing seasons. Str272 applications increased soil bacterial diversity and affected the bacterial composition in the potato tuberosphere. The most pronounced differences were observed between the untreated control and the treatments in which the antagonist had been applied in three or four consecutive years. The differences remained similar until the following growing season. Bacterial composition after repeated antagonist applications was associated with lower common scab severity. The antagonist applications had no or only slight effect on the number or abundance of OTUs belonging to Actinobacteria or Streptomyces, and no differences in quantities of pathogenic Streptomyces populations were detected by qPCR. This indicates that suppression of common scab by Str272 may not be based on direct effect on the common scab pathogens but is more likely to be associated with the alterations of the soil bacterial community. The most abundant bacteria phyla in the potato tuberosphere were Actinobacteria, Proteobacteria and Acidobacteria. However, the OTUs responding greatest to the antagonist treatments belonged to Bacterioidetes and Gemmatimonadetes. Results indicate that repeated applications of Str272 can change the bacterial community in the potato tuberosphere and lead to development of soil that is suppressive against potato common scab for several growing seasons after the last application.

The interactions between the phloem-limited pathogen ‘Candidatus Liberibacter solanacearum’ haplotype C and carrot (Daucus carota subsp. sativus) were studied at 4, 5, and 9 weeks postinoculation (wpi), by combining dual RNA-Seq results with data on bacterial colonization and observations of the plant phenotype. In the infected plants, genes involved in jasmonate biosynthesis, salicylate signaling, pathogen-associated molecular pattern- and effector-triggered immunity, and production of pathogenesis-related proteins were up-regulated. At 4 wpi, terpenoid synthesis-related genes were up-regulated, presumably as a response to the psyllid feeding, whereas at 5 and 9 wpi, genes involved in both the terpenoid and flavonoid production were down-regulated and phenylpropanoid genes were up-regulated. Chloroplast-related gene expression was down-regulated, in concordance with the observed yellowing of the infected plant leaves. Both the RNA-Seq data and electron microscopy suggested callose accumulation in the infected phloem vessels, likely to impair the transport of photosynthates, while phloem regeneration was suggested by the formation of new sieve cells and the upregulation of cell wall–related gene expression. The ‘Ca. L. solanacearum’ genes involved in replication, transcription, and translation were expressed at high levels at 4 and 5 wpi, whereas, at 9 wpi, the Flp pilus genes were highly expressed, suggesting adherence and reduced mobility of the bacteria. The ‘Ca. L. solanacearum’ genes encoding ATP and C4-dicarboxylate uptake were differentially expressed between the early and late infection stages, suggesting a change in the dependence on different host-derived energy sources. HPE1 effector and salicylate hydroxylase were expressed, presumably to suppress host cell death and salicylic acid-dependent defenses during the infection. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Publisher Copyright: © 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature. Oriented cell divisions are crucial throughout plant development to define the final size and shape of organs and tissues. As most of the tissues in mature roots and stems are derived from vascular tissues, studying cell proliferation in the vascular cell lineage is of great importance. Although perturbations of vascular development are often visible already at the whole plant macroscopic phenotype level, a more detailed characterization of the vascular anatomy, cellular organization, and differentiation status of specific vascular cell types can provide insights into which pathway or developmental program is affected. In particular, defects in the frequency or orientation of cell divisions can be reliably identified from the number of vascular cell files. Here, we provide a detailed description of the different clearing, staining, and imaging techniques that allow precise phenotypic analysis of vascular tissues in different organs of the model plant Arabidopsis thaliana throughout development, including the quantification of cell file numbers, differentiation status of vascular cell types, and expression of reporter genes.

Plant-soil microbe interactions are determined by plant characters. Sexual dimorphism in root development, nitrogen (N) assimilation and resource allocation have been studied in different environments. However, how dioecious plants affect soil microbial communities in natural forests, particularly in low precipitation regions, is still poorly known. In this study, natural Populus euphratica forests were investigated in three arid regions. We hypothesized that males and females impose sex-specific impacts on physiochemical traits of soil, microbial communities and N-cycling processes. We discovered only little sex effect on most physiochemical traits, and bacterial and fungal communities in top soil (0-20 cm) in the three studied forests. However, the sex effect was greater in deep soil. Compared with fungi, the structure and composition of bacterial communities were affected more by plant sex in the rhizosphere and bulk soil. Sex indirectly affected N-cycling processes through a negative impact on the soil water content. Expressions of AOA, AOB, nifH, nirS and nirK in the rhizosphere soil were significantly affected by sex, forest site and their interactions. Proteobacteria, Actinobacteria and Firmicutes in the rhizosphere and bulk soils of P. euphratica males showed more significant effects on ammoxidation, N fixation, denitrification and protease activities when compared to females. The results suggest that sexual differences in shaping bacterial communities and affecting N-cycling processes are greater when the soil becomes drier. Thus, low precipitation causes intense sex differences in the nitrogen uptake and use efficiency. Our study highlights the importance of sexual effects on shaping specific microbial communities and N-cycling processes.

Tree stems have been identified as sources of volatile organic compounds (VOCs) that play important roles in tree defence and atmospheric chemistry. Yet, we lack understanding on the magnitude and environmental drivers of stem VOC emissions in various forest ecosystems. Due to the increasing importance of extreme drought, we studied drought effects on the VOC emissions from mature Scots pine (Pinus sylvestris L.) stems. We measured monoterpenes, acetone, acetaldehyde and methanol emissions with custom-made stem chambers, online PTR-MS and adsorbent sampling in a drought-prone forest over the hot-dry summer of 2018 and compared the emission rates and dynamics between trees in naturally dry conditions and under long-term irrigation (drought release). The pine stems were significant monoterpene sources. The stem monoterpene emissions potentially originated from resin, based on their similar monoterpene spectra. The emission dynamics of all VOCs followed temperature at a daily scale, but monoterpene and acetaldehyde emission rates decreased nonlinearly with drought over the summer. Despite the dry conditions, large peaks of monoterpene, acetaldehyde and acetone emissions occurred in late summer potentially due to abiotic or biotic stressors. Our results highlight the potential importance of stem emissions in the ecosystem VOC budget, encouraging further studies in diverse environments.

Thaumatin-like proteins (TLPs) are involved in the plant defense response against pathogens, and most of them exhibit antifungal activity. However, the role of TLPs in pathogen-induced defense responses in spruce is not fully understood. In this study, four TLP genes encoding thaumatin-like protein, designated as PlTLP1–4, were isolated and identified from Picea likiangensis needles. Sequence analysis showed that PlTLP1, PlTLP3, and PlTLP4 contained 16 conserved cysteine residues, while PlTLP2 had only 10 conserved cysteine residues. qPCR analysis showed that PlTLPs were expressed in all tissues tested, PlTLP1, PlTLP3, and PlTLP4 had the highest expression levels in young fruits, while PlTLP2 had the highest expression levels in roots. In addition, the expression levels of four PlTLPs were significantly upregulated during infection by Lophodermium piceae. Four recombinant PlTLPs expressed in Escherichia coli exhibited obvious β-1,3-glucanase activity. The antifungal activity assay showed that four recombinant PlTLPs had significant inhibitory effects on the mycelial growth of L. piceae, Fusarium proliferatum, Botrytis cinerea, and Roussoella doimaesalongensis. Microscopic observation revealed that the recombinant PlTLP1–4 induced the morphological changes of the mycelia of L. piceae, and the recombinant PlTLP2 and PlTLP3 induced the morphological changes of the mycelia of F. proliferatum and R. doimaesalongensis, while all the recombinant PlTLPs had no obvious negative effect on the morphology of B. cinerea mycelium. These results suggest that PlTLP genes may play an important role in the defense response of P. likiangensis against L. piceae invasion.

Jasmonic acid (JA) and salicylic acid (SA) regulate stomatal closure, preventing pathogen invasion into plants. However, to what extent abscisic acid (ABA), SA and JA interact, and what the roles of SA and JA are in stomatal responses to environmental cues, remains unclear. Here, by using intact plant gas-exchange measurements in JA and SA single and double mutants, we show that stomatal responsiveness to CO2, light intensity, ABA, high vapor pressure deficit and ozone either did not or, for some stimuli only, very slightly depended upon JA and SA biosynthesis and signaling mutants, including dde2, sid2, coi1, jai1, myc2 and npr1 alleles. Although the stomata in the mutants studied clearly responded to ABA, CO2, light and ozone, ABA-triggered stomatal closure in npr1-1 was slightly accelerated compared with the wild type. Stomatal reopening after ozone pulses was quicker in the coi1-16 mutant than in the wild type. In intact Arabidopsis plants, spraying with methyl-JA led to only a modest reduction in stomatal conductance 80 min after treatment, whereas ABA and CO2 induced pronounced stomatal closure within minutes. We could not document a reduction of stomatal conductance after spraying with SA. Coronatine-induced stomatal opening was initiated slowly after 1.5-2.0 h, and reached a maximum by 3 h after spraying intact plants. Our results suggest that ABA, CO2 and light are major regulators of rapid guard cell signaling, whereas JA and SA could play only minor roles in the whole-plant stomatal response to environmental cues in Arabidopsis and Solanum lycopersicum (tomato).

The peatlands of the West Siberian Lowlands, comprising the largest pristine peatland area of the world, have not previously been covered by continuous measurement and monitoring programs. The response of peatlands to climate change occurs over several decades. This paper summarizes the results of peatland carbon balance studies collected over ten years at the Mukhrino field station (Mukhrino FS, MFS) operating in the Middle Taiga Zone of Western Siberia. A multiscale approach was applied for the investigations of peatland carbon cycling. Carbon dioxide fluxes at the local scale studied using the chamber method showed net accumulation with rates from 110, to 57.8 gC m−2 at the Sphagnum hollow site. Net CO2 fluxes at the pine-dwarf shrubs-Sphagnum ridge varied from negative (−32.1 gC m−2 in 2019) to positive (13.4 gC m−2 in 2017). The cumulative May-August net ecosystem exchange (NEE) from eddy-covariance (EC) measurements at the ecosystem scale was −202 gC m−2 in 2015, due to the impact of photosynthesis of pine trees which was not registered by the chamber method. The net annual accumulation of carbon in the live part of mosses was estimated at 24–190 gC m−2 depending on the Sphagnum moss species. Long-term carbon accumulation rates obtained by radiocarbon analysis ranged from 28.5 to 57.2 gC m−2 yr−1, with local extremes of up to 176.2 gC m−2 yr−1. The obtained estimates of various carbon fluxes using EC and chamber methods, the accounting for Sphagnum growth and decomposition, and long-term peat accumulation provided information about the functioning of the peatland ecosystems at different spatial and temporal scales. Multiscale carbon flux monitoring reveals useful new information for forecasting the response of northern peatland carbon cycles to climatic changes.

Witches' broom of birch (Betula spp.) caused by Taphrina betulina is an understudied disease that causes the formation of woody tumours, from which ectopic axillary buds and branches grow to form a broom-like structure. We have addressed two aspects of this disease using naturally infected mature trees in the field. Broom symptoms offer a convenient means of scoring susceptibility in the field. Variation in broom symptom presentation suggests possible variation in resistance against witches' broom disease. We tracked the local distribution of susceptible individuals among 721 trees at 159 independent sites. The analysis supports the hypothesis that there was genetic resistance segregating in these birch populations. Anatomical changes in broom symptom bearing branches of European silver birch (Betula pendula) were also addressed by comparing sections of tissues from three locations in the same branch, which were normal, swollen in infected tissue adjacent to a tumour, and inside a tumour. Examination of tumours revealed disorganized and swollen xylem, expanded secondary phloem and expanded periderm. Swollen tissues newly infected from spreading disease adjacent to tumours exhibited enhanced growth only in secondary phloem and the periderm, which also exhibited distortions. This finding suggests that tumour formation and possibly pathogen colonization may initiate in these tissues.

Primocane raspberry (Rubus idaeus L.) may produce crop on the lower part of the cane during the second year of its' growth cycle, if the canes are retained over winter. We aimed to evaluate the floricane cropping potential and berry quality in seven primocane raspberry cultivars, which are too late for fall cropping in the Northern growing conditions. Long cane plants of these cultivars were produced, overwintered, and grown like floricane cultivars in a high tunnel. Harvesting floricane crop started in mid-June, and the length of the summer cropping season was between eight to 10 weeks. 'Joan J' and 'Imara' were the earliest cultivars, and 'Autumn Bliss' was the latest. Total floricane yield in most cultivars was between 1.7 to 2.0 kg/cane. 'Autumn Treasure' yielded significantly less, and 'Autumn Bliss' produced 2.5 kg berries /cane. The average berry size was 5 g, except in 'Autumn Bliss' and 'Autumn Treasure', which had smaller berries. The highest sugar content was observed in 'Kwanza' berries and the lowest in 'Polka'. High throughput phenotyping proved an effective tool for determining berry shape and color. 'Kwanza' berry color was the reddest, yellowest and lightest of all cultivars. Based on our results, growing primocane raspberry cultivars as long cane plants for production of summer crop is more reliable than growing them as fall cropping in short growing season conditions. The cultivars 'Imara' and 'Kweli' were the most promising in our experiment; they had a high yield potential and excellent berry quality.

Faba bean is a widely adapted and high-yielding legume cultivated for its protein-rich seeds1. However, the seeds accumulate the anti-nutritional pyrimidine glucosides vicine and convicine, which can cause haemolytic anaemia—favism—in the 400 million individuals genetically predisposed by a deficiency in glucose-6-phosphate dehydrogenase2. Here, we identify the first enzyme associated with vicine and convicine biosynthesis, which we name VC1. We show that VC1 co-locates with the major QTL for vicine and convicine content and that the expression of VC1 correlates highly with vicine content across tissues. We also show that low-vicine varieties express a version of VC1 carrying a small, frame-shift insertion, and that overexpression of wild-type VC1 leads to an increase in vicine levels. VC1 encodes a functional GTP cyclohydrolase II, an enzyme normally involved in riboflavin biosynthesis from the purine GTP. Through feeding studies, we demonstrate that GTP is a precursor of vicine both in faba bean and in the distantly related plant bitter gourd. Our results reveal an unexpected biosynthetic origin for vicine and convicine and pave the way for the development of faba bean cultivars that are free from these anti-nutrients, providing a safe and sustainable source of dietary protein.

The accurate estimation of an increase in forest stand biomass has remained a challenge. Traditionally, in situ measurements are done by inventorying a number of trees and their biometric parameters such as diameter at the breast height (DBH) and height, sometimes these are complemented by carbon (C) content studies. Here we present the estimation of net primary productivity (NPP) over a two years period (2019–2020) at a 25-year-old Scots pine stand. Research was based on allometric equations made by direct biomass analysis (tree extraction) and carbon content estimations in individual components of sampled trees, combined with a series of stem diameter increments recorded by a network of band dendrometers. Site-specific allometric equations were obtained using two different approaches: using the whole tree biomass vs DBH (M1), and total dry biomass-derived as a sum of the results from individual tree components’ biomass vs DBH (M2). Moreover, equations for similar forest stands from the literature were used for comparison. Gross primary productivity (GPP) estimated from the eddy-covariance measurements allowed the calculation of carbon use efficiency (CUE = NPP/GPP). The two investigated years differed in terms of the sum and patterns of precipitation distribution, with a moderately dry year of 2019 that followed the extremely dry 2018, and the relatively average year of 2020. As expected, a higher increase in biomass was recorded in 2020 compared to 2019, as determined by both allometric equations based on in situ and literature data. For the former approach, annual NPP estimates reached ca. 2.0–2.1 t C ha−1 in 2019 and 2.6–2.7 t C ha−1 in 2020 depending on the “in situ equations” (M1-M2) used, while literature-derived equations for the same site resulted in NPP values ca. 20–30% lower. CUE was higher in 2020, which resulted from a higher NPP total than in 2019, with lower summer and spring GPP in 2020. However, the CUE values were lower than those reported in the literature for comparable temperate forest stands. A thorough analysis of the low CUE value would require a full interpretation of interrelated physiological responses to extreme conditions.

Young moso bamboo shoots are a popular seasonal food and an important source of income for farmers, with value for cultivation estimated at $30,000 per hectare. Bamboo also has great environmental importance and its unique physiology is of scientific interest. A rare and valuable phenomenon has recently appeared where a large number of adjacent buds within a single moso bamboo rhizome have grown into shoots. Although of practical importance for the production of edible shoots, such occurrences have not been scientifically studied, due to their rarity. Analysis of collected reports from enhanced shoot production events in China showed no evidence that enhanced shoot development was heritable. We report the analysis of the rhizosphere microbiome from a rhizome with 18 shoots, compared to rhizomes having one or no shoots as controls. The community of prokaryotes, but not fungi, correlated with the shoot number. Burkholderia was the most abundant genus, which was negatively correlated with rhizome shoot number, while Clostridia and Ktedonobacteria were positively correlated. Two Burkholderia strains were isolated and their plant-growth promoting activity was tested. The isolated Burkholderia strains attenuated the growth of bamboo seedlings. These data provide the first study on excessive shoot development in bamboo, which will facilitate hypothesis building for future studies.