Similar to other cropping systems, few walnut cultivars are used as scion in commercial production. Germplasm collections can be used to diversify cultivar options and hold potential for improving crop productivity, disease resistance and stress tolerance. In this study we explored the anatomical and biochemical bases of photosynthetic capacity in 11 J. regia accessions in the USDA-ARS National Clonal Germplasm Repository. Net assimilation rate (An) differed significantly among accessions and was greater in those from lower latitudes coincident with increases in stomatal and mesophyll conductance, leaf thickness, mesophyll porosity and gas-phase diffusion, and leaf nitrogen, and lower leaf mass and stomatal density. High CO2-saturated assimilation rates led to increases in An under limiting conditions. Greater An was found in lower latitude accessions native to climates with more frost-free days, greater precipitation seasonality, and lower temperature seasonality. As expected, water stress consistently impaired photosynthesis with the highest % reductions in three lower latitude accessions (A3, A5, and A9), which had the highest An under well-watered conditions. However, An for A3 and A5 remained amongst the highest under dehydration. J. regia accessions, which have leaf structural traits and biochemistry that enhance photosynthesis, could be used as commercial scions or breeding parents to enhance productivity.

Cold acclimation in plants is a complex phenomenon involving numerous stress-responsive transcriptional and metabolic pathways. Existing gene expression studies have primarily addressed cold acclimation responses in herbaceous plants, and few have focused on perennial evergreens, such as conifers, that survive extremely low temperatures during winter. Relative to Arabidopsis leaves, the main transcriptional response of Norway spruce (Picea abies (L.) H. Karst) needles exposed to cold was delayed, and this delay was associated with slower development of freezing tolerance. Despite this difference in timing, our results indicate that, similar to herbaceous species, Norway spruce principally utilizes early response transcription factors (TFs) of the APETALA 2/ethylene-responsive element binding factor (AP2/ERF) superfamily and NAM (no apical meristem)/ATAF (Arabidopsis Transcription Factors)/CUC (cup shaped cotyledon) (NACs). The needles and root of Norway spruce showed contrasting results, in keeping with their different metabolic and developmental states. Regulatory network analysis identified conserved TFs, including a root-specific bHLH101 homolog, and other members of the same TF family with a pervasive role in cold regulation, such as homologs of ICE1 and AKS3, and also homologs of the NAC (anac47 and anac28) and AP2/ERF superfamilies (DREB2 and ERF3), providing new functional insights into cold stress response strategies in Norway spruce.

Oxylipins are lipid-derived molecules that are ubiquitous in eukaryotes and whose functions in plant physiology have been widely reported. They appear to play a major role in plant immunity by orchestrating reactive oxygen species (ROS) and hormone-dependent signalling pathways. The present work focuses on the specific case of fatty acid hydroperoxides (HPOs). Although some studies report their potential use as exogenous biocontrol agents for plant protection, evaluation of their efficiency in planta is lacking and no information is available about their mechanism of action. In this work, the potential of 13(S)-hydroperoxyoctadeca-(9Z,11E)-dienoic acid (13-HPOD) and 13(S)-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic acid (13-HPOT), as plant defence elicitors and the underlying mechanism of action are investigated. Arabidopsis thaliana leaf resistance to Botrytis cinerea was observed after root application with HPOs. They also activate early immunity-related defence responses, like ROS. As previous studies have demonstrated their ability to interact with plant plasma membranes (PPM), we have further investigated the effects of HPOs on biomimetic PPM structure using complementary biophysics tools. Results show that HPO insertion into PPM impacts its global structure without solubilizing it. Relationship between biological assays and biophysical analysis suggests that lipid amphiphilic elicitors that directly act on membrane lipids might trigger early plant defence events

Salt stress is a major limiting factor that severely affects the survival and growth of crops. It is important to understand the salt tolerance ability of Brassica napus and explore the underlying related genetic resources. We used a high-throughput phenotyping platform to quantify 2,111 image-based traits (i-traits) of a natural population under 3 different salt stress conditions and an intervarietal substitution line (ISL) population under 9 different stress conditions to monitor and evaluate the salt stress tolerance of B. napus over time. We finally identified 928 high-quality i-traits associated with the salt stress tolerance of B. napus. Moreover, we mapped the salt stress-related loci in the natural population via a genome-wide association study (GWAS) and performed a linkage analysis associated with the ISL population, respectively. The results revealed 234 candidate genes associated with salt stress response, and two novel candidate genes, BnCKX5 and BnERF3, were experimentally verified to regulate the salt stress tolerance of B. napus. This study demonstrates the feasibility of using high-throughput phenotyping-based QTL mapping to accurately and comprehensively quantify i-traits associated with B. napus. The mapped loci could be used for genomics-assisted breeding to genetically improve the salt stress tolerance of B. napus.

Dolichols (Dols), ubiquitous components of living organisms, are indispensable for cell survival. In plants, as well as other eukaryotes, Dols are crucial for posttranslational protein glycosylation, aberration of which leads to fatal metabolic disorders in humans and male sterility in plants. Until now, the mechanisms underlying Dol accumulation remain elusive. In this report, we have analyzed the natural variation of the accumulation of Dols and six other isoprenoids between more than 120 Arabidopsis thaliana accessions. Subsequently, by combining QTL and GWAS approaches, we have identified several candidate genes involved in the accumulation of Dols, polyprenols, plastoquinone, and phytosterols. The role of two genes implicated in the accumulation of major Dols in Arabidopsis – the AT2G17570 gene encoding a long searched for cis-prenyltransferase (CPT3) and the AT1G52460 gene encoding an alpha-beta hydrolase (ABH) – is experimentally confirmed. These data will help to generate Dol-enriched plants which might serve as a remedy for Dol-deficiency in humans.

The concentration and homeostasis of intracellular phosphate (Pi) are crucial for sustaining cell metabolism and growth. During short-term Pi starvation, intracellular Pi is maintained relatively constant at the expense of vacuolar Pi. After the vacuolar stored Pi is exhausted, the plant cells induce the synthesis of intracellular acid phosphatase (APase) to recycle Pi from expendable organic phosphate (Po). In this study, the expression, enzymatic activity and subcellular localization of ACID PHOSPHATASE 1 (OsACP1) were determined. OsACP1 expression is specifically induced in almost all cell types of leaves and roots under Pi stress conditions. OsACP1 encodes an acid phosphatase with broad Po substrates and localizes in the endoplasmic reticulum (ER) and Golgi apparatus (GA). Phylogenic analysis demonstrates that OsACP1 has a similar structure with human acid phosphatase PHOSPHO1. Overexpression or mutation of OsACP1 affected Po degradation and utilization, which further influenced plant growth and productivity under both Pi-sufficient and Pi-deficient conditions. Moreover, overexpression of OsACP1 significantly affected intracellular Pi homeostasis and Pi starvation signalling. We concluded that OsACP1 is an active acid phosphatase that regulates rice growth under Pi stress conditions by recycling Pi from Po in the ER and GA.

When grown under cool temperature, winter annuals upregulate photosynthetic capacity as well as freezing tolerance. Here, the role of three cold-induced C-repeat-Binding Factor (CBF1–3) transcription factors in photosynthetic upregulation and freezing tolerance was examined in two Arabidopsis thaliana ecotypes originating from Italy (IT) or Sweden (SW), and their corresponding CBF1–3-deficient mutant lines it:cbf123 and sw:cbf123. Photosynthetic, morphological, and freezing-tolerance phenotypes as well as gene expression profiles were characterized in plants grown from seedling stage under different combinations of light level and temperature. Under high light and cool growth temperature (HLC), a greater role of CBF1–3 in IT versus SW was evident from both phenotypic and transcriptomic data, especially with respect to photosynthetic upregulation and freezing tolerance of whole plants. Overall, features of SW were consistent with a different approach to HLC acclimation than seen in IT, and an ability of SW to reach the new homeostasis through involvement of transcriptional controls other than CBF1–3. These results provide tools and direction for further mechanistic analysis of the transcriptional control of approaches to cold acclimation suitable for either persistence through brief cold spells or for maximization of productivity in environments with continuous low temperatures.

To explore diversity in cold hardiness mechanisms, high resolution magnetic resonance imaging (MRI) was used to visualize freezing behaviors in wintering flower buds of Daphne kamtschatica var. jezoensis, which have no bud scales surrounding well-developed florets. MRI images showed that anthers remained stably supercooled to -14 ∼ -21°C or lower whilst most other tissues froze by -7°C. Freezing of some anthers detected in MRI images at ∼ -21°C corresponded with numerous low temperature exotherms and also with the “all-or-nothing” type of anther injuries. In ovules/pistils, only embryo sacs remained supercooled at -7°C or lower, but slowly dehydrated during further cooling. Cryomicroscopic observation revealed ice formation in the cavities of calyx tubes and pistils but detected no ice in embryo sacs or in anthers. The distribution of ice nucleation activity in floral tissues corroborated the tissue freezing behaviors. Filaments likely work as the ice blocking barrier that prevents ice intrusion from extracellularly frozen calyx tubes to connecting unfrozen anthers. Unique freezing behaviors were demonstrated in Daphne flower buds: preferential freezing avoidance in male and female gametophytes and their surrounding tissues (by stable supercooling in anthers and by supercooling with slow dehydration in embryo sacs) whilst the remaining tissues tolerate extracellular freezing.

Xylem embolism resistance varies across species influencing drought tolerance, yet little is known about the determinants of the embolism resistance of an individual conduit. Here we conducted an experiment using the optical vulnerability method to test whether individual conduits have a specific water potential threshold for embolism formation and whether pre-existing embolism in neighbouring conduits alters this threshold. Observations were made on a diverse sample of angiosperm and conifer species through a cycle of dehydration, rehydration and subsequent dehydration to death. Upon rehydration after the formation of embolism, no refilling was observed. When little pre-existing embolism was present, xylem conduits had a conserved, individual, embolism resistance threshold that varied across the population of conduits. The consequence of a variable conduit-specific embolism threshold is that a small degree of pre-existing embolism in the xylem results in an apparently more resistant xylem in a subsequent dehydration, particularly in angiosperms with vessels. While our results suggest that pit membranes separating xylem conduits are critical for maintaining a conserved individual embolism threshold for given conduit when little pre-exisiting embolism is present, as the percentage of embolized conduits increases, gas movement, local pressure differences, and connectivity between conduits increasingly contribute to embolism spread.

Xylem is a main road in plant long-distance communication. Through xylem plants transport water, minerals and myriad of signaling molecules. With the onset during early embryogenesis, the development of xylem tissues relays on hormone gradients, activity of unique transcription factors, distribution of mobile miRNAs and receptor-ligand pathways. These regulatory mechanisms are often interconnected and all together contribute to the plasticity of water conducting tissue. Remarkably, root xylem carries water to all above-ground organs and therefore influences all aspects of plant growth. Because of the global warming and increasing water deficit, we need to come up with solutions for the crops of the future. It is clear that structure of water conducting elements directly impacts water transport within the plant. Among plant pathogens- vascular wilts attacking xylem -are the most harmful. Our knowledge about xylem anatomy and rewiring ability could bring the solutions against these diseases. In this review we summarize the recent findings on the molecular mechanisms of xylem formation with a special attention to the cellular changes, and cell wall rearrangements that are necessary to create functional capillaries. We emphasize the impact of abiotic factors and pathogens on xylem plasticity and discuss multidisciplinary approach to model xylem in crops.

Known elicitors of plant defenses against eggs of herbivorous insects are low-molecular-weight organic compounds associated with the eggs. However, previous studies provided evidence that also proteinaceous compounds present in secretion associated with eggs of the herbivorous sawfly Diprion pini can elicit defensive responses in Pinus sylvestris. Pine responses induced by the proteinaceous secretion are known to result in enhanced emission of (E)-β-farnesene, which attracts egg parasitoids killing the eggs. Here, we aimed to identify the defense-eliciting protein and elucidate its function. After isolating the defense-eliciting protein from D. pini egg secretion by ultrafiltration and gel electrophoresis, we identified it by MALDI-ToF mass spectrometry as an annexin-like protein, which we named “diprionin”. Further GC-MS analyses showed that pine needles treated with heterologously expressed diprionin released enhanced quantities of (E)-β-farnesene. Our bioassays confirmed attractiveness of diprionin-treated pine to egg parasitoids. Expression of several pine candidate genes involved in terpene biosynthesis and regulation of ROS homeostasis was similarly affected by diprionin and natural sawfly egg deposition. However, the two treatments had different effects on expression of pathogenesis related genes (PR1, PR5). Diprionin is the first egg-associated proteinaceous elicitor of indirect plant defense against insect eggs described so far.

The Antarctic green alga Chlamydomonas sp. UWO241 is an obligate psychrophile that thrives in the cold (4-6°C) but is unable to survive at temperatures ≥18°C. Little is known how exposure to heat affects its physiology or whether it mounts a heat stress response in a manner comparable to mesophiles. Here, we dissect the responses of UWO241 to temperature stress by examining its growth, primary metabolome and transcriptome under steady-state low temperature and heat stress conditions. In comparison with Chlamydomonas reinhardtii, UWO241 constitutively accumulates metabolites and proteins commonly considered as stress markers, including soluble sugars, antioxidants, polyamines, and heat shock proteins to ensure efficient protein folding at low temperatures. We propose that this permanent stress metabolism is an adaptive advantage to life at extreme conditions. A shift from 4°C to a non-permissive temperature of 24°C alters the UWO241 primary metabolome and transcriptome, but growth of UWO241 at higher permissive temperatures (10°C and 15°C) does not provide enhanced heat protection. UWO241 also fails to induce the accumulation of HSPs when exposed to heat, suggesting that it has lost the ability to fine-tune its heat stress response. Our work adds to the growing body of research on temperature stress in psychrophiles, many of which are threatened by climate change.

The flagellin epitope flg22, a pathogen-associated molecular pattern (PAMP), binds to the receptor-like kinase FLAGELLIN SENSING2 (FLS2), and triggers Ca2+ influx across the plasma membrane (PM). The flg22-induced increases in cytosolic Ca2+ concentration ([Ca2+]i) (FICA) play a crucial role in plant innate immunity. It’s well established that the receptor FLS2 and the key downstream component, reactive oxygen species (ROS) burst, undergoes sensitivity adaptation after flg22 stimulation, referred to as desensitization and resensitization, to prevent over responses to pathogens. However, whether FICA also mount adaptation mechanisms to ensure appropriate and efficient responses against pathogens remains poorly understood. Here, we carried out detailed analyses of [Ca2+]i increases upon two successive flg22 treatments, recorded and characterized, for the first time, rapid desensitization but slow resensitization of FICA in Arabidopsis thaliana. Pharmacological analyses showed that the rapid desensitization might be synergistically regulated by ligand-induced FLS2 endocytosis as well as the PM depolarization. The recovery of desensitized FICA might require to de novo FLS2 protein synthesis. FICA resensitization appeared significantly slower than FLS2 protein recovery, suggesting additional regulatory mechanisms of other components, such as flg22-related Ca2+ permeable channels. Taken together, we have carefully defined the FICA sensitivity adaptation, which will facilitate further molecular and genetic dissection of the Ca2+-mediated adaptive mechanisms in PAMP-triggered immunity.

Recent research has shown that plants can distinguish genetically-related individuals from strangers (kin recognition) and exhibit more cooperative behaviours towards these more related individuals (kin discrimination). The first evidence for this was found when Cakile edentula plants growing with half-sibs allocated relatively less biomass to roots than plants growing with unrelated individuals, indicating that kin recognition can reduce the intensity of competition (Dudley & File, 2007). Since then, kin discrimination has been shown to result in reduced competition for soil resources (Semchenko, Saar, & Lepik, 2014), light (Crepy & Casal, 2015) and pollinators (Torices, Gómez, & Pannell, 2018). On the other hand, allelopathy, plants producing chemical compounds that negatively affect performance of neighbour plants, has also been widely documented (Inderjit & Duke, 2003) and shown to profoundly affect local species coexistence and plant community structure (Meiners, Kong, Ladwig, Pisula, & Lang, 2012). In crops allelopathy can also be beneficial in suppressing weeds (Macías, Mejías, & Molinillo, 2019). In the current issue, Xu, Cheng, Kong, and Meiners (2021) published the first study to show that kin discrimination can also affect the balance between direct competition for resources and allelopathy, and this together may lead to improved weed suppression in rice.

Human activity and natural processes have led to widespread dissemination of metals and metalloids, many of which are toxic and have a negative impact on agronomic production. Roots, as the first point of contact, are essential in endowing plants with tolerance to excess metal(loid) in the soil. The most important root responses include: adaptation of transport processes that affect uptake, efflux and long distance transport of metal(loid)s; metal(loid) detoxification within root cells via conjugation to thiol rich compounds and subsequent sequestration in the vacuole; plasticity in root architecture; the presence of bacteria and fungi in the rhizosphere that impact on metal(loid) bioavailability; the role of root exudates. In this review we will provide details on these processes and assess their relevance for the detoxification of arsenic, cadmium, mercury and zinc. Furthermore, we will assess if any of these methodologies has been tested in field conditions and whether they are effective in terms of improving crop metal(loid) tolerance.

Plants transitioned from an aquatic to a terrestrial lifestyle during their evolution. On land, fluctuations on water availability in the environment became one of the major problems they encountered. The appearance of morpho-physiological adaptations to cope with and tolerate water loss from the cells was undeniably useful to survive on dry land. Some of these adaptations, such as carbon concentrating mechanisms (CCMs), desiccation tolerance (DT) and root impermeabilization, appeared in multiple plant lineages. Despite being crucial for evolution on land, it has been unclear how these adaptations convergently evolved in the various plant lineages. Recent advances on whole genome and transcriptome sequencing are revealing that co-option of genes and gene regulatory networks (GRNs) is a common feature underlying the convergent evolution of these adaptations. In this review we address how the study of CCMs and DT have provided insight into convergent evolution of GRNs underlying plant adaptation to dry environments, and how these insights could be applied to currently emerging understanding of evolution of root impermeabilization through different barrier cell types. We discuss examples of co-option, conservation, and innovation of genes and GRNs at the cell, tissue and organ levels revealed by recent phylogenomic (comparative genomic) and comparative transcriptomic studies.

The biosynthesis of anthocyanins has been shown to be influenced by light quality. However, the molecular mechanisms underlying the light-mediated regulation of fruit anthocyanin biosynthesis are not well understood. In this study, we analyzed the effects of supplemental red and blue light on the anthocyanin biosynthesis in non-climacteric bilberry (Vaccinium myrtillus L.). After six days of continuous irradiation during fruit ripening, both red and blue light elevated concentration of total anthocyanins, but especially red light promoted accumulation of delphinidins. Transcriptomic analysis of ripening berries showed that both light treatments up-regulated all the major anthocyanin structural genes, the key regulatory MYB transcription factors and abscisic acid (ABA) biosynthetic genes. However, higher induction of specific genes of anthocyanin and delphinidin biosynthesis alongside ABA signal perception and metabolism were found in red light. The difference in red and blue light signaling was found in NCED, ABA receptor PYL and catabolic ABA-8’hydroxylase gene expression. Red light also up-regulated expression of SNARE domain transporters, which may indicate involvement of these proteins in vesicular trafficking of anthocyanins during fruit ripening. Our results suggest differential signal transduction and transport mechanisms between red and blue light in in ABA- regulated anthocyanin and delphinidin biosynthesis during non-climacteric fruit ripening.

Translocation of metabolites between different plant species provides important hints in understanding the fate of bioactive root exudates. In the present study, targeted and untargeted mass spectrometry-based metabolomics was applied to elucidate the transfer of bioactive compounds between rye and several crops and weed species. Our results demonstrated that benzoxinoids (BXs) synthesized by rye were taken up by roots of neighboring plant species and translocated into their shoots. Furthermore, we showed roots of the rye plant took up compounds originating from neighboring plants. Among the compounds taken up by rye roots, wogonin was detected in the rye shoot, which indicates the root-to-shoot translocation of this compound. Elucidating the transfer of bioactive compounds between plants is essential for understanding plant-plant interactions, developing natural pesticides and understanding their modes of action.