During the later stages of seed maturation, two key adaptive traits are acquired that contribute to seed lifespan and dispersal, longevity and dormancy. The seed-specific heat shock transcription factor A9 is an important hub gene in the transcriptional network of late seed maturation. Here we demonstrate that HSFA9 plays a role in thermotolerance rather than in ex situ seed conservation. Storage of hsfa9 seeds of Medicago truncatula and Arabidopsis had comparable lifespan at moderate storage relative humidity (RH), whereas at high RH, hsfa9 seeds lost their viability much faster than wild type seeds. Furthermore, we show that in M. truncatula, Mthsfa9 seeds acquired more dormancy during late maturation than wild type. Transient expression of MtHSFA9 in hairy roots and transcriptome analysis of Mthsfa9 Tnt1 insertion mutants identified a deregulation of genes involved in ABA biosynthesis, catabolism and signalling. Consistent with these results, Mthsfa9 seeds exhibited increased ABA levels and higher sensitivity to ABA. These data suggest that in legumes, HSFA9 acts as a negative regulator of the depth of seed dormancy during seed development via the modulation of hormonal balance.

CO 2-induced chloroplast movement was reported in the monograph by Gustav Senn in 1908: unilateral CO 2 supply to the one cell-layered moss leaves induced the positively CO 2-tactic periclinal arrangement of chloroplasts. However, from the modern criteria, several experimental settings are unacceptable. Here, using a model moss plant Physcomitrium patens, we examined basic features of chloroplast CO 2-tactic relocation with a modernized experimental system. The CO 2 relocation was light-dependent and especially the CO 2 relocation in red light was substantially dependent on photosynthetic activity. Between the cytoskeletons responsible for chloroplast movement of P. patens, the microfilament mainly worked for CO 2 relocation, but the microtubule-based movement was insensitive to CO 2. The CO 2 relocation was induced not only by air with and without CO 2 but also by the more realistic difference in CO 2 concentration between the two sides. In the leaves placed on the surface of a gel sheet, chloroplasts avoided the gel side and positioned in the air facing surface. This was also shown to be photosynthesis dependent. Based on these observations, we propose a working hypothesis that the threshold light intensity between the light-accumulation and -avoidance responses of the photorelocation would be increased by CO 2, resulting in the CO 2-tactic relocation of chloroplast.

Plant growth depends on the diurnal regulation of cellular processes, but it is not well understood if and how transcriptional regulation controls diurnal fluctuations at the protein-level. Here we report a high-resolution Arabidopsis thaliana (Arabidopsis) leaf rosette proteome acquired over a 12 h light : 12 h dark diurnal cycle and the phosphoproteome immediately before and after the light-to-dark and dark-to-light transitions. We quantified nearly 5000 proteins and 800 phosphoproteins, of which 288 fluctuated in their abundance and 226 fluctuated in their phosphorylation status. Of the phosphoproteins, 60% were quantified for changes in protein abundance. This revealed six proteins involved in nitrogen and hormone metabolism that had concurrent changes in both protein abundance and phosphorylation status. The diurnal proteome and phosphoproteome changes involve proteins in key cellular processes, including protein translation, light perception, photosynthesis, metabolism and transport. The phosphoproteome at the light-dark transitions revealed the dynamics at phosphorylation sites in either anticipation of or response to a change in light regime. Phosphorylation site motif analyses implicate casein kinase II and calcium/calmodulin dependent kinases among the primary light-dark transition kinases. The comparative analysis of the diurnal proteome and diurnal and circadian transcriptome established how mRNA and protein accumulation intersect in leaves during the diurnal cycle of the plant.

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.

Drought-related tree mortality is increasing globally, but the sequence of events leading to it remains poorly understood. To identify such sequence, we used a 2016 tree mortality event in the semi-arid pine forest of Yatir were dendrometry and sap flow measurements were carried out in 31 trees, of which seven died. A comparative analysis revealed three stages leading to mortality. First, a decrease in tree diameter in all dying trees, but not in the living ones, eight months ‘prior to the visual signs of mortality’ (PVSM; e.g., brown needles). Second, a decay to near zero in the diurnal stem swelling/shrinkage dynamics, reflecting the loss of stem radial water flow in the dying trees, six months PVSM. Third, cessation of stem sap flow three months PVSM. Eventual mortality could therefore be detected long before visual signs are observed, and the three stages identified here demonstrated the differential effects of drought on stem growth, water storage capabilities, and soil water uptake. The results indicated that breakdown of radial stem water flow and phloem functionality is a critical element in defining the ‘point of no return’ in the sequence of events leading to mortality of mature trees.

Nicotinamide-adenine dinucleotide (NAD) is involved in redox homeostasis and acts as a substrate for NADases, including poly (ADP-ribose) polymerases (PARPs) that add poly (ADP-ribose) polymers to proteins and DNA, and sirtuins that deacetylate proteins. Nicotinamide, a biproduct of NADases increases circadian period in both plants and animals. In mammals, the effect of nicotinamide on circadian period might be mediated by the PARPs and sirtuins because thy directly bind to core circadian oscillator genes. We have investigated whether PARPs and sirtuins contribute to the regulation of the circadian oscillator in Arabidopsis. We found no evidence that PARPs and sirtuins regulate the circadian oscillator of Arabidopsis or are involved in the response to nicotinamide. RNA-seq analysis indicated that PARPs regulate the expression of only a few genes, including FLOWERING LOCUS C. However, we found profound effects of reduced sirtuin 1 expression on gene expression during the day but not at night, and an embryo lethal phenotype in knockouts. Our results demonstrate that PARPS and sirtuins are not associated with NAD regulation of the circadian oscillator and that sirtuin 1 is associated with daytime regulation of gene expression.

Plant vacuoles serve as the primary intracellular compartments for phosphorus (P) storage and play a central role to maintain P homeostasis. The Oryza sativa (rice) genome contains three genes that encode SPX (SYG1/PHO81/XPR1)-MFS (Major Facility Superfamily) proteins (OsSPX-MFS1, 2, 3). OsSPX-MFS1 and OsSPX-MFS3 were shown previously to have vacuolar phosphate (Pi) transporter activities, but the physiological role of the three transporters under varying P conditions and under field grown conditions for a crop plant is not known. To address this knowledge gap, we generated single, double, and triple mutants (7 mutants with at least two lines of each) for the three rice Os SPX-MFS genes. All the mutants except osspx-mfs2 display lower vacuolar Pi concentrations and all Os SPX-MFSs overexpression plant lines display higher Pi accumulation, demonstrating that all three OsSPX-MFSs are vacuolar Pi influx transporters. OsSPX-MFS3 plays the dominant role based on the phenotypes of three single mutants in terms of growth, vacuolar and tissue Pi concentrations. OsSPX-MFS2 is the weakest and only functions as vacuole Pi sequestration under osspx-mfs1/3 background. The vacuolar Pi sequestration was severely impaired in osspx-mfs1/ 3 and osspx-mfs1/2/ 3, which led to Pi toxicity and subsequently increased Pi allocation to aerial organs. High Pi in the panicle result in necrotic symptoms on husks and impaired panicle and grain development in osspx-mfs1/ 3 and osspx-mfs1/2/ 3 mutant lines. The mutation in the weak vacuolar Pi transporter OsSPX-MFS2 resulted more stable yield compared to the wildtype under low P field conditions. The results suggest that alteration of vacuolar Pi sequestration may be a novel effective strategy to improve rice (crop) tolerance to low phosphorus field conditions and maintain yield.

Leaf stomatal density is known to covary with leaf vein density. However, the functional underpinning of this relation, and how it scales to whole-plant water transport anatomy, is still unresolved. We hypothesized that the balance of water exchange between the vapour phase (in stomata) and liquid phase (in vessels) depends on the consistent scaling between the summed stomatal areas and xylem cross-sectional areas, both at the whole-plant and single-leaf level. This predicted size-covariation should be driven by the covariation of numbers of stomata and terminal vessels. We examined the relationships of stomatal traits and xylem anatomical traits from the entire plant to individual leaves across seedlings of 53 European woody angiosperm species. There was strong and convergent scaling between total stomatal area and stem xylem area per plant and between leaf total stomatal area and midvein xylem area per leaf across all the species, irrespective of variation in leaf habit, growth-form or relative growth rate (RGR). Moreover, strong scaling was found between stomatal number and terminal vessel number while not in their respective average areas. Our findings have broad implications for integrating xylem architecture and stomatal distribution, and deepen our understanding of the design rules of plants’ water transport network.

Effects of K deficiency have been investigated for several decades and recently, progress has been made in identifying metabolomics signatures thereby offering potential to monitor the K status of crops in the field. However, effects of low K conditions could also be due to the antagonism with other nutrients like calcium (Ca) and the well-known biomarker of K deficiency, putrescine, could be a response to Ca/K imbalance rather than K deficiency. We carried out experiments in sunflower grown at either low or high K, at high or low Ca, with or without putrescine added to the nutrient solution. Using metabolomics and proteomics analysis, we show that a significant part of the low-K response such as lower photosynthesis and N assimilation, is due to calcium and can be suppressed by low Ca conditions. Putrescine addition tends to restore photosynthesis and N assimilation but but aggravates the impact of low-K conditions on catabolism. We conclude that (i) effects of K deficiency can be partly alleviated by the use of low Ca and not only by K fertilization, and (ii) in addition to its role as a metabolite, putrescine participates in the regulation of the content in enzymes involved in carbon primary metabolism.

Brassica crops include various edible vegetable and plant oil crops, and their production is limited by low temperature beyond their tolerant capability. The key regulators of low-temperature resistance in Brassica remain largely unexplored. To identify post-transcriptional regulators of plant response to low temperature, we performed small RNA profiling, and found that 16 known miRNAs were responsive to cold treatment in Brassica rapa. The cold response of seven of those miRNAs were further confirmed by qRT-PCR and/or northern blotting analyses. In parallel, a genome-wide association study of 220 accessions of Brassica napus identified four candidate MIRNA genes, all of which were cold-responsive, at the loci associated with low temperature resistance. Specifically, these large-scale data analyses revealed a link between miR1885 and the plant response to low temperature in both B. rapa and B. napus. Using 5′ rapid amplification of cDNA ends approach, we validated that miR1885 can cleave its putative target transcripts, Bn.TIR.A09 and Bn.TNL.A03, in B. napus. Furthermore, overexpression of miR1885 in Semi-winter type B. napus decreased the mRNA abundance of Bn.TIR.A09 and Bn.TNL.A03, resulting in increased sensitivity to low temperature. Knocking down of miR1885 in Spring type B. napus led to increased mRNA abundance of its targets and improved rapeseed tolerance to low temperature. Together, our results suggested that the loci of miR1885 and its targets could be potential candidates for the molecular breeding of low temperature-tolerant Spring type Brassica crops.

β-Glucosidase is validated as an elicitor for early immune responses in plants and it was detected in the salivary glands of Frankliniella occidentalis in previous research. Seven differentially expressed genes encoding β-Glucosidase were obtained by comparing the transcriptomes of F. occidentalis adults grown under two different CO 2 concentrations (800 ppm vs. 400 ppm), which might be associated with the differences in the interaction between F. occidentalis adults and its host plant, Phaseolus vulgaris under different CO 2 levels. To verify this speculation, changes in defense responses based on the production and elimination of reactive oxygen species (ROS) in P. vulgaris leaves treated with three levels of β-Glucosidase activity under ambient CO 2 (aCO 2) and elevated CO 2 (eCO 2) were measured in this study. The results showed that both leaves infested with thrips and those sprayed with the pure β-Glucosidase solution showed significant increases in ROS levels under aCO 2 and eCO 2, and the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were increased correspondingly, while in leaves infested with FoβGlu-1-silenced thrips, the ROS levels and activities of these enzymes did not change significantly during the first 12 hours of injury regardless of CO 2 level. Besides, significantly higher levels of ROS and lower activities of SOD, POD and CAT in injured leaves under eCO 2 compared to aCO 2 were noticed, which would negatively affect P. vulgaris leaves and facilitate thrips damage.

Phloem sap transport, velocity and allocation have been proposed to play a role in physiological limitations of crop yield, along with photosynthetic activity or water use efficiency. Although there is clear evidence that carbon allocation to grains effectively drives yield in cereals like wheat (as reflected by the harvest index), the influence of phloem transport rate and velocity is less clear. Here, we took advantage of previously published data on yield, respiration, carbon isotope composition, nitrogen content and water consumption in winter wheat cultivars grown across several sites with or without irrigation, to express grain production in terms of phloem sucrose transport and compare with xylem water transport. Our results suggest that phloem sucrose transport rate follows the same relationship with phloem N transport regardless of irrigation conditions and cultivars, and seems to depend mostly on grain weight (i.e. mg per grain). When compared to xylem sap water movement, phloem sap velocity (in m s -1) was 5.8 to 7.7 times lower. Depending on the assumption made for phloem sap sucrose concentration, either phloem sap velocity or its proportionality coefficient to xylem velocity change little with environmental conditions. Taken as a whole, phloem transport from leaves to grains seems to be homeostatic within a narrow range of values and following relationships with other plant physiological parameters across cultivars and conditions. This suggests that phloem transport per se is not a limitation for yield in wheat but rather, is controlled to sustain grain filling.

Utilizing phosphate more efficiently is crucial for sustainable crop production. Highly efficient rice (Oryza sativa) cultivars have been identified and this study aims to identify metabolic markers associated with P utilization efficiency. P deficiency generally reduced leaf P concentrations and CO2 assimilation rates but efficient cultivars were reducing leaf P concentrations further than inefficient ones while maintaining similar CO2 assimilation rates. Adaptive changes in carbon metabolism were detected but equally in efficient and inefficient cultivar groups. Groups furthermore did not differ with respect to partial substitutions of phospholipids by sulfo- and galactolipids. Metabolites significantly more abundant in the efficient group, such as sinapate, benzoate and glucoronate, were related to antioxidant defense and may help alleviating oxidative stress caused by P deficiency. Sugar alcohols ribitol and threitol were another marker metabolite for higher phosphate efficiency as were several amino acids, especially threonine. Since these metabolites are not known to be associated with P deficiency, they may provide novel clues for the selection of more P efficient genotypes. In conclusion, metabolite signatures detected here were not related to phosphate metabolism but rather helped P efficient lines to keep vital processes functional under the adverse conditions of P starvation.

Gastrodia elata, a fully mycoheterotrophic orchid without photosynthetic ability, only grows symbiotically with the fungus Armillaria. The mechanism of carbon distribution in this mycoheterotrophy is unknown. We detected high sucrose concentrations in all stages of Gastrodia tubers, suggesting sucrose may be the major sugar transported between fungus and orchid. Thick symplasm-isolated wall interfaces in colonized and adjacent large cells implied involvement of sucrose importers. Two sucrose transporter (SUT)-like genes, GeSUT4 and GeSUT3, were identified that are highly expressed in young Armillaria-colonized tubers. Yeast complementation and radio tracer experiments confirmed that GeSUT4 functioned as a high-affinity sucrose-specific proton-dependent importer. Plasma-membrane/tonoplast localization of GeSUT4-GFP fusions, and high RNA expression of GeSUT4 in symbiotic and large cells indicated that GeSUT4 likely functions in active sucrose transport for intercellular allocation and intracellular homeostasis. Transgenic Arabidopsis overexpressing GeSUT4 had larger leaves but sensitive to excess sucrose and roots were colonized with fewer mutualistic Bacillus, supporting the role of GeSUT4 in regulating sugar allocation. This is not only the first documented carbon import system in a mycoheterotrophic interaction, but also highlights the evolutionary importance of sucrose transporters for regulation of carbon flow in all types of plant-microbe interactions.

Control of plant virus diseases largely depends on the induced plant defense achieved by the external application of synthetic chemical inducers with the ability to modify defense-signaling pathways. However, most of the molecular mechanisms underlying these chemical inducers remain unknown. Here, we developed a lentinan-loaded hydrogel with a core-shell structure and discovered how it protects plants from different virus infections. The hydrogel was synthesized by adding a chitosan shell on the surface of the polyanion sodium alginate-Ca 2+-lentinan (LNT) hydrogel (SL-gel) to form a CSL-gel. CSL-gels exhibit the capacity to prolong the stable release of lentinan and promote Ca 2+ release. Application of CSL-gels on the root of plants induces broad-spectrum resistance against TMV, TRV, PVX and TuMV). RNA-seq analysis identified that the calmodulin-like protein 19 gene ( NbCML19) is upregulated by the sustained release of Ca 2+ from the CSL-gel, and silencing and overexpression of NbCML19 alter the susceptibility and resistance of tobacco to TMV. Our findings provide evidence that this novel and synthetic CSL-gel strongly inhibits the infection of plant viruses by the sustainable release of LNT and Ca 2+. This study uncovers a novel mode of action by which CSL-gels trigger NbCML19 expression through the stable and sustained release of Ca 2+.

Photosynthesis is especially sensitive to environmental conditions and the composition of the photosynthetic apparatus can be modulated in response to environmental change, a process termed photosynthetic acclimation. Previously, we identified a role for a cytosolic fumarase, FUM2 in acclimation to low temperature in Arabidopsis thaliana. Mutant lines lacking FUM2 were unable to acclimate their photosynthetic apparatus to cold. Here, using gas exchange measurements and metabolite assays of acclimating and non-acclimating plants, we show that acclimation to low temperature results in a change in the distribution of photosynthetically fixed carbon to different storage pools during the day. Proteomic analysis of wild-type Col-0 Arabidopsis and of a fum2 mutant which was unable to acclimate to cold indicates that extensive changes occurring in response to cold are affected in the mutant. Metabolic and proteomic data were used to parameterise metabolic models. Using an approach called flux sampling, we show how the relative export of triose phosphate and 3-phsphoglycerate provides a signal of the chloroplast redox state that could underly photosynthetic acclimation to cold.

The mechanisms by which herbivores induce plant defenses are well studied. However, how specialized herbivores suppress plant resistance is still poorly understood. Here, we discovered a rice (Oryza sativa) leucine-rich repeat receptor-like kinase, OsLRR-RLK2, which is induced upon attack by gravid females of a specialist piercing-sucking herbivore, the brown planthopper (BPH, Nilaparvata lugens). Silencing OsLRR-RLK2 decreases the constitutive activity of mitogen-activated protein kinase (OsMPK6) and alters BPH-induced transcript levels of several defense-related WRKY transcription factors. Moreover, silencing OsLRR-RLK2 reduces BPH-induction of jasmonic acid and ethylene but promotes the biosynthesis of both elicited salicylic acid and H2O2; silencing also enhances the production of volatiles emitted from rice plants infested with gravid BPH females. These changes decrease BPH preference and performance in the glasshouse and the field. Our study identifies OsLRR-RLK2 as a major susceptibility factor of rice against BPH. It is likely to be employed by BPH to suppress host plant defenses for its own benefit via signaling crosstalk and/or changing the plant’s defense-related signaling profile.

Aphids often carry facultative symbionts to achieve diverse advantages. Serratia symbiotica, one of facultative endosymbionts, increases aphid tolerance to heat. However, whether it benefits aphid colonization on host plants is yet to be determined. In the current study, we found that Acyrthosiphon pisum harboring S. symbiotica had longer feeding duration on Medicago truncatula than Serratia-free aphids. Contrastingly, Serratia-free aphids triggered higher accumulation of ROS, jasmonic acid and salicylic acid responsive genes and cytosolic Ca2+ elevations than Serratia-infected aphids. Transcriptomic analysis of salivary glands indicated that a histidine-rich Ca2+-binding protein-like gene (ApHRC) was expressed more highly in the salivary gland of Serratia-infected aphids than that of Serratia-free aphids. Once ApHRC was silenced, Serratia-infected aphids also displayed shorter phloem-feeding duration and caused Ca2+ elevation and ROS accumulation in plants. Our results suggest that ApHRC, a potential effector up-regulated by S. symbiotica in the salivary glands, evaded plant defense response by suppressing Ca2+ elevation and ROS accumulation, allowing colonization of aphids. This study has provided a revolutionary insight into how facultative symbionts facilitate aphid colonization and adaption to host plants.