Nicotiana benthamiana is predominantly distributed in arid habitats across northern Australia. However, none of six geographically isolated accessions shows obvious xerophytic morphological features. To investigate how these tender-looking plants withstand drought, we examined their responses to water deprivation, assessed phenotypic, physiological, and cellular responses, and analysed cuticular wax composition and wax biosynthesis gene expression profiles. Results showed that the Central Australia (CA) accession, globally known as a research tool, has evolved a drought escape strategy with early vigour, short life cycle, and weak, water loss-limiting responses. By contrast, a northern Queensland (NQ) accession responded to drought by slowing growth, inhibiting flowering, increasing leaf cuticle thickness, and altering cuticular wax composition. Under water stress, NQ increased the heat stability and water impermeability of its cuticle by extending the carbon backbone of cuticular long-chain alkanes from c. 25 to 33. This correlated with rapid upregulation of at least five wax biosynthesis genes. In CA, the alkane chain lengths (c. 25) and gene expression profiles remained largely unaltered. This study highlights complex genetic and environmental control over cuticle composition and provides evidence for divergence into at least two fundamentally different drought response strategies within the N. benthamiana species in < 1 million years.

Understanding how widespread species adapt to variation in abiotic conditions across their ranges is fundamental to ecology. Insight may come from studying how among-population variation (APV) in the common garden corresponds with the environmental conditions of source populations. However, there are no such studies comparing native vs non-native populations across multiple life stages. We examined APV in the performance and functional traits of 59 Conyza canadensis populations, in response to drought, across large aridity gradients in the native (North America) and non-native (Eurasia) ranges in three experiments. Our treatment (dry vs wet) was applied at the recruitment, juvenile, and adult life stages. We found contrasting patterns of APV in drought responses between the two ranges. In the native range, plant performance was less reduced by drought in populations from xeric than mesic habitats, but such relationship was not apparent for non-native populations. These range-specific patterns were consistent across the life stages. The weak adaptive responses of non-native populations indicate that they can become highly abundant even without complete local adaptation to abiotic environments and suggest that long-established invaders may still be evolving to the abiotic environment. These findings may explain lag times in invasions and raise concern about future expansions.

BACKGROUND: Drought is considered the main environmental factor restricting apple production and thus the development of the apple industry. Rootstocks play an important role in enhancing the drought tolerance of apple plants. Studies of the physiology have demonstrated that \'ZC9-3\' is a strong drought-resistant rootstock, whereas \'Jizhen-2\' is a weak drought-resistant rootstock. However, the metabolites in these two apple rootstock varieties that respond to drought stress have not yet been characterized, and the molecular mechanisms underlying their responses to drought stress remain unclear.
RESULTS: In this study, the physiological and molecular mechanisms underlying differences in the drought resistance of \'Jizhen-2\' (drought-sensitive) and \'ZC9-3\' (drought-resistant) apple rootstocks were explored. Under drought stress, the relative water content of the leaves was maintained at higher levels in \'ZC9-3\' than in \'Jizhen-2\', and the photosynthetic, antioxidant, and osmoregulatory capacities of \'ZC9-3\' were stronger than those of \'Jizhen-2\'. Metabolome analysis revealed a total of 95 and 156 differentially accumulated metabolites in \'Jizhen-2\' and \'ZC9-3\' under drought stress, respectively. The up-regulated metabolites in the two cultivars were mainly amino acids and derivatives. Transcriptome analysis revealed that there were more differentially expressed genes and transcription factors in \'ZC9-3\' than in \'Jizhen-2\' throughout the drought treatment. Metabolomic and transcriptomic analysis revealed that amino acid biosynthesis pathways play key roles in mediating drought resistance in apple rootstocks. A total of 13 metabolites, including L-α-aminoadipate, L-homoserine, L-threonine, L-isoleucine, L-valine, L-leucine, (2S)-2-isopropylmalate, anthranilate, L-tryptophan, L-phenylalanine, L-tyrosine, L-glutamate, and L-proline, play an important role in the difference in drought resistance between \'ZC9-3\' and \'Jizhen-2\'. In addition, 13 genes encoding O-acetylserine-(thiol)-lyase, S-adenosylmethionine synthetase, ketol-acid isomeroreductase, dihydroxyacid dehydratase, isopropylmalate isomerase, branched-chain aminotransferase, pyruvate kinase, 3-dehydroquinate dehydratase/shikimate 5-dehydrogenase, N-acetylglutamate-5-P-reductase, and pyrroline-5-carboxylate synthetase positively regulate the response of \'ZC9-3\' to drought stress.
CONCLUSIONS: This study enhances our understanding of the response of apple rootstocks to drought stress at the physiological, metabolic, and transcriptional levels and provides key insights that will aid the cultivation of drought-resistant apple rootstock cultivars. Especially, it identifies key metabolites and genes underlying the drought resistance of apple rootstocks.

UNASSIGNED: Red clover (Trifolium pratense) is a protein-rich, short-lived perennial forage crop that can achieve high yields, but suffers increasingly from drought in different cultivation areas. Breeding for increased adaptation to drought is becoming essential, but at this stage it is unclear which traits breeders should target to phenotype responses to drought that allow them to identify the most promising red clover genotypes. In this study, we assessed how prolonged periods of drought affected plant growth in field conditions, and which traits could be used to distinguish better adapted plant material.
UNASSIGNED: A diverse panel of 395 red clover accessions was evaluated during two growing seasons. We simulated 6-to-8-week drought periods during two consecutive summers, using mobile rain-out shelters, while an irrigated control field was established in an adjacent parcel. Plant growth was monitored throughout both growing seasons using multiple flights with a drone equipped with RGB and thermal sensors. At various observation moments throughout both growing seasons, we measured canopy cover (CC) and canopy height (CH). The crop water stress index (CWSI) was determined at two moments, during or shortly after the drought event.
UNASSIGNED: Manual and UAV-derived measurements for CH were well correlated, indicating that UAV-derived measurements can be reliably used in red clover. In both years, CC, CH and CWSI were affected by drought, with measurable growth reductions by the end of the drought periods, and during the recovery phase. We found that the end of the drought treatment and the recovery phase of approximately 20 days after drought were suitable periods to phenotype drought responses and to distinguish among genotypes.
UNASSIGNED: Multifactorial analysis of accession responses revealed interactions of the maturity type with drought responses, which suggests the presence of two independent strategies in red clover: \'drought tolerance\' and \'drought recovery\'. We further found that a large proportion of the accessions able to perform well under well-watered conditions were also the ones that were less affected by drought. The results of this investigation are interpreted in view of the development of breeding for adaptation to drought in red clover.

The responses of rare plants to environmental stressors will determine their potential to adapt to a rapidly changing climate. We used a common garden approach to evaluate how six populations of the annual San Diego thornmint (Acanthomintha ilicifolia Lamiaceae; listed as endangered in the state of California and as threatened by the US Fish and Wildlife Service) from across the species range respond in terms of growth (biomass, height, and width) and reproduction (seed production, floral production, and next generation seed viability) to experimental differences in water availability. We found a significant irrigation-by-population interaction on the aboveground growth, wherein the differences in the magnitude and direction of treatment did not correlate directly with climate variables in natural populations. With respect to reproduction, the low-irrigation treatment produced more seeds per plant, more reproductive individuals, and a larger proportion of viable seed in most, but not all, populations. The seed production and the effect of irrigation on seed production correlated positively with rainfall at wild source populations. These results suggest that Acanthomintha ilicifolia responds to water limitation by creating more and higher-quality seed, and that plants locally adapted to a higher annual rainfall show a greater plasticity to differences in water availability than plants adapted to a lower annual rainfall, a finding that can inform the in situ demographic management and ex situ collection strategy for Acanthomintha ilicifolia and other rare California annuals.

Abscisic acid (ABA) plays an important role in plant adaptation to water deficits, but its role in regulating root growth (primary root elongation and lateral root number) during different drought-phases remains unclear. Here, we exposed wild-type (WT) and ABA-deficient (not) tomato plants to three continuous drought-phases (moderate drying: day 0-21; severe drying: day 22-47 and re-watering: day 48-51). It was found that WT increased primary root growth during moderate drying; maintained more lateral roots, and greater primary root and total root length under severe drying; and produced more roots after re-watering. After RNA-Seq analysis, we found that the auxin-related genes in root showed different expression patterns between WT and not under drying or re-watering. Further, exogenous supply of IAA partially recovered the root growth of ABA-deficient not plants under three continuous drought-phases. Our results suggested that ABA regulation of tomato root growth during soil drying and recovery can involve auxin response.

Plants from arid environments display covarying traits to survive or resist drought. Plant drought resistance and ability to survive long periods of low soil water availability should involve leaf phenology coordination with leaf and stem functional traits related to water status. This study tested correlations between phenology and functional traits involved in plant water status regulation in 10 Sonoran Desert tree species with contrasting phenology. Species seasonal variation in plant water status was defined by calculating their relative positions along the iso/anisohydric regulation continuum based on their hydroscape areas (HA)-a metric derived from the relationship between predawn and midday water potentials-and stomatal and hydraulic traits. Additionally, functional traits associated with plant water status regulation, including lamina vessel hydraulic diameter (DHL), stem-specific density (SSD) and leaf mass per area (LMA) were quantified per species. To characterize leaf phenology, leaf longevity (LL) and canopy foliage duration (FD) were determined. Hydroscape area was strongly correlated with FD but not with leaf longevity (LL); HA was significantly associated with SSD and leaf hydraulic traits (DHL, LMA) but not with stem hydraulic traits (vulnerability index, relative conductivity); and FD was strongly correlated with LMA and SSD. Leaf physiological characteristics affected leaf phenology when it was described as canopy FD better than when described as LL. Stem and leaf structure and hydraulic functions were not only relevant for categorizing species along the iso/anisohydric continuum but also allowed identifying different strategies of desert trees within the \'fast-slow\' plant economics spectrum. The results in this study pinpoint the set of evolutionary pressures that shape the Sonoran Desert Scrub physiognomy.

Bark water vapour conductance (gbark) is a rarely considered functional trait. However, for the few tree species measured to date, it appears high enough to create stem water deficits associated with mortality during droughts, when access to water is limited. I tested whether gbark correlates with stem water deficit during drought conditions in two datasets of tropical trees: one of saplings in forest understories during an annual dry season and one of potted saplings in a shadehouse during extreme drought conditions. Among all 14 populations of eight species measured, gbark varied more than 10-fold (0.86-12.98 mmol m-2 s-1). In the forest understories, gbark was highly correlated with stem water deficit among four deciduous species, but not among evergreen species that likely maintained access to soil water. In the shadehouse, gbark was positively correlated with stem water deficit and mortality among all six species. Overall, tree species with higher gbark suffer higher stem water deficit when soil water is unavailable. Incorporating gbark into soil-plant-atmosphere hydrodynamic models may improve projections of plant mortality under drought conditions.

Physio-biochemical adaptations of wheat landraces may have great importance in their growth, survival and yield under drought stress. Here, we evaluated the effects of drought stress on some defense systems of wheat cultivar \"Sistan\" (drought-sensitive) and landrace \"Bolani\" (drought-tolerant). Under drought stress, Bolani plants showed lower increases in hydrogen peroxide content compared to Sistan ones, which was accompanied with significant decrease in malondialdehyde and electrolyte leakage indices. Increasing the transcript levels and activity of enzymatic and non-enzymatic antioxidants along with phenylpropanoid metabolites improved relative tolerance to drought-induced oxidative stress, particularly in Bolani plants, results which may be confirmed by a significant decrease in the damage indices. In the phenylpropanoid pathway, the biosynthetic pathway of total phenol, flavonoids and anthocyanins was more active than lignin-biosynthetic pathway, which could early respond to drought stress. These results may be confirmed by their negative significant correlations with damage indices as well as a non-significant correlation of lignin with most enzymatic and non-enzymatic antioxidants in plants. Lower decrease of chlorophyll (Chl) and carotenoid contents in Bolani plants compared to Sistan ones indicated the relative stability of photosynthetic pigments under drought stress. Our results suggested that integrating metabolic pathways could coordinately alleviate oxidative stress that can lead to introducing suitable genetic sources for drought tolerance.

Pentatricopeptide-repeat (PPR) proteins were identified as a type of nucleus coding protein that is composed of multiple tandem repeats. It has been reported that PPR genes play an important role in RNA editing, plant growth and development, and abiotic stresses in plants. However, the functions of PPR proteins remain largely unknown in soybean. In this study, 179 DYW subgroup PPR genes were identified in soybean genome (Glycine max Wm82.a2.v1). Chromosomal location analysis indicated that DYW subgroup PPR genes were mapped to all 20 chromosomes. Phylogenetic relationship analysis revealed that DYW subgroup PPR genes were categorized into three distinct Clusters (I to III). Gene structure analysis showed that most PPR genes were featured by a lack of intron. Gene duplication analysis demonstrated 30 PPR genes (15 pairs; ~35.7%) were segmentally duplicated among Cluster I PPR genes. Furthermore, we validated the mRNA expression of three genes that were highly up-regulated in soybean drought- and salt-induced transcriptome database and found that the expression levels of GmPPR4 were induced under salt and drought stresses. Under drought stress condition, GmPPR4-overexpressing (GmPPR4-OE) plants showed delayed leaf rolling; higher content of proline (Pro); and lower contents of H2O2, O2- and malondialdehyde (MDA) compared with the empty vector (EV)-control plants. GmPPR4-OE plants exhibited increased transcripts of several drought-inducible genes compared with EV-control plants. Our results provided a comprehensive analysis of the DYW subgroup PPR genes and an insight for improving the drought tolerance in soybean.