SUPPRESSOR of MAX2 1 (SMAX1) and SMAX1-LIKE2 (SMXL2) Negatively Regulate Drought Resistance in Arabidopsis thaliana.

Recent investigations in Arabidopsis thaliana suggest that SUPPRESSOR of MORE AXILLARY GROWTH 2 1 (SMAX1) and SMAX1-LIKE2 (SMXL2) are negative regulators of karrikin (KAR) and strigolactone (SL) signaling during plant growth and development, but their functions in drought resistance and related mechanisms of action remain unclear. To understand the roles and mechanisms of SMAX1 and SMXL2 in drought resistance, we investigated the drought-resistance phenotypes and transcriptome profiles of smax1 smxl2 (s1,2) double-mutant plants in response to drought stress. The s1,2 mutant plants showed enhanced drought-resistance and lower leaf water loss when compared with wild-type (WT) plants. Transcriptome comparison of rosette leaves from the s1,2 mutant and the WT under normal and dehydration conditions suggested that the mechanism related to cuticle formation was involved in drought resistance. This possibility was supported by enhanced cuticle formation in the rosette leaves of the s1,2 mutant. We also found that the s1,2 mutant plants were more sensitive to abscisic acid in assays of stomatal closure, cotyledon opening, chlorophyll degradation and growth inhibition, and they showed a higher reactive oxygen species detoxification capacity than WT plants. In addition, the s1,2 mutant plants had longer root hairs and a higher root-to-shoot ratio than the WT plants, suggesting that the mutant had a greater capacity for water absorption than the WT. Taken together, our results indicate that SMAX1 and SMXL2 negatively regulate drought resistance, and disruption of these KAR- and SL-signaling-related genes may therefore provide a novel means for improving crop drought resistance.

Karrikin Receptor KAI2 Coordinates Salt Tolerance Mechanisms in Arabidopsis thaliana.

Plants activate a myriad of signaling cascades to tailor adaptive responses under environmental stresses, such as salinity. While the roles of exogenous karrikins (KARs) in salt stress mitigation are well comprehended, genetic evidence of KAR signaling during salinity responses in plants remains unresolved. Here, we explore the functions of the possible KAR receptor KARRIKIN-INSENSITIVE2 (KAI2) in Arabidopsis thaliana tolerance to salt stress by investigating comparative responses of wild-type (WT) and kai2-mutant plants under a gradient of NaCl. Defects in KAI2 functions resulted in delayed and inhibited cotyledon opening in kai2 seeds compared with WT seeds, suggesting that KAI2 played an important role in enhancing seed germination under salinity. Salt-stressed kai2 plants displayed more phenotypic aberrations, biomass reduction, water loss and oxidative damage than WT plants. kai2 shoots accumulated significantly more Na+ and thus had a lower K+/Na+ ratio, than WT, indicating severe ion toxicity in salt-stressed kai2 plants. Accordingly, kai2 plants displayed a lower expression of genes associated with Na+ homeostasis, such as SALT OVERLY SENSITIVE (SOS) 1, SOS2, HIGH-AFFINITY POTASSIUM TRANSPORTER 1;1 (HKT1;1) and CATION-HYDROGEN EXCHANGER 1 (NHX1) than WT plants. WT plants maintained a better glutathione level, glutathione-related redox status and antioxidant enzyme activities relative to kai2 plants, implying KAI2's function in oxidative stress mitigation in response to salinity. kai2 shoots had lower expression levels of genes involved in the biosynthesis of strigolactones (SLs), salicylic acid and jasmonic acid and the signaling of abscisic acid and SLs than those of WT plants, indicating interactive functions of KAI2 signaling with other hormone signaling in modulating plant responses to salinity. Collectively, these results underpin the likely roles of KAI2 in the alleviation of salinity effects in plants by regulating several physiological and biochemical mechanisms involved in ionic and osmotic balance, oxidative stress tolerance and hormonal crosstalk.

The Karrikin Receptor Karrikin Insensitive2 Positively Regulates Heat Stress Tolerance in Arabidopsis thaliana.

In this study, we investigated the potential role of the karrikin receptor KARRIKIN INSENSITIVE2 (KAI2) in the response of Arabidopsis seedlings to high-temperature stress. We performed phenotypic, physiological and transcriptome analyses of Arabidopsis kai2 mutants and wild-type (WT) plants under control (kai2_C and WT_C, respectively) and 6- and 24-h heat stress conditions (kai2_H6, kai2_H24, WT_H6 and WT_H24, respectively) to understand the basis for KAI2-regulated heat stress tolerance. We discovered that the kai2 mutants exhibited hypersensitivity to high-temperature stress relative to WT plants, which might be associated with a more highly increased leaf surface temperature and cell membrane damage in kai2 mutant plants. Next, we performed comparative transcriptome analysis of kai2_C, kai2_H6, kai2_H24, WT_C, WT_H6 and WT_H24 to identify transcriptome differences between WT and kai2 mutants in response to heat stress. K-mean clustering of normalized gene expression separated the investigated genotypes into three clusters based on heat-treated and non-treated control conditions. Within each cluster, the kai2 mutants were separated from WT plants, implying that kai2 mutants exhibited distinct transcriptome profiles relative to WT plants. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed a repression in 'misfolded protein binding', 'heat shock protein binding', 'unfolded protein binding' and 'protein processing in endoplasmic reticulum' pathways, which was consistent with the downregulation of several genes encoding heat shock proteins and heat shock transcription factors in the kai2 mutant versus WT plants under control and heat stress conditions. Our findings suggest that chemical or genetic manipulation of KAI2 signaling may provide a novel way to improve heat tolerance in plants.

The histidine phosphotransfer AHP4 plays a negative role in Arabidopsis plant response to drought.

Cytokinin plays an important role in plant stress responses via a multistep signaling pathway, involving the histidine phosphotransfer proteins (HPs). In Arabidopsis thaliana, the AHP2, AHP3 and AHP5 proteins are known to affect drought responses; however, the role of AHP4 in drought adaptation remains undetermined. In the present study, using a loss-of-function approach we showed that AHP4 possesses an important role in the response of Arabidopsis to drought. This is evidenced by the higher survival rates of ahp4 than wild-type (WT) plants under drought conditions, which is accompanied by the downregulated AHP4 expression in WT during periods of dehydration. Comparative transcriptome analysis of ahp4 and WT plants revealed AHP4-mediated expression of several dehydration- and/or abscisic acid-responsive genes involved in modulation of various physiological and biochemical processes important for plant drought acclimation. In comparison with WT, ahp4 plants showed increased wax crystal accumulation in stems, thicker cuticles in leaves, greater sensitivity to exogenous abscisic acid at germination, narrow stomatal apertures, heightened leaf temperatures during dehydration, and longer root length under osmotic stress. In addition, ahp4 plants showed greater photosynthetic efficiency, lower levels of reactive oxygen species, reduced electrolyte leakage and lipid peroxidation, and increased anthocyanin contents under drought, when compared with WT. These differences displayed in ahp4 plants are likely due to upregulation of genes that encode enzymes involved in reactive oxygen species scavenging and non-enzymatic antioxidant metabolism. Overall, our findings suggest that AHP4 plays a crucial role in plant drought adaptation.

Modulation of osmoprotection and antioxidant defense by exogenously applied acetate enhances cadmium stress tolerance in lentil seedlings.

To examine the potential role of acetate in conferring cadmium (Cd) stress tolerance in lentil (Lens culinaris), several phenotypical and physio-biochemical properties have been examined in Cd-stressed lentil seedlings following acetate applications. Acetate treatment inhibited the translocation of Cd from roots to shoots, which resulted in a minimal reduction in photosynthetic pigment contents. Additionally, acetate-treated lentil showed higher shoot (1.1 and 11.72%) and root (4.98 and 30.64%) dry weights compared with acetate-non-treated plants under low-Cd and high-Cd concentrations, respectively. Concurrently, acetate treatments increase osmoprotection under low-Cd stress through proline accumulation (24.69%), as well as enhancement of antioxidant defense by increasing ascorbic acid content (239.13%) and catalase activity (148.51%) under high-Cd stress. Acetate-induced antioxidant defense resulted in a significant diminution in hydrogen peroxide, malondialdehyde and electrolyte leakage in Cd-stressed lentil seedlings. Our results indicated that acetate application mitigated oxidative stress-induced damage by modulating antioxidant defense and osmoprotection, and reducing root-to-shoot Cd transport. These findings indicate an important contribution of acetate in mitigating the Cd toxicity during growth and development of lentil seedlings, and suggest that the exogenous applications of acetate could be an economical and new avenue for controlling heavy metal-caused damage in lentil, and potentially in many other crops.

KARRIKIN UPREGULATED F-BOX 1 negatively regulates drought tolerance in Arabidopsis.

The karrikin (KAR) receptor and several related signaling components have been identified by forward genetic screening, but only a few studies have reported on upstream and downstream KAR signaling components and their roles in drought tolerance. Here, we characterized the functions of KAR UPREGULATED F-BOX 1 (KUF1) in drought tolerance using a reverse genetics approach in Arabidopsis (Arabidopsis thaliana). We observed that kuf1 mutant plants were more tolerant to drought stress than wild-type (WT) plants. To clarify the mechanisms by which KUF1 negatively regulates drought tolerance, we performed physiological, transcriptome, and morphological analyses. We found that kuf1 plants limited leaf water loss by reducing stomatal aperture and cuticular permeability. In addition, kuf1 plants showed increased sensitivity of stomatal closure, seed germination, primary root growth, and leaf senescence to abscisic acid (ABA). Genome-wide transcriptome comparisons of kuf1 and WT rosette leaves before and after dehydration showed that the differences in various drought tolerance-related traits were accompanied by differences in the expression of genes associated with stomatal closure (e.g. OPEN STOMATA 1), lipid and fatty acid metabolism (e.g. WAX ESTER SYNTHASE), and ABA responsiveness (e.g. ABA-RESPONSIVE ELEMENT 3). The kuf1 mutant plants had higher root/shoot ratios and root hair densities than WT plants, suggesting that they could absorb more water than WT plants. Together, these results demonstrate that KUF1 negatively regulates drought tolerance by modulating various physiological traits, morphological adjustments, and ABA responses and that the genetic manipulation of KUF1 in crops is a potential means of enhancing their drought tolerance.

Insights into the gene and protein structures of the CaSWEET family members in chickpea (Cicer arietinum), and their gene expression patterns in different organs under various stress and abscisic acid treatments.

'Sugars Will Eventually be Exported Transporters' (SWEETs) are a group of sugar transporters that play crucial roles in various biological processes, particularly plant stress responses. However, no information is available yet for the CaSWEET family in chickpea. Here, we identified all putative CaSWEET members in chickpea, and obtained their major characteristics, including physicochemical patterns, chromosomal distribution, subcellular localization, gene organization, conserved motifs and three-dimensional protein structures. Subsequently, we explored available transcriptome data to compare spatiotemporal transcript abundance of CaSWEET genes in various major organs. Finally, we studied the changes in their transcript levels in leaves and/or roots following dehydration and exogenous abscisic acid treatments using RT-qPCR to obtain valuable information underlying their potential roles in chickpea responses to water-stress conditions. Our results provide the first insights into the characteristics of the CaSWEET family members and a foundation for further functional characterizations of selected candidate genes for genetic engineering of chickpea.

Defective cytokinin signaling reprograms lipid and flavonoid gene-to-metabolite networks to mitigate high salinity in Arabidopsis.

Cytokinin (CK) in plants regulates both developmental processes and adaptation to environmental stresses. Arabidopsis histidine phosphotransfer ahp2,3,5 and type-B Arabidopsis response regulator arr1,10,12 triple mutants are almost completely defective in CK signaling, and the ahp2,3,5 mutant was reported to be salt tolerant. Here, we demonstrate that the arr1,10,12 mutant is also more tolerant to salt stress than wild-type (WT) plants. A comprehensive metabolite profiling coupled with transcriptome analysis of the ahp2,3,5 and arr1,10,12 mutants was conducted to elucidate the salt tolerance mechanisms mediated by CK signaling. Numerous primary (e.g., sugars, amino acids, and lipids) and secondary (e.g., flavonoids and sterols) metabolites accumulated in these mutants under nonsaline and saline conditions, suggesting that both prestress and poststress accumulations of stress-related metabolites contribute to improved salt tolerance in CK-signaling mutants. Specifically, the levels of sugars (e.g., trehalose and galactinol), amino acids (e.g., branched-chain amino acids and γ-aminobutyric acid), anthocyanins, sterols, and unsaturated triacylglycerols were higher in the mutant plants than in WT plants. Notably, the reprograming of flavonoid and lipid pools was highly coordinated and concomitant with the changes in transcriptional levels, indicating that these metabolic pathways are transcriptionally regulated by CK signaling. The discovery of the regulatory role of CK signaling on membrane lipid reprogramming provides a greater understanding of CK-mediated salt tolerance in plants. This knowledge will contribute to the development of salt-tolerant crops with the ability to withstand salinity as a key driver to ensure global food security in the era of climate crisis.

Strigolactones regulate arsenate uptake, vacuolar-sequestration and antioxidant defense responses to resist arsenic toxicity in rice roots.

We explored genetic evidence for strigolactones' role in rice tolerance to arsenate-stress. Comparative analyses of roots of wild-type (WT) and strigolactone-deficient mutants d10 and d17 in response to sodium arsenate (Na2AsO4) revealed differential growth inhibition [WT (11.28%) vs. d10 (19.76%) and d17 (18.03%)], biomass reduction [(WT (33.65%) vs. d10 (74.86%) and d17 (60.65%)] and membrane damage (WT < d10 and d17) at 250 µM Na2AsO4. Microscopic and biochemical analyses showed that roots of WT accumulated lower levels of arsenic and oxidative stress indicators like reactive oxygen species and malondialdehyde than those of strigolactone-deficient mutants. qRT-PCR data indicated lower expression levels of genes (OsPT1, OsPT2, OsPT4 and OsPT8) encoding phosphate-transporters in WT roots than mutant roots, explaining the decreased arsenate and phosphate uptake by WT roots. Increased levels of glutathione and OsPCS1 and OsABCC1 transcripts indicated an efficient vacuolar-sequestration of arsenic in WT roots. Furthermore, higher activities (transcript levels) of SOD (OsCuZnSOD1 and OsCuZnSOD2), APX (OsAPX1 and OsAPX2) and CAT (OsCATA) corresponded to lower oxidative damage in WT roots compared with strigolactone-mutant roots. Collectively, these results highlight that strigolactones are involved in arsenic-stress mitigation by regulating arsenate-uptake, glutathione-biosynthesis, vacuolar-sequestration of arsenic and antioxidant defense responses in rice roots.

Resequencing of 672 Native Rice Accessions to Explore Genetic Diversity and Trait Associations in Vietnam.

BACKGROUND: Vietnam possesses a vast diversity of rice landraces due to its geographical situation, latitudinal range, and a variety of ecosystems. This genetic diversity constitutes a highly valuable resource at a time when the highest rice production areas in the low-lying Mekong and Red River Deltas are enduring increasing threats from climate changes, particularly in rainfall and temperature patterns. RESULTS: We analysed 672 Vietnamese rice genomes, 616 newly sequenced, that encompass the range of rice varieties grown in the diverse ecosystems found throughout Vietnam. We described four Japonica and five Indica subpopulations within Vietnam likely adapted to the region of origin. We compared the population structure and genetic diversity of these Vietnamese rice genomes to the 3000 genomes of Asian cultivated rice. The named Indica-5 (I5) subpopulation was expanded in Vietnam and contained lowland Indica accessions, which had very low shared ancestry with accessions from any other subpopulation and were previously overlooked as admixtures. We scored phenotypic measurements for nineteen traits and identified 453 unique genotype-phenotype significant associations comprising twenty-one QTLs (quantitative trait loci). The strongest associations were observed for grain size traits, while weaker associations were observed for a range of characteristics, including panicle length, heading date and leaf width. CONCLUSIONS: We showed how the rice diversity within Vietnam relates to the wider Asian rice diversity by using a number of approaches to provide a clear picture of the novel diversity present within Vietnam, mainly around the Indica-5 subpopulation. Our results highlight differences in genome composition and trait associations among traditional Vietnamese rice accessions, which are likely the product of adaption to multiple environmental conditions and regional preferences in a very diverse country. Our results highlighted traits and their associated genomic regions that are a potential source of novel loci and alleles to breed a new generation of low input sustainable and climate resilient rice.