Using seasonal physiological and biochemical responses to select forest components adapted to soybean and corn intercropping.

Given the increasing utilization of forest components in integration systems worldwide, coupled with the growing demand for food in regions facing water restrictions, this study aims to evaluate how physiological and biochemical parameters contribute to the diversification of adaptive mechanisms among native species and eucalyptus genotypes intercropped with soybean or corn. The native tree species Anadenanthera macrocarpa and Dipteryx alata, and the eucalyptus genotypes Urograndis I-144 and Urocam VM01, were grown in soybean and corn intercropping areas and evaluated in fall, winter, spring, and summer. The study evaluated morning water potential, chloroplast pigment concentration, gas exchange, cell damage, and antioxidant enzyme activity. Intercropped with soybean, development the of A. macrocarpa improved through instantaneous water use efficiency, energy use by the electron transport chain, chloroplast pigments, and catalase enzyme activity. On the other hand, A. macrocarpa when, intercropped with corn, despite increasing energy absorption by the reaction center, there is a need for non-photochemical dissipation and in the activity of the enzymes superoxide dismutase and ascorbate peroxidase in response to water and oxidative deficits. In D. alata, the physiological and biochemical responses were not influenced by intercropping but by seasons, with increased chloroplast pigments in fall and electron transport in summer. However, in corn intercropping, the dissipation of excess energy allowed leaf acclimatization. The I-144 and VM01 genotypes also showed no significant differences between intercrops. The results describe photosynthetic and biochemical challenges in the native species A. macrocarpa intercropped with corn, such as a greater need for enzymatic and non-enzymatic defense mechanisms in response to more negative water potential. In D. alata, the challenges are present in both intercrops due to improved mechanisms to protect the photosynthetic apparatus. The survival of the I-144 genotype may be inefficient in both intercrops under prolonged drought conditions, as it modifies the photosystem; in contrast, genotype VM01 was the most adapted to the system for using captured energy, reducing water loss and being resilient.

Leaf starch degradation by ß-amylase ZmBAM8 influences drought tolerance in maize.

As a typical C4 plant and important crop worldwide, maize is susceptible to drought. In maize, transitory starch (TS) turnover occurs in the vascular bundle sheath of leaves, differing from that in Arabidopsis (a C3 plant). This process, particularly its role in drought tolerance and the key starch-hydrolyzing enzymes involved, is not fully understood. We discovered that the expression of the ß-amylase (BAM) gene ZmBAM8 is highly upregulated in the drought-tolerant inbred line Chang7-2t. Inspired by this finding, we systematically investigated TS degradation in maize lines, including Chang7-2t, Chang7-2, B104, and ZmBAM8 overexpression (OE) and knockout (KO) lines. We found that ZmBAM8 was significantly induced in the vascular bundle sheath by drought, osmotic stress, and abscisic acid. The stress-induced gene expression and chloroplast localization of ZmBAM8 align with the tissue and subcellular sites where TS turnover occurs. The recombinant ZmBAM8 was capable of effectively hydrolyzing leaf starch. Under drought conditions, the leaf starch in ZmBAM8-OE plants substantially decreased under light, while that in ZmBAM8-KO plants did not decrease. Compared with ZmBAM8-KO plants, ZmBAM8-OE plants exhibited increased drought tolerance. Our study provides insights into the significance of leaf starch degradation in C4 crops and contributes to the development of drought-resistant maize.

CRISPR/dCas13(Rx) Derived RNA N6-methyladenosine (m6A) Dynamic Modification in Plant.

N6-methyladenosine (m6A) is the most prevalent internal modification of mRNA and plays an important role in regulating plant growth. However, there is still a lack of effective tools to precisely modify m6A sites of individual transcripts in plants. Here, programmable m6A editing tools are developed by combining CRISPR/dCas13(Rx) with the methyltransferase GhMTA (Targeted RNA Methylation Editor, TME) or the demethyltransferase GhALKBH10 (Targeted RNA Demethylation Editor, TDE). These editors enable efficient deposition or removal of m6A modifications at targeted sites of endo-transcripts GhECA1 and GhDi19 within a broad editing window ranging from 0 to 46 nt. TDE editor significantly decreases m6A levels by 24%-76%, while the TME editor increases m6A enrichment, ranging from 1.37- to 2.51-fold. Furthermore, installation and removal of m6A modifications play opposing roles in regulating GhECA1 and GhDi19 mRNA transcripts, which may be attributed to the fact that their m6A sites are located in different regions of the genes. Most importantly, targeting the GhDi19 transcript with TME editor plants results in a significant increase in root length and enhanced drought resistance. Collectively, these m6A editors can be applied to study the function of specific m6A modifications and have the potential for future applications in crop improvement.

Osmotolerant plant growth promoting bacteria mitigate adverse effects of drought stress on wheat growth.

Drought stress represents a major constraint with significant impacts on wheat crop globally. The use of plant growth-promoting bacteria (PGPB) has emerged as a promising strategy to alleviate the detrimental impacts of water stress and enhance plant development. We investigated 24 strains from diverse ecosystems, assessed for PGP traits and tolerance ability to abiotic stresses (drought, salinity, temperature, pH, heavy metals, pollutants, herbicides, and fungicides). The most effective bacterial strains Providencia vermicola ME1, Pantoea agglomerans Pa, Pseudomonas knackmussi MR6, and Bacillus sp D13 were chosen. Furthermore, these strains exhibited PGP activities under osmotic stress (0, 10, 20, and 30% PEG-6000). The impact of these osmotolerant PGPBs on wheat (Triticum durum L.) growth under drought stress was assessed at two plant growth stages. In an in vitro wheat seed germination experiment, bacterial inoculation significantly enhanced germination parameters. In pot experiments, the potential of these bacteria was evaluated in wheat plants under three treatments: Well-watered (100% field capacity), moderate stress (50% FC), and severe stress (25% FC). Results showed a significant decline in wheat growth parameters under increasing water stress for uninoculated seedlings. In contrast, bacterial inoculation mitigated these adverse effects, significantly improving morphological parameters and chlorophyll pigment contents under the stress conditions. While malondialdehyde (lipid peroxidation) and proline contents increased significantly with drought intensity, they decreased after bacterial inoculation. The antioxidant enzyme activities (GPX, CAT, and SOD) in plants decreased after bacterial inoculation. The increased root colonization capacity observed under water stress was attributed to their ability to favorable adaptations in a stressful environment. This study highlighted the potential of selected PGPB to alleviate water stress effects on wheat, promoting practical applications aimed at enhancing crop resilience under conditions of water shortage.

Comprehensive analysis of B3 family genes in pearl millet (Pennisetum glaucum) and the negative regulator role of PgRAV-04 in drought tolerance.

Introduction: Members of the plant-specific B3 transcription factor superfamily play crucial roles in various plant growth and developmental processes. Despite numerous valuable studies on B3 genes in other species, little is known about the B3 superfamily in pearl millet. Methods and results: Here, through comparative genomic analysis, we identified 70 B3 proteins in pearl millet and categorized them into four subfamilies based on phylogenetic affiliations: ARF, RAV, LAV, and REM. We also mapped the chromosomal locations of these proteins and analyzed their gene structures, conserved motifs, and gene duplication events, providing new insights into their potential functional interactions. Using transcriptomic sequencing and real-time quantitative PCR, we determined that most PgB3 genes exhibit upregulated expression under drought and high-temperature stresses, indicating their involvement in stress response regulation. To delve deeper into the abiotic stress roles of the B3 family, we focused on a specific gene within the RAV subfamily, PgRAV-04, cloning it and overexpressing it in tobacco. PgRAV-04 overexpression led to increased drought sensitivity in the transgenic plants due to decreased proline levels and peroxidase activity. Discussion: This study not only adds to the existing body of knowledge on the B3 family's characteristics but also advances our functional understanding of the PgB3 genes in pearl millet, reinforcing the significance of these factors in stress adaptation mechanisms.

Characterization of bZIP Transcription Factors in Transcriptome of Chrysanthemum mongolicum and Roles of CmbZIP9 in Drought Stress Resistance.

bZIP transcription factors play important roles in regulating plant development and stress responses. Although bZIPs have been identified in many plant species, there is little information on the bZIPs in Chrysanthemum. In this study, bZIP TFs were identified from the leaf transcriptome of C. mongolicum, a plant naturally tolerant to drought. A total of 28 full-length bZIP family members were identified from the leaf transcriptome of C. mongolicum and were divided into five subfamilies based on their phylogenetic relationships with the bZIPs from Arabidopsis. Ten conserved motifs were detected among the bZIP proteins of C. mongolicum. Subcellular localization assays revealed that most of the CmbZIPs were predicted to be localized in the nucleus. A novel bZIP gene, designated as CmbZIP9, was cloned based on a sequence of the data of the C. mongolicum transcriptome and was overexpressed in tobacco. The results indicated that the overexpression of CmbZIP9 reduced the malondialdehyde (MDA) content and increased the peroxidase (POD) and superoxide dismutase (SOD) activities as well as the expression levels of stress-related genes under drought stress, thus enhancing the drought tolerance of transgenic tobacco lines. These results provide a theoretical basis for further exploring the functions of the bZIP family genes and lay a foundation for stress resistance improvement in chrysanthemums in the future.

The transmembrane domain of the rice small protein OsS1Fa1 is responsible for subcellular localization and drought tolerance.

OsS1Fa1, a homologue of spinach S1Fa, is a small protein in rice that contains four distinct conserved motifs and participates in drought tolerance. However, the biological functions of these conserved motifs have not been characterized to date. Therefore, we investigated the roles of these conserved domains in the localization and cellular function of OsS1Fa1. We analysed the subcellular localization of OsS1Fa1 using confocal laser scanning microscopy (CLSM), following particle bombardment and bacterial infiltration. An E. coli in vivo reconstituted sumoylation assay was conducted to investigate sumoylation of OsS1Fa1. We characterized the function of the transmembrane domain of OsS1Fa1 in drought tolerance using transgenic Arabidopsis plants. Fluorescence analysis showed that OsS1Fa1 localized to the nuclear and cytoplasmic membranes. Mutation and cell fractionation analyses revealed that the membrane localization domain determined the subcellular localization of OsS1Fa1. The rice homologue OsS1Fa2 and Arabidopsis orthologs AtS1Fa1, AtS1Fa2, and AtS1Fa3 also exhibited similar localization patterns as OsS1Fa1. Sumoylation analysis demonstrated that OsS1Fa1 was conjugated with the small ubiquitin-related modifier (SUMO). Transgenic analysis showed that overexpression of OsS1Fa1(TMm1), a mutant form of the transmembrane domain of OsS1Fa1, in Arabidopsis did not enhance drought stress tolerance, whereas OsS1Fa1 overexpression improved the drought tolerance of transgenic Arabidopsis. Our data indicate that rice and Arabidopsis S1Fa1 proteins localize in the nuclear and cytoplasmic membranes, and that transmembrane domain determines subcellular localization and plays an important role in drought stress tolerance.

A resilient mutualistic interaction between cucumber mosaic virus and its natural host to adapt to an excess zinc environment and drought stress.

A perennial pseudometallophyte Arabidopsis halleri is frequently infected with cucumber mosaic virus (CMV) in its natural habitat. The purpose of this study was to characterize the effect of CMV infection on the environmental adaptation of its natural host A. halleri. The CMV(Ho) strain isolated from A. halleri was inoculated into clonal virus-free A. halleri plants, and a unique plant-virus system consisting of CMV(Ho) and its natural wild plant host was established. In a control environment with ambient zinc supplementation, CMV(Ho) infection retarded growth in the above-ground part of host plants but conferred strong drought tolerance. On the other hand, in an excess zinc environment, simulating a natural edaphic environment of A halleri, host plants hyperaccumulated zinc and CMV(Ho) infection did not cause any symptoms to host plants while conferring mild drought tolerance. We also demonstrated in Nicotiana benthamiana as another host that similar effects were induced by the combination of excess zinc and CMV(Ho) infection. Transcriptomic analysis indicated that the host plant recognized CMV(Ho) as a mutualistic symbiont rather than a parasitic pathogen. These results suggest a resilient mutualistic interaction between CMV(Ho) and its natural host A. halleri in its natural habitat.

PvMYB60 gene, a candidate for drought tolerance improvement in common bean in a climate change context.

BACKGROUND: Common bean (Phaseolus vulgaris) is one of the main nutritional resources in the world, and a low environmental impact source of protein. However, the majority of its cultivation areas are affected by drought and this scenario is only expected to worsen with climate change. Stomatal closure is one of the most important plant responses to drought and the MYB60 transcription factor is among the key elements regulating stomatal aperture. If targeting and mutating the MYB60 gene of common bean would be a valuable strategy to establish more drought-tolerant beans was therefore investigated. RESULTS: The MYB60 gene of common bean, with orthology to the Arabidopsis AtMYB60 gene, was found to have conserved regions with MYB60 typical motifs and architecture. Stomata-specific expression of PvMYB60 was further confirmed by q-RT PCR on organs containing stomata, and stomata-enriched leaf fractions. Further, function of PvMYB60 in promoting stomata aperture was confirmed by complementing the defective phenotype of a previously described Arabidopsis myb60-1 mutant. CONCLUSIONS: Our study finally points PvMYB60 as a potential target for obtaining more drought-tolerant common beans in the present context of climate change which would further greatly contribute to food security particularly in drought-prone countries.

Exopolysaccharide is required by Paraburkholderia phytofirmans PsJN to confer drought-stress tolerance in pea.

Paraburkholderia phytofirmans PsJN is a plant symbiotic bacterium that can colonize a broad spectrum of plant hosts and frequently shows beneficial effects on plant growth. Exopolysaccharide (EPS) is known to be important in plant-bacteria interactions. Previously, we reported that EPS is required for PsJN to survive from drought stress and colonize in pea (Pisum sativum) under drought condition. However, whether EPS is necessary for PsJN to promote plant growth remains unknown. In this work, a comparative study was conducted between the wild-type PsJN and its ∆bceQ mutant that lacks EPS to investigate the role of EPS in PsJN to confer drought-stress tolerance on pea plant. Our results showed that wild type PsJN, but not the ∆bceQ mutant, promoted pea seed germination and seedlings growth under drought stress. Pea plants inoculated with the wild type PsJN had a higher level of drought tolerance, as shown by a better vegetative growth and enhanced nodule formation, than plants inoculated with the ∆bceQ mutant. Moreover, EPS plays a role in the plant colonization under drought stress, because the ∆bceQ mutant was unable to colonize pea seeds and roots as effectively as the wild type PsJN. Further, expression of the EPS biosynthesis genes in the bceOVN operon of the wild type PsJN was induced by the presence of glucose. Overall, this study demonstrated that PsJN can promote pea plant growth under drought conditions and EPS is required for PsJN to confer beneficial effects to host plant.

Overexpression of PagLOL1b improves drought tolerance through increasing water use efficiency and reactive oxygen species scavenging in transgenic poplar.

LESION SIMULATING DISEASE1 (LSD) family genes play a key role in plant response to abiotic and biotic stress. However, their functions in the resistance of tree to drought stress are still largely not clear. Here, five LSD family genes in poplar genome were identified. Phylogenetic and collinear relationship analysis showed that they belonged to LSD, LSD-one-like 1 (LOL1) and LSD-one-like 2 (LOL2) subfamilies, and experienced two segmental duplication events. PagLSDs were highly conserved in gene structure, and all PagLSDs contained at least two LSD domains. Expression pattern and cis-acting element analyses showed that PagLSDs were widely expressed in different organs, significantly induced by polyethylene glycol, and possessed a great number of plant growth, development, plant hormones, and biotic and abiotic stress elements in their promoter regions. Further physiological experiments with transgenic poplar plants revealed that overexpression of PagLOL1b significantly enhanced the drought tolerance of transgenic plants. The improved drought tolerance was closely associated with the significant increase in stomatal closure, water use efficiency, antioxidant enzyme gene expression and antioxidant enzyme activity in transgenic plants. The results in our study imply that PagLOL1b has great potential in the engineering of new tree varieties resistant to drought stress.

Integrating genetic assortment and molecular insights for climate-resilient breeding to unravel drought tolerance in cotton.

This study addresses the challenges posed by rainfall variability, leading to water deficits during critical stages of crop growth, resulting in a drastic reduction of cotton yield. In a comprehensive evaluation, thirty cotton genotypes, including five Gossypium arboreum (wild) and twenty-five Gossypium hirsutum (cultivated), were grown under rainfed and irrigated conditions. Drought tolerance indices (DTI) were evaluated, categorizing genotypes based on their resilience. Further, in-vitro screening at the seedling stage (20 days) under PEG-induced drought identified tolerant genotypes exhibiting elevated levels of free proline (19.07±5.31 mg.g-100fr.wt.), amino acids (34.59±6.51 mg.g-100fr.wt.), soluble proteins (13.73±2.65 mg.g-1fr.wt.), and glycine betaine (10.95±3.62 mg.g-100fr.wt.), in their leaves, positively correlating (p<0.001) with relative water content (87.70±3.45 %). Molecular analysis using drought-specific simple sequence repeats (SSR) markers revealed significant genetic variability in a cotton genotypes, with lower observed and higher expected heterozygosity. F statistics exposed a higher level of gene flow corresponding to low differentiation among populations. Among the genotypes group, wild GAM-67 and cultivated Deviraj emerged as the most potent, exhibiting the higher DTI and diverse gene pools. Study exhibited higher total gene diversity in drought-tolerant wild GAM-67 (0.8501) and greater expected heterozygosity (0.626) and gene flow (0.6731) in cultivated Deviraj, underlining their robust genetic adaptability to drought conditions. The integrated approach of field evaluations, in-vitro screening, and molecular analyses explained substantial genetic diversity, with the identification of promising genotypes displaying higher drought tolerance indices, elevated levels of stress-related biochemical osmolytes, and pronounced genetic adaptability, thereby contributing to the advancement of breeding initiatives for enhanced drought resilience in cotton.

Physiological traits, crop growth, and grain quality of quinoa in response to deficit irrigation and planting methods.

Climate change has become a concern, emphasizing the need for the development of crops tolerant to drought. Therefore, this study is designed to explore the physiological characteristics of quinoa that enable it to thrive under drought and other extreme stress conditions by investigating the combined effects of irrigation water levels (100%, 75%, and 50% of quinoa's water requirements, WR as I1, I2 and I3) and different planting methods (basin, on-ridge, and in-furrow as P1, P2 and P3) on quinoa's physiological traits and gas exchange. Results showed that quinoa's yield is lowest with on-ridge planting and highest in the in-furrow planting method. Notably, the seed protein concentrations in I2 and I3 did not significantly differ but they were 25% higher than those obtained in I1, which highlighted the possibility of using a more effective irrigation method without compromising the seed quality. On the other hand, protein yield (PY) was lowest in P2 (mean of I1 and I2 as 257 kg ha-1) and highest in P3 (mean of I1 and I2 as 394 kg ha-1, 53% higher). Interestingly, PY values were not significantly different in I1 and I2, but they were lower significantly in I3 by 28%, 27% and 20% in P1, P2, and P3, respectively. Essential plant characteristics including plant height, stem diameter, and panicle number were 6.1-16.7%, 6.4-24.5%, and 18.4-36.5% lower, respectively, in I2 and I3 than those in I1. The highest Leaf Area Index (LAI) value (5.34) was recorded in the in-furrow planting and I1, while the lowest value was observed in the on-ridge planting method and I3 (3.47). In I3, leaf temperature increased by an average of 2.5-3 oC, particularly during the anthesis stage. The results also showed that at a similar leaf water potential (LWP) higher yield and dry matter were obtained in the in-furrow planting compared to those obtained in the basin and on-ridge planting methods. The highest stomatal conductance (gs) value was observed within the in-furrow planting method and full irrigation (I1P3), while the lowest values were obtained in the on-ridge and 50%WR (I3P2). Finally, photosynthesis rate (An) reduction with diminishing LWP was mild, providing insights into quinoa's adaptability to drought. In conclusion, considering the thorough evaluation of all the measured parameters, the study suggests using the in-furrow planting method with a 75%WR as the best approach for growing quinoa in arid and semi-arid regions to enhance production and resource efficiency.

Mitigation of Drought Stress for Quinoa (Chenopodium quinoa Willd.) Varieties Using Woodchip Biochar-Amended Soil.

Drought stress deteriorates agro-ecosystems and poses a significant threat to crop productivity and food security. Soil amended with biochar has been suggested to mitigate water stress, but there is limited knowledge about how biochar affects the physiology and vegetative growth of quinoa plants under soil water deficits. We grew three quinoa (Chenopodium quinoa Willd.) varieties, Titicaca (V1), Quipu (V2), and UAFQ7 (V3) in sandy loam soil without (B0) and with 2% woodchip biochar (B2) under drought conditions. The drought resulted in significant growth differences between the varieties. V3 performed vegetatively better, producing 46% more leaves, 28% more branches, and 25% more leaf area than the other two varieties. Conversely, V2 displayed significantly higher yield-contributing traits, with 16% increment in panicle length and 50% more subpanicles compared to the other varieties. Woodchip biochar application significantly enhanced the root development (i.e., root biomass, length, surface, and projected area) and plant growth (i.e., plant height, leaf area, and absolute growth rate). Biochar significantly enhanced root growth, especially fresh and dry weights, by 122% and 127%, respectively. However, biochar application may lead to a trade-off between vegetative growth and panicle development under drought stress as shown for V3 grown in soil with woodchip biochar. However, V3B2 produced longer roots and more biomass. Collectively, we suggest exploring the effects of woodchip biochar addition to the soil on the varietal physiological responses such as stomatal regulations and mechanisms behind the increased quinoa yield under water stress conditions.

Morphological Differentiation, Yield, and Cutting Time of Lolium multiflorum L. under Acid Soil Conditions in Highlands.

Livestock production in the basins of the northern macro-region of Peru has as its primary source pastures of Lolium multiflorum L. 'Cajamarquino ecotype' (ryegrass CE) in monoculture, or in association with white clover Ladino variety, for feeding. The objective of this research work was the morphological characterisation, yield evaluation, and cutting time evaluation of two local genotypes (LM-58 and LM-43) of Lolium multiflorum L. in six locations. An ANOVA was performed to compare fixed effects and interaction. It was determined that the LM-58 genotype is intermediate, growing semi-erect, with a dark green colouring and 0.8 cm broadleaf, and can reach an average stem length of 46 cm, up to 1.6 cm. day-1, achieving fourth-leaf growth at 28 days under appropriate management conditions. Despite the differentiated characteristics, according to BLASTn evaluation, the ITS1 sequences showed a greater than 99.9% similar identification to Lolium multiflorum L., characterising it as such. It was determined that the LM-58 genotype outperforms LM-43, achieving a forage yield of 4.49 Mg. ha-1, a seed production of 259.23 kg. ha-1, and an average of 13.48% crude protein (CP). The best biomass yield (49.10 Mg. ha-1.yr-1) is reached at 60 days; however, at 30 days, there is a high level of CP (14.84%) and there are no differences in the annual protein production at the cutting age of 60 and 45 days. With the results of the present study, LM-58 from a selection and crossbreeding of 680 ryegrass EC accessions emerges as an elite genotype adapted to the conditions of the northern high Andean zone of Peru.

ZmDST44 Gene Is a Positive Regulator in Plant Drought Stress Tolerance.

Improving drought tolerance in plants is essential for increasing crop yields under water-limited conditions. In this study, we investigated the functional role of the maize gene ZmDST44, which is targeted by the miRNA ZmmiR139. Our results indicate that ZmmiR139 regulates ZmDST44 by cleaving its mRNA, as confirmed by inverse expression patterns and 5'-RACE analysis. Overexpression of ZmDST44 in Arabidopsis, rice, and maize resulted in significant enhancements in drought tolerance. Transgenic plants exhibited reduced malondialdehyde (MDA) levels, increased proline accumulation, and upregulation of drought-responsive genes compared to wild-type plants. Transgenic Arabidopsis and rice showed improved drought resistance and higher post-drought recovery rates, and transgenic maize displayed lower sensitivity to drought stress. These findings suggest that ZmDST44 acts as a positive regulator of drought tolerance across different plant species and holds promise for developing drought-resistant crops through genetic engineering.

Uncovering novel genes for drought stress in rice at germination stage using genome wide association study.

Introduction: Breeding rice with drought tolerance for harsh environments is crucial for agricultural sustainability. Understanding the genetic underpinnings of drought tolerance is vital for developing resilient rice varieties. Genome-wide association studies (GWAS) have emerged as pivotal tools in unravelling the complex genetic architecture of traits like drought tolerance, capitalizing on the natural genetic diversity within rice germplasm collections. Methods: In this study, a comprehensive panel of 210 rice varieties was phenotyped over ten days in controlled conditions, subjected to simulated drought stress using 20% PEG 6000 in petri dishes. Throughout the stress period, crucial traits such as germination percentage (GP), germination rate index (GRI), mean germination time (MGT), and seedling percentage (SP) were meticulously monitored. Results: The GWAS analysis uncovered a total of 38 QTLs associated with drought tolerance traits, including novel loci like qMGT-5.2, qSP-3, qSP7.2, and qGP-5.2. Additionally, RNA-seq analysis identified ten genes with significant expression differences under drought stress conditions. Notably, haplotype analysis pinpointed elite haplotypes in specific genes linked to heightened drought tolerance. Discussion: Overall, this study underscores the importance of GWAS in validating known genes while unearthing novel loci to enrich the genetic resources for enhancing drought tolerance in rice breeding programs.

The ATP-binding cassette transporter subfamily G member 4 mediates cuticular hydrocarbon transport to regulate drought tolerance in Acyrthosiphon pisum.

ABC transporters are a highly conserved membrane protein class that promote the transport of substances across membranes. Under drought conditions, insects primarily regulate the content of cuticular hydrocarbon (CHC) to retain water and prevent evaporative loss. Involvement of ABC transporter protein G (ABCG) subfamily genes in insect CHC transport has been relatively understudied. In this study, we demonstrated that ABCG4 gene in Acyrthosiphon pisum (ApABCG4) is involved in CHC transport and affects drought tolerance by regulating CHC accumulation. ApABCG4 is strongly expressed in the abdominal cuticle and embryonic stages of A. pisum. Effective silencing of ApABCG4 was achieved using RNAi, and the silencing duration was analyzed. ApABCG4 silencing resulted in a significant decrease in the total and component contents of the CHC and cuticular waxy coatings of A. pisum. Nevertheless, the internal hydrocarbon content remained unchanged. The lack of cuticular hydrocarbons significantly reduced the drought tolerance of A. pisum, shortening its survival time under drought stress. Drought stress caused significant upregulation of ApABCG4. Molecular docking showed that ApABCG4 has a high binding affinity for nine n-alkanes of CHC through electrostatic interactions. These results indicate that ApABCG4 is a novel RNAi target with key applications in aphid biological control.

Cytokinin oxidase2-deficient mutants improve panicle and grain architecture through cytokinin accumulation and enhance drought tolerance in indica rice.

KEY MESSAGE: The Osckx2 mutant accumulates cytokinin thereby enhancing panicle branching, grain yield, and drought tolerance, marked by improved survival rate, membrane integrity, and photosynthetic function. Cytokinins (CKs) are multifaceted hormones that regulate growth, development, and stress responses in plants. Cytokinins have been implicated in improved panicle architecture and grain yield; however, they are inactivated by the enzyme cytokinin oxidase (CKX). In this study, we developed a cytokinin oxidase 2 (Osckx2)-deficient mutant using CRISPR/Cas9 gene editing in indica rice and assessed its function under water-deficit and salinity conditions. Loss of OsCKX2 function increased grain number, secondary panicle branching, and overall grain yield through improved cytokinin content in the panicle tissue. Under drought conditions, the Osckx2 mutant conserved more water and demonstrated improved water-saving traits. Through reduced transpiration, Osckx2 mutants showed an improved survival response than the wild type to unset dehydration stress. Further, Osckx2 maintained chloroplast and membrane integrity and showed significantly improved photosynthetic function under drought conditions through enhanced antioxidant protection systems. The OsCKX2 function negatively affects panicle grain number and drought tolerance, with no discernible impact in response to salinity. The finding suggests the utility of the beneficial Osckx2 allele in breeding to develop climate-resilient, high-yielding cultivars for future food security.