CsCuAO1 Associated with CsAMADH1 Confers Drought Tolerance by Modulating GABA Levels in Tea Plants.

Our previous study showed that COPPER-CONTAINING AMINE OXIDASE (CuAO) and AMINOALDEHYDE DEHYDROGENASE (AMADH) could regulate the accumulation of γ-aminobutyric acid (GABA) in tea through the polyamine degradation pathway. However, their biological function in drought tolerance has not been determined. In this study, Camellia sinensis (Cs) CsCuAO1 associated with CsAMADH1 conferred drought tolerance, which modulated GABA levels in tea plants. The results showed that exogenous GABA spraying effectively alleviated the drought-induced physical damage. Arabidopsis lines overexpressing CsCuAO1 and CsAMADH1 exhibited enhanced resistance to drought, which promoted the synthesis of GABA and putrescine by stimulating reactive oxygen species' scavenging capacity and stomatal movement. However, the suppression of CsCuAO1 or CsAMADH1 in tea plants resulted in increased sensitivity to drought treatment. Moreover, co-overexpressing plants increased GABA accumulation both in an Agrobacterium-mediated Nicotiana benthamiana transient assay and transgenic Arabidopsis plants. In addition, a GABA transporter gene, CsGAT1, was identified, whose expression was strongly correlated with GABA accumulation levels in different tissues under drought stress. Taken together, CsCuAO1 and CsAMADH1 were involved in the response to drought stress through a dynamic GABA-putrescine balance. Our data will contribute to the characterization of GABA's biological functions in response to environmental stresses in plants.

Cultivar-specific dynamics: unravelling rhizosphere microbiome responses to water deficit stress in potato cultivars.

BACKGROUND: Growing evidence suggests that soil microbes can improve plant fitness under drought. However, in potato, the world's most important non-cereal crop, the role of the rhizosphere microbiome under drought has been poorly studied. Using a cultivation independent metabarcoding approach, we examined the rhizosphere microbiome of two potato cultivars with different drought tolerance as a function of water regime (continuous versus reduced watering) and manipulation of soil microbial diversity (i.e., natural (NSM), vs. disturbed (DSM) soil microbiome). RESULTS: Water regime and soil pre-treatment showed a significant interaction with bacterial community composition of the sensitive (HERBST) but not the resistant cultivar (MONI). Overall, MONI had a moderate response to the treatments and its rhizosphere selected Rhizobiales under reduced watering in NSM soil, whereas Bradyrhizobium, Ammoniphilus, Symbiobacterium and unclassified Hydrogenedensaceae in DSM soil. In contrast, HERBST response to the treatments was more pronounced. Notably, in NSM soil treated with reduced watering, the root endophytic fungus Falciphora and many Actinobacteriota members (Streptomyces, Glycomyces, Marmoricola, Aeromicrobium, Mycobacterium and others) were largely represented. However, DSM soil treatment resulted in no fungal taxa and fewer enrichment of these Actinobacteriota under reduced watering. Moreover, the number of bacterial core amplicon sequence variants (core ASVs) was more consistent in MONI regardless of soil pre-treatment and water regimes as opposed to HERBST, in which a marked reduction of core ASVs was observed in DSM soil. CONCLUSIONS: Besides the influence of soil conditions, our results indicate a strong cultivar-dependent relationship between the rhizosphere microbiome of potato cultivars and their capacity to respond to perturbations such as reduced soil moisture. Our study highlights the importance of integrating soil conditions and plant genetic variability as key factors in future breeding programs aiming to develop drought resistance in a major food crop like potato. Elucidating the molecular mechanisms how plants recruit microbes from soil which help to mitigate plant stress and to identify key microbial taxa, which harbour the respective traits might therefore be an important topic for future research.

StHAB1, a negative regulatory factor in abscisic acid signaling, plays crucial roles in potato drought tolerance and shoot branching.

Abscisic acid (ABA) is critical in drought tolerance and plant growth. Group A protein type 2C phosphatases (PP2Cs) are negative regulators of ABA signaling and plant adaptation to stress. Knowledge about the functions of potato group A PP2Cs is limited. Here, we report that the potato group A PP2C StHAB1 is broadly expressed in potato plants and strongly induced by ABA and drought. Suppression of StHAB1 enhanced potato ABA sensitivity and drought tolerance, whereas overexpression of the dominant mutant StHAB1G276D compromised ABA sensitivity and drought tolerance. StHAB1 interacts with almost all ABA receptors and the Snf1-Related Kinase OST1. Suppressing StHAB1 and overexpressing StHAB1G276D alter potato growth morphology; notably, overexpression of StHAB1G276D causes excessive shoot branching. RNA-sequencing analyses identified that the auxin efflux carrier genes StPIN3, StPIN5, and StPIN8 were up-regulated in StHAB1G276D-overexpressing axillary buds. Correspondingly, the auxin concentration was reduced in StHAB1G276D-overexpressing axillary buds, consistent with the role of auxin in repressing lateral branch outgrowth. The expression of BRANCHED1s (StBRC1a and StBRC1b) was unchanged in StHAB1G276D-overexpressing axillary buds, suggesting that StHAB1G276D overexpression does not cause axillary bud outgrowth via regulation of BRC1 expression. Our findings demonstrate that StHAB1 is vital in potato drought tolerance and shoot branching.

RNA-seq profiling in leaf tissues of two soybean (Glycine max [L.] Merr.) cultivars that show contrasting responses to drought stress during early developmental stages.

Drought stress is the major environment constraint on soybean yield, and a variety of pathways underlie drought tolerance mechanisms. Transcriptomic profiling of two soybean cultivars, drought-tolerant SS2-2 and drought-sensitive Taekwang, was performed under normal and drought conditions to identify genes involved in drought tolerance. This revealed large differences in water loss during drought treatment. Genes involved in signaling, lipid metabolism, phosphorylation, and gene regulation were overrepresented among genes that were differentially expressed between cultivars and between treatments in each cultivar. The analysis revealed transcription factors from six families, including WRKYs and NACs, showed significant SS2-2-specific upregulation. Genes involved in stress defense pathways, including MAPK signaling, Ca2+ signaling, ROS scavenging, and NBS-LRR, were also identified. Expression of non-specific phospholipases, phospholipase D, and PHOSPHATIDYL INOSITOL MONOPHOSPHATE 5 KINASE (PIP5K), which act in the lipid-signaling pathway, was greatly increased in SS2-2. The roles of PIP5K in drought stress tolerance were confirmed in Arabidopsis thaliana. Arabidopsis pip5k mutants had significantly lower survival rates under drought stress than wild-type plants. This study identified additional elements in the mechanisms used by plants to protect themselves from drought stress and provides valuable information for the development of drought-tolerant soybean cultivars. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-023-01385-1.

Modifying the Expression of Cysteine Protease Gene PCP Affects Pollen Development, Germination and Plant Drought Tolerance in Maize.

Cysteine proteases (CPs) are vital proteolytic enzymes that play critical roles in various plant processes. However, the particular functions of CPs in maize remain largely unknown. We recently identified a pollen-specific CP (named PCP), which highly accumulated on the surface of maize pollen. Here, we reported that PCP played an important role in pollen germination and drought response in maize. Overexpression of PCP inhibited pollen germination, while mutation of PCP promoted pollen germination to some extent. Furthermore, we observed that germinal apertures of pollen grains in the PCP-overexpression transgenic lines were excessively covered, whereas this phenomenon was not observed in the wild type (WT), suggesting that PCP regulated pollen germination by affecting the germinal aperture structure. In addition, overexpression of PCP enhanced drought tolerance in maize plants, along with the increased activities of the antioxidant enzymes and the decreased numbers of the root cortical cells. Conversely, mutation of PCP significantly impaired drought tolerance. These results may aid in clarifying the precise functions of CPs in maize and contribute to the development of drought-tolerant maize materials.

Drought resistance evaluation of sugar beet germplasms by response of phenotypic indicators.

Sugar beet is a main sugar crop worldwide that often faces drought stress. The identification of drought tolerance of sugar beet germplasms is beneficial for breeding, but the research about it has been rarely reported. In this study, the drought tolerance of germplasms 92005-1, 94002-2 and 92021-1-1 was tested under simulated conditions. Seven days and 9% PEG treatment were the optimal conditions for evaluation, under which more phenotypic indicators showed significant difference in drought tolerance coefficient. The objective weighting and membership function method were established for evaluating the drought tolerance of different sugar beet germplasms. Drought stress decreased the biomass of leaves and roots of sugar beet germplasms. The drought-sensitive germplasm responded faster for leaf weight, root weight, plant height and root length. These indicators declined more significantly under long-term and severe stress. Increasing the root-shoot ratio and proline content were universal strategies of sugar beet germplasms to overcome drought stress. The drought-tolerant germplasms held higher peroxidase activity and better ability to scavenge reactive oxygen for preventing the damage.

Population growth of Tetranychus truncatus (Acari: Tetranychidae) on different drought-tolerant potato cultivars.

Tetranychus truncatus Ehara (Acari: Tetranychidae) has become one of the major phytophagous pests in China in recent years, and is found on a wide range of host plants. However, little information is available on the population performance of this arthropod pest on potatoes. In this study, we explored the population growth of T. truncatus on two drought-tolerant potato (Solanum tuberosum L.) cultivars under laboratory conditions using the age-stage, two-sex life table. Tetranychus truncatus completed its entire life history on both potato cultivars tested, Holland 15 and Longshu 10. There was no significant difference between two potato cultivars in developmental duration. Tetranychus truncatus had shorter adult longevity (20.61 days), adult female longevity (20.41 days), and total female longevity (33.66 days) on Longshu 10 than Holland 15 (21.16 days, 21.19 days, and 34.38 days, respectively). However, it exhibited a higher preadult survival rate, higher fecundity (F = 88.32 eggs per female), and relatively higher population parameters when reared on Longshu 10 than on Holland 15 (F = 75.70 eggs per female). Growth projection also showed that the population size of T. truncatus on Longshu 10 (expand 750-fold) was larger than that on Holland 15 (expand 273-fold) after 60 days. Our results demonstrate that the drought-sensitive potato variety, Holland 15, is relatively resistant to T. truncatus compared with the drought-tolerant variety, Longshu 10, and suggest that T. truncatus exhibited a trade-off between longevity and reproduction on both potato cultivars. Our findings provide information on population prediction, which may aid the management of this pest mite species of potatoes.

Melatonin and arbuscular mycorrhizal fungi synergistically improve drought toleration in kiwifruit seedlings by increasing mycorrhizal colonization and nutrient uptake.

Kiwifruit is a vine fruit tree that is vulnerable to water deficiency due to its shallow root system and large leaves. Although mycorrhizal inoculation and melatonin application has been proved to improve plants drought tolerance, their interaction effects are still unclear. In this study, arbuscular mycorrhizal (AM) fungi incubation and melatonin (MT) irrigation were applied to kiwifruit seedlings alone or in combination to investigate their effect on drought tolerance. The results revealed that AM had more effect on promoting root biomass, water use efficiency, and uptake of nitrogen, phosphorus and iron. While MT was more effective in promoting shoot biomass and antioxidant enzyme activities to remove reactive oxygen species accumulation. Moreover, MT supplementary significantly increased the AM colonization, spore density and hyphal length density in roots. Therefore, combined application of AM fungi and MT had additive effects on improvement biomass accumulation, increasing chlorophyll content, photosynthetic efficiency, catalase activity, and decreasing malondialdehyde accumulation under drought stress, thus promoting plant growth and alleviating the drought damage to plant. These results provide guidance for AM and MT combined application to improve abiotic resistance in plants.

Nanobiotechnological Approaches to Enhance Drought Tolerance in Catharanthus roseus Plants Using Salicylic Acid in Bulk and Nanoform.

Drought has a detrimental effect on crop production, affecting economically important plants' growth rates and development. Catharanthus roseus is an important medicinal plant that produces many pharmacologically active compounds, some of which have significant antitumor activity. The effect of bulk salicylic acid (SA) and salicylic acid nanoparticles (SA-NPs) were evaluated on water-stressed Catharanthus roseus plants. The results showed that SA and SA-NPs alleviated the negative effects of drought in the treated plants by increasing their shoot and root weights, relative water content, leaf area index, chlorophyll content, and total alkaloids percentage. From the results, a low concentration (0.05 mM) of SA-NPs exerted positive effects on the treated plants, while the best results of the bulk SA were recorded after using the highest concentration (0.1 mM). Both treatments increased the expression level of WRKY1, WRKY2, WRKY40, LEA, and MYC2 genes, while the mRNA level of MPKK1 and MPK6 did not show a significant change. This study discussed the importance of SA-NPs in the induction of drought stress tolerance even when used in low concentrations, in contrast to bulk SA, which exerts significant results only at higher concentrations.

Comprehensive analysis of the BES1 gene family and its expression under abiotic stress and hormone treatment in Populus trichocarpa.

The BRI1 EMS SUPPRESSOR 1/BRASSINAZOLE RESISTANT 1 (BES1/BZR1) plays a vital role in plant growth and development and stress responses, but there are few studies on poplar BES1 genes. In this study, we identified 14 BES1 genes in the Populus trichocarpa genome and analyzed the expression under hormone treatment and abiotic stress. The PtrBES1 genes were classified into seven subgroups (I-VII) through phylogenetic analysis. All the paralogous gene pairs were shown to be subjected to expansion by segment duplication and purification selection during the PtrBES1 family evolution. Promoter cis-element analysis showed that the PtrBES1 promoter contains stress related cis-elements including ABRE-motif, MBS and TC-rich elements. Quantitative real time reverse transcription PCR (RT-qPCR) analysis showed that the PtrBES1 genes were upregulated upon NaCl, Polyethylene glycol 6000 (PEG6000) stress as well as the major stress hormone abscisic acid (ABA) treatment. Under the three treatments, PtrBES1-7 showed high expression levels in leaves and roots. Physiological experiments showed that the overexpression PtrBES1-7 line could enhance tolerance to drought stress in P. trichocarpa by improving the ability to scavenge ROS (reactive oxygen species). This is specifically reflected in the fact that the overexpression line contains less ROS (O2- and H2O2) and more antioxidant enzymes (1.42 times SOD and 1.5 times POD) than the control line. The preliminary results of this study provided a solid basis for the future functional studies of the BES1 gene family in P. trichocarpa.

Genome-Wide Approach to Identify Quantitative Trait Loci for Drought Tolerance in Tetraploid Potato (Solanum tuberosum L.).

Drought represents a major abiotic stress factor negatively affecting growth, yield and tuber quality of potatoes. Quantitative trait locus (QTL) analyses were performed in cultivated potatoes for drought tolerance index DRYM (deviation of relative starch yield from the experimental median), tuber starch content, tuber starch yield, tuber fresh weight, selected transcripts and metabolites under control and drought stress conditions. Eight genomic regions of major interest for drought tolerance were identified, three representing standalone DRYM QTL. Candidate genes, e.g., from signaling pathways for ethylene, abscisic acid and brassinosteroids, and genes encoding cell wall remodeling enzymes were identified within DRYM QTL. Co-localizations of DRYM QTL and QTL for tuber starch content, tuber starch yield and tuber fresh weight with underlying genes of the carbohydrate metabolism were observed. Overlaps of DRYM QTL with metabolite QTL for ribitol or galactinol may indicate trade-offs between starch and compatible solute biosynthesis. Expression QTL confirmed the drought stress relevance of selected transcripts by overlaps with DRYM QTL. Bulked segregant analyses combined with next-generation sequencing (BSAseq) were used to identify mutations in genes under the DRYM QTL on linkage group 3. Future analyses of identified genes for drought tolerance will give a better insight into drought tolerance in potatoes.

Spatially Resolved Metabolomics and Lipidomics Reveal Salinity and Drought-Tolerant Mechanisms of Cottonseeds.

In the current era of global climate change, environmental stresses, especially drought and salt, have impaired the growth and productivity of crops, e.g., cotton. Understanding the mechanisms of plants' adaptation to these abiotic stresses is crucial to breed stress-tolerant crop species. In the present study, integrated metabolomics, lipidomics, and mass spectrometry imaging (MSI) were used to discover the spatial distribution of differential metabolites and lipids in two cottonseed cultivars with contrasting drought and salt tolerance properties. Seventeen differential metabolites and 125 differential lipids were identified. Their possible roles in augmenting stress tolerance were illustrated, which were involved in reactive oxygen species scavenging, osmotic adjustment, and cell membrane structure reconstruction. MSI analysis provided a visualization of nine differential lipids and four differential metabolites in cottonseeds with varied abundances and distributions. The results may help understand cottonseeds' convictive metabolic and lipidomic regulatory networks in coping with salinity and drought stresses and give new insights into the stress-tolerance traits relevant to other crops.

Response to water deficit of semi-desert wild potato Solanum kurtzianum genotypes collected from different altitudes.

Drought-sensitive crops are threatened as a consequence of limited available water due to climate change. The cultivated potato (Solanum tuberosum) is susceptible to drought and within its wild relative species, Solanum kurtzianum is the Argentinian wild potato species best adapted to arid conditions. However, its physiological responses to water deficit (WD) are still missing. Within the distribution of S. kurtzianum, genotypes could be adapted to differential precipitation regimes. The aim of this work was to evaluate responses of three S. kurtzianum genotypes collected at 1100 (G1), 1900 (G2) and 2100 m a.s.l. (G3) to moderate and severe WD. Treatments were imposed since flowering and lasted 36 days. Yield components, morpho-physiological and biochemical responses; and phenotypic plasticity were evaluated. The three genotypes presented mechanisms to tolerate both WD treatments. G1 presented the lowest yield reduction under moderate WD, mainly through a rapid stomatal closure and a modest vegetative growth. The differences among genotypes suggest that local adaptation is taking place within its natural habitat. Also, G2 presented environmentally induced shifts in plasticity for stomatal length and carotenoids, suggesting that phenotypic plasticity has a role in acclimation of plants to WD until selection works.

Unravelling Differences in Candidate Genes for Drought Tolerance in Potato (Solanum tuberosum L.) by Use of New Functional Microsatellite Markers.

Potato is regarded as drought sensitive and most vulnerable to climate changes. Its cultivation in drought prone regions or under conditions of more frequent drought periods, especially in subtropical areas, requires intensive research to improve drought tolerance in order to guarantee high yields under limited water supplies. A candidate gene approach was used to develop functional simple sequence repeat (SSR) markers for association studies in potato with the aim to enhance breeding for drought tolerance. SSR primer combinations, mostly surrounding interrupted complex and compound repeats, were derived from 103 candidate genes for drought tolerance. Validation of the SSRs was performed in an association panel representing 34 mainly starch potato cultivars. Seventy-five out of 154 SSR primer combinations (49%) resulted in polymorphic, highly reproducible banding patterns with polymorphic information content (PIC) values between 0.11 and 0.90. Five SSR markers identified allelic differences between the potato cultivars that showed significant associations with drought sensitivity. In all cases, the group of drought-sensitive cultivars showed predominantly an additional allele, indicating that selection against these alleles by marker-assisted breeding might confer drought tolerance. Further studies of these differences in the candidate genes will elucidate their role for an improved performance of potatoes under water-limited conditions.

Potato CYCLING DOF FACTOR 1 and its lncRNA counterpart StFLORE link tuber development and drought response.

Plants regulate their reproductive cycles under the influence of environmental cues, such as day length, temperature and water availability. In Solanum tuberosum (potato), vegetative reproduction via tuberization is known to be regulated by photoperiod, in a very similar way to flowering. The central clock output transcription factor CYCLING DOF FACTOR 1 (StCDF1) was shown to regulate tuberization. We now show that StCDF1, together with a long non-coding RNA (lncRNA) counterpart, named StFLORE, also regulates water loss through affecting stomatal growth and diurnal opening. Both natural and CRISPR-Cas9 mutations in the StFLORE transcript produce plants with increased sensitivity to water-limiting conditions. Conversely, elevated expression of StFLORE, both by the overexpression of StFLORE or by the downregulation of StCDF1, results in an increased tolerance to drought through reducing water loss. Although StFLORE appears to act as a natural antisense transcript, it is in turn regulated by the StCDF1 transcription factor. We further show that StCDF1 is a non-redundant regulator of tuberization that affects the expression of two other members of the potato StCDF gene family, as well as StCO genes, through binding to a canonical sequence motif. Taken together, we demonstrate that the StCDF1-StFLORE locus is important for vegetative reproduction and water homeostasis, both of which are important traits for potato plant breeding.

Exogenous Melatonin Modulates the Physiological and Biochemical Mechanisms of Drought Tolerance in Tartary Buckwheat (Fagopyrum tataricum (L.) Gaertn).

Tartary buckwheat is one of the nutritious minor cereals and is grown in high-cold mountainous areas of arid and semi-arid zones where drought is a common phenomenon, potentially reducing the growth and yield. Melatonin, which is an amphiphilic low molecular weight compound, has been proven to exert significant effects in plants, under abiotic stresses, but its role in the Tartary buckwheat under drought stress remains unexplored. We evaluated the influence of melatonin supplementation on plant morphology and different physiological activities, to enhance tolerance to posed drought stress by scavenging reactive oxygen species (ROS) and alleviating lipid peroxidation. Drought stress decreased the plant growth and biomass production compared to the control. Drought also decreased Chl a, b, and the Fv/Fm ratio by 54%, 70%, and 8%, respectively, which was associated with the disorganized stomatal properties. Under drought stress, H2O2, O2•-, and malondialdehyde (MDA) contents increased by 2.30, 2.43, and 2.22-folds, respectively, which caused oxidative stress. In contrast, proline and soluble sugar content were increased by 84% and 39%, respectively. However, exogenous melatonin (100 µM) could improve plant growth by preventing ROS-induced oxidative damage by increasing photosynthesis, enzymatic antioxidants (superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase), secondary metabolites like phenylalanine ammonialyase, phenolics, and flavonoids, total antioxidant scavenging (free radical DPPH scavenging), and maintaining relative water content and osmoregulation substances under water stress. Therefore, our study suggested that exogenous melatonin could accelerate drought resistance by enhancing photosynthesis and antioxidant defense in Tartary buckwheat plants.

Overexpression of a novel E3 ubiquitin ligase gene from Coptis chinensis Franch enhances drought tolerance in transgenic tobacco.

Drought stress has a significant effect on the growth, physiology and biochemistry of medicinal plants. SDIR1 (Salt- and Drought-Induced Ring Finger1), a C3H2C3-type RING-finger E3 ubiquitin ligase gene plays an important role in the stress response of various plants. However, the role of this gene is not clear in Coptis chinensis. In this study, the CcSDIR1 gene was cloned from C. chinensis using RACE and RT-PCR. Sequence analysis revealed that CcSDIR1 had an open reading frame of 840 bp that encodes 279 amino acids with a theoretical molecular weight about 31 kDa and pI value of 5.65 and shared conserved domains with other plants. On comparison with the wild-type plants, overexpression of CcSDIR1 in transgenic tobaccos increased drought tolerance and showed better growth performance. However, lower malondialdehyde contents and high antioxidant enzyme activities were observed in transgenic tobacco plants compared to wild-type plants. In addition, Evans blue staining showed high cell viability of transgenic lines under drought stress. These results suggest that CcSDIR1 regulates various responses to drought stress by increasing antioxidant enzyme activities and reducing oxidative damage. From the study results, the CcSDIR1 gene will be very useful for drought stress research in plants.

Safflower (Carthamus tinctorius) Biochemical Properties, Yield, and Oil Content Affected by 24-Epibrassinosteroid and Genotype under Drought Stress.

The steroid hormones, including brassinosteroids, regulate plant growth under stress. It is hypothesized that 24-epibrassinosteroids (24-EBR) can affect safflower (Carthamus tinctorius) biochemical properties, crop yield, and oil content under drought stress. The objective of our study was to determine the response of three safflower genotypes (Goldasht, Faraman, and Sina) to exogenous 24-EBR (0 and 10-7 M) under drought stress, including 85, 65, and 45% of field capacity in 2015. Stress decreased chlorophyll-a, chlorophyll-b, total chlorophyll, carotenoid, relative water content (RWC), seed yield, and oil percentage. The activities of superoxide dismutase (SOD), catalase (CAT), polyphenol oxidase (PPO), and proline contents increased in response to either drought stress or 24-EBR. Genotypes behaved significantly different under stress. 24-EBR significantly increased plant chlorophyll contents and oil percentage, and it significantly reduced the malondialdehyde (MDA) content via increasing the proline and carotenoid contents under stress. 24-EBR can increase safflower oil and seed yield under drought stress.

Molybdenum induces alterations in the glycerolipidome that confer drought tolerance in wheat.

Molybdenum (Mo), which is an essential microelement for plant growth, plays important roles in multiple metabolic and physiological processes, including responses to drought and cold stress in wheat. Lipids also have crucial roles in plant adaptions to abiotic stresses. The aim of this study was to use glycerolipidomic and transcriptomic analyses to determine the changes in lipids induced by Mo that are associated with Mo-enhanced drought tolerance in wheat. Mo treatments increased the transcript levels of genes involved in fatty acid and glycerolipid biosynthesis and desaturation, but suppressed the expression of genes involved in oxylipin production. Wheat plants supplemented with Mo displayed higher contents of monogalactosyldiacyglycerol (MGDG), digalactosyldoacylglycerol (DGDG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and phosphatidylcholine (PC) with increased levels of unsaturation. The levels of MGDG, DGDG, PG, and PC increased under PEG-simulated drought (PSD), and the magnitude of the responses varied in the presence and absence of Mo. Mo increased the accumulation of the most abundant glycerolipid species of C36:6, C34:4, and C34:3 by increasing the expression of genes related to desaturation under PSD, and this contributed to maintaining the fluidity of membranes. In addition, Mo attenuated the decreases in the ratios of DGDG/MGDG and PC/PE that were observed under PSD. These changes in lipids in Mo-treated wheat would contribute to maintaining the integrity of membranes and to protecting the photosynthetic apparatus, thus acting together to enhance drought tolerance.