Sex-specific phosphorus acquisition strategies and cycling in dioecious Populus euphratica forests along a natural water availability gradient.

Soil phosphorus (P) availability affects plant growth and distribution. However, it is still unknown how sex-specific variation in functional traits affects plants' P acquisition and soil P transformation. On wet sites, female poplars had a greater specific root length (SRL), and a higher diversity of arbuscular mycorrhizal fungi (AMF) and phosphate-solubilizing bacteria (PSB). Male poplars living on wet sites increased the abundance of AMF and PSB communities and enhanced moderately labile and highly resistant organic P mineralisation via increased phosphatase activity. In contrast, on the dry site, the abundance and diversity of AMF and PSB communities increased in females, enhancing moderately labile and highly resistant organic P mineralisation via elevating phosphatase activities. Males maintained greater SRL and promoted Ca-P mobilisation via the release of root carboxylic acids and rhizosphere acidification on the dry site. The AMF community diversity followed a similar pattern as that of the PSB community when altering the P availability of different-sex plants. Our results indicated that organic P and Ca-P are the major sources of plant-available P in natural P. euphratica forests. Seasonal shifts and geographic locations affected the share of organic and inorganic P pools, and AMF and PSB diversities, ultimately altering sex-specific P acquisition strategies of plants.

Male, female, and mixed-sex poplar plantations support divergent soil microbial communities.

Males and females of dioecious plants have sex-specific adaptations to diverse habitats. The effects of inter- and intrasexual interactions in poplar plantations on composition, structure, and function of soil microbiota have not been explored in degraded areas. We conducted a series of greenhouse and field experiments to investigate how belowground competition, soil microbial communities, and seasonal variation nitrogen content differ among female, male, and mixed-sex Populus cathayana plantations. In the greenhouse experiment, female neighbors suppressed the growth of males under optimal nitrogen conditions. However, male neighbors enhanced stable isotope ratio of nitrogen (δ15 N) of females under intersexual competition. In the field, the root length density, root area density, and biomass of fine roots were lower in female plantations than in male or mixed-sex plantations. Bacterial networks of female, male, and mixed-sex plantations were characterized by different composition of hub nodes, including connectors, modules, and network hubs. The sex composition of plantations altered bacterial and fungal community structures according to Bray-Curtis distances, with 44% and 65% of variance explained by the root biomass, respectively. The total soil nitrogen content of mixed-sex plantation was higher than that in female plantation in spring and summer. The mixed-sex plantation also had a higher ß-1,4-N-acetyl-glucosaminidase activity in summer and a higher nitrification rate in autumn than the other two plantations. The seasonal soil N content, nitrification rate, and root distribution traits demonstrated spatiotemporal niche separation in the mixed-sex plantation. We argue that a strong female-female competition and limited nitrogen content could strongly impede plant growth and reduce the resistance of monosex plantations to climate change and the mixed-sex plantations constitutes a promising way to restore degraded land.

Allelochemicals and soil microorganisms jointly mediate sex-specific belowground interactions in dioecious Populus cathayana.

Little is known about how sex differences in root zone characteristics, such as contents of allelochemicals and soil microbial composition, mediate intra- and intersexual interactions in dioecious plants. We examined the processes and mechanisms of sex-specific belowground interactions mediated by allelochemicals and soil microorganisms in Populus cathayana females and males in replicated 30-yr-old experimental stands in situ and in a series of controlled experiments. Female roots released a greater amount and more diverse phenolic allelochemicals into the soil environment, resulting in growth inhibition of the same sex neighbors and deterioration of the community of soil microorganisms. When grown with males, the growth of females was consistently enhanced, especially the root growth. Compared with female monocultures, the presence of males reduced the total phenolic accumulation in the soil, resulting in a shift from allelopathic inhibition to chemical facilitation. This association was enhanced by a favorable soil bacterial community and increased bacterial diversity, and it induced changes in the orientation of female roots. Our study highlighted a novel mechanism that enhances female performance by males through alterations in the allelochemical content and soil microbial composition. The possibility to improve productivity by chemical mediation provides novel opportunities for managing plantations of dioecious plants.

Ammonium and nitrate affect sexually different responses to salt stress in Populus cathayana.

Nitrogen (N) fertilization is a promising approach to improve salt tolerance. However, it is poorly known how plant sex and inorganic N alter salt stress-induced Na+ uptake, distribution and tolerance. This study employed Populus cathayana Rehder females and males to examine sex-related mechanisms of salt tolerance under nitrate (NO3 - ) and ammonium (NH4 + ) nutrition. Males had a higher root Na+ efflux, lower root-to-shoot translocation of Na+ , and higher K+ /Na+ , which enhanced salt tolerance under both N forms compared to females. On the other hand, decreased root Na+ efflux and K+ retention, and an increased ratio of Na+ in leaves relative to shoots in females caused greater salt sensitivity. Females receiving NH4 + rather than NO3 - had greater net root Na+ uptake, K+ efflux, and translocation to the shoots, especially in leaves. In contrast, males receiving NO3 - rather than NH4 + had increased Na+ translocation to the shoots, especially in the bark, which may narrow the difference in leaf damage by salt stress between N forms despite a higher shoot Na+ accumulation and lower root Na+ efflux. Genes related to cell wall synthesis, K+ and Na+ transporters, and denaturized protein scavenging in the barks showed differential expression between females and males in response to salt stress under both N forms. These results suggested that the regulation of N forms in salt stress tolerance was sex-dependent, which was related to the maintenance of the K+ /Na+ ratio in tissues, the ability of Na+ translocation to the shoots, and the transcriptional regulation of bark cell wall and proteolysis profiles.

Nitrogen addition alleviated sexual differences in responses to cadmium toxicity by regulating the antioxidant system and root characteristics, and inhibiting Cd translocation in mulberry seedlings.

Cadmium (Cd) toxicity and nitrogen (N) deposition are two major environmental stresses which can affect plant growth. It's less clear that how the combined Cd accumulation and N deposition affect the male and female plants of dioecious species. The aim of the present study was to detect sex-specific responses to Cd stress and simulated N deposition in one-year-old male, female and hermaphrodite seedlings of Morus alba. Changes in morphology, physiology, root architecture and biomass of the three sex types of mulberry seedlings were determined. The results showed that Cd toxicity caused limited growth, impaired photosynthetic apparatus and decreased gas exchange rates with significant sex-specific differences. Mulberry was found to deploy detoxification mechanisms to avoid or tolerate toxic Cd effects through the activation of the antioxidant system, increasing proline and non-protein thiol contents, translocating Cd into different plant parts and decreasing biomass. Females displayed a low tolerance to high Cd and were more sensitive to Cd stress. Simulated N deposition alleviated the negative effects of Cd on leaves and decreased sex-specific differences in the three kinds of mulberry seedlings, but N fertilizer did not affect the total biomass. The N-stimulated increasing in proline and non-protein thiol contents might play a crucial role in resisting the damage caused by Cd stress, and the three kinds of mulberry seedlings had slightly different ways of improving Cd tolerance by N deposition. Sexual differences in Cd accumulation are correlated with root architecture. This study provides evidence for the utilization of mulberry to treat Cd-contaminated soils under N deposition.

Sex-specific strategies of phosphorus (P) acquisition in Populus cathayana as affected by soil P availability and distribution.

Soil phosphorus (P) availability and its distribution influence plant growth and productivity, but how they affect the growth dynamics and sex-specific P acquisition strategies of dioecious plant species is poorly understood. In this study, the impact of soil P availability and its distribution on dioecious Populus cathayana was characterized. P. cathayana males and females were grown under three levels of P supply, and with homogeneous or heterogeneous P distribution. Females had a greater total root length, specific root length (SRL), biomass and foliar P concentration under high P supply. Under P deficiency, males had a smaller root system than females but a greater exudation of soil acid phosphatase, and a higher colonization rate and arbuscular mycorrhizal hyphal biomass, suggesting a better capacity to mine P and a stronger association with arbuscular mycorrhizal fungi to forage P. Heterogeneous P distribution enhanced growth and root length density (RLD) in females. Female root proliferation in P-rich patches was related to increased foliar P assimilation. Localized P application for increasing P availability did not enhance the biomass accumulation and the morphological plasticity of roots in males, but it raised hyphal biomass. The findings herein indicate that sex-specific strategies in P acquisition relate to root morphology, root exudation and mycorrhizal symbioses, and they may contribute to sex-specific resource utilization patterns and niche segregation.

Asymmetric pruning reveals how organ connectivity alters the functional balance between leaves and roots of Chinese fir.

The functional balance between leaves and roots is believed to be mediated by the specific location of shoots and roots, i.e. differences in transport distances and degrees of organ connectivity. However, it remains unknown whether the adaptive responses of trees to biomass removal depend on the relative orientation of leaf and root pruning. Here, we applied five pruning treatments to saplings of Cunninghamia lanceolata (Chinese fir) under field and glasshouse conditions, namely no pruning (control), half of lateral branches pruned, half of lateral roots pruned, half of the branches and roots pruned on the same side of the plant, and half of the branches and roots pruned on opposite sides of the plant. The effects of pruning on the growth, carbon storage and allocation, and physiology of leaves and fine roots on the same and opposite sides of the plant were investigated. Compared with the effect of root-pruning on leaves, fine roots were more limited by carbon availability and their physiological activity was more strongly reduced by shoot pruning, especially when branches on the same side of the plant were removed. Pruning of branches and roots on the opposite side of the plant resulted in the lowest carbon assimilation rates and growth among all treatments. The results of a stable-isotope labeling indicated that less C was distributed to fine roots from the leaves on the opposite side of the plant compared to those on the same side, but N allocation from roots to leaves depended less on the relative root and leaf orientation. The results collectively indicate that the functional responses of C. lanceolata to pruning are not only determined by the source-sink balance model but are also related to interactions between leaves and fine roots. We argue that the connectivity among lateral branches and roots depends on their relative orientation, which is therefore critical for the functional balance between leaves and fine roots.

Physiological responses of Elaeocarpus glabripetalus seedlings exposed to simulated acid rain and cadmium.

Combined effects of cadmium (Cd) and acid rain on physiological characteristics in Eleocarpus glabripetalus seedlings were investigated under controlled conditions. The single Cd treatment and the combined Cd and acid rain treatment increased growth at low Cd concentrations, while decreased growth and photosynthesis at high Cd2+ concentrations. A low Cd2+ concentration (50 mg kg-1) combined with different acid rain treatments increased the seedling biomass. A high Cd2+ concentration (100 mg kg-1) under different acid rain treatments significantly decreased the biomass, the Fe content, chlorophyll fluorescence and photosynthetic parameters. Relative electric conductivity, malondialdehyde (MDA) content and peroxidase (POD) activity were increased while the reduced glutathione (GSH) content and catalase (CAT) activity were significantly lower at high Cd2+ concentration under acid rain. The results indicated that the combination of a high concentration of Cd2+ and acid rain aggravated the toxic effect of Cd2+ or acid rain alone on the growth and physiological parameters of E. glabripetalus due to serious damage to the chloroplast structure. These results provide novel insights into the combined effects of Cd2+and acid rain on woody plants and might also serve as a guide to evaluate forest restoration and biological safety in areas with Cd2+and acid rain pollution.

Diversity of indoor fungi as revealed by DNA metabarcoding.

In the present study, we conducted DNA metabarcoding (the nuclear ITS2 region) for indoor fungal samples originating from two nursery schools with a suspected mould problem (sampling before and after renovation), from two university buildings, and from an old farmhouse. Good-quality sequences were obtained, and the results showed that DNA metabarcoding provides high resolution in fungal identification. The pooled proportions of sequences representing filamentous ascomycetes, filamentous basidiomycetes, yeasts, and other fungi equalled 62.3%, 8.0%, 28.3%, and 1.4%, respectively, and the total number of fungal genera found during the study was 585. When comparing fungal diversities and taxonomic composition between different types of buildings, no obvious pattern was detected. The average pairwise values of SørensenChao indices that were used to compare similarities for taxon composition between samples among the samples from the two university buildings, two nurseries, and farmhouse equaled 0.693, 0.736, 0.852, 0.928, and 0.981, respectively, while the mean similarity index for all samples was 0.864. We discovered that making explicit conclusions on the relationship between the indoor air quality and mycoflora is complicated by the lack of appropriate indicators for air quality and by the occurrence of wide spatial and temporal changes in diversity and compositions among samples.

iTRAQ-based quantitative proteomic analysis gives insight into sexually different metabolic processes of poplars under nitrogen and phosphorus deficiencies.

Male and female poplars (Populus cathayana Rehd.) respond differently to nitrogen (N) and phosphorus (P) deficiencies. In this study, an iTRAQ-based quantitative proteomic analysis was performed. N and P deficiencies caused 189 and 144 proteins to change in abundance in males and 244 and 464 in females, respectively. Compared to N- and P-deficient males, both N- and P-deficient females showed a wider range of changes in proteins that are involved in amino acid, carbohydrate and protein metabolism, and the sexual differences were significant. When comparing the effects of N- and P-deficiencies, N-deficient females expressed more changes in proteins that are involved in stress responses and gene expression regulation, while P-deficient females showed more changes in proteins that are involved in energy and lipid metabolism, stress responses and gene expression regulation. The quantitative RT-PCR analysis of stress-related proteins showed that males have a better expression correlation between mRNA and protein levels than do females. This study shows that P. cathayana females are more sensitive and have more rapid metabolic mechanisms when responding to N and P deficiencies than do males, and P deficiency has a wider range of effects on females than does N deficiency.

Identifying sex in non-fertile individuals of the moss Drepanocladus turgescens (Bryophyta: Amblystegiaceae) using a novel molecular approach.

Sex identification before sexual maturity is notoriously difficult in plants with separate sexes, but is crucial to address many life history related issues. To study the performance of the two sexes in the rarely sexually reproducing dioecious moss Drepanocladus turgescens a molecular sex marker is needed. The female-targeting marker previously developed for D. trifarius and D. lycopodioides amplifies for a few D. turgescens males, which can thus not be distinguished from females. In a significant addition to the earlier developed method we sequenced the portion successfully amplified by the primers PT-3f and PT-3r for six females and three males. Differences between males and females were revealed at five sequence positions. Examination of a total of fourteen females and seven marker amplifying males confirm that females and such males differ consistently at these positions. The usefulness of a previous protocol for moss sex identification is thus extended to another dioecious moss by the addition of a step where a portion of the sex-correlated region is sequenced.

Sexual competition and N supply interactively affect the dimorphism and competiveness of opposite sexes in Populus cathayana.

Several important dioecious species show sexual spatial segregation (SSS) along environmental gradients that have significant ecological effect on terrestrial ecosystem. However, little attention has been paid to understanding of how males and females respond to environmental gradients and sexual competition. We compared eco-physiological parameters of males and females of Populus cathayana under different sexual competition patterns and nitrogen (N) supply levels. We found that males and females interacting with the same or opposite sex showed significant differences in biomass partition, photosynthetic capacity, carbon (C) and N metabolism, and leaf ultrastructure, and that the sexual differences to competition were importantly driven by N supply. The intersexual competition was enhanced under high N, while the intrasexual competition among females was amplified under low N. Under high N, the intersexual competition stimulated the growth of the females and negatively affected the males. In contrast, under low N, the males exposed to intrasexual competition had the highest tolerance, whereas females exposed to intrasexual competition showed the lowest adaptation among all competition patterns. Sexual competition patterns and N supply levels significantly affected the sexual dimorphism and competitiveness, which may play an important role in spatial segregation of P. cathayana populations.

No evidence of sexual niche partitioning in a dioecious moss with rare sexual reproduction.

BACKGROUND AND AIMS: Roughly half of the species of bryophytes have separate sexes (dioecious) and half are hermaphroditic (monoecious). This variation has major consequences for the ecology and evolution of the different species. In some sexually reproducing dioecious bryophytes, sex ratio has been shown to vary with environmental conditions. This study focuses on the dioecious wetland moss Drepanocladus trifarius, which rarely produces sexual branches or sporophytes and lacks apparent secondary sex characteristics, and examines whether genetic sexes exhibit different habitat preferences, i.e. whether sexual niche partitioning occurs. METHODS: A total of 277 shoots of D. trifarius were randomly sampled at 214 locations and 12 environmental factors were quantified at each site. Sex was assigned to the individual shoots collected in the natural environments, regardless of their reproductive status, using a specifically designed molecular marker associated with female sex. KEY RESULTS: Male and female shoots did not differ in shoot biomass, the sexes were randomly distributed with respect to each other, and environmental conditions at male and female sampling locations did not differ. Collectively, this demonstrates a lack of sexual niche segregation. Adult genetic sex ratio was female-biased, with 2·8 females for every male individual. CONCLUSIONS: The results show that although the sexes of D. trifarius did not differ with regard to annual growth, spatial distribution or habitat requirements, the genetic sex ratio was nevertheless significantly female-biased. This supports the notion that factors other than sex-related differences in reproductive costs and sexual dimorphism can also drive the evolution of biased sex ratios in plants.

Transcriptional profiling in dioecious plant Populus cathayana reveals potential and sex-related molecular adaptations to solar UV-B radiation.

There is increasing evidence confirming that dioecious plants have evolved a series of sex-related physiological mechanisms allowing acclimation to environmental stresses, but few studies have attempted to thoroughly uncover the potential molecular regulation underlying these responses. Recently, we found in the dioecious plant, Populus cathayana, that males possess a greater tolerance to enhanced UV-B radiation than do females. To continue this work further, comparative transcriptome analyses were carried out. A total of 362 and 296 differentially expressed genes (DEGs) were detected in males and females, respectively. Similarly, as shown in previous studies, these DEGs were involved in a set of conserved functions and pathways related to UV-B stress, regardless of the sex. On the other hand, sex-related responses via transcriptome remodeling were detected, as changes in sex-related gene expression occurred in some pathways. For example, many DEGs involved in amino acid metabolism were mainly upregulated in males but downregulated in females. In some pathways, DEGs were expressed predominantly or exclusively in one sex. This may directly contribute to sex-related physiological responses. Taken together, our results not only provide strong evidence that males and females have adopted sex-related molecular strategies in response to solar UV-B radiation but also suggest many new insights, in particular into molecular regulation, via sex-biased gene expression. We conclude that these results can further enhance the sustainable exploitation and management of P. cathayana plants.

Male poplars have a stronger ability to balance growth and carbohydrate accumulation than do females in response to a short-term potassium deficiency.

Potassium (K) deficiency influences plant performance, such as ion uptake and carbohydrate transport. However, little is known about differences between males and females in response to K deficiency. In this study, dry matter accumulation, photosynthetic capacity, allocation patterns of K(+) , Na(+) and carbohydrates, and ultrastructural changes in males and females of Populus cathayana exposed to K deficiency were investigated. The results indicated that males maintained a significantly higher K(+) content and K(+) /Na(+) ratio in leaves and stems than did females under K deficiency. Moreover, K deficiency significantly increased the sucrose content of females, whereas no significant effect on males was detected. In addition, a comparative analysis showed that males allocated more resources to roots, while females allocated more to leaves, which resulted in sexually different root/shoot (R/S) ratios. Transmission electron microscopic (TEM) observations showed that males suffered fewer injuries than did females. These results suggested that males have a better ability to cope with K deficiency. In addition, the combined effects of salinity and K deficiency on poplars were studied. The results indicated that salt stress aggravates the negative effects caused by K deficiency. Taken together, our study provided evidence for gender-specific strategies in ion and carbohydrate allocation in poplars exposed to a short-term K deficiency. In leaves and stems, the lower K(+) accumulation inhibited sucrose translocation and resulted in a decreased R/S ratio, which may contribute to males having a stronger ability to balance growth and carbohydrate accumulation when compared with females.

Selenium availability in tea: Unraveling the role of microbiota assembly and functions.

Tea (Camellia sinensis (L.) O. Kuntze) plants have a strong ability to accumulate selenium (Se). However, the question of how tea plants affect Se availability has received little attention. In this study, five tea cultivars, including Soubei (SB), Aolǜ (AL), Longjing43 (LJ), Zhaori (ZR) and Fenglǜ (FL), were chosen for the study. Quantitative Microbial Ecology Chip and high-throughput sequencing were used to explore the effects of five tea cultivars on soil functions, microbial community structures and Se availability. The results showed that the total soil Se content in the FL garden was lower compared to LJ and SB gardens, whereas available Se was highest in the FL garden. Based on the Bray-Curtis distances, tea cultivar was the main factor affecting bacterial and fungal community structures. The abundance of functional genes concerning carbon, nitrogen, phosphorus and sulfur cycling processes varied among tea gardens. The higher soil NH4+ and NO3- contents, and higher abundance of functional genes like nifH, amoA1 and narG, whereas lower total nitrogen in the FL garden than in the AL and LJ tea gardens demonstrated that the FL tea plants induced microbes to accelerate soil nitrogen cycling processes. Dominant microbes that positively related with functional genes like nifH, narG, and amoA1 were also positively related with the available Se content. In conclusion, tea cultivars could regulate soil functions through affecting microbial community structures and then affecting the soil Se availability. The soil nitrogen cycle processes are suggested to be closely related with Se transformation in tea gardens.

Belowground ecological interactions in dioecious plants: why do opposites attract but similar ones repel?

Dioecious plant species exhibit sexual dimorphism in various aspects, including morphology, physiology, life history, and behavior, potentially influencing sex-specific interactions. While it is generally accepted that intersexual interactions in dioecious species are less intense compared with intrasexual interactions, the mechanisms underlying belowground facilitation in intersexual combinations remain less understood. Here, we explore these mechanisms, which encompass resource complementarity, mycorrhizal fungal networks, root exudate-mediated belowground chemical communication, as well as plant-soil feedback. We address the reason for the lack of consistency in the strength of inter- and intrasexual interactions. We also propose that a comprehensive understanding of the potential positive consequences of sex-specific interactions can contribute to maintaining ecological equilibrium, conserving biodiversity, and enhancing the productivity of agroforestry.

Populus cathayana males are less affected than females by excess manganese: comparative proteomic and physiological analyses.

The comprehension of sexually different responses in dioecious plants to excess manganese (Mn) stress requires molecular explanation. Physiological and proteomic changes in leaves of Populus cathayana males and females were analyzed after 4 wk of exposure to Mn stress. Under excess Mn conditions, shoot height and photosynthesis decreased more in females than in males. Females also showed severe browning and subcellular damage, higher Mn2+ absorption, and different antioxidant enzyme activities compared with males. There were ten differently regulated protein spots induced by excess Mn stress. They were mainly related to photosynthesis, ROS cleaning, and cell signaling associated to ROS, plant cell death, heat shock, cell defense and rescue, and gene expression and regulation. Variation in protein expression between the sexes clearly showed that males have evolved more efficient photosynthesis capacity, more stable gene expression and regulation, and better cell defense and rescue to prevent further injury under excess Mn stress.

Reciprocal grafting separates the roles of the root and shoot in sex-related drought responses in Populus cathayana males and females.

Drought stress responses and sensitivity of dioecious plants, such as Populus cathayana Rehd., are determined by different mechanisms in each sex. In general, males tend to be more resistant while females are more sensitive. Here, we used reciprocal grafting between males and females to determine the relative importance of roots and shoots when plants are exposed to drought stress. Total dry matter accumulation (DMA), photosynthetic capacity, long-term water-use efficiency (Δ), water potential and ultrastructure of mesophyll cells were evaluated to determine the different roles of root and shoot in sex-related drought responses. Plants with male roots were found to be more resistant and less sensitive to water stress than those with female roots under drought conditions. On the contrary, plants with female shoots grew better than those with male shoots under well-watered conditions. These results indicated that the sensitivity of males and females to water stress is primarily influenced by root processes, while under well-watered conditions sexual differences in growth are primarily driven by shoot processes. Furthermore, grafting female shoot scion onto male rootstock was proved to be an effective mean to improve resistance to water stress in P. cathayana females.

Sex-specific responses of Populus yunnanensis exposed to elevated CO2 and salinity.

Populus yunnanensis Dode., a native dioecious woody plant in southwestern China, was employed as a model species to study sex-specific morphological, physiological and biochemical responses to elevated CO2 and salinity. To investigate the effects of elevated CO2 , salinity and their combination, the cuttings were exposed to two CO2 regimes (ambient CO2 and double ambient CO2 ) and two salt treatments in growth chambers. Males exhibited greater downregulation of net photosynthesis rate (Anet ) and carboxylation efficiency (CE) than females at elevated CO2 , whereas these sexual differences were lessened under salt stress. On the other hand, salinity induced a higher decrease in Anet and CE, more growth inhibition and leaf Cl(-) accumulation and more damage to cell organelles in females than in males, whereas the sexual differences in photosynthesis and growth were lessened at elevated CO2 . Moreover, elevated CO2 exacerbated membrane lipid peroxidation and organelle damage in females but not in males under salt stress. Our results indicated that: (1) females are more sensitive and suffer from greater negative effects than do males under salt stress, and elevated CO2 lessens the sexual differences in photosynthesis and growth under salt stress; (2) elevated CO2 tends to aggravate the negative effects of salinity in females; and (3) sex-specific reactions under the combination of elevated CO2 and salinity are distinct from single-stress responses. Therefore, these results provide evidence for different adaptive responses between plants of different sexes exposed to elevated CO2 and salinity.