Higher soil salinity causes more physiological stress in female of Populus cathayana cuttings

Dioecious plant species represent an major component of terrestrial ecosystems. Little is known about sex-specific responses to soil salinity. Populus cathayana Rehd, which is a dioecious, deciduous tree, was employed as a test species in our study. In a semi-controlled environment, physiological responses to salinity were investigated in male and female P. cathayana cuttings, which were subjected to two salt regimes: 0 and 80 mM NaCl added to the Hoagland’s solution for one month growth. Relative to the control, the saline treatment significantly decreased net photosynthetic rate (P_n), transpiration (E), stomatal conductance (g_s), carotenoids (Caro), chlorophyll a (Chl a), total chlorophyll (TC) and catalase (CAT) activity, but increased Na⁺, Ca²⁺, K⁺, malondialdehyde (MDA) content, superoxide dismutase (SOD) activity and carbon isotope composition (δ¹³C) in both sexes. Different sensitivity to saline conditions between males and females was detected. With higher soil salinity, females exhibited lower P_n, Chl a, TC, Chl a/b, Ca²⁺, Ca²⁺/Na⁺, K⁺/Na⁺, SOD and CAT activities but higher Na⁺ and MDA content than males. However, there were no significant differences in these traits (except for SOD and CAT activities) detected in the control group. Our results indicated that males may be more tolerate to salinity than females, with females having lower gas exchanges, chlorophyll pigments, antioxidant enzyme activities, K⁺/Na⁺ ratio and water use efficiency (WUE) than males.     

Effect of arbuscular mycorrhizal inoculation on water status and photosynthesis of Populus cathayana males and females under water stress

Drought is one of the most serious environmental limitations for poplar growth. Although the ways in which plants deal with water stress and the effects of arbuscular mycorrhizal (AM) formation have been well documented, little is known about how the male and female plants of Populus cathayana respond to drought and AM formation. We also aimed to investigate the potential role of AM fungi in maintaining gender balance. We tested the impact of drought and AM formation on water status and photosynthesis. The results suggested that both sexes showed similar responses to water stress: drought decreased the growth of stem length (GSL), growth of ground diameter (GGD), relative water content (RWC), increased the relative electrolyte leakage (REL), and limited the photosynthesis and chlorophyll fluorescence indexes. However, the responses of the two sexes to drought and AM formation differed to some extent. AM formation had positive effects on RWC, photosynthesis and the intrinsic water use efficiency (WUEi) but negative effects on the REL of males and females, especially under drought. AM formation enhanced the maximum quantum yield of photosystem II (PSII) (Fv/Fm), the actual quantum yield of PSII (ΦPSII), non‐photochemical quenching (qN) and photochemical quenching (qP) under drought conditions, and had no significant effects under well‐watered conditions except on the qP of males. Principal component analysis showed that males were significantly more drought tolerant than females, and AM formation enhanced drought tolerance, particularly among males, which suggested that AM fungi are beneficial for ecological stability and for P. cathayana survival under drought conditions.     

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.     

Nitrogen deposition limits photosynthetic response to elevated CO2 differentially in a dioecious species

Sexual dimorphisms of dioecious plants are important in controlling and maintaining sex ratios under changing climate environments. Yet, little is known about sex-specific responses to elevated CO₂ with soil nitrogen (N) deposition. To investigate sex-related physiological and biochemical responses to elevated CO₂ with N deposition, Populus cathayana Rehd. was employed as a model species. The cuttings were subjected to two CO₂ regimes (350 and 700???mol?mol^?1) with two N levels (0 and 5?g?N?m^?2?year^?1). Our results showed that elevated CO₂ and N deposition separately increased the total number of leaves, leaf area (LA), leaf mass, net photosynthetic rate (P _n), light saturated photosynthetic rate (P _max), chlorophyll a (Chl a), and chlorophyll a to chlorophyll b ratio (Chl a/b) in both males and females of P. cathayana. However, the effects on LA, leaf mass, P _n, P _max, Chl a and Chl a/b were weakened under the combined treatment of elevated CO₂ and N deposition. Males had higher leaf mass, P _n, P _max, apparent quantum yield (??), carboxylation efficiency (CE), Chl a, Chl a/b, leaf N, and root carbon to N ratio (C/N) than did females under elevated CO₂ with N deposition. In contrast to males, females had significantly higher levels of soluble sugars in leaves and greater starch accumulation in roots and stems under the same condition. The results of the present work imply that P. cathayana females are more responsive and suffer from greater negative effects on growth and photosynthetic capacity than do males when grown under elevated CO₂ with soil N deposition.     

Salt stress induced sex-related spatial heterogeneity of gas exchange rates over the leaf surface in Populus cathayana Rehd.

The sex-related spatial heterogeneity of gas exchange rates over the leaf surface under salt stress was investigated in the dioecious species, Populus cathayana Rehd. Cuttings were subjected to two salt regimes: 0 and 75 mM NaCl added to the Hoagland solution, the control and the treatment group, respectively. Measurements of gas exchange parameters were taken from over 40 sites on the surfaces of representative ‘non-stressed’ and ‘salt-treated’ leaves which had the same insertion point for two sexual cuttings. Compared to the control group, the treatment group showed a significant decrease in the mean values of the following: water use efficiency (WUE), Chlorophyll a (Chl a) concentration, chlorophyll b (Chl b) concentration, concentration of carotenoids (Caro), total chlorophyll concentration (TC) in two sexes, and net photosynthesis rate (P _n), stomatal conductance (g _s), and stomatal length/width ratio (SR) in females. Also, in the treatment group, females exhibited lower WUE, P _n, g _s, E, Chl a, Chl b, TC, and SR than males. Comparison of contour maps showed that the net photosynthesis rate decreased gradually from apical to basal zones over the leaf surface occurred in the two sexes under natural conditions, but under salt stress, the opposite trend was found in females only. The results suggest that the heterogeneity pattern of the gas exchange parameters in response to salt stress between the two sexes is quite different due to different strategies employed by males and females to maintain the photosynthesis rate under salt stress. This heterogeneity phenomenon under salt stress may mainly be attributed to the chlorophyll pigments in males and the stomatal apertures in 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 (A_net) 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 A_net 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.     

Leaf gas exchange,water relations,nutrient content and growth in citrus and olive seedlings under salinity

The effects of salinity on growth, leaf nutrient content, water relations, gas exchange parameters and chlorophyll fluorescence were studied in six-month-old seedlings of citrus (Citrus limonia Osbeck) and rooted cuttings of olive (Olea europaea L. cv. Arbequina). Citrus and olive were grown in a greenhouse and watered with half strength Hoagland’s solution plus 0 or 50 mM NaCl for citrus, or plus 0 or 100 mM NaCl for olive. Salinity increased Cl⁻ and Na⁺ content in leaves and roots in both species and reduced total plant dry mass, net photosynthetic rate and stomatal conductance. Decreased growth and gas exchange was apparently due to a toxic effect of Cl⁻ and/or Na⁺ and not due to osmotic stress since both species were able to osmotically adjust to maintain pressure potential higher than in non-salinized leaves. Internal CO₂ concentration in the mesophyll was not reduced in either species. Salinity decreased leaf chlorophyll a content only in citrus.     

Effect of increased alkalinity on Na+ and K+ contents, lipid peroxidation and antioxidative enzymes in two populations of Populus cathayana

We compared two populations of Populus cathayana Rehder, originating from altitudes 2 840 m and 1 450 m, to determine whether trees from different altitudes exhibit different tolerance to alkalinity. The tree cuttings were exposed to nutrient solutions with pH 7.9, 8.8, 9.8 and 10.4 and the salt concentration 200 mM. Na⁺ and K⁺ contents, and Na⁺/K⁺ ratios in leaves and roots were greatly affected by pH values. At pH 10.4, the Na⁺/K⁺ ratios in both leaves and roots sharply dropped in the higher altitude population but were maintained at higher levels in the lower altitude population. The patterns of pH-induced changes in contents of malondialdehyde and free proline, and antioxidative enzyme activities indicated that the higher altitude population exhibits greater tolerance to alkalinity stress than does the lower altitude population.     

Sex‐related adaptive responses to interaction of drought and salinity in Populus yunnanensis

We used Populus yunnanensis Dode., a native dioecious species in southwestern China, as a model species to study morphological, physiological, biochemical and ultrastructural responses to drought, salinity and their combination. Females exhibited more growth inhibition, gas exchange rate depression and reactive oxygen species (ROS) accumulation; higher lipid peroxide levels, lower osmotic adjustment capacity and ascorbate–glutathione cycle enzyme activities; and more damage to cell organelles than did males under drought, salinity and especially under their combination. In addition, we found sex‐specific responses in total chlorophyll content (TC), carotenoid concentration and carbon isotope composition under different osmotic stresses. Our results indicated that: (1) females are more sensitive and suffer from greater negative effects than do males under drought, salinity and especially under their combination; (2) sexual differences in adaptive responses to drought, salinity and their combination are context dependent; and (3) sex‐specific reactions under a combination of stresses are distinct from single‐stress responses. Thus, these results provide evidence for adaptive differentiation between sexes in responses to osmotic stresses and in the sensitivity to environmental change.     

Differences in growth and physiological traits of two poplars originating from different altitudes as affected by UV‐B radiation and nutrient availability

Cuttings of Populus kangdingensis and Populus cathayana originating from altitudes of 3500 and 1500 m in southwestern China, respectively, were grown for one growing season in the field under ambient or ambient plus supplemental ultraviolet‐B (UV‐B) radiation with two levels of nutrients. In both species, enhanced UV‐B radiation significantly increased UV‐B absorbing compounds and guaiacol peroxidase (GPX) activity, while no significant effects were observed in photosynthetic pigments and proline content. On the other hand, cuttings grown with high‐nutrient availability had larger leaf area, higher total biomass and GPX activity as well as higher water use efficiency (WUE) (as measured by stable carbon isotope composition, δ¹³C) when compared with low‐nutrient conditions, while UV‐B absorbing compounds and ascorbic acid (AsA) content significantly decreased. Differences in responses to enhanced UV‐B radiation and nutrient availability were observed between the two species. Nutrient‐induced increases in chlorophyll a, chlorophyll b and total chlorophyll as well as in carotenoids were greater in P. kangdingensis than in P. cathayana. In P. cathayana, enhanced UV‐B radiation significantly decreased leaf area and total biomass, while it significantly increased WUE and ascorbate peroxidase (APX). In contrast, such changes were not observed in P. kangdingensis. In addition, the effects of enhanced UV‐B radiation on leaf area, total biomass and UV‐B absorbing compounds were closely related to the nutrient status. Our results indicated that P. kangdingensis, which originates from the altitude of 3500 m and is apparently adapted to low‐nutrient and high‐UV‐B habitats, exhibits better tolerance to enhanced UV‐B radiation and greater growth under low‐nutrient availability than does P. cathayana originating from the altitude of 1500 m.     

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.     

Comparative physiological,ultrastructural and proteomic analyses reveal sexual differences in the responses of Populus cathayana under drought stress

Drought is a major abiotic stress, limiting the survival and growth of young plants. However, little is known about sex‐dependent responses to drought at the proteome level. In this study, we carried out investigations on comparative proteomics, combined with physiological and organelle structure analyses, in males and females of Populus cathayana Rehd. Three‐month‐old poplar cuttings were treated at 30% of field capacity and at 100% field capacity as a control in a greenhouse for 40 days. Drought greatly inhibited plant growth, damaged the photosynthetic system and destructed the structures of chloroplasts, mitochondria and cellular membranes. However, males suffered less from the adverse effects of drought than did females. Using 2‐DE, 563 spots were detected, of which 64 spots displayed significant drought effect and 44 spots displayed a significant sex by drought interaction effect. The results suggest that the different responses to drought stress detected between the sexes have a close relationship to the changes in the expression of sex‐dependent proteins, including, e.g. photosynthesis‐related proteins, homeostasis‐related proteins and stress response proteins. These proteins could contribute to a physiological advantage under drought, giving potential clues for understanding sexual differences in the performance of plants in different environments.     

Foliar application of the triterpene derivative 24-methylen-elemo-lanosta-8,24-dien-3-one alleviates salt toxicity in grapevine

This work focused on the effect triterpene derivative 24-methylen-elemo-lanosta-8,24-dien-3-one (F3) on the induction of salt stress tolerance of the Moroccan grapevine cv. “Doukkali”. Hardwood cuttings of the grapevine from a homogeneous plant material collected in the field were grown in hydroponic medium under different salt concentrations and treated with 50 or 100 µg ml^?1 of F3. Salt stress affected several physiological and biochemical parameters including relative water content, chlorophyll a and b content, peroxidase, and polyphenol oxidase activities, which decreased along with time. Meanwhile, proline, proteins, soluble sugars, H₂O₂, and carotenoid content, as well as phenolic compound content increased, suggesting an evidence of tolerance of this local variety to salinity. An exogenous supply of the triterpenic product increased all these parameters under normal conditions. In addition, F3 at low dose was found to be successful in lowering Na⁺ content and alleviating the inhibitory effects of salt stress on relative water content as well as on chlorophyll a and b.     

Alleviation of salt stress in Lotus glaber by Glomus intraradices

Lotus glaber is a glycophytic, perennial legume from Europe that occurs widely in saline habitats. We evaluated the effect of mycorrhizal fungus colonization on the response to salt stress of two genotypes of L. glaber differing in their tolerance to salinity. The experiment consisted of a randomized block design with two factors: (1) mycorrhizal fungus treatments (with or without AM fungus) and (2) two salinity levels of 0 and 200 mM NaCl. Our results indicated that Glomus intraradices established a more efficient symbiosis with the tolerant than with the sensitive genotype. G. intraradices improved growth of L. glaber plants under saline conditions. They showed higher values of net growth, shoot/root and K⁺/Na⁺ ratios, and protein concentrations than controls. Tolerant AM plants also showed higher chlorophyll levels than non-AM ones. Prevention of Na⁺ accumulation in the plant and enhancement of K⁺ concentrations in roots observed in this work could be part of the general mechanism of salt stress alleviation of L. glaber by G. intraradices.     

Growth,ionic response,and gene expression of shoots and roots of perennial ryegrass under salinity stress

Growth, ionic responses, and expression of candidate genes to salinity stress were examined in two perennial ryegrass accessions differing in salinity tolerance. The salinity tolerant (PI265349) and sensitive accessions (PI231595) were subjected to 75-mM NaCl for 14 days in a growth chamber. Across two accessions, salinity stress increased shoot dry weight and concentrations of malondialdehyde (MDA) and Na⁺ in the shoots and roots, but decreased shoot Ca²⁺ and root K⁺ concentrations. Salinity stress also increased root expressions of SOS1, PIP1, and TIP1. Plant height and chlorophyll content were unaffected by salinity stress in the tolerant accession but significantly decreased in the sensitive accession. Shoot MDA content did not change in the tolerant accession but increased in the sensitive accession. A more dramatic increase in Na⁺ was found in the roots of the sensitive accession. Relative to the control, salinity stress reduced expression of SOS1, NHX1, PIP1, and TIP1 in the shoots but increased expression of these genes in the roots of the tolerant accession. Expression levels of SOS1 increased in the roots and expression of NHX1 increased in the shoots but decreased in the roots of the sensitive accession under salinity stress. A decline in PIP1 expression in the shoots and dramatic increases in TIP expression in both shoots and roots were found in the sensitive accession under salinity stress. The results suggested maintenance of plant growth and leaf chlorophyll content, lesser Na⁺ accumulation in the roots, and lower lipid peroxidation in the shoots which could be associated with salinity tolerance. The decreased expressions of SOS1, NHX1, and TIP1 in the shoots, and increased expressions of NHX1 and PIP1 in the roots might also be related to salinity tolerance in perennial ryegrass.     

Responses to nonaeration and/or salinity stress in hydroponically cultured <Emphasis Type="Italic">Populus nigra</Emphasis> and <Emphasis Type="Italic">Populus alba</Emphasis> cuttings

Growth, photosynthesis, and Na⁺, K⁺, Ca²⁺, and Mg²⁺ distributions were examined in two-year-old hydroponically cultured Populus nigra and Populus alba cuttings exposed to salt stress (0, 50, or 100 mM NaCl) for four or six weeks and to nonaeration stress for one or three weeks, followed by a three-week aeration period in 2/5 Hoagland solution. Salt stress with 100 mM NaCl totally inhibited height increase in P. nigra cuttings. Combined salinity and nonaeration inhibited height increase to a greater degree than either stress alone in both species. Simple salt stress did not affect diameter increase in P. alba, whereas combined high salinity (100 mM NaCl) and nonaeration inhibited diameter increase. Growth and biomass accumulation were more sensitive to salt stress in P. nigra cuttings than in P. alba, although P. alba showed a more rapid decrease in photosynthesis in response to nonaeration stress. Ion distributions in the leaves and roots differed between species. P. alba was superior to P. nigra in terms of Na⁺ exclusion capacity, such that most of the absorbed Na⁺ was confined to the root system, with little reaching the leaves. The distributions of K⁺, Ca²⁺, and Mg²⁺ in the leaves and roots of each species under the two stressors were also analyzed. The lower Na⁺/K⁺ ratio in leaves indicated that P. alba was more tolerant to salt stress than P. nigra.     

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.     

The over-expression of Chrysanthemum crassum CcSOS1 improves the salinity tolerance of chrysanthemum

Soil salinity represents a major constraint on plant growth. Here, we report that the over-expression of the Chrysanthemum crassum plasma membrane Na⁺/H⁺ antiporter gene CcSOS1, driven by the CaMV 35S promoter, improved the salinity tolerance of chrysanthemum ‘Jinba’. In salinity-stressed transgenic plants, both the proportion of the leaf area suffering damage and the electrical conductivity of the leaf were lower in the transgenic lines than in salinity-stressed wild type plants. After a 6 day exposure to 200 mM NaCl, the leaf content of both chlorophyll (a+b) and proline was higher in the transgenic than in the wild type plants. The activity of both superoxide dismutase and peroxidase was higher in the transgenic than in the wild type plants throughout the period of NaCl stress. The transgenic plants had a stronger control over the ingress of Na⁺ into the plant, particularly with respect to the youngest leaves, and so maintained a more favorable K⁺/Na⁺ ratio. The result suggests that a possible strategy for improving the salinity tolerance of chrysanthemum could target the restriction of Na⁺ accumulation. This study is the first to report the transgenic expression of a Na⁺ efflux carrier in chrysanthemum.