Effect of Arbuscular Mycorrhizal Fungi and Rock Phosphate on Growth, Physiology, and Biochemistry of Carob under Water Stress and after Rehydration in Vermicompost-Amended Soil.

In the Mediterranean region, reforestation programs record failures following successive drought periods. The use of different plant-growth-promoting amendments and the understanding of drought-induced physiological and biochemical responses of carob will contribute to the reforestation program's success. In this study, the effects of arbuscular-mycorrhizal-fungi (AMF), vermicompost (VC), and rock phosphate (RP) on carob seedlings under drought stress (DS) and recovery (REC) conditions were evaluated. A greenhouse experiment was conducted with carob seedlings grown in the presence of AMF, VC, and RP, applied alone or in combination under well-watered (WW), DS (by stopping irrigation for 12 days), and recovery (REC) conditions. The obtained results indicated that the triple combination (AMF + VC + RP) presented the highest improvement in water potential, photosynthetic pigment content, stomatal conductance, and chlorophyll fluorescence compared to the controls under DS and after REC. In addition, this combination resulted in improved tolerance of carob seedlings to DS and a high potential for rapid recovery after rehydration due to a high accumulation of sugars, proteins, and antioxidant enzymes. In summary, the results underline the importance of inoculating carob with AMF in combination with (in)-organic amendments in improving its tolerance to DS and its recovery performances.

The Effect of Chitosan on Plant Physiology, Wound Response, and Fruit Quality of Tomato.

In agriculture, chitosan has become popular as a metabolic enhancer; however, no deep information has been obtained yet regarding its mechanisms on vegetative tissues. This work was conducted to test the impact of chitosan applied at different plant growth stages on plant development, physiology, and response to wounding as well as fruit shape and composition. Five concentrations of chitosan were tested on tomato. The most effective chitosan doses that increased leaf number, leaf area, plant biomass, and stomatal conductance were 0.75 and 1 mg mL-1. Chitosan (1 mg mL-1) applied as foliar spray increased the levels of jasmonoyl-isoleucine and abscisic acid in wounded roots. The application of this dose at vegetative and flowering stages increased chlorophyll fluorescence (Fv/Fm) values, whereas application at the fruit maturation stage reduced the Fv/Fm values. This decline was positively correlated with fruit shape and negatively correlated with the pH and the content of soluble sugars, lycopene, total flavonoids, and nitrogen in fruits. Moreover, the levels of primary metabolites derived from glycolysis, such as inositol phosphate, lactic acid, and ascorbic acid, increased in response to treatment of plants with 1 mg mL-1- chitosan. Thus, chitosan application affects various plant processes by influencing stomata aperture, cell division and expansion, fruit maturation, mineral assimilation, and defense responses.

Assemblage of indigenous arbuscular mycorrhizal fungi and green waste compost enhance drought stress tolerance in carob (Ceratonia siliqua L.) trees.

In the current study, an eco-friendly management technology to improve young carob (Ceratonia siliqua L.) tree tolerance to water deficit was set up by using single or combined treatments of arbuscular mycorrhizal fungi (AMF) and/or compost (C). Two groups of young carob have been installed: (i) carob cultivated under well-watered conditions (WW; 70% field capacity (FC)) and (ii) where the plants were drought-stressed (DS; 35% FC) during 2, 4, 6, and 8 months. The effect of used biofertilizers on the course of growth, physiological (photosynthetic traits, water status, osmolytes, and mineral content), and biochemical (hydrogen peroxide (H2O2), oxidative damage to lipids (malondialdehyde (MDA), and membrane stability (MS)) traits in response to short- and long-term droughts were assessed. The dual application of AMF and C (C + AMF) boosted growth, physiological and biochemical parameters, and nutrient uptake in carob under WW and DS. After eight months, C + AMF significantly enhanced stomatal conductance by 20%, maximum photochemical efficiency of PSII by 7%, leaf water potential by 23%, chlorophyll and carotenoid by 40%, plant uptake of mineral nutrients (P by 75%, N by 46%, K+ by 35%, and Ca2+ by 40%), concentrations of soluble sugar by 40%, and protein content by 44% than controls under DS conditions. Notably, C + AMF reduced the accumulation of H2O2 and MDA content to a greater degree and increased MS. In contrast, enzyme activities (superoxide dismutase, catalase, peroxidase, and polyphenoloxidase) significantly increased in C + AMF plants under DS. Overall, our findings suggest that the pairing of C + AMF can mediate superior drought tolerance in young carob trees by increasing leaf stomatal conductance, cellular water content, higher solute concentration, and defense response against oxidative damage during the prolonged period of DS.

Potential of Native Arbuscular Mycorrhizal Fungi, Rhizobia, and/or Green Compost as Alfalfa (Medicago sativa) Enhancers under Salinity.

Salinity is one of the devastating abiotic stresses that cause reductions in agricultural production. The increased salinization affects alfalfa growth, metabolism, and rhizobium capacity for symbiotic N2 fixation negatively. This study was undertaken to investigate the efficiency of green compost (C; made from green waste), arbuscular mycorrhizal fungi (M; field-sourced native consortium), and/or rhizobium (R; a salt-tolerant rhizobium strain) individually or in combination as an effective strategy to improve alfalfa productivity under non-saline and high-saline (120 mM NaCl) conditions. In addition, we aimed to understand the agro-physiological and metabolic basis as well as glomalin content in the soil of biofertilizers-induced salt tolerance in alfalfa. Here, we show that mycorrhizal infection was enhanced after MR inoculation, while C application decreased it significantly. Salinity reduced growth, physiological functioning, and protein concentration, but the antioxidant system has been activated. Application of the selected biofertilizers, especially C alone or combined with M and/or R improved alfalfa tolerance. The tri-combination CMR mitigated the negative effects of high salinity by stimulating plant growth, roots and nodules dry matters, mineral uptake (P, N, and K), antioxidant system, synthesis of compatible solutes, and soil glomalin content, sustaining photosynthesis-related performance and decreasing Na+ and Cl- accumulation, lipid peroxidation, H2O2 content, and electrolyte leakage.

Arbuscular Mycorrhizal Fungi Mediate Drought Tolerance and Recovery in Two Contrasting Carob (Ceratonia siliqua L.) Ecotypes by Regulating Stomatal, Water Relations, and (In)Organic Adjustments.

Irregular precipitation and drought caused an increase in tree mortality rates in multiple forest biomes with alterations in both ecosystem services and carbon balance. Carob (Ceratonia siliqua) growth and production in arid and semi-arid ecosystems are likely affected by climate change-induced droughts. Understanding the physiological responses of drought-induced early-stage tree death and strategies to enhance drought tolerance and optimize growth will help tree improvement programs. Mycorrhizal inoculation has a pronounced impact on plant growth, water absorption, mineral nutrition, and protection from abiotic stresses. However, a better understanding of these complex interconnected cellular processes and arbuscular mycorrhizal fungi (AMF)-mediated mechanisms regulating drought tolerance in plants will enhance its potential application as an efficient approach for bio-amelioration of stresses. The objectives of this work were to elucidate the different effects of autochthone AMF on inorganic solute and water content uptakes, organic adjustments (sugar and proteins content), leaf gas exchange (stomatal conductance and efficiency of photosystems I and II), and oxidative damage of two contrasting ecotypes of carob seedlings: coastal (southern ecotype (SE)) and in-land (northern ecotype (NE)) under control (C), drought (by cessation of irrigation for 15 days (15D)), and recovery (R) conditions. Our findings showed that AMF promoted growth, nutrient content, and physiological and biochemical parameters in plants of both ecotypes during C, 15D, and R conditions. After four days of recovery, stomatal conductance (gs), the maximum photochemical efficiency of PSII (Fv/Fm), water content, and plant uptake of mineral nutrients (P, K, Na, and Ca) were significantly higher in shoots of mycorrhizal (AM) than non-mycorrhizal (NM) control plants. Consequently, AMF reduced to a greater degree the accumulation of hydrogen peroxide (H2O2) and oxidative damage to lipid (malondialdehyde (MDA)) content in AM than NM plants in NE and SE, after recovery. Altogether, our findings suggest that AMF can play a role in drought resistance of carob trees at an early stage by increasing the inorganic solutes (P, K, Na, and Ca), water content uptake, organic solutes (soluble sugars and protein content), stomatal conductance, and defense response against oxidative damage during re-watering after drought stress.

Foliar Application of Chitosan Increases Tomato Growth and Influences Mycorrhization and Expression of Endochitinase-Encoding Genes.

Nowadays, applying bio-organic fertilizer (e.g., chitosan, Ch) or integrating beneficial microorganisms (e.g., arbuscular mycorrhizal fungi, AMF) are among the successful strategies to promote plant growth. Here, the effect of two application modes of Ch (foliar spray or root treatment) and Ch-derived nanoparticles (NPs) on tomato plants colonized with the AMF Rhizophagus irregularis were analyzed, thereby focusing on plant biomass, flowering and mycorrhization. An increase of shoot biomass and flower number was observed in arbuscular mycorrhizal (AM) plants sprayed with Ch. The interaction with AMF, however, was reduced as shown by decreased mycorrhization rates and AM-specific gene expression. To get insights into Ch effect on mycorrhization, levels of sugars, jasmonates, abscisic acid, and the expression of two chitinase-encoding genes were determined in mycorrhizal roots. Ch had no effect on sugar and phytohormone levels, but the reduced mycorrhization was correlated with down- and upregulated expression of Chi3 and Chi9, respectively. In contrast, application of NPs to leaves and Ch applied to the soil did not show any effect, neither on mycorrhization rate nor on growth of mycorrhizal plants. Concluding, Ch application to leaves enhanced plant growth and flowering and reduced interaction with AMF, whereas root treatment did not affect these parameters.

Uprising the antioxidant power of Argania spinosa L. callus through abiotic elicitation.

This study was carried out in order to investigate the ability of tissues of Argania spinosa (L.) to undergo unlimited cell divisions by triggering their proliferative potential via callogenesis. Axenic cultures were efficiently established using axillary buds cultured on half-strength Murashige and Skoog (MS) medium after 20min of surface sterilization with sodium hypochlorite 6% (v/v). The highest callus rate was achieved with 1.0mgL-1 of naphthaleneacetic acid (NAA) and 1.0mgL-1 of 2,4-dichlorophenoxyacetic acid (2,4D) or similarly with 0.01mgL-1 of 6-benzylaminopurine (BAP) and 1.0mgL-1 of 2,4D at pH of 5.8, under dark conditions. The results of this study show also a significant increase in the callus's antioxidant power under abiotic pressure induced by NaCl. Catalase (CAT), peroxidase (PO), and superoxide dismutase (SOD) activities were significantly triggered, which protected the cells from the stimulated oxidative stress, under hydrogen peroxide (H2O2) significant release. This reaction favors subsequently the tissue recover process linked to the low abundance of polyphenol oxidase (PPO) activity and malondialdehyde (MDA) content. This work proves the efficiency of salt stress in boosting the argan cell's antioxidant status, which could be commercially applied in the field of cells regenerative therapy.

An Assessment of Genetic Diversity and Drought Tolerance in Argan Tree (Argania spinosa) Populations: Potential for the Development of Improved Drought Tolerance.

The argan tree (Argania spinosa) occurs in a restricted area of Southwestern Morocco characterized by low water availability and high evapotranspirative demand. Despite the adaptation of the argan tree to drought stress, the extent of the argan forest has declined markedly due to increased aridity, land use changes and the expansion of olive cultivation. The oil of the argan seed is used for cooking and as the basis for numerous cosmetics. The identification of argan tree varieties with enhanced drought tolerance may minimize the economic losses associated with the decline of the argan forest and constrain the spread of desertification. In this study we collected argan ecotypes from four contrasting habitats and grew them under identical controlled environment conditions to investigate their response to drought. Leaf gas exchange analysis indicated that the argan ecotypes showed a high degree of adaptation to drought stress, maintaining photosynthetic activity at low levels of foliar water content and co-ordinating photosynthesis, stomatal behavior and metabolism. The stomata of the argan trees were highly sensitive to increased leaf to air vapor pressure deficit, representing an adaptation to growth in an arid environment where potential evapotranspiration is high. However, despite originating in contrasting environments, the four argan ecotypes exhibited similar gas exchange characteristics under both fully irrigated and water deficit conditions. Population genetic analyses using microsatellite markers indicated a high degree of relatedness between the four ecotypes; indicative of both artificial selection and the transport of ecotypes between different provinces throughout centuries of management of the argan forest. The majority of genetic variation across the four populations (71%) was observed between individuals, suggesting that improvement of argan is possible. Phenotypic screening of physiological responses to drought may prove effective in identifying individuals and then developing varieties with enhanced drought tolerance to enable the maintenance of argan production as climate change results in more frequent and severe drought events in Northern Africa.

Abscisic Acid Induces Rapid Reductions in Mesophyll Conductance to Carbon Dioxide.

The rate of photosynthesis (A) of plants exposed to water deficit is a function of stomatal (gs) and mesophyll (gm) conductance determining the availability of CO2 at the site of carboxylation within the chloroplast. Mesophyll conductance often represents the greatest impediment to photosynthetic uptake of CO2, and a crucial determinant of the photosynthetic effects of drought. Abscisic acid (ABA) plays a fundamental role in signalling and co-ordination of plant responses to drought; however, the effect of ABA on gm is not well-defined. Rose, cherry, olive and poplar were exposed to exogenous ABA and their leaf gas exchange parameters recorded over a four hour period. Application with ABA induced reductions in values of A, gs and gm in all four species. Reduced gm occurred within one hour of ABA treatment in three of the four analysed species; indicating that the effect of ABA on gm occurs on a shorter timescale than previously considered. These declines in gm values associated with ABA were not the result of physical changes in leaf properties due to altered turgor affecting movement of CO2, or caused by a reduction in the sub-stomatal concentration of CO2 (Ci). Increased [ABA] likely induces biochemical changes in the properties of the interface between the sub-stomatal air-space and mesophyll layer through the actions of cooporins to regulate the transport of CO2. The results of this study provide further evidence that gm is highly responsive to fluctuations in the external environment, and stress signals such as ABA induce co-ordinated modifications of both gs and gm in the regulation of photosynthesis.

On the use of leaf spectral indices to assess water status and photosynthetic limitations in Olea europaea L. during water-stress and recovery.

Diffusional limitations to photosynthesis, relative water content (RWC), pigment concentrations and their association with reflectance indices were studied in olive (Olea europaea) saplings subjected to water-stress and re-watering. RWC decreased sharply as drought progressed. Following rewatering, RWC gradually increased to pre-stress values. Photosynthesis (A), stomatal conductance (gs), mesophyll conductance (gm), total conductance (gt), photochemical reflectance index (PRI), water index (WI) and relative depth index (RDI) closely followed RWC. In contrast, carotenoid concentration, the carotenoid to chlorophyll ratio, water content reflectance index (WCRI) and structural independent pigment index (SIPI) showed an opposite trend to that of RWC. Photosynthesis scaled linearly with leaf conductance to CO2; however, A measured under non-photorespiratory conditions (A1%O2) was approximately two times greater than A measured at 21% [O2], indicating that photorespiration likely increased in response to drought. A1%O2 also significantly correlated with leaf conductance parameters. These relationships were apparent in saturation type curves, indicating that under non-photorespiratory conditions, CO2 conductance was not the major limitations to A. PRI was significant correlated with RWC. PRI was also very sensitive to pigment concentrations and photosynthesis, and significantly tracked all CO2 conductance parameters. WI, RDI and WCRI were all significantly correlated with RWC, and most notably to leaf transpiration. Overall, PRI correlated more closely with carotenoid concentration than SIPI; whereas WI tracked leaf transpiration more effectively than RDI and WCRI. This study clearly demonstrates that PRI and WI can be used for the fast detection of physiological traits of olive trees subjected to water-stress.

Partial root zone drying: regulation of photosynthetic limitations and antioxidant enzymatic activities in young olive (Olea europaea) saplings.

The effect of partial root drying (PRD) irrigation on split-root olive (Olea europaea L. cv Picholine marocaine) saplings was investigated. An irrigated control and two PRD regimes were applied (control: irrigation applied on both sides of the root system to keep the soil water content close to field capacity; PRD(50): irrigation applied at 50% of the control amount on one side of the root system and irrigation withheld from the other side, with irrigation regimes switched between the sides of the root system every 2 weeks; and PRD(100): irrigation applied at 100% of the control amount on one side and irrigation withheld on the other side, with irrigation regimes switched between the sides of the root system every 2 weeks. Only saplings in the PRD(50) regime were subjected to water-deficit irrigation. The PRD treatments significantly affected water relations and vegetative growth throughout the growing season. Predawn leaf water potential and relative water content differed significantly between the PRD(50) and PRD(100) saplings, leading to reduced stomatal conductance, carbon assimilation, shoot length and leaf number in PRD(50) saplings. However, the PRD(50) water-deficit treatment did not affect the capacity of the saplings to assimilate CO(2). Activities of superoxide dismutase, soluble and insoluble peroxidase (POX) and polyphenol oxidase were up-regulated by the PRD(50) and PRD(100) treatments compared with control values. The higher activities of both soluble and insoluble POX observed in PRD(50) saplings may reflect the greater inhibitory effect of this treatment on vegetative growth. Up-regulation of the detoxifying systems in the PRD(100) and PRD(50) saplings may have provided protection mechanisms against irreversible damage to the photosynthetic machinery, thereby allowing the photosynthetic apparatus to function and preventing the development of severe water stress. We also measured CO(2) assimilation rate/internal leaf CO(2) concentration (A/C(i)) after exposing the attached leaves to very low [CO(2)] (approximately 50 micromol mol(-1)) to force stomatal opening. These results confirmed that, under conditions of moderate water stress, the sum of the diffusional resistances (i.e., stomatal and mesophyll resistances) sets the limit to photosynthetic rates. Assessing photosynthetic capacity without removing the diffusional limitations may lead to an overestimation of the biochemical limitations to photosynthesis in sclerophyllous plants.

[Pigmented villonodular synovitis]. / Synovite villonodulaire.

OBJECTIVE: To evaluate clinical, radiological and histological characteristics as well as outcome of pigmented villonodular synovitis (PVNS) in 14 patients. METHODS: This retrospective study examined data from 14 cases of pathologically-confirmed PVNS over a 10-year period (1990-1999). RESULTS: All 14 cases (9 women and 5 men) of PVNS involved the knee. Mean age at diagnosis was 32.7 years. The principal functional signs were mechanical pain and disability. All patients had radiographs (normal for 8 patients); 3 had arthrography, which found diffuse cyst formations, 2 had MRI, which showed images characteristic of PVNS, and 4 had arthroscopy. All patients underwent complete surgical excision, and 4 also had osmic acid synoviorthesis. Recurrence occurred in 4 patients, within 10 months on average. CONCLUSION: MRI and arthroscopy improve the early management of pigmented villonodular synovitis.

Tuberculous sacroiliitis. Four cases.

Tuberculous sacroiliitis with no local abscesses or tuberculosis at other sites occurred in four patients, three men and one woman with a mean age of 42 years. Slow progression characterized this uncommon variant of joint tuberculosis. Two patients reported contact with family members known to have tuberculosis. Mean time from symptom onset to presentation was 14 months. Surgical biopsy showed epithelioid and giant-cell granulomas with caseous necrosis; culturing on Lowenstein-Jensen medium recovered the tubercle bacillus. Treatment was with antitubercular drugs, and protection from weight bearing for 4 weeks. These four cases illustrate the slowly progressive course of the clinical and radiological manifestations and underline the diagnostic difficulties met in the early stages of tuberculous sacroiliitis without tuberculosis at other sites. Antitubercular treatment can restore joint function provided the diagnosis is made before radiological destruction occurs.