Light-induced fluorescence decay during the greening of normal and lincomycin-treated maize leaves

Light-induced fluorescence decay was examined during the greening of control and lincomycintreated maize (Zea mays L.) leaves. Assuming that this decay to a first approximation is the result of two parallel first-order reactions, the fluorescence induction curves were linearized on the logarithm plot and the parameters were determined. The variable fluorescence increased, and the parameters of the two linear sections of the fluorescence decay—that is, the kinetics of the induction curves—changed during the greening of the control leaves. Lincomycin treatment caused some chlorophyll deficiency and the lowering of the chlorophyll a/b ratio, changed the fluorescence emission spectra and the effect of Mg²⁺ on the regulation of the excitation energy distribution. The structure of the thylakoids and the kinetics of the fluorescence decay were also changed in the treated leaves. The possible relationship between the change of the kinetics of the fluorescence decay and the change of spillover during greening and after lincomycin treatment is discussed.Abbreviations LHC light-harvesting complex - Chl chlorophyll - LM lincomycin - PS photosystem - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea     

Fluorescence emission spectra of Photosystem I,Photosystem II and the light-harvesting chlorophyll a/b complex of higher plants

Fluorescence emission spectra excited at 514 and 633 nm were measured at ?196 °C on dark-grown bean leaves which had been partially greened by a repetitive series of brief xenon flashes. Excitation at 514 nm resulted in a greater relative enrichment of the 730 nm emission band of Photosystem I than was obtained with 633 nm excitation. The difference spectrum between the 514 nm excited fluorescence and the 633 nm excited fluorescence was taken to be representative of a pure Photosystem I emission spectrum at ?196 °C. It was estimated from an extrapolation of low temperature emission spectra taken from a series of flashed leaves of different chlorophyll content that the emission from Photosystem II at 730 nm was 12% of the peak emission at 694 nm. Using this estimate, the pure Photosystem I emission spectrum was subtracted from the measured emission spectrum of a flashed leaf to give an emission spectrum representative of pure Photosystem II fluorescence at ?196 °C. Emission spectra were also measured on flashed leaves which had been illuminated for several hours in continuous light. Appreciable amounts of the light-harvesting chlorophyll a/b protein, which has a low temperature fluorescence emission maximum at 682 nm, accumulate during greening in continuous light. The emission spectra of Photosystem I and Photosystem II were subtracted from the measured emission spectrum of such a leaf to obtain the emission spectrum of the light-harvesting chlorophyll a/b protein at ?196 °C.     

Light-harvesting chlorophyll a-b complex requirement for regulation of Photosystem II photochemistry by non-photochemical quenching

Recently, it has been suggested (Horton et al. 1992) that aggregation of the light-harvesting a-b complex (LHC II) in vitro reflects the processes which occur in vivo during fluorescence induction and related to the major non-photochemical quenching (qE). Therefore the requirement of this chlorophyll a-b containing protein complex to produce qN was investigated by comparison of two barley mutants either lacking (chlorina f2) or depressed (chlorina¹⁰⁴) in LHC II to the wild-type and pea leaves submitted to intermittent light (IL) and during their greening in continuous light. It was observed that qN was photoinduced in the absence of LHC II, i.e. in IL grown pea leaves and the barley mutants. Nevertheless, in these leaves qN had no (IL, peas) or little (barley mutants) inhibitory effect on the photochemical efficiency of Q_A reduction measured by flash dosage response curves of the chlorophyll fluorescence yield increase induced by a single turn-over flash During greening in continuous light of IL pea leaves, an inhibitory effect on Q_A photoreduction associated to qN developed as Photosystem II antenna size increased with LHC II synthesis. Utilizing data from the literature on connectivity between PS II units versus antenna size, the following hypothesis is put forward to explain the results summarized above. qN can occur in the core antenna or Reaction Center of a fraction of PS II units and these units will not exhibit variable fluorescence. Other PS II units are quenched indirectly through PS II-PS II exciton transfer which develops as the proportion of connected PS II units increases through LHC II synthesis.     

Spectral and kinetic parameters of phosphorescence of triplet chlorophyll a in the photosynthetic apparatus of plants

Spectral and kinetic parameters and quantum yield of IR phosphorescence accompanying radiative deactivation of the chlorophyll a (Chl a) triplet state were compared in pigment solutions, greening and mature plant leaves, isolated chloroplasts, and thalluses of macrophytic marine algae. On the early stages of greening just after the Shibata shift, phosphorescence is determined by the bulk Chl a molecules. According to phosphorescence measurement, the quantum yield of triplet state formation is not less than 25%. Further greening leads to a strong decrease in the phosphorescence yield. In mature leaves developing under normal irradiation conditions, the phosphorescence yield declined 1000-fold. This parameter is stable in leaves of different plant species. Three spectral forms of phosphorescence-emitting chlorophyll were revealed in the mature photosynthetic apparatus with the main emission maxima at 955, 975, and 995 nm and lifetimes ～1.9, ～1.5, and 1.1–1.3 ms. In the excitation spectra of chlorophyll phosphorescence measured in thalluses of macrophytic green and red algae, the absorption bands of Chl a and accessory pigments — carotenoids, Chl b, and phycobilins — were observed. These data suggest that phosphorescence is emitted by triplet chlorophyll molecules that are not quenched by carotenoids and correspond to short wavelength forms of Chl a coupled to the normal light harvesting pigment complex. The concentration of the phosphorescence-emitting chlorophyll molecules in chloroplasts and the contribution of these molecules to chlorophyll fluorescence were estimated. Spectral and kinetic parameters of the phosphorescence corresponding to the long wavelength fluorescence band at 737 nm were evaluated. The data indicate that phosphorescence provides unique information on the photophysics of pigment molecules, molecular organization of the photosynthetic apparatus, and mechanisms and efficiency of photodynamic stress in plants.     

Fluorescence emission spectra of photosystem I, photosystem II and the light-harvesting chlorophyll a/b complex of higher plants

Fluorescence emission spectra excited at 514 and 633 nm were measured at -196 degrees C on dark-grown bean leaves which had been partially greened by a repetitive series of brief xenon flashes. Excitation at 514 nm resulted in a greater relative enrichment of the 730 nm emission band of Photosystem I than was obtained with 633 nm excitation. The difference spectrum between the 514 nm excited fluorescence and the 633 nm excited fluorescence was taken to be representative of a pure Photosystem I emission spectrum at -196 degrees C. It was estimated from an extrapolation of low temperature emission spectra taken from a series of flashed leaves of different chlorophyll content that the emission from Photosystem II at 730 nm was 12% of the peak emission at 694 nm. Using this estimate, the pure Photosystem I emission spectrum was subtracted from the measured emission spectrum of a flashed leaf to give an emission spectrum representative of pure Photosystem II fluorescence at -196 degrees C. Emission spectra were also measured on flashed leaves which had been illuminated for several hours in continuous light. Appreciable amounts of the light-harvesting chlorophyll a/b protein, which has a low temperature fluorescence emission maximum at 682 nm, accumulate during greening in continuous light. The emission spectra of Photosystem I and Photosystem II were subtracted from the measured emission spectrum of such a leaf to obtain the emission spectrum of the light-harvesting chlorophyll a/b protein at -196 degrees C.     

Development of the Primary Photochemical Apparatus of Photosynthesis during Greening of Etiolated Bean Leaves

Seven-day-old dark-grown bean leaves were greened under continuous light. The amount of chlorophyll, the ratio of chlorophyll a to chlorophyll b, the O₂ evolving capacity and the primary photochemical activities of Photosystem I and Photosystem II were measured on the leaves after various times of greening. The primary photochemical activities were measured as the photo-oxidation of P₇₀₀, the photoreduction of C-550, and the photo-oxidation of cytochrome b₅₅₉ in intact leaves frozen to −196 C. The results indicate that the reaction centers of Photosystem I and Photosystem II begin to appear within the first few minutes and that Photosystem II reaction centers accumulate more rapidly than Photosystem I reaction centers during the first few hours of greening. The very early appearances of the primary photochemical activity of Photosystem II was also confirmed by light-induced fluorescence yield measurements at −196 C.     

The Early Stages of Photosystem II Assembly Monitored by Measurements of Fluorescence Lifetime, Fluorescence Induction and Isoelectric Focusing of Chlorophyll-Proteins in Barley Etiochloroplasts

The relationship between functional and structural aspects ofPSII formation during greening of etiolated barley leaves hasbeen investigated using fluorescence lifetime measurements,fluorescence kinetics analysis and analysis of chlorophyll-proteincomplexes by IEF-PAGE. Two phases of different character couldbe distinguished in the course of the greening process in dark-grownplants. An early phase covering the first 3–4 h afterthe onset of illumination and a late phase covering the subsequentgreening. During the first phase the formation of PSII reactioncenters and their minor antenna components was observed as manifestedby the IEF-PAGE polypeptide pattern. This was accompanied byshortening of the slow and middle components of the fluorescencelifetime, as well as by the rapid drop in the amplitude of theslow component. A room temperature emission band at 676 nm wasassociated with uncoupled chlorophyll and with the slow fluorescencelifetime component during the first hours of greening. Duringthe late greening phase peripheral light-harvesting complexesof PSII were formed concomitantly to an increase in lifetimeand amplitude of the fast component and to a further decreasein the lifetime of the middle component. The gradual increasein PSII complexity during both phases of greening was also manifestedby changes in proportion and kinetic properties of PSII     

Discrete character of the development of the photosynthetic apparatus in greening barley leaves

Biogenesis of the photosynthetic apparatus in greening etiolated leaves of barley (Hordeum vulgare L) was investigated by an approach permitting investigation of this process under conditions that minimize differences in plastid development. Distributions of barley leaves greening for 24 h as to chlorophyll content and of chloroplast grana as to number of thylakoids were shown to be of a multimodal character. The shape of time-course curves of chlorophyll accumulation in local sites of greening etiolated leaves was of a stepped or (at the end of greening) undulated character. The stepwise accumulation of chlorophyll was accompanied by wave-like changes in chlorophyll b/a ratio, intensity of low-temperature chlorophyll fluorescence and photosynthetic activity with minima at the time points of transition to accelerated chlorophyll accumulation. It is assumed that (1) development of the photosynthetic apparatus in local sites of greening etiolated leaves occurs stepwise, from one steady level to another, but not as gradually as is generally accepted, and (2) every separate step in development of the photosynthetic apparatus seems to begin with formation of photosystem cores and to end with the synthesis of light-harvesting complexes. This revised version was published online in June 2006 with corrections to the Cover Date.     

Excitation kinetics of chlorophyll fluorescence during light-induced greening and establishment of photosynthetic activity of barley seedlings

Excitation kinetics based on feedback regulation of chlorophyll (Chl) fluorescence of leaves measured with the chlorophyll fluorometer, FluoroMeter Modul (FMM), are presented. These kinetics showed the variation of excitation light (laser power, LP) regulated by the feedback mechanism of the FMM, an intelligent Chl fluorometer with embedded computer, which maintains the fluorescence response constant during the 300-s transient between the dark- and lightadapted state of photosynthesis. The excitation kinetics exhibited a rise of LP with different time constants and fluctuations leading to a type of steady state. The variation of excitation kinetics were demonstrated using the example of primary leaves of etiolated barley seedlings (Hordeum vulgare L. cv. Barke) during 48 h of greening in the light with gradual accumulation of Chl and development of photosynthetic activity. The excitation kinetics showed a fast rise followed by a short plateau at ca. 30 s and finally a slow constant increase up to 300 s. Only in the case of 2 h of greening in the light, the curve reached a stable steady state after 75 s followed by a slight decline. The final LP value (at 300 s of illumination) increased up to 12 h of greening and decreased with longer greening times. The active feedback mechanism of the FMM adjusted the excitation light during the measurement to the actual photosynthetic capacity of the individual leaf sample. In this way, the illumination with excessive light was avoided. The novel excitation kinetics can be used to characterize health, stress, disease, and/or product quality of plant material.     

The chlorophyll fluorescence ratio F690/F730 in leaves of different chlorophyll content

The red laser-induced chlorophyll-fluorescence induction kinetics of predarkened leaf samples were registered simultaneously in the 690 and 730 nm regions i.e., in the region of the two chlorophyll fluorescence emission maxima. From the induction kinetics the chlorophyll fluorescence ratio F690/F730 was calculated. The ratio F690/F730 shows to be dependent on the chlorophyll content of leaves. It is significantly higher in needles of damaged spruces (values of 0.45–0.9) than in normal green needles of healthy trees (values of 0.35–0.5). During development and greening of maple leaves the ratio F690/F730 decreases with increasing chlorophyll content. Determination of the ratio F690/F730 can be a suitable method of monitoring changes in chlorophyll content in a non-destructive way in the same leaves during development or the yellowish-green discolouration of needles of damaged spruces in the Black Forest with the typical tree decline symptoms.Abbreviations F690/F730 ratio of the fluorescence yield at the two fluorescence-emission maxima in the 690 and 730 nm regions - F_m maximum fluorescence - F_s steady-state fluorescence     

The development of antenna complexes of barley (Hordeum vulgare cv. Akcent) under different light conditions as judged from the analysis of 77 K chlorophyll a fluorescence spectra

Using 77 K chlorophyll a (Chl a) fluorescence spectra in vivo, the development was studied of Photosystems II (PS II) and I (PS I) during greening of barley under intermittent light followed by continuous light at low (LI, 50 μmol m⁻² s⁻¹) and high (HI, 1000 μmol m⁻² s⁻¹) irradiances. The greening at HI intermittent light was accompanied with significantly reduced fluorescence intensity from Chl b excitation for both PS II (F685) and PS I (F743), in comparison with LI plants, indicating that assembly of light-harvesting complexes (LHC) of both photosystems was affected to a similar degree. During greening at continuous HI, a slower increase of emission from Chl b excitation in PS II as compared with PS I was observed, indicating a preferred reduction in the accumulation of LHC II. The following characteristics of 77 K Chl a fluorescence spectra documented the photoprotective function of an elevated content of carotenoids in HI leaves: (1) a pronounced suppression of Soret region of excitation spectra (410–450 nm) in comparison with the red region (670–690 nm) during the early stage of greening indicated a strongly reduced excitation energy transfer from carotenoids to the Chl a fluorescing forms within PS I and PS II; (2) changes in the shape of the excitation band of Chl b and carotenoids (460–490 nm) during greening under continuous light confirmed that the energy transfer from carotenoids to Chl a within PS II remained lower as compared with the LI plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

Development of Photochemical Activity and the Appearance of the High Potential Form of Cytochrome b-559 in Greening Barley Seedlings

The development of photochemical activity during the greening of dark-grown barley seedlings (Hordeum vulgare L. cv. Svalöfs Bonus) was studied in relation to the formation of the high potential form of cytochrome b-559 (cytochrome b-559_HP). Photosynthetic oxygen evolution from leaves was detected at 30 minutes of illumination. The rate of oxygen evolution per gram fresh weight of leaf was as high at 2 to 2.5 hours of greening as at 24 hours or in fully greened leaves. On a chlorophyll basis, the photosynthetic rate at 90 minutes of greening was 80-fold greater than the rate at 45 hours. It is concluded that the majority of photosynthetic units are functional at an early stage of greening, and that chlorophyll synthesis during greening serves to increase the size of the units.     

Physiological, anatomical and phenotypical effects of a cadmium stress in different-aged chlorophyllian organs of Myriophyllum alterniflorum DC (Haloragaceae)

Cadmium stress on aquatic macrophytes is currently few documented and no studies were conducted on clonal populations in controlled conditions. Our aim is to assess the effect of cadmium stress on Myriophyllum alterniflorum from physiological, anatomical and phenotypical parameters among different developmental stages. An experiment was carried out in order to test the hypothesis that a high external dose of Cd affects differentially young and old leaves of watermilfoil in terms of (1) anatomical and biological parameters, (2) physiological parameters and (3) that these differences result from the overall level of oxidative stress. Cadmium stress strongly affects the morphology of chlorophyllian organs and decreases the xylem-vessel number and diameter in matures leaves. Moreover we first report a new cadmium-induced anatomical feature: a double endodermis with separated layers which limits water and gaseous losses in mature leaves. From a physiological point of view, cadmium induces at all developmental stages a reduction in chlorophyll contents, a malondialdehyde production indicating a drastic lipoperoxydation, an enhanced production of reactive-oxygen-species scavengers (free proline and in lesser parts carotenoids) and a perturbation in hydric status in M. alterniflorum clones. The results obtained support the initial hypotheses. Indeed, it should be noted that cadmium stress mainly disturbs mature-leave structure and function, increasing their ageing and senescence. This mechanism probably contributes to limit the cadmium impact on young and growing parts as cadmium is known to preferentially accumulate in senescent leaves.     

Chloroplast Biogenesis: XXII. Contribution of Short Wavelength and Long Wavelength Protochlorophyll Species to the Greening of Higher Plants

The contribution of short and long wavelength membrane-bound fluorescing protochlorophyll species to the over-all process of chlorophyll formation was assessed during photoperiodic growth. Protochlorophyll forms were monitored spectrofluorometrically at 77 K during the first six light and dark cycles in homogenates of cucumber (Cucumis sativus L.) cotyledons grown under a 14-hour light/10-hour dark photoperiodic regime, and in cotyledons developing in complete darkness. In the etiolated tissue, short wavelength protochlorophyll having a broad emission maximum between 630 and 640 nm appeared within 24 hours after sowing. Subsequently, the long wavelength species fluorescing at 657 nm appeared, and accumulated rapidly. This resulted in the preponderance of the long wavelength species which characterizes the protochlorophyll profile of etiolated tissues. The forms of protochlorophyll present in etiolated cucumber cotyledons resembled those in etiolated bean leaves in their absorption, fluorescence, and phototransformability. A different pattern of protochlorophyll accumulation was observed during the dark cycles of photoperiodic greening. The short wavelength species appeared within 24 hours after sowing. Subsequently, the long wavelength form accumulated and disappeared. The long wavelength to short wavelength protochlorophyll emission intensity ratio reached a maximum (~3:1) during the second dark cycle, then declined during subsequent dark cycles. Short wavelength species were continuously present in the light and dark. Primary corn and bean leaves exhibited a similar pattern of protochlorophyll accumulation. In cucumber cotyledons, both the short and long wavelengths species appeared to be directly phototransformable at all stages of photoperiodic development. It thus appears that whereas the long wavelength protochlorophyll species is the major chlorophyll precursor during primary photoconversion in older etiolated tissues, both long wavelength and short wavelength species seem to contribute to chlorophyll formation during greening under natural photoperiodic conditions.     

Development of photosynthetic apparatus and respiration in pea seedlings during greening as influenced by toxic concentration of lead

Detached leaves of 14 day-old dark-grown pea seedlings were immersed with their cut ends either in water (control) or in 20 mM Pb(NO₃)₂ solution. They were exposed to continuous illumination during 24 and 48 h. The formation of PSII primary photochemistry in thylakoids was determined in vivo by measuring changes in values of parameters of chlorophyll a fast fluorescence kinetics: Fo, Fm, Fv, Fv/Fm and t 1/2. The amount of lead accumulation in leaves, content of chlorophylls and carotenoids and rates of CO₂ uptake in light and evolution in darkness (Pn-net photosynthesis and DR - dark respiration respectively) were determined. It has been found that with the exception of Fo, values of Fv, Fm and Fv/Fm were reduced by Pb²⁺. The values of t 1/2 were significantly larger in Pb²⁺ treated leaves. Decrease in the chlorophyll a fluorescence parameters was paralleled with the strong inhibition by this metal the biosynthesis of chlorophyll a and b but less of the carotenoids. Pb²⁺ drastically reduced Pn but had a stimulatory action on DR after 24 h and small inhibition of DR after 48 h exposure of leaves to this metal. As a consequence, after 48 h of greening the ratio of DR/Pn of control leaves was 0.45 whereas in Pb²⁺ treated leaves 2.7. It is proposed that DR in leaves plays a protective role against damage of Pn by Pb²⁺. Protection can be due to the supply the respiratory derived reductant and ATP to carry out cell metabolism upon reduced photosynthesis.     

Photosynthetic responses of Oryza sativa L. seedlings to cadmium stress: physiological,biochemical and ultrastructural analyses

In the present study, photosynthetic responses induced by cadmium stress in chlorophyll biosynthesis, photochemical activities, the stability of thylakoid membranes chlorophyll-protein complexes and the chloroplast ultrastructure of the cereal crop Oryza sativa L. were characterized. Cadmium inhibited the biosynthesis of chlorophyll by interfering with activity of δ-aminolevulinic acid dehydratase in the rice seedlings. For the photochemical activities analyses, the extent of the decrease in photosystem II activity was much greater than that in the PS I activity. The variations in the chlorophyll a fluorescence parameters also indicated that cadmium toxicity drastically affected the photochemistry of PS II. Biochemical analyses by BN-PAGE and protein immunoblot showed that cadmium toxicity considerably affected the stability of PS II-core, cytb ₆ /f, RuBisCO, PSI + LHCI and LHCII (Trimeric). We observed the rate of the thylakoid membranes protein degradation, was mainly at the level of RbcL, PsaA, Lhca1 and D1. In addition, the damages to chloroplast structure and thylakoid stacking analyzed by transmission electron microscopy were indicative of general disarray in the photosynthetic functions exerted by cadmium toxicity. These results are valuable for understanding the biological consequences of heavy metals contamination particularly in soils devoted to organic agriculture.     

Detection of the photoactive protochlorophyllide-protein complex in the light during the greening of barley.

A photoactive protochlorophyllide-protein complex with absorbance and fluorescence maxima at 648 and 653 nm was detected in greening barley leaves without any re-darkening. The variations of the amplitudes of the absorbance and the fluorescence of the photoactive protochlorophyllide with greening time at two different light intensities indicate a close relationship between the rate of chlorophyll synthesis and the amount of the complex during the first hours. The chlorophyllide resulting from photoreduction during greening has an absorbance maximum at 684 nm, which shifts towards a shorter wavelength within a few seconds, indicating rapid liberation of the pigment from the enzyme. We conclude that chlorophyll accumulation proceeds through continuous regeneration and phototransformation of the photoactive complex.     

Photoinhibition in vivo of Photosystem II reactions during developmentof the photosystems of wheat seedlings

Photoinhibition of O₂ evolution and reactions leading to millisecond-delayed light emission (ms-DLE) of chlorophyll by illumination of leaves with excess white light were investigated in wheat seedlings greened for different times in a special chamber with constant conditions (20°C; CO₂ and humidity). A sharp reduction in initial and steady state rates of O₂ evolution and in the intensity of different components of ms-DLE under excess light on the stage of lag-phase of chlorophyll biosynthesis (4–6h of greening) were observed. An increasing stability of the oxygen-evolving process and ms-DLE of chlorophyll during formation of the thylakoid membrane photosystems (12–24 h of greening) was shown. Rifampicin did not influence the stability of oxygen evolution whereas cycloheximide led to the intensification of photoinhibition of the initial and steady-state rates of oxygen evolution under the inhibitory light action. The early stages of photosystems formation during short time of greening of etiolated seedlings were more sensitive to the action of inhibitory light, possibly due to a weak interaction of the oxygen-evolving system components and connection with reaction centers of Photosystem II.     

Greening of etiolated bean leaves in far red light

Eight-day-old dark-grown bean leaves were greened by prolonged irradiation with far red light. Growth, chlorophyll content, oxygen-evolving capacity, photophosphorylation capacity, chloroplast structure (by electron microscopy), and in vivo forms of chlorophyll (by low temperature absorption and derivative spectroscopy on intact leaves) were followed during the greening process. Chlorophyll a accumulated slowly but continuously during the 7 days of the experiment (each day consisted of 12 hours of far red light and 12 hours of darkness). Chlorophyll b was not detected until the 5th day. The capacity for oxygen evolution and photophosphorylation began at about the 2nd day. Electron microscopy showed little formation of grana during the 7 days but rather unfused stacks of primary thylakoids. The thylakoids would fuse to give grana if the leaves were placed subsequently in white light. The low temperature spectroscopy of intact leaves showed that the chlorophyll a was differentiated into three forms with absorption maxima near 670, 677, and 683 nanometers at −196 C during the first few hours and that these forms accumulated throughout the greening process. Small amounts of two longer wavelength forms with maxima near 690 and 698 nanometers appeared at about the same time as photosynthetic activity.     

Bioremediation of adverse impact of cadmium toxicity on Cassia italica Mill by arbuscular mycorrhizal fungi

Cassia italica Mill is an important medicinal plant within the family Fabaceae. Pot experiment was conducted to evaluate cadmium stress induced changes in physiological and biochemical attributes in C. italica with and without arbuscular mycorrhizal fungi (AMF). Cadmium stressed plant showed reduced chlorophyll pigment and protein content while AMF inoculation enhanced the chlorophyll and protein content considerably. AMF also ameliorated the cadmium stress induced reduction in total chlorophyll and protein contents by 19.30% and 38.29%, respectively. Cadmium stress enhanced lipid peroxidation while AMF inoculation reduced lipid peroxidation considerably. Increase in proline and phenol content was observed due to cadmium stress and AMF inoculation caused a further increase in proline and phenol content ensuring better growth under stressed conditions. AMF alone also enhanced proline and phenol content. Activity of antioxidant enzymes enhanced under cadmium treatment and AMF inoculation further enhanced their activity thereby strengthening the antioxidant system. Enhanced activities of antioxidants and increased accumulation of osmolytes help plants to avoid damaging impact of oxidative damage. The research has shown that AMF inoculation mitigated the negative impact of stress by reducing the lipid peroxidation and enhancing the antioxidant activity. The present study strongly supports employing AMF as the biological mean for enhancing the cadmium stress tolerance of C. italica.