T. K. Golovko

Effect of cadmium on growth and respiration of barley plants grown under two temperature regimes.

We studied cadmium effect on the respiratory pathways ratio in the organs of barley (Hordeum distichum L., cv. Novichok) plants grown in water culture at two temperature regimes. Mineral nutrients were supplied daily in exponentially increasing amounts in order to provide for steady-state growth. CdSO4 (30, 60, or 100 μmol/l) was added to nutrient solution at a single time in the beginning of the exponential growth period (19 days after germination). In further 6 days, the relative growth rate and biomass accumulation declined stronger with the increase in the cadmium concentration in plants grown at 13/8°C (day/night) than at 21/17°C (day/night). Cadmium suppressed root respiration (down to 60% of control) stronger than leaf respiration, and the roots manifested a higher sensitivity to the inhibitor of alternative oxidase, salicylhydroxamic acid. The respiratory pathways ratio in the roots occurred against the background of activated lipid peroxidation (POL). The highest POL activity and the highest proportion of alternative respiration pathway (AP) (up to 46% of total respiration) were observed in the roots in the presence of the highest cadmium concentration (100 μM) under lower temperature (13/8°C). Thus, high cadmium concentrations affected strongly the total rate of respiration and respiratory pathways ratio. Growth temperature modulated Cd effects on respiration. AP activation is one of the mechanisms for maintenance of root cell homeostasis under cadmium-induced stress.

Pigment Apparatus in Ajuga reptans Plants as Affected by Adaptation to Light Growth Conditions

Mechanisms of adaptation of the photosynthetic apparatus at the level of pigment complex in a shade-tolerant bugle plant (Ajuga reptans L.) grown at full solar irradiation in an open plot were studied. In “sun” plants, the content of photosynthetic pigments decreased markedly as compared to “shade” plants grown under a forest canopy at 5–10% of the full solar irradiation. In leaves of sun plants, the portion of β-carotene and lutein in the carotenoid spectrum was higher than in shade plant leaves, antheraxanthin and zeaxanthin were present, and de-epoxidation of violaxanthin was by an order of magnitude higher in sun plant leaves reaching 40%. The data obtained indicate the role of the violaxanthin cycle in the protection of photosynthetic apparatus in a shade-tolerant plant against destruction under excessive irradiation.

Light regulation of mitochondrial alternative oxidase pathway during greening of etiolated wheat seedlings.

This study deals with effects of de-etiolation (48h) of spring wheat (Triticum aestivum L., var. Irgina) seedlings on differential expression of AOX1 genes, levels of AOX protein and the alternative respiratory pathway (AP) capacity. As a result of exposure to continuous irradiation of dark-grown wheat seedlings, the respiratory activity and AP capacity in leaves significantly increased during the first 6h of studies. Expression of AOX1a was up-regulated by light and proved consistent with changes in the AP capacity. Effects on expression of AOX1c were less pronounced. Immunoblot analysis showed three distinct bands of AOX with molecular weights of 34, 36 and 38kDa, with no significant changes in the relative levels during de-etiolation. The lack of a clear correlation between AOX protein amount, AOX1a expression, and AP capacity suggests post-translational control of the enzyme activation. The AOX1a suppression and a decrease in the AP capacity correlated with the sugar pool depletion after 24h of the de-etiolation, which may mean a possible substrate dependence of the AOX activity in the green cells. More efficient malate oxidation by mitochondria as well as the higher AOX capacity during the first 6h of de-etiolation was detected, whereas respiration and AOX capacity with exogenous NADH and glycine increased after 6 and 24h, respectively. We conclude that AOX plays an important role during development of an actively photosynthesizing cell, and can rapidly adapt to changes in metabolism and photosynthesis.

Functional plasticity of photosynthetic apparatus and its resistance to photoinhibition in Plantago media

Morphological and functional characteristics of Plantago media L. leaves were compared for plants growing at different light regimes on limestone outcrops in Southern Timan (62°45′N, 55°49′E). The plants grown in open areas under exposure to full sunlight had small leaves with low pigment content and high specific leaf weight; these leaves exhibited high photosynthetic capacity and elevated water use efficiency at high irradiance. The maximum photochemical activity of photosystem II (Fv/Fm) in leaves of sun plants remained at the level of about 0.8 throughout the day. The photosynthetic apparatus of sun plants was resistant to excess photosynthetically active radiation, mostly due to non-photochemical quenching of chlorophyll fluorescence (qN). This quenching was promoted by elevated deepoxiation of violaxanthin cycle pigments. Accumulation of zeaxanthin, a photoprotective pigment in sun plant leaves was observed already in the morning hours. The plant leaves grown in the shade of dense herbage were significantly larger than the sun leaves, with pigment content 1.5–2.0 times greater than in sun leaves; these leaves had low qN values and did not need extensive deepoxidation of violaxanthin cycle pigments. The data reveal the morphophysiological plasticity of plantain plants in relation to lighting regime. Environmental conditions can facilitate the formation of the ecotype with photosynthetic apparatus resistant to photoinhibition. Owing to this adjustment, hoary plantain plants are capable of surviving in ecotopes with high insolation.

Developmental changes in energy dissipation in etiolated wheat seedlings during the greening process

We studied the developmental changes in photosynthetic and respiration rates and thermal dissipation processes connected with chloroplasts and mitochondria activity in etiolated wheat (Triticum aestivum L., var. Irgina) seedlings during the greening process. Etioplasts gradually developed into mature chloroplasts under continuous light [190 μmol(photon) m−2 s−1] for 48 h in 5-day-dark-grown seedlings. The net photosynthetic rate of irradiated leaves became positive after 6 h of illumination and increased further. The first two hours of de-etiolation were characterized by low values of maximum (Fv/Fm) and actual photochemical efficiency of photosystem II (PSII) and by a coefficient of photochemical quenching in leaves. Fv/Fm reached 0.8 by the end of 24 h-light period. During greening, energy-dependent component of nonphotochemical quenching of chlorophyll fluorescence, violaxanthin cycle (VXC) operation, and lipoperoxidation activity changed in a similar way. Values of these parameters were the highest at the later phase of de-etiolation (4–12 h of illumination). The respiration rate increased significantly after 2 h of greening and it was the highest after 4–6 h of illumination. It was caused by an increase in alternative respiration (AP) capacity. The strong, positive linear correlation was revealed between AP capacity and heat production in greening tissues. These results indicated that VXC in chloroplasts and AP in mitochondria were intensified as energy-dissipating systems at the later stage of greening (after 4 h), when most of prolamellar bodies converted into thylakoids, and they showed the greatest activity until the photosynthetic machinery was almost completely developed.

Violaxanthin cycle pigment de-epoxidation and thermal dissipation of light energy in three boreal species of evergreen conifer plants

We studied carotenoid composition and chlorophyll fluorescence in two-year-old needles from Siberian spruce (Picea obovata (L.) Karst.), Siberian fir (Abies sibirica L.), and common juniper (Juniperus communis L.). The highest values of maximum PSII photochemical activity (Fv/Fm) equaling 0.82–0.85 were observed in July–September. The decrease in Fv/Fm in December–March was more pronounced in juniper (down to 0.15) than in spruce and fir (0.45–0.50). In May, we observed a nearly complete recovery in maximum PSII photochemical activity in fir and spruce (0.72–0.77), while in juniper, the Fv/Fm value was notably lower (0.65–0.67). The amount of thermal dissipation of energy absorbed by PSII LHC did not exceed 30% in summer and equaled 60–90% in winter and early spring. The carotenoid pool consisted mainly of xanthophylls, among which lutein (70%), neoxanthin (7–10%), and a violaxanthin cycle (VXC) component — violaxanthin (3–15%) were constantly present. The accumulation of two other VXC pigments—zeaxanthin and antheraxanthin, was noted in December–March. In July, these xanthophylls were not identified. We discovered a direct connection between VXC pigment de-epoxidation level and light energy thermal dissipation in boreal conifer leaves. Such association reflects the non-species-specific character of the mechanism for quenching zeaxanthin-dependent nonphotochemical chlorophyll fluorescence in PSII LHC in winter and spring.

The Effect of Light Regime on Source–Sink Relations in the Shade-Enduring Ajuga reptans Plant

A quantitative approach to the evaluation of source–sink relations in Ajuga reptans plants grown under the forest canopy (shade plants) and on an open plot (sun plants) was worked out in terms of growth characteristics, CO2 exchange, and carbon balance. Shade plants developed leaves with the relative and specific areas twice exceeding those of sun plants. Sun plants assimilated more carbon, using a significant part of it for the development of numerous runners. During a day, shade and sun plants produced 0.03 and 0.67 g of substrate, respectively. At the same time, forest (shade) plants spent 48% of assimilates for the respiration, in comparison with plants from the open plot that spent almost 70% of assimilates for respiration as they were greater in size. It was concluded, that light controls source–sink relations, which is a way of realization of the life strategy and a coordination mechanism of functional integrity of the plant organism. Light not only controls photosynthesis (source activity) but morphophysiological characteristics of plants with their hierarchical structure of sinks too.

Adaptive changes in pigment complex of Pinus sylvestris needles upon cold acclimation

We studied seasonal changes in the content and ratio between photosynthetic pigments in one-yearold needles of Scotch pine (Pinus sylvestris L.) growing in Central Yakutia. Maximum accumulation of chlorophylls in developed young needles occurred in July when light and temperature conditions were favorable. In this period, the needles were notable for a relatively high level of β-carotene and neoxanthin and a reduced content of lutein and the pigments of violaxanthin cycle (VXC). In the course of autumn hardening, the content of chlorophylls decreased two times. Total content of carotenoids remained the same, but pigment composition considerably changed when plants progressed from a vegetating to frost-resistant state. We revealed time and temperature ranges of variation for individual carotenoids. In the beginning of hardening at reduced and low abovezero temperatures, the content of β-carotene in the needles decreased, the pigment-protein complexes (PPC) became enriched in lutein, the pigment pool of VXC gradually increased, and the content of neoxanthin transiently rose. When average daily air temperature further decreased to near- zero values, the content of zeaxanthin sharply rose. In winter, high levels of lutein and zeaxanthin were maintained. Main changes in pigment complex of the needles of P. sylvestris were completed before the coming of steady below-zero temperatures. The obtained data suggested that, upon seasonal decrease in temperature in early stages of hardening, a decrease in the level of chlorophyll promotes a reduction in the quantity of absorbed radiant energy. Apparently, this is accompanied by activation of the role of lutein and neoxanthin that perform specific photoprotective functions in antenna PPC associated with a gradual decrease in plants’ ability to quench singlet energy of excited chlorophyll. Accumulation of zeaxanthin as a result of inhibition of back reaction of epoxidation at near-zero temperatures creates necessary prerequisites for turning on the mechanisms of steady dissipation of absorbed light energy, which do not depend on transmembrane proton gradient of thylakoids. At the same time, zeaxanthin can perform antioxidant functions both in PPC and in the lipid phase of thylakoid membranes. The obtained data point to an adaptive nature of the observed reactions and a specific role of individual pigments in structural and functional reorganization of photosynthetic machinery in the course of development of frost-resistance in the needles.

Activities of respiratory pathways and the pool of nonstructural carbohydrates in greening leaves of spring wheat seedlings

Seedlings of spring wheat (Triticum aestivum L.) were used to study the dynamics of leaf respiration, the respiratory pathway ratio, and relation of activities of these pathways to the content of soluble carbohydrates in the leaf during greening of seedlings for 48 h under continuous photosynthetically active light (190 μmol/(m2 s)). Changes in leaf respiration during de-etiolation were closely related to modulation of the alternative respiratory pathway (AP) activity. The rate of cytochrome respiratory pathway (CP) depended directly on the carbohydrate content and growth rate. These relations suggest that the substrate regulation of CP activity during greening is mediated by the energy needs for growth and is effectively regulated by the mechanism of respiratory control. The highest rates of AP were observed after a 6-h exposure of seedlings to light. The proportion of CP/AP at this stage was close to unity. The temporal pattern of AP activity during de-etiolation was independent on the content of soluble carbohydrates. Hence, in addition to substrate regulation of AP, there are other intricate mechanisms of AP involvement. Our results are in accordance to the state that the alternative respiratory pathway participates in maintaining homeostasis in phototrophic cells during development of the photosynthetic function.

Photoprotective Mechanisms in Photosystem II of Ephedra monosperma during Development of Frost Tolerance

Dissipation of light energy absorbed by photosystem II (PSII) in assimilating shoots of an evergreen shrub Ephedra monosperma was investigated during its transition from the vegetative to frost-tolerant state under natural conditions of Central Yakutia. The dynamics of modulated chlorophyll fluorescence and carotenoid content was analyzed during seasonal decrease in ambient temperature. The seasonal cooling was accompanied by a stepwise decrease in photochemical activity of PSII (Fv/Fm = (Fm − F0)/Fm). The decrease in Fv/Fm occurred from the beginning of September to the end of October, when the temperature was lowered from 10 to −8°C. During winter period the residual activity of PSII was retained at about 30% of the summer values. The seasonal decrease in temperature was accompanied by a significant stimulation of pH-independent dissipative processes in reaction centers and antenna of PSII. The increase in energy losses was paralleled by a proportional increase in zeaxanthin content on the background of decreasing content of violaxanthin and β-carotene as possible zeaxanthin precursors. At the same time, inhibition of light-induced non-photochemical quenching in the PSII antenna was observed. The results suggest that principal photoprotective mechanisms during seasonal lowering of temperature are: (1) inactivation of PSII and dissipation of excitation energy in PSII reaction centers and (2) zeaxanthin-mediated energy dissipation in the antenna complexes. The first mechanism seems to prevail at early stages of seasonal cooling, whereas both mechanisms are recruited from the onset of sustained freezing temperatures.