Thylakoid ultrastructural variations in chlorophyll-deficient wheat: aberrations or structural acclimation?

MAIN CONCLUSION: A structural re-modeling of the thylakoid system, including granum size and regularity, occurs in chlorophyll-deficient wheat mutants affected by photosynthetic membrane over-reduction. In the chloroplast of land plants, the thylakoid system is defined by appressed grana stacks and unstacked stroma lamellae. This study focuses on the variations of the grana organization occurring in outdoor-grown wheat mutants characterized by low chlorophyll content and a tendency for photosynthetic membrane over-reduction. Triticum aestivum ANK-32A and Triticum durum ANDW-7B were compared to their corresponding WT lines, NS67 and LD222, respectively. Electron micrographs of chloroplasts were used to calculate grana ultrastructural parameters. Photosynthetic parameters were obtained by modulated chlorophyll fluorescence and applying Light Curves (LC) and Rapid Light Curves (RLC) protocols. For each photosynthetic parameter, the difference Δ(RLC-LC) was calculated to evaluate the flexible response to light in the examined lines. In the mutants, fewer and smaller disks formed grana stacks characterized by a marked increase in lateral and cross-sectional irregularity, both negatively correlated with the number of layers per granum. A relationship was found between membrane over-reduction and granum structural irregularity. The possible acclimative significance of a greater proportion of stroma-exposed grana domains in relieving the excess electron pressure on PSI is discussed.

Silicon nanoparticles: Comprehensive review on biogenic synthesis and applications in agriculture.

Recent advancements in nanotechnology have opened new advances in agriculture. Among other nanoparticles, silicon nanoparticles (SiNPs), due to their unique physiological characteristics and structural properties, offer a significant advantage as nanofertilizers, nanopesticides, nanozeolite and targeted delivery systems in agriculture. Silicon nanoparticles are well known to improve plant growth under normal and stressful environments. Nanosilicon has been reported to enhance plant stress tolerance against various environmental stress and is considered a non-toxic and proficient alternative to control plant diseases. However, a few studies depicted the phytotoxic effects of SiNPs on specific plants. Therefore, there is a need for comprehensive research, mainly on the interaction mechanism between NPs and host plants to unravel the hidden facts about silicon nanoparticles in agriculture. The present review illustrates the potential role of silicon nanoparticles in improving plant resistance to combat different environmental (abiotic and biotic) stresses and the underlying mechanisms involved. Furthermore, our review focuses on providing the overview of various methods exploited in the biogenic synthesis of silicon nanoparticles. However, certain limitations exist in synthesizing the well-characterized SiNPs on a laboratory scale. To bridge this gap, in the last section of the review, we discussed the possible use of the machine learning approach in future as an effective, less labour-intensive and time-consuming method for silicon nanoparticle synthesis. The existing research gaps from our perspective and future research directions for utilizing SiNPs in sustainable agriculture development have also been highlighted.

Diversity of responses to nitrogen deficiency in distinct wheat genotypes reveals the role of alternative electron flows in photoprotection.

Nitrogen (N) deficiency represents an important limiting factor affecting photosynthetic productivity and the yields of crop plants. Significant reported differences in N use efficiency between the crop species and genotypes provide a good background for the studies of diversity of photosynthetic and photoprotective responses associated with nitrogen deficiency. Using distinct wheat (Triticum aestivum L.) genotypes with previously observed contrasting responses to nitrogen nutrition (cv. Enola and cv. Slomer), we performed advanced analyses of CO2 assimilation, PSII, and PSI photochemistry, also focusing on the heterogeneity of the stress responses in the different leaf levels. Our results confirmed the loss of photosynthetic capacity and enhanced more in lower positions. Non-stomatal limitation of photosynthesis was well reflected by the changes in PSII and PSI photochemistry, including the parameters derived from the fast-fluorescence kinetics. Low photosynthesis in N-deprived leaves, especially in lower positions, was associated with a significant decrease in the activity of alternative electron flows. The exception was the cyclic electron flow around PSI that was enhanced in most of the samples with a low photosynthetic rate. We observed significant genotype-specific responses. An old genotype Slomer with a lower CO2 assimilation rate demonstrated enhanced alternative electron flow and photorespiration capacity. In contrast, a modern, highly productive genotype Enola responded to decreased photosynthesis by a significant increase in nonphotochemical dissipation and cyclic electron flow. Our results illustrate the importance of alternative electron flows for eliminating the excitation pressure at the PSII acceptor side. The decrease in capacity of electron acceptors was balanced by the structural and functional changes of the components of the electron transport chain, leading to a decline of linear electron transport to prevent the overreduction of the PSI acceptor side and related photooxidative damage of photosynthetic structures in leaves exposed to nitrogen deficiency.

The different patterns of post-heat stress responses in wheat genotypes: the role of the transthylakoid proton gradient in efficient recovery of leaf photosynthetic capacity.

The frequency and severity of heat waves are expected to increase in the near future, with a significant impact on physiological functions and yield of crop plants. In this study, we assessed the residual post-heat stress effects on photosynthetic responses of six diverse winter wheat (Triticum sp.) genotypes, differing in country of origin, taxonomy and ploidy (tetraploids vs. hexaploids). After 5 days of elevated temperatures (up to 38 °C), the photosynthetic parameters recorded on the first day of recovery (R1) as well as after the next 4-5 days of the recovery (R2) were compared to those of the control plants (C) grown under moderate temperatures. Based on the values of CO2 assimilation rate (A) and the maximum rates of carboxylation (VCmax) in R1, we identified that the hexaploid (HEX) and tetraploid (TET) species clearly differed in the strength of their response to heat stress. Next, the analyses of gas exchange, simultaneous measurements of PSI and PSII photochemistry and the measurements of electrochromic bandshift (ECS) have consistently shown that photosynthetic and photoprotective functions in leaves of TET genotypes were almost fully recovered in R2, whereas the recovery of photosynthetic and photoprotective functions in the HEX group in R2 was still rather low. A poor recovery was associated with an overly reduced acceptor side of photosystem I as well as high values of the electric membrane potential (Δψ component of the proton motive force, pmf) in the chloroplast. On the other hand, a good recovery of photosynthetic capacity and photoprotective functions was clearly associated with an enhanced ΔpH component of the pmf, thus demonstrating a key role of efficient regulation of proton transport to ensure buildup of the transthylakoid proton gradient needed for photosynthesis restoration after high-temperature episodes.

Electron and proton transport in wheat exposed to salt stress: is the increase of the thylakoid membrane proton conductivity responsible for decreasing the photosynthetic activity in sensitive genotypes?

Effects of salinity caused by 150 mM NaCl on primary photochemical reactions and some physiological and biochemical parameters (K+/Na+ ratio, soluble sugars, proline, MDA) have been studied in five Triticum aestivum L. genotypes with contrasting salt tolerance. It was found that 150 mM NaCl significantly decreased the photosynthetic efficiency of two sensitive genotypes. The K+/Na+ ratio decreased in all genotypes exposed to salinity stress when compared with the control. Salinity stress also caused lipid peroxidation and accumulation of soluble sugars and proline. The amounts of soluble sugars and proline were higher in tolerant genotypes than sensitive ones, and lipid peroxidation was higher in sensitive genotypes. The noninvasive measurements of photosynthesis-related parameters indicated the genotype-dependent effects of salinity stress on the photosynthetic apparatus. The significant decrease of chlorophyll content (SPAD values) or adverse effects on photosynthetic functions at the PSII level (measured by the chlorophyll fluorescence parameters) were observed in the two sensitive genotypes only. Although the information obtained by different fast noninvasive techniques were consistent, the correlation analyses identified the highest correlation of the noninvasive records with MDA, K+/Na+ ratio, and free proline content. The lower correlation levels were found for chlorophyll content (SPAD) and Fv/Fm values derived from chlorophyll fluorescence. Performance index (PIabs) derived from fast fluorescence kinetics, and F735/F685 ratio correlated well with MDA and Na+ content. The most promising were the results of linear electron flow measured by MultispeQ sensor, in which we found a highly significant correlation with all parameters assessed. Moreover, the noninvasive simultaneous measurements of chlorophyll fluorescence and electrochromic band shift using this sensor indicated the apparent proton leakage at the thylakoid membranes resulting in a high proton conductivity (gH+), present in sensitive genotypes only. The possible consequences for the photosynthetic functions and the photoprotection are discussed.

Photosynthesis research under climate change.

Increasing global population and climate change uncertainties have compelled increased photosynthetic efficiency and yields to ensure food security over the coming decades. Potentially, genetic manipulation and minimization of carbon or energy losses can be ideal to boost photosynthetic efficiency or crop productivity. Despite significant efforts, limited success has been achieved. There is a need for thorough improvement in key photosynthetic limiting factors, such as stomatal conductance, mesophyll conductance, biochemical capacity combined with Rubisco, the Calvin-Benson cycle, thylakoid membrane electron transport, nonphotochemical quenching, and carbon metabolism or fixation pathways. In addition, the mechanistic basis for the enhancement in photosynthetic adaptation to environmental variables such as light intensity, temperature and elevated CO2 requires further investigation. This review sheds light on strategies to improve plant photosynthesis by targeting these intrinsic photosynthetic limitations and external environmental factors.

Physiological and molecular mechanisms of metal accumulation in hyperaccumulator plants.

Most of the heavy metals (HMs), and metals/metalloids are released into the nature either by natural phenomenon or anthropogenic activities. Being sessile organisms, plants are constantly exposed to HMs in the environment. The metal non-hyperaccumulating plants are susceptible to excess metal concentrations. They tend to sequester metals in their root vacuoles by forming complexes with metal ligands, as a detoxification strategy. In contrast, the metal-hyperaccumulating plants have adaptive intrinsic regulatory mechanisms to hyperaccumulate or sequester excess amounts of HMs into their above-ground tissues rather than accumulating them in roots. They have unique abilities to successfully carry out normal physiological functions without showing any visible stress symptoms unlike metal non-hyperaccumulators. The unique abilities of accumulating excess metals in hyperaccumulators partly owes to constitutive overexpression of metal transporters and ability to quickly translocate HMs from root to shoot. Various metal ligands also play key roles in metal hyperaccumulating plants. These metal hyperaccumulating plants can be used in metal contaminated sites to clean-up soils. Exploiting the knowledge of natural populations of metal hyperaccumulators complemented with cutting-edge biotechnological tools can be useful in the future. The present review highlights the recent developments in physiological and molecular mechanisms of metal accumulation of hyperaccumulator plants in the lights of metal ligands and transporters. The contrasting mechanisms of metal accumulation between hyperaccumulators and non-hyperaccumulators are thoroughly compared. Moreover, uses of different metal hyperaccumulators for phytoremediation purposes are also discussed in detail.

Phenotyping of isogenic chlorophyll-less bread and durum wheat mutant lines in relation to photoprotection and photosynthetic capacity.

In our experiments, we examined high light responses and photosynthetic capacity of chlorophyll-less isogenic mutant lines of hexaploid bread wheat (Triticum aestivum L.) and tetraploid durum wheat (Triticum durum L.) in comparison to parental lines representing the wild type (WT), in two growth phases and two environments. In young plants, we observed a typical yellow-green phenotype with low chlorophyll content, significantly lower CO2 assimilation rate, elevated chlorophyll a-to-b ratio and insufficient regulation of linear electron transport. In the mutants grown in a moderate light in the growth chamber, a typical "chlorina" phenotype almost disappeared or, at least, was significantly alleviated in later growth stages, including the values of CO2 assimilation and the majority of the measured parameters related to photoprotective responses. On the other hand, in the case of the mutant lines grown in direct sunlight and fluctuating environment, the chlorophyll-less phenotype was evident also in latter growth phases. The chlorophyll-less phenotype was more severe in the durum wheat mutant lines compared to the bread wheat. For example, the durum wheat mutant lines grown outdoors expressed lower flexibility of photoprotective responses, including lower non-photochemical quenching and low rate of cyclic electron flow compared to WT or bread wheat mutants. Based on the analyses, we have identified a set of parameters providing information on the specific photosynthetic traits typical for the chlorophyll-less phenotype. Thus, the proposed way of phenotyping may serve for efficient selection of mutant genotypes for future research or screening activities. As a general result, we observed that the decrease of the chlorophyll content due to mutation was always associated with improper regulation of linear electron transport and a limited ability to prevent over-reduction of PSI acceptor side, regardless of the genotype, environment, and growth stage. This can partly explain why the low chlorophyll mutants were not successful in the evolution of higher plants, despite the photosynthetic capacity observed is high enough and they are fully competitive with wild-type plants in non-fluctuating controlled environment.

Wheat plant selection for high yields entailed improvement of leaf anatomical and biochemical traits including tolerance to non-optimal temperature conditions.

Assessment of photosynthetic traits and temperature tolerance was performed on field-grown modern genotype (MG), and the local landrace (LR) of wheat (Triticum aestivum L.) as well as the wild relative species (Aegilops cylindrica Host.). The comparison was based on measurements of the gas exchange (A/ci, light and temperature response curves), slow and fast chlorophyll fluorescence kinetics, and some growth and leaf parameters. In MG, we observed the highest CO2 assimilation rate [Formula: see text] electron transport rate (Jmax) and maximum carboxylation rate [Formula: see text]. The Aegilops leaves had substantially lower values of all photosynthetic parameters; this fact correlated with its lower biomass production. The mesophyll conductance was almost the same in Aegilops and MG, despite the significant differences in leaf phenotype. In contrary, in LR with a higher dry mass per leaf area, the half mesophyll conductance (gm) values indicated more limited CO2 diffusion. In Aegilops, we found much lower carboxylation capacity; this can be attributed mainly to thin leaves and lower Rubisco activity. The difference in CO2 assimilation rate between MG and others was diminished because of its higher mitochondrial respiration activity indicating more intense metabolism. Assessment of temperature response showed lower temperature optimum and a narrow ecological valence (i.e., the range determining the tolerance limits of a species to an environmental factor) in Aegilops. In addition, analysis of photosynthetic thermostability identified the LR as the most sensitive. Our results support the idea that the selection for high yields was accompanied by the increase of photosynthetic productivity through unintentional improvement of leaf anatomical and biochemical traits including tolerance to non-optimal temperature conditions.

Melatonin alleviates low PS I-limited carbon assimilation under elevated CO2 and enhances the cold tolerance of offspring in chlorophyll b-deficient mutant wheat.

Melatonin is involved in the regulation of carbohydrate metabolism and induction of cold tolerance in plants. The objective of this study was to investigate the roles of melatonin in modulation of carbon assimilation of wild-type wheat and the Chl b-deficient mutant ANK32B in response to elevated CO2 concentration ([CO2 ]) and the transgenerational effects of application of exogenous melatonin (hereafter identified as melatonin priming) on the cold tolerance in offspring. The results showed that the melatonin priming enhanced the carbon assimilation in ANK32B under elevated [CO2 ], via boosting the activities of ATPase and sucrose synthesis and maintaining a relatively higher level of total chlorophyll concentration in leaves. In addition, melatonin priming in maternal plants at grain filling promoted the seed germination in offspring by accelerating the starch degradation and improved the cold tolerance of seedlings through activating the antioxidant enzymes and enhancing the photosynthetic electron transport efficiency. These findings suggest the important roles of melatonin in plant response to future climate change, indicating that the melatonin priming at grain filling in maternal plants could be an effective approach to improve cold tolerance of wheat offspring at seedling stage.

Comparison on Photosynthesis and Antioxidant Defense Systems in Wheat with Different Ploidy Levels and Octoploid Triticale.

To investigate the evolutionary differences of wheat with different ploidy levels and octoploid Triticale, photosynthetic capacity, and antioxidant defenses system were compared within and between diploid, tetraploid and hexaploid wheat, and octoploid Triticale seedlings. The results showed that seed germination rate, chlorophyll content, and photochemical activity of photosystems, and the activities of antioxidative enzymes in hexaploid wheat and octoploid Triticale were significantly higher than in diploid and tetraploid wheat. Compared to other two wheat species and octoploid Triticale, hexaploid wheat presented lower levels of reactive oxygen species (ROS). Furthermore, we found that the levels of photosystem II reaction center protein D1, light-harvesting complex II b4 (CP29), and D subunit of photosystem I (PsaD) in diploid wheat were significantly lower compared with hexaploid wheat and octoploid Triticale. Taken together, we concluded that hexaploid wheat and octoploid Triticale have higher photosynthetic capacities and better antioxidant systems. These findings indicate that different ploidy levels of chromosome probably play an important regulatory role in photosystems and antioxidative systems of plants.

Bioactive Phytochemicals and Antioxidant Properties of the Grains and Sprouts of Colored Wheat Genotypes.

The grains and sprouts of colored wheat genotypes (having blue, purple and yellow colored grains) contain specific anthocyanidins, such as pelargonidin and cyanidin derivatives, that produce beneficial health effects. The objective of the presented study is to compare the antioxidant capacity and contents of bioactive phytochemicals in grains and sprouts of wheat genotypes that differ in grain color. The methods α, α-diphenyl-ß-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activities, together with spectrophotometrical and high-performance thin-layer chromatography (HPTLC) methods, were used to study the presence of total phenolics, flavonoids, anthocyanins and anthocyanidins (pelargonidin, peonidin, cyanidin, delphinidin) content. It was predicted that the sprouts of all colored wheat genotypes would have significantly higher total flavonoids, total phenolics, anthocyanidin levels and antioxidant activity than the grains. The correlation results between antioxidant activity and contents of bioactive phytochemicals in grains and sprouts of colored wheat genotypes have shown a high correlation for cyanidin and pelargonidin, especially in grains, as well as quercetin in sprouts. It was found that total anthocyanin, quercetin and pelargonidin contents were significantly higher in the sprouts of the purple wheat genotypes than in the blue or yellow wheat genotypes. Delphinidin was detected at a higher level in the grains than in the sprouts of the blue wheat genotypes. Peonidin was present at very low quantities in the grains of all colored wheat genotypes. The sprouts of the purple wheat genotypes, among the colored wheat genotypes, had the highest pelargonidin, cyanidin and quercetin contents and, therefore, can be a promising source for functional food use.

Anthocyanins of Coloured Wheat Genotypes in Specific Response to SalStress.

The present study investigated the effect of salt stress on the development of adaptive responses and growth parameters of different coloured wheat genotypes. The different coloured wheat genotypes have revealed variation in the anthocyanin content, which may affect the development of adaptive responses under increasing salinity stress. In the early stage of treatment with salt at a lower NaCl concentration (100 mM), anthocyanins and proline accumulate, which shows rapid development of the stress reaction. A dose-dependent increase in flavonol content was observed for wheat genotypes with more intense purple-blue pigmentation after treatment with 150 mM and 200 mM NaCl. The content of Na⁺ and K⁺ obtained at different levels of salinity based on dry weight (DW) was more than 3 times greater than the control, with a significant increase of both ions under salt stress. Overall, our results demonstrated that coloured wheat genotypes with high anthocyanin content are able to maintain significantly higher dry matter production after salt stress treatment.

Lettuce flavonoids screening and phenotyping by chlorophyll fluorescence excitation ratio.

MAIN CONCLUSION: Environmentally induced variation and the genotypic differences in flavonoid and phenolic content in lettuce can be reliably detected using the appropriate parameters derived from the records of rapid non-invasive fluorescence technique. The chlorophyll fluorescence excitation ratio method was designed as a rapid and non-invasive tool to estimate the content of UV-absorbing phenolic compounds in plants. Using this technique, we have assessed the dynamics of accumulation of flavonoids related to developmental changes and environmental effects. Moreover, we have tested appropriateness of the method to identify the genotypic differences and fluctuations in total phenolics and flavonoid content in lettuce. Six green and two red genotypes of lettuce (Lactuca sativa L.) grown in pots were exposed to two different environments for 50 days: direct sunlight (UV-exposed) and greenhouse conditions (low UV). The indices based on the measurements of chlorophyll fluorescence after red, green and UV excitation indicated increase of the content of UV-absorbing compounds and anthocyanins in the epidermis of lettuce leaves. In similar, the biochemical analyses performed at the end of the experiment confirmed significantly higher total phenolic and flavonoid content in lettuce plants exposed to direct sun compared to greenhouse conditions and in red compared to green genotypes. As the correlation between the standard fluorescence indices and the biochemical records was negatively influenced by the presence of red genotypes, we proposed the use of a new parameter named Modified Flavonoid Index (MFI) taking into an account both absorbance changes due to flavonol and anthocyanin content, for which the correlation with flavonoid and phenolic content was relatively good. Thus, our results confirmed that the fluorescence excitation ratio method is useful for identifying the major differences in phenolic and flavonoid content in lettuce plants and it can be used for high-throughput pre-screening and phenotyping of leafy vegetables in research and breeding applications towards improvement of vegetable health effects.

Frequently asked questions about chlorophyll fluorescence, the sequel.

Using chlorophyll (Chl) a fluorescence many aspects of the photosynthetic apparatus can be studied, both in vitro and, noninvasively, in vivo. Complementary techniques can help to interpret changes in the Chl a fluorescence kinetics. Kalaji et al. (Photosynth Res 122:121-158, 2014a) addressed several questions about instruments, methods and applications based on Chl a fluorescence. Here, additional Chl a fluorescence-related topics are discussed again in a question and answer format. Examples are the effect of connectivity on photochemical quenching, the correction of F V /F M values for PSI fluorescence, the energy partitioning concept, the interpretation of the complementary area, probing the donor side of PSII, the assignment of bands of 77 K fluorescence emission spectra to fluorescence emitters, the relationship between prompt and delayed fluorescence, potential problems when sampling tree canopies, the use of fluorescence parameters in QTL studies, the use of Chl a fluorescence in biosensor applications and the application of neural network approaches for the analysis of fluorescence measurements. The answers draw on knowledge from different Chl a fluorescence analysis domains, yielding in several cases new insights.

High temperature specifically affects the photoprotective responses of chlorophyll b-deficient wheat mutant lines.

The effects of high temperature on CO2 assimilation rate, processes associated with photosynthetic electron and proton transport, as well as photoprotective responses, were studied in chlorophyll b-deficient mutant lines (ANK-32A and ANK-32B) and wild type (WT) of wheat (Triticum aestivum L.). Despite the low chlorophyll content and chlorophyll a-to-b ratio, the non-stressed mutant plants had the similar level of CO2 assimilation and photosynthetic responses as WT. However, in ANK mutant plants exposed to prolonged high temperature episode (42 °C for ~10 h), we observed lower CO2 assimilation compared to WT, especially when a high CO2 supply was provided. In all heat-exposed plants, we found approximately the same level of PSII photoinhibition, but the decrease in content of photooxidizable PSI was higher in ANK mutant plants compared to WT. The PSI damage can be well explained by the level of overreduction of PSI acceptor side observed in plants exposed to high temperature, which was, in turn, the result of the insufficient transthylakoid proton gradient associated with low non-photochemical quenching and lack of ability to downregulate the linear electron transport to keep the reduction state of PSI acceptor side low enough. Compared to WT, the ANK mutant lines had lower capacity to drive the cyclic electron transport around PSI in moderate and high light; it confirms the protective role of cyclic electron transport for the protection of PSI against photoinhibition. Our results, however, also suggest that the inactivation of PSI in heat stress conditions can be the protective mechanism against photooxidative damage of chloroplast and cell structures.

The Contribution of Buckwheat Genetic Resources to Health and Dietary Diversity.

Despite several reports on the beneficial effects of buckwheat in prevention of human diseases, little attention has been devoted to the variability of biochemical and physiological traits in different buckwheat genetic resources. This review describes the biochemical evaluation of buckwheat genetic resources and the identification of elite genotypes for plant breeding and exploitation. The various types of bioactive compounds present in different varieties provide basic background information needed for the efficient production of buckwheat foods with added value. In this review, we will provide an integrated view of the biochemistry of bioactive compounds of buckwheat plants of different origin, especially of fagopyrin, proteins and amino acids, as well as of other phenolic compounds including rutin and chlorogenic acid. In addition to the genetic background, the effect of different growth conditions is discussed. The health effects of fagopyrin, phenolic acids, specific proteins and rutin are also presented.

Repetitive light pulse-induced photoinhibition of photosystem I severely affects CO2 assimilation and photoprotection in wheat leaves.

It was previously found that photosystem I (PSI) photoinhibition represents mostly irreversible damage with a slow recovery; however, its physiological significance has not been sufficiently characterized. The aim of the study was to assess the effect of PSI photoinhibition on photosynthesis in vivo. The inactivation of PSI was done by a series of short light saturation pulses applied by fluorimeter in darkness (every 10 s for 15 min), which led to decrease of both PSI (~60 %) and photosystem II (PSII) (~15 %) photochemical activity. No PSI recovery was observed within 2 days, whereas the PSII was fully recovered. Strongly limited PSI electron transport led to an imbalance between PSII and PSI photochemistry, with a high excitation pressure on PSII acceptor side and low oxidation of the PSI donor side. Low and delayed light-induced NPQ and P700(+) rise in inactivated samples indicated a decrease in formation of transthylakoid proton gradient (ΔpH), which was confirmed also by analysis of electrochromic bandshift (ECSt) records. In parallel with photochemical parameters, the CO2 assimilation was also strongly inhibited, more in low light (~70 %) than in high light (~45 %); the decrease was not caused by stomatal closure. PSI electron transport limited the CO2 assimilation at low to moderate light intensities, but it seems not to be directly responsible for a low CO2 assimilation at high light. In this regard, the possible effects of PSI photoinhibition on the redox signaling in chloroplast and its role in downregulation of Calvin cycle activity are discussed.

Low PSI content limits the photoprotection of PSI and PSII in early growth stages of chlorophyll b-deficient wheat mutant lines.

In vivo analyses of electron and proton transport-related processes as well as photoprotective responses were carried out at different stages of growth in chlorophyll b (Chl b)-deficient mutant lines (ANK-32A and ANK-32B) and wild type (WT) of wheat (Triticum aestivum L.). In addition to a high Chl a-b ratio, ANK mutants had a lower content of photo-oxidizable photosystem I (PSI, P m), and several parameters indicated a low PSI/PSII ratio. Moreover, simultaneous measurements of Chl fluorescence and P700 indicated a shift of balance between redox poise of the PSII acceptor side and the PSII donor side, with preferential reduction of the plastoquinone pool, resulting in an over reduced PSI acceptor side (high Φ NA values). This was the probable reason for PSI inactivation observed in the ANK mutants, but not in WT. In later growth phases, we observed partial relief of "chlorina symptoms," toward WT. Measurements of ΔA 520 decay confirmed that, in early growth stages, the ANK mutants with low PSI content had a limited capacity to build up the transthylakoid proton gradient (ΔpH) needed to trigger non-photochemical quenching (NPQ) and to regulate the electron transport by cytochrome b 6/f. Later, the increase in the PSI/PSII ratio enabled ANK mutants to reach full NPQ, but neither over reduction of the PSI acceptor side nor PSI photoinactivation due to imbalance between the activity of PSII and PSI was mitigated. Thus, our results support the crucial role of proper regulation of linear electron transport in the protection of PSI against photoinhibition. Moreover, the ANK mutants of wheat showing the dynamic developmental changes in the PSI/PSII ratio are presented here as very useful models for further studies.

The application of multiplex fluorimetric sensor for the analysis of flavonoids content in the medicinal herbs family Asteraceae, Lamiaceae, Rosaceae.

BACKGROUND: The aim of our research work was to quantify total flavonoid contents in the leaves of 13 plant species family Asteraceae, 8 representatives of family Lamiaceae and 9 plant species belonging to family Rosaceae, using the multiplex fluorimetric sensor. Fluorescence was measured using optical fluorescence apparatus Multiplex(R) 3 (Force-A, France) for non-destructive flavonoids estimation. The content of total flavonoids was estimated by FLAV index (expressed in relative units), that is deduced from flavonoids UV absorbing properties. RESULTS: Among observed plant species, the highest amount of total flavonoids has been found in leaves of Helianthus multiflorus (1.65 RU) and Echinops ritro (1.27 RU), Rudbeckia fulgida (1.13 RU) belonging to the family Asteraceae. Lowest flavonoid content has been observed in the leaves of marigold (Calendula officinalis) (0.14 RU) also belonging to family Asteraceae. The highest content of flavonoids among experimental plants of family Rosaceae has been estimated in the leaves of Rosa canina (1.18 RU) and among plant species of family Lamiaceae in the leaves of Coleus blumei (0.90 RU). CONCLUSIONS: This research work was done as pre-screening of flavonoids content in the leaves of plant species belonging to family Asteraceae, Lamiaceae and Rosaceae. Results indicated that statistically significant differences (P > 0.05) in flavonoids content were observed not only between families, but also among individual plant species within one family.