Anjana Jajoo

Chlorophyll a fluorescence study revealing effects of high salt stress on Photosystem II in wheat leaves.

In order to study the effects of high salt stress on PS II in detached wheat (Triticum aestivum) leaves, the seedlings were grown in Knop solution and temperature was 20 +/- 2 degrees C. Detached leaves were exposed to high salt stress (0.1-0.5 M NaCl) for 1 h in dark and Chl a fluorescence induction kinetics was measured. Various parameters like Fv/Fm, ABS/RC, ETo/TRo, performance index and area over the florescence curve were measured and the energy pipeline model was deduced in response to salt stress. Our results show that the damage caused due to high salt stress is more prominent at the donor side rather than the acceptor side of PS II. Moreover the effects of high salt stress are largely reversible, as the acceptor side damage is completely recovered (approximately 100%) while the recovery of the donor side is less than 85%. Based on our results we suggest that in response to high salt stress, the donor side of PS II is affected more as compared to the acceptor side of PS II.

Frequently asked questions about in vivo chlorophyll fluorescence: practical issues

The aim of this educational review is to provide practical information on the hardware, methodology, and the hands on application of chlorophyll (Chl) a fluorescence technology. We present the paper in a question and answer format like frequently asked questions. Although nearly all information on the application of Chl a fluorescence can be found in the literature, it is not always easily accessible. This paper is primarily aimed at scientists who have some experience with the application of Chl a fluorescence but are still in the process of discovering what it all means and how it can be used. Topics discussed are (among other things) the kind of information that can be obtained using different fluorescence techniques, the interpretation of Chl a fluorescence signals, specific applications of these techniques, and practical advice on different subjects, such as on the length of dark adaptation before measurement of the Chl a fluorescence transient. The paper also provides the physiological background for some of the applied procedures. It also serves as a source of reference for experienced scientists.

Chlorophyll a fluorescence as a tool to monitor physiological status of plants under abiotic stress conditions

Plants living under natural conditions are exposed to many adverse factors that interfere with the photosynthetic process, leading to declines in growth, development, and yield. The recent development of Chlorophyll a fluorescence (ChlF) represents a potentially valuable new approach to study the photochemical efficiency of leaves. Specifically, the analysis of fluorescence signals provides detailed information on the status and function of Photosystem II (PSII) reaction centers, light-harvesting antenna complexes, and both the donor and acceptor sides of PSII. Here, we review the results of fast ChlF analyses of photosynthetic responses to environmental stresses, and discuss the potential scientific and practical applications of this innovative methodology. The recent availability of portable devices has significantly expanded the potential utilization of ChlF techniques, especially for the purposes of crop phenotyping and monitoring.

PHOTOSYNTHESIS: RESPONSE TO HIGH TEMPERATURE STRESS

Global warming has led to increased temperature of the earth which is a major abiotic stress posing a serious threat to the plants. Photosynthesis is amongst the plant cell functions that is highly sensitive to high temperature stress and is often inhibited before other cell functions are impaired. The primary sites of targets of high temperature stress are Photosystem II (PSII), ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) while Cytochrome b559 (Cytb559) and plastoquinone (PQ) are also affected. As compared to PSII, PSI is stable at higher temperatures. ROS production, generation of heat shock proteins, production of secondary metabolites are some of the consequences of high temperature stress. In this review we have summarized the physiological, biochemical and molecular aspects of high temperature stress on the process of photosynthesis, as well as the tolerance and adaptive mechanisms involved.

Analysis of high temperature stress on the dynamics of antenna size and reducing side heterogeneity of Photosystem II in wheat leaves (Triticum aestivum)

This study demonstrates the effect of high temperature stress on the heterogeneous behavior of PSII in Wheat (Triticum aestivum) leaves. Photosystem II in green plant chloroplasts displays heterogeneity both in the composition of its light harvesting antenna i.e. on the basis of antenna size (α, β and γ centers) and in the ability to reduce the plastoquinone pool i.e. the reducing side of the reaction centers (Q(B)-reducing centers and Q(B)-non-reducing centers). Detached wheat leaves were subjected to high temperature stress of 35°C, 40°C and 45°C. The chlorophyll a (Chl a) fluorescence transient were recorded in vivo with high time resolution and analyzed according to JIP test which can quantify PS II behavior using Plant efficiency analyzer (PEA). Other than PEA, Biolyzer HP-3 software was used to evaluate different types of heterogeneity in wheat leaves. The results revealed that at high temperature, there was a change in the relative amounts of PSII α, β and γ centers. As judged from the complementary area growth curve, it seemed that with increasing temperature the PSII(β) and PSII(γ) centers increased at the expense of PSII(α) centers. The reducing side heterogeneity was also affected as shown by an increase in the number of Q(B)-non-reducing centers at high temperatures. The reversibility of high temperature induced damage on PSII heterogeneity was also studied. Antenna size heterogeneity was recovered fully up to 40°C while reducing side heterogeneity showed partial recovery at 40°C. An irreversible damage to both the types of heterogeneity was observed at 45°C. The work is a significant contribution to understand the basic mechanism involved in the adaptation of crop plants to stress conditions.

Impact of increasing Ultraviolet-B (UV-B) radiation on photosynthetic processes.

Increased UV-B radiation on the earths surface due to depletion of stratospheric ozone layer is one of the changes of current climate-change pattern. The deleterious effects of UV-B radiation on photosynthesis and photosynthetic productivity of plants are reviewed. Perusal of relevant literature reveals that UV-B radiation inflicts damage to the photosynthetic apparatus of green plants at multiple sites. The sites of damage include oxygen evolving complex, D1/D2 reaction center proteins and other components on the donor and acceptor sides of PS II. The radiation inactivates light harvesting complex II and alters gene expression for synthesis of PS II reaction center proteins. Mn cluster of water oxidation complex is the most important primary target of UV-B stress whereas D1 and D2 proteins, quinone molecules and cytochrome b are the subsequent targets of UV-B. In addition, photosynthetic carbon reduction is also sensitive to UV-B radiation which has a direct effect on the activity and content of Rubisco. Some indirect effects of UV-B radiation include changes in photosynthetic pigments, stomatal conductance and leaf and canopy morphology. The failure of protective mechanisms makes PS II further vulnerable to the UV-B radiation. Reactive oxygen species are involved in UV-B induced responses in plants, both as signaling and damaging agents. Exclusion of ambient UV components under field conditions results in the enhancement of the rate of photosynthesis, PS II efficiency and subsequently increases the biomass accumulation and crop yield. It is concluded that predicted future increase in UV-B irradiation will have significant impact on the photosynthetic efficiency and the productivity of higher plants.

Towards a critical understanding of the photosystem II repair mechanism and its regulation during stress conditions

Photosystem II (PSII) is vulnerable to high light (HL) illumination resulting in photoinhibition. In addition to photoprotection mechanisms, plants have developed an efficient PSII repair mechanism to save themselves from irreversible damage to PSII under abiotic stresses including HL illumination. The phosphorylation/dephosphorylation cycle along with subsequent degradation of photodamaged D1 protein to be replaced by the insertion of a newly synthesized copy of D1 into the PSII complex, is the core function of the PSII repair cycle. The exact mechanism of this process is still under discussion. We describe the recent progress in identifying the kinases, phosphatases and proteases, and in understanding their involvement in the maintenance of thylakoid structure and the quality control of proteins by PSII repair cycle during photoinhibition.

Quality Control of Photosystem II THYLAKOID UNSTACKING IS NECESSARY TO AVOID FURTHER DAMAGE TO THE D1 PROTEIN AND TO FACILITATE D1 DEGRADATION UNDER LIGHT STRESS IN SPINACH THYLAKOIDS

Photosystem II is vulnerable to light damage. The reaction center-binding D1 protein is impaired during excessive illumination and is degraded and removed from photosystem II. Using isolated spinach thylakoids, we investigated the relationship between light-induced unstacking of thylakoids and damage to the D1 protein. Under light stress, thylakoids were expected to become unstacked so that the photodamaged photosystem II complexes in the grana and the proteases could move on the thylakoids for repair. Excessive light induced irreversible unstacking of thylakoids. By comparing the effects of light stress on stacked and unstacked thylakoids, photoinhibition of photosystem II was found to be more prominent in stacked thylakoids than in unstacked thylakoids. In accordance with this finding, EPR spin trapping measurements demonstrated higher production of hydroxyl radicals in stacked thylakoids than in unstacked thylakoids. We propose that unstacking of thylakoids has a crucial role in avoiding further damage to the D1 protein and facilitating degradation of the photodamaged D1 protein under light stress.

Photodamage of iron–sulphur clusters in photosystem I induces non-photochemical energy dissipation

Photosystem I (PSI) uses light energy and electrons supplied by photosystem II (PSII) to reduce NADP+ to NADPH. PSI is very tolerant of excess light but extremely sensitive to excess electrons from PSII. It has been assumed that PSI is protected from photoinhibition by strict control of the intersystem electron transfer chain (ETC). Here we demonstrate that the iron–sulphur (FeS) clusters of PSI are more sensitive to high light stress than previously anticipated, but PSI with damaged FeS clusters still functions as a non-photochemical photoprotective energy quencher (PSI-NPQ). Upon photoinhibition of PSI, the highly reduced ETC further triggers thylakoid phosphorylation-based mechanisms that increase energy flow towards PSI. It is concluded that the sensitivity of FeS clusters provides an additional photoprotective mechanism that is able to downregulate PSII, based on PSI quenching and protein phosphorylation.

Changes in PS II heterogeneity in response to osmotic and ionic stress in wheat leaves (Triticum aestivum)

High salt stress involves ionic stress as well as osmotic stress. In this work we have tried to differentiate between the ionic and osmotic effects of salt stress on the basis of their ability to cause changes in PS II heterogeneity. PS II heterogeneity is found to vary with environmental conditions. Osmotic stress caused no change in the QB reducing side heterogeneity and a reversible change in antenna heterogeneity. The number of QB non-reducing centers increased under ionic stress but were unaffected by osmotic stress. On the other hand ionic stress led to a partially irreversible change in QB reducing side heterogeneity and a reversible change in antenna heterogeneity. In response to both ionic and osmotic effect, there is conversion of active PS IIα centres to inactive PSIIβ and γ centres.