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Several food dyes are known to induce amyloid fibrillation when interacting with proteins. Here, we studied the role of sunset yellow (SY) in the amyloid fibrillation of hen egg white lysozyme (HEWL) and characterized the changes using spectroscopy techniques. Turbidity results showed that SY dye induces aggregation in HEWL in concentrations dependent manner. The aggregation induced by SY dye is kinetically very fast, no lag phase was detected, and the kinetics process follows an isodesmic kinetics pathway. The SY-dye induce aggregates have cross-β secondary structure confirmed by far-UV CD measurements. The effect of salts and solvents was also seen on SY-induced aggregates. Turbidity, far-UV CD, and kinetics results suggest that certain concentrations of NaCl and (NH

The central cell withstands heat stress better than the egg and antipodal cells. Insilco analysis of transcriptomic data identified several heat responsive genes which are central cell specific. Crop damage due to heat stress (HS) is a major cause of yield lost. Plants are particularly susceptible to negative effects of HS during gametophyte development and fertilization. Extensive studies have been performed on the male gametophyte under HS, but how the female gametophyte copes with HS is largely unknown. To learn how the different cell types of the female gametophyte reacts to HS, we studied unfertilized CDC123::H2B:YFP ovules. We found that the YFP-specific florescent signal persisted in the central cell during HS significantly more than the egg cell. We also found that the fluorescent signal persistence was the lowest in the antipodal cells. This finding suggests that the reaction of the female gametophyte to HS is rather unique and differentially mediated according to the cell’s identity. In addition, mining through published transcriptomic datasets we found that several important heat stress responsive genes which are extremely upregulated during HS (more than 64-fold) are specifically expressed in the CC but not in the EC. Further research such as comparative transcriptomics and cell biology will likely shed more light on the phenomena reported here and increase our basic understandings about the ways sexual reproduction processes are affected by heat stress.

Plant responses to salinity have been extensively studied over the last decades. Despite the vast accumulated knowledge, the ways Arabidopsis lateral roots (LR) cope with lethal salinity has not been fully resolved. Here we compared the primary root (PR) and the LR responses during events leading to lethal salinity (NaCl 200 mM) in Arabidopsis. We found that the PR and young LR responded differently to lethal salinity: While the PR died, emerging and young LR’s remained strikingly viable. Moreover, “age acquired salt tolerance” (AAST) was observed in the PR. During the 2 days after germination (DAG) the PR was highly sensitive, but at 8 DAG there was a significant increase in the PR cell survival. Nevertheless, the young LR exhibited an opposite pattern and completely lost its salinity tolerance, as it elongated beyond 400 µm. Examination of several cell death signatures investigated in the young LR showed no signs of an active programmed cell death (PCD) during lethal salinity. However, Autophagic PCD (A-PCD) but not apoptosis-like PCD (AL-PCD) was found to be activated in the PR during the high salinity conditions. We further found that salinity induced NADPH oxidase activated ROS, which were more highly distributed in the young LR compared to the PR, is required for the improved viability of the LR during lethal salinity conditions. Our data demonstrated a position-dependent resistance of Arabidopsis young LR to high salinity. This response can lead to identification of novel salt stress coping mechanisms needed by agriculture during the soil salinization challenge.

Earlier, we reported that Arabidopsis young lateral roots (LR) exhibited improved lethal salinity tolerance as compared with the primary root (PR). We have shown that cell death processes which take place in the PR during salt stress are postponed in the LR. Still, very little is known about the regulation of cell survival mechanisms in the LR during salinity stress. Here we used transgenic Arabidopsis plants expressing Cyclin B1;1:GUS, to further study the responses to salinity in the PR and LR positions. We found strong Cyclin B1;1:GUS activity in young budding LR of salt stressed and stress recovered plants. The Cyclin B1;1:GUS activity dropped significantly in long LR and was almost completely abolished in the PR. Our data provides another line of evidence that position-dependent response occurs in Arabidopsis roots during lethal salinity. The possible roles Cyclin B1;1 plays in the young LR during the response to lethal salinity are discussed.

Absorption of excess excitation energy induces overproduction of singlet oxygen (1O2) in plants. The major sources of singlet oxygen production are chlorophyll and its intermediates located in the chloroplast. Over-accumulation of the chlorophyll biosynthetic intermediate protochlorophyllide by the exogenous application of 5-aminolevulinic acid (ALA), the precursor of tetrapyrrole, induced singlet oxygen production in the plastidic membranes. Over-expression of protochlorophyllide oxidoreductase C (PORC) in Arabidopsis thaliana resulted in efficient light-induced photo-transformation of protochlorophyllide to chlorophyllide that limited the accumulation of protochlorophyllide. Consequently, the 1O2 generation decreased in the PORC overexpressors (PORCx) and their cell death was minimal. Conversely, porC-2 over-accumulated protochlorophyllide in response to ALA treatment and generated higher amounts of 1O2 in light and had highest cell death as monitored by Evans blue staining. The protoplasts isolated from PORCx plants, when treated with ALA, generated minimal amounts of 1O2 as revealed by singlet oxygen sensor green (SOSG) fluorescence emission from chloroplasts. Conversely, the protoplasts of porC-2 mutants under identical conditions generated the maximum SOSG fluorescence in their chloroplasts and cytosol surrounding the chloroplasts most likely due to the leakage from the organelle. The membrane blebbing, a hallmark of programmed cell death, was clearly visible in WT and porC-2 protoplasts. Similarly, the nick end labelling (TUNEL) assay revealed nicks in the DNA. The TUNEL-positive nuclei after 30 min of light exposure were highest in porC-2 and lowest in PORCx protoplasts. The results demonstrate that higher amounts of singlet oxygen produced in the chloroplasts play an important role in programmed cell death.

In this paper we provide evidence for involvement of chloroplast as alternate organelle for initiating PCD in plants under light and abiotic stress. In animals, mitochondria are the major source of reactive oxygen species (ROS) and key executioner of programmed cell death (PCD). In plants, however, the primary site of generation of ROS is chloroplast and yet its involvement in PCD has not been worked out in details. We found by Evans blue staining that salt (150 mM NaCl)-treated protoplasts obtained from green seedlings had higher rate of cell death than protoplasts obtained from etiolated seedlings. This indicated that cell death induced by NaCl is accentuated by light. Imposition of salt-stress to green protoplasts generated H2O2. Known hallmarks of PCD i.e., blebbing of cell membrane, loabing in nucleus, nick in DNA were observed in light-exposed salt-treated protoplasts and seedlings. TUNEL-FACS assay demonstrate several DNA nicks in the salt-treated green protoplasts exposed to light. Conversely, salt-treated etiolated protoplasts kept in dark had only a few TUNEL-positive nuclei. Similarly, a substantial numbers of TUNEL positive nuclei were observed in green seedlings due to salt treatment in light. However, salt-treated etiolated seedlings kept in dark had very few TUNEL positive nuclei. Addition of Caspase 3 inhibitor (DAVD-CHO) rescued (~50 %) green protoplasts from salt-stress induced cell death suggesting an involvement of apoptosis like PCD (AL-PCD). Ultra structure studies of chloroplast, mitochondria and nucleus from the leaves obtained from salt treated seedlings at the time point that showed PCD signature, resulted to severe granal de-stacking in chloroplasts while structural integrity of mitochondria was maintained. These studies demonstrate the photo-modulation of salinity-induced PCD in photosynthetic tissues is mainly executed by chloroplasts.

Anhydrobiosis is an adaptive strategy of certain organisms or specialised propagules to survive in the absence of water while programmed cell death (PCD) is a finely tuned cellular process of the selective elimination of targeted cell during developmental programme and perturbed biotic and abiotic conditions. Particularly during water stress both the strategies serve single purpose i.e., survival indicating PCD may also function as an adaptive process under certain conditions. During stress conditions PCD cause targeted cells death in order to keep the homeostatic balance required for the organism survival, whereas anhydrobiosis suspends cellular metabolic functions mimicking a state similar to death until reestablishment of the favourable conditions. Anhydrobiosis is commonly observed among organisms that have ability to revive their metabolism on rehydration after removal of all or almost all cellular water without damage. This feature is widely represented in terrestrial cyanobacteria and bryophytes where it is very common in both vegetative and reproductive stages of life-cycle. In the course of evolution, with the development of advanced vascular system in higher plants, anhydrobiosis was gradually lost from the vegetative phase of life-cycle. Though it is retained in resurrection plants that primarily belong to thallophytes and a small group of vascular angiosperm, it can be mostly found restricted in orthodox seeds of higher plants. On the contrary, PCD is a common process in all eukaryotes from unicellular to multicellular organisms including higher plants and mammals. In this review we discuss physiological and biochemical commonalities and differences between anhydrobiosis and PCD.

Anhydrobiosis is a well-known phenomenon represented in a range of plant, animal and microbial taxa. It is a unique faculty of some organisms to survive in the absence of even traceable water in their cells. As water is essential for life and unequivocally important for all the biological reactions and molecular integrity of cellular components, surviving without water is nature’s toughest puzzle. Initial research in this direction to understand the overall mechanism and dynamics of cellular water was done several decades ago with primary focus on characterizing cellular water at different intensities of dehydration using biophysical tools that lead to the establishment (/development) of a correlation of water-binding ability of cell constituents with the extent of anhydrobiosis. Unfortunately, in due course, the research focus had gradually changed and lost this initial direction. We believe a precise characterization of cellular water during dehydrated conditions is still pertinent in the current era of multidisciplinary research. Here cellular water characterized employing biophysical tools can be very precisely correlated with ongoing metabolic activities and other molecular regulatory events that in turn help in characterizing anhydrobiosis. This review intends to revive earlier approaches that were used to characterize the state of cellular water. This could help modern research in anhydrobiosis to correlate ongoing molecular events and/or adaptations during any change in the hydration level very precisely with the particular state of cell water.

Programmed cell death (PCD) is an integral cellular program by which targeted cells culminate to demise under certain developmental and pathological conditions. It is essential for controlling cell number, removing unwanted diseased or damaged cells and maintaining the cellular homeostasis. The details of PCD process has been very well elucidated and characterized in animals but similar understanding of the process in plants has not been achieved rather the field is still in its infancy that sees some sporadic reports every now and then. The plants have 2 energy generating sub-cellular organelles- mitochondria and chloroplasts unlike animals that just have mitochondria. The presence of chloroplast as an additional energy transducing and ROS generating compartment in a plant cell inclines to advocate the involvement of chloroplasts in PCD execution process. As chloroplasts are supposed to be progenies of unicellular photosynthetic organisms that evolved as a result of endosymbiosis, the possibility of retaining some of the components involved in bacterial PCD by chloroplasts cannot be ruled out. Despite several excellent reviews on PCD in plants, there is a void on an update of information at a place on the regulation of PCD by chloroplast. This review has been written to provide an update on the information supporting the involvement of chloroplast in PCD process and the possible future course of the field.

Cancer is currently the second biggest cause of death in the Western world. Cancer cells escape the normal process of programmed cell death i.e., fail to die on schedule. The ability of cancer cells to avoid programmed cell death and continue to proliferate is one of the fundamental hallmarks of cancer and is a major target of cancer therapy development. Universities and research institutes are playing a major role in progress of cancer research. The aim of this study is to attract graduates of different disciplines towards cancer research and bring together researchers from different disciplines with an interest in the role of programmed cell death in cancer therapy and exploitation of programmed cell death research for therapeutic targeting of cancer. In spite of this, it is of broad interest to make a bridge or to start collaborations in between basic researchers and medical oncologists as well as for pharmaceutical companies i.e., aim of this study is to bridging the gap between knowledge and its action or application.