Suprasanna Penna

Building stress tolerance through over-producing trehalose in transgenic plants

Trehalose is a rare sugar with unique abilities to protect biomolecules from environmental stresses and is present in many bacteria, fungi and some desiccation-tolerant higher plants. Increasing trehalose accumulation in crop plants could improve drought and salinity tolerance. Transgenic plants have been developed with trehalose biosynthetic genes--a recent study on the stress-inducible overexpression of the bifunctional TPSP fusion gene in transgenic rice could offer novel strategies for improving abiotic stress tolerance in crop plants.

Abiotic Stress Responses in Plants: Present and Future

Drought, cold, high-salinity and heat are major abiotic stresses that severely reduce the yield of food crops worldwide. Traditional plant breeding approaches to improve abiotic stress tolerance of crops had limited success due to multigenic nature of stress tolerance. In the last decade, molecular techniques have been used to understand the mechanisms by which plants perceive environmental signals and further their transmission to cellular machinery to activate adaptive responses. This knowledge is critical for the development of rational breeding and transgenic strategies to impart stress tolerance in crops. Studies on physiological and molecular mechanisms of abiotic stress tolerance have led to characterisation of a number of genes associated with stress adaptation. Techniques like microarrays have proven to be invaluable in generating a list of stress-related genes. Some of these genes are specific for a particular stress while others are shared between various stresses. Interestingly, a number of genes are shared in abiotic and biotic stress responses. This highlights the complexity of stress response and adaptation in plants. There is a whole cascade of genes involved in abiotic stress tolerance; starting from stress perception to transcriptional activation of downstream genes leading to stress adaptation and tolerance. A number of these genes have been discovered but we still do not have the complete list with all interactions. There is also significant number of genes with unknown functions found to be regulated by abiotic stresses. Understanding the function of these genes and their interaction with other known genes to effect stress adaptation is required.

Positive selectable marker genes for routine plant transformation

SummaryPlant genetic transformation technologies rely upon the selection and recovery of transformed cells. Selectable marker genes used so far have been either antibiotic resistance genes or herbicide tolerance genes. There is a need to apply alternative principles of selection, as more transgenic traits have to be incorporated into a transgenic crop and because of concern that the use of conventional marker genes may pose a threat to humans and the environment. New classes of marker genes are now available, conferring metabolic advantage of the transgenic cells over the non-transformed cells. The new selection systems, as described in this review, are being used with success and superior performance over the traditional marker systems.

Thiourea, a ROS Scavenger, Regulates Source-to-Sink Relationship to Enhance Crop Yield and Oil Content in Brassica juncea (L.)

In the present agricultural scenario, the major thrust is to increase crop productivity so as to ensure sustainability. In an earlier study, foliar application of thiourea (TU; a non physiological thiol based ROS scavenger) has been demonstrated to enhance the stress tolerance and yield of different crops under field condition. Towards this endeavor, present work deals with the effect of TU on photosynthetic efficiency and source-to-sink relationship of Indian mustard (Brassica juncea) for understanding its mode of action. The application of TU increased the efficiency of both PSI and PSII photosystems and vegetative growth of plant. The comparative analysis of sucrose to starch ratio and expression level of sugar transporters confirmed the higher source and sink strength in response to TU treatment. The biochemical evidence in support of this was derived from higher activities of sucrose phosphate synthase and fructose-1,6-bis-phosphatase at source; and sucrose synthase and different classes of invertases at both source and sink. This indicated an overall increase in photoassimilate level at sink. An additional contribution through pod photosynthesis was confirmed through the analysis of phosphoenol pyruvate carboxylase enzyme activity and level of organic acids. The increased photoassimilate level was also co-ordinated with acetyl coA carboxylase mediated oil biosynthesis. All these changes were ultimately reflected in the form of 10 and 20% increase in total yield and oil content, respectively under TU treatment as compared to control. Additionally, no change was observed in oil composition of seeds derived from TU treated plants. The study thus signifies the co-ordinated regulation of key steps of photosynthesis and source-to-sink relationship through the external application of TU resulting in increased crop yield and oil content.

Multifaceted roles of aquaporins as molecular conduits in plant responses to abiotic stresses.

Abstract Abiotic stress has become a challenge to food security due to occurrences of climate change and environmental degradation. Plants initiate molecular, cellular and physiological changes to respond and adapt to various types of abiotic stress. Understanding of plant response mechanisms will aid in strategies aimed at improving stress tolerance in crop plants. One of the most common and early symptoms associated with these stresses is the disturbance in plant–water homeostasis, which is regulated by a group of proteins called “aquaporins”. Aquaporins constitute a small family of proteins which are classified further on the basis of their localization, such as plasma membrane intrinsic proteins, tonoplast intrinsic proteins, nodulin26-like intrinsic proteins (initially identified in symbiosomes of legumes but also found in the plasma membrane and endoplasmic reticulum), small basic intrinsic proteins localized in ER (endoplasmic reticulum) and X intrinsic proteins present in plasma membrane. Apart from water, aquaporins are also known to transport CO2, H2O2, urea, ammonia, silicic acid, arsenite and wide range of small uncharged solutes. Besides, aquaporins also function to modulate abiotic stress-induced signaling. Such kind of versatile functions has made aquaporins a suitable candidate for development of transgenic plants with increased tolerance toward different abiotic stress. Toward this endeavor, the present review describes the versatile functions of aquaporins in water uptake, nutrient balancing, long-distance signal transfer, nutrient/heavy metal acquisition and seed development. Various functional genomic studies showing the potential of specific aquaporin isoforms for enhancing plant abiotic stress tolerance are summarized and future research directions are given to design stress-tolerant crops.

Thiourea orchestrates regulation of redox state and antioxidant responses to reduce the NaCl-induced oxidative damage in Indian mustard (Brassica juncea (L.) Czern.).

Thiourea (TU) has been found to enhance the stress tolerance of plants in our earlier field trials. In the present study, the TU mediated effect on the redox and antioxidant responses were studied in response to salinity (NaCl) stress in Indian mustard (Brassica juncea (L.) Czern.) seedlings. Biochemical analyses of reactive oxygen species (ROS) and lipid peroxidation revealed that TU supplementation to NaCl brought down their levels to near control values as compared to that of NaCl stress. These positive effects could be correlated to the significant increases in the 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity, in the levels of reduced glutathione (GSH) and GSH/GSSG (reduced/oxidized glutathione) ratio and in the activities of superoxide dismutase (SOD; EC 1.1.5.1.1) and glutathione reductase (GR; EC 1.6.4.2) in NaCl+TU treatment as compared to that of NaCl treatment. Further, TU supplementation allowed plants to avoid an over-accumulation of pyridine nucleotides, to stimulate alternative pathways (through higher glycolate oxidase activity; EC 1.1.3.15) for channeling reducing equivalents and thus, to maintain the redox state to near control levels. These positive responses were also linked to an increased energy utilization (analyzed in terms of ATP/ADP ratio) and presumably to an early signaling of the stress through stimulated activity of ascorbate oxidase (EC 1.10.3.3), an important component of stress signaling. A significant reduction observed in the level of sodium ion (Na(+)) accumulation indicated that TU mediated tolerance is attributable to salt avoidance. Thus, the present study suggested that TU treatment regulated redox and antioxidant machinery to reduce the NaCl-induced oxidative stress.

Differential Osmotic Adjustment to Iso-osmotic NaCl and PEG Stress in the in vitro Cultures of Sesuvium portulacastrum (L.) L.

The influence of iso-osmotic (−0.7 MPa) NaCl and PEG stress on growth, osmotic adjustment and antioxidant defense mechanisms was investigated in the in vitro cultures of Sesuvium portulacastrum (L.) L. The decreased relative growth rate (RGR) and water content of PEG-stressed calli in comparison to NaCl was found to be correlated with differences observed in the energy expenditure for the maintenance of osmotic balance. Osmotic adjustment in the NaCl-stressed calli favored higher accumulation of saline ions and soluble sugars, whereas PEG-stressed calli confirmed increased levels of organic osmolytes (proline, glycine betaine and soluble sugars). Permeability of Na+ ions across the membrane revealed increased relative electrolytic leakage (REL) in NaCl-stressed calli, however non-penetrating and highly viscous solution of PEG amplified the peroxidation of membrane lipids. Increased activities of superoxide dismutase and catalase displayed efficient removal of toxic reactive oxygen species in comparison to ascorbate peroxidase in the calli exposed to iso-osmotic stress. These findings suggest that differential tolerance potential to iso-osmotic NaCl and PEG stress in terms of osmotic adjustment appears to be the prime defense mechanism of Sesuvium for its survival under iso-osmotic stress conditions at the expense of reduced growth and water content.

Allyl isothiocyanate enhances shelf life of minimally processed shredded cabbage

The effect of allyl isothiocyanate (AITC), in combination with low temperature (10°C) storage on post harvest quality of minimally processed shredded cabbage was investigated. An optimum concentration of 0.05μL/mL AITC was found to be effective in maintaining the microbial and sensory quality of the product for a period of 12days. Inhibition of browning was shown to result from a down-regulation (1.4-fold) of phenylalanine ammonia lyase (PAL) gene expression and a consequent decrease in PAL enzyme activity and o-quinone content. In the untreated control samples, PAL activity increased following up-regulation in PAL gene expression that could be linearly correlated with enhanced o-quinone formation and browning. The efficacy of AITC in extending the shelf life of minimally processed shredded cabbage and its role in down-regulation of PAL gene expression resulting in browning inhibition in the product is reported here for the first time.

Salt-induced stress responses of Brassica (Brassica juncea L.) genotypes

The effects of salt stress were evaluated in two Brassica juncea cultivars, Varuna and TM2, in relation to growth, osmolyte accumulation and antioxidant enzymes at 50, 100 and 200 mmol l−1 NaCl concentration. In general, significant reduction in shoot and root growth and relative water content was observed under salt stress.Among the varieties, extent of reduction for these parameters was more pronounced and significant in the case of Varuna than TM2. Significant accumulation of glycine betaine was observed in Varuna and TM2 in 100 and 200 mmol l−1 salt stress treatments than control but the magnitude of increase was significantly higher in TM2. In general, salt stress led to increased activities of Superoxid dismutase, Catalase and Guaiacol peroxidase. The SOD activity was significantly higher in Varuna than TM2. Catalase activity showed an increase up to 100 mmol l−1 NaCl concentration whereas it decreased at 200 mmol l−1 in both the cultivars. Based on the results, TM2 appears to have higher salt tolerance than Varuna. It is also suggestive that osmotic adjustment and antioxidant enzymes play an important role in contributing to salinity tolerance in Brassica juncea L.

Aliphatic glucosinolate synthesis and gene expression changes in gamma-irradiated cabbage.

Glucosinolates, found principally in the plant order Brassicales, are modulated by different post-harvest processing operations. Among these, ionizing radiation, a non-thermal process, has gained considerable interest for ensuring food security and safety. In gamma-irradiated cabbage, enhanced sinigrin, a major glucosinolate, has been reported. However, the molecular basis of such a radiation induced effect is not known. Herein, the effect of radiation processing on the expression of glucosinolate biosynthetic genes was investigated. RT-PCR based expression analysis of seven glucosinolate biosynthetic pathway genes (MYB28, CYP79F1, CYP83A1, SUR1, UGT74B1, SOT18 and TGG1) showed that CYP83A1, MYB28, UGT74B1, CYP79F1 and SUR1 were up-regulated in irradiated cabbage. The content of jasmonates, signalling molecules involved in glucosinolate induction was, however, unaffected in irradiated cabbage suggesting their non-involvement in glucosinolate induction during radiation processing. This is the first report on the effect of gamma irradiation on the expression of glucosinolate biosynthetic genes in vegetables.