Transgenic Approaches For Abiotic Stress Tolerance In Plants

1. Transgenic Approaches For Abiotic Stress Tolerance In Plants

2. Contents • Abiotic stress • Different abiotic stress creates water deficit • Problems caused by water deficit • Transgenic approach to improve stress tolerance • Single antion genes • Osmoprotectants • Detoxifying genes • LEA proteins • Transporter genes • Multifunctional genes for lipid biosynthesis • Heat shock protein genes • Regulatory genes • Signal transduction genes • Cold induced proteins • Conclusion • Refrences

3. Abiotic Stress • The negative impact of non-living factors on the living organisms in a specific environment. • The most basic stressors include: high winds, extreme temperatures,  drought, flood, and other natural disasters, such as tornados and wildfires and lesser known includes poor edaphic conditions like rock content and pH, high radiation, compaction, contamination. • Stress triggers a wide range of plant responses: - Altered gene expression - Cellular metabolism - Changes in growth rate and crop yield

4. EFFECCT OF ABIOTIC STREES ON PLANTS

5. Different abiotic stresses create a water deficit • Drought, is by definition, a water deficit stress, because the environmental conditions either reduce the soil water potential or increase the leaf water potential due to hot, dry, or windy conditions. • High sat conditions can result in water deficit because the soil water potential is decreased, making it more difficult for root to extract water from the soil. • High ambient temprature cause increased water loss by evaporation. • Freezing temprature also cause osmotic stress because of the formation of ice crystals in extracellular space. These various cause of water deficit result in efflux of cellular water, leading to plasmolysis and eventually death.

6. Problems Caused By Water Deficit • Inhibits photosynthesis via its effect on thylakoid membrane. • Leads to increase in concentration of toxic ions. • Loss of protective hydration shell around vulnerable molecules.

8. Transgenic Approach to Improve Stress Tolerance • Stress-induced gene expression can be broadly categorized into three groups: • genes encoding proteins with known enzymatic or structural functions, • proteins with as yet unknown functions, and • regulatory proteins. • Initial attempts to develop transgenics (mainly tobacco) for abiotic stress tolerance involved ‘‘single action genes’’ i.e., genes responsible for modification of a single metabolite that would confer increased tolerance to salt or drought stress

9. Single Action Genes • Osmoprotectants: • These fall into two categories: sugars and sugar alcohols, and zwitter ion compounds. Sugars include sugar alcohols such as mannitol, sorbitol, pinitol and oligosaccharides such as trehalose and fructans. The latter class includes amino acids such as proline, and quaternary ammonium compounds such as glycine betaine. • Basic strategy for engineering resistance to water deficit stress have therefore focused on production of osmoprotectants as a mechanism for overcoming the osmotic stress generated by water deficit. • This requires the determination of the biosynthetic pathways for various osmoprotectants, isolation of relevant genes, and appropriate engineering of constructs to target gene expression and protein destination.

11. Osmoproctentants have two roles: protection of vulnerable molecules and osmotic adjustment. Drought toleramce by expressing these molecules can improve plant survival but may not increase yields to viable levels

12. Detoxifying genes • In most of the aerobic organisms, there is a need to effectively eliminate reactive oxygen species (ROS) generated as a result of environmental stresses. • In order to control the level of ROS and protect the cells from oxidative injury, plants have developed a complex antioxidant defense system to scavenge the ROS. • These antioxidant systems include various enzymes and non-enzymatic metabolites that may also play a significant role in ROS signaling in plants. • For example transgenic tobacco coding for superoxide dismutase in cytosol, mitochondria, chloroplast have been generated • Ascobateperoxidase , glutathione reductase and glutathione peroxidase have been transformed into Arabidopsis and tobacco plants and shown to have some impact on various abiotic stresses such as heat, cold and salinity.

13. Reactive oxygen species (ROS)

14. Late embryogenesis abundant (LEA) proteins • Another category of high molecular weight proteins that are abundant during late embryogenesis and accumulate during seed desiccation and in response to water stress • Constitutive overexpression of the HVA1, a group 3 LEA protein from barley conferred tolerance to soil water deficit and salt stress in transgenic rice plants • The group 1 LEA proteins are predicted to have enhanced water-binding capacity, while the group 5 LEA proteins are thought to sequester ions during water loss. • Also play a role in anti aggregation of enzymes under dessication and freezing stresses.

15. Transporter Genes • Important strategy for achieving greater tolerance to abiotic stress is to help plants to re-establish homeostasis under stressful environments, restoring both ionic and osmotic homeostasis. • A number of abiotic stress tolerant transgenic plants have been produced by increasing the cellular levels of proteins (such as vacuolar antiporter proteins) that control the transport functions. • For example, transgenic melonand tomato plants expressing the HAL1 gene showed a certain level of salt tolerance as a result of retaining more K+ than the control plants under salinity stress.

16. Multifunctional genes for lipid biosynthesis • Transgenic approaches also aim to improve photosynthesis under abiotic stress conditions through changes in the lipid biochemistry of the membranes. • Adaptation of living cells to chilling temperatures is a function of alteration in the membrane lipid composition by increased fatty acid unsaturation. • Genetically engineered tobacco plants over-expressing chloroplast glycerol-3-phosphate acyltransferase (GPAT) gene (involved in phosphatidyl glycerol fatty acid desaturation) from squash (Cucurbita maxima) and A. thaliana showed an increase in the number of unsaturated fatty acids and a corresponding decrease in the chilling sensitivity.

17. Heat shock protein genes • The heat shock response, the increased transcription of a set of genes in response to heat or other toxic agent exposure is a highly conserved biological response, occurring in all organism. • The response is mediated by heat shock transcription factor (HSF) which is present in a monomeric, non-DNA binding form in unstressed cells and is activated by stress to a trimeric form which can bind to promoters of heat shock genes. • The induction of genes encoding heat shock proteins (Hsps) is one of the most prominent responses observed at the molecular level of organisms exposed to high temperature • Genetic engineering for increased thermo-tolerance by enhancing heat shock protein synthesis in plants has been achieved in a number of plant species

18. Regulatory genes • Many genes that respond to multiple stresses like dehydration and low temperature at the transcriptional level are also induced by ABA, which protects the cell from dehydration • In order to restore the cellular function and make plants more tolerant to stress, transferring a single gene encoding a single specific stress protein may not be sufficient to reach the required tolerance levels • To overcome such constraints, enhancing tolerance towards multiple stresses by a gene encoding a stress inducible transcription factor that regulates a number of other genes is a promising approach • The CBF1 cDNA when introduced into tomato (Lycopersiconesculentum) under the control of aCaMV35S promoter improved tolerance to chilling, drought and salt stress but exhibited dwarf phenotype and reduction in fruit set and seed number

19. Signal transduction genes • Genes involved in stress signal sensing and a cascade of stress-signaling in A. thaliana has been of recent research interest. • Abiotic stress signaling in plants involves receptor-coupled phospho-relay, phosphoionositol- induced Ca2+ changes, mitogen activated protein kinase (MAPK) cascade, and transcriptional activation of stress responsive genes • One of the merits for the manipulation of signaling factors is that they can control a broad range of downstream events that can result in superior tolerance for multiple aspects • Alteration of these signal transduction components is an approach to reduce the sensitivity of cells to stress conditions, or such that a low level of constitutive expression of stress genes is induced. • Overexpression of functionally conserved At-DBF2 (homolog of yeast DBf2 kinase) showed striking multiple stress tolerance in Arabidopsis plants

20. Cold induced proteins • During the period of acclimation, plants produce a number of cold induced proteins that are assumed to play a role in the subsequent cold resistance. • These are encoded by class of genes designated as cold-responsive (COR) genes according to their patterns of expression. • Overexpression of ectopic expression of these cold induced proteins could therefore be a possible route to the specific engineering of cold or freezing stress tolerance

21. Conclusion • Crops experience a number of different abiotic stress, several of these cause the two major problems: water deficit and oxidative stress. Two general strategies for engineering tolerance to abiotic stress in plants are therefore possible. Some measures of tolerance to water deficit stress can be provided by the synthesis of compatible solutes. On the other hand, expressing enzymes involved in protection against ROS can combat oxidative stress. In some cases, single gene mechanism for tolerating specific stresses can be deployed (eg. Salt stress and cold stress). However, the overriding theme is that abiotic stress induce complex reactions from plants, and that optimal protection may well involve several genes.

22. Refrences • Slater Adrian, Scott W. Nigel “Plant biotechnology- the genetic manipulatin of plants” oxford university press Second Edition 2008 “strategies for engineering stress tolerance”212-234 • MathurPoojaBhatnagar Sharma k. kiran “Transgenic approaches for abiotic stress tolerance in plants:retrospect and prospects” 2007 • Alia H, Sakamoto A, Murata N (1998) Enhancement of tolerance of Arabidopsis to high temperature by genetic engineering of the synthesis of glycine betaine. Plant J 16:155–161 • Allen RD (1995) Dissection of oxidative stress tolerance using transgenic plants. Plant Physiol 107:1049–1054 • Murata N, Ishizaki-Nishizawa O, Higashi S, Hayashi S, Tasaka Y, Nishida I (1992) Genetically engineered alteration in the chilling sensitivity of plants. Nature 356:710–713