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Transgenic Approach for Abiotic Stress Tolerance No. Characteristics features of plant stress protein Stress Protein Induction agent Characteristics features 1 HSP (heat shock Mostly by high protein) temperature stress, also by water stress, salt stress, low temperature stress, in some cases also by abscisic acid Classified aslow molecular weight HSPs and high molecular weight HSPs, highly conserved amino acid sequence, nucleotide sequence of the corresponding genes is also conserved, stress-inducible of hs genes governed by HSEs (heat shock elements), selected HSPs are shown to act as chaperons 2 Osmotic stress proteins such as WSPs (water stress proteins) and SSPs (salt stress proteins) Varied molecular weights and cellular locations, these proteins are mostly the enzymes involved in diverse functions such as production of different osmolytes, protein degradation, signal transduction events, gene regulation and transport. Roles of some WSPs is not well defined (i.e. such as for dehydrins, late embryogenesis, abundant proteins) Mostly by low water availability and salt stress, also induced by ABA Characteristics features of plant stress protein No. Stress Protein Induction agent Characteristics features 3 ANPs (anaerobic proteins) Mostly by anaerobic stress (caused by flooding or submergence stress) Discovered initially in maize and later shown tin o be universally present, most of the ANPs have been shown to be the enzymes of the fermentative or the glycolytic pathway, most genes encoding ANPs contain . AREs (anaerobic response elements) in their promoters 4 Cold stress proteins such as COR (coldregulated proteins) Mostly by low temperature stress, also by osmotic, oxidative stress and ABA Conserved proteins of varied sizes and functions, most COR genes contain specific nucleotide sequence that stimulate transcription in response to low temperature Stress Induced gene expression 1.Genes encoding proteins with known enzymatic or structural functions 2.Protein with as yet unknown functions 3.Regulatory protein Single action gene Osmoprotectant genes: Proline, Gly/Bet, Sugar polyols Detoxifying genes: SOD, PX Late embryogenesis abundant genes: LEA Transporter genes: Aquapurin, Ion transporter Multifunctional genes for lipid biosynthesis:GPAT (glycerol-3-phosphate acyltransferase), FAD7 (Fatty acid desaturase) Heat shock genes: HS Regulatory Genes Transcription factors genes CBF/DREB, ABF, HSF, bZIP, MYC/MYB Signal transduction genes a. Osmocensors (AtHK-1) b. Phospholipid cleaving enzymes (PLD) c. Second messengers (Ptd-OH, ROS) d. MAP kinases, Ca+ censor (SOS-3) e. Calcium –dependent protein kinases (CDPKs) Abiotic stress tolerance 1.Increase cellular level of osmotically-active solute (proline, glycinebetaine, mannitol, trehalose, fructans) 2.Increase levels of osmolytes 3.It is mediated by a number of biochemical reactions /physiological processes (a multi genic trait) 4.It can be augmented by pyramiding different stress-responsive genes Osmolyte and Compatible Solutes Gene gpat Protein Glycerol 3phosphate acyltransferase Source Cucurbita maxima, Arabidopsis thaliana Cellular role(s) Fatty acid unsaturation mtlD Mannitol 1phosphate dehydrogenase Superoxide dismutase Betaine aldehyde dehydrogenase Eschericia coli Manitol biosynthesis Nicotiana plumbaginifolia Eschericia coli Superoxide dismutase Glycinebetaine dismutase sod Bet-B Osmolyte and Compatible Solutes Gene Bet-A Source Eschericia coli Sac-B Protein Choline dehydrogenase Pyroline 5carboxylase synthase Levan sucrase Hva-1 LEA protein Tps-1 Trehalose 6phosphate synthase Hordeum vulgare Arabidopsis thaliana p5cs V. aconitifolia Cellular role(s) Glycinebetaine dismutase Proline biosynthesis Baccilus subtilis Fructan biosynthesis - Trehalose biosynthesis Osmolyte and Compatible Solutes Gene Protein Cod-A/Cod-1/Cox Choline oxidase Source Arthrobacter globiformis afp Antifreeze protein Synthetic (AFP) Imt-1 Myo-inositol-omethyl transferase Betaine dehydrogenase Inhibit ice growth and recrystallization Messembryanthe D-ononitol mum crystallinum biosynthesis BADH Spinach Cellular role(s) Glycinebetaine biosynthesis Glycinebetaine byosynthesis Osmolyte and Compatible Solutes Gene Ect-A, Ect-B, Ect-C Ots-A, Ots-B Protein L-2,4diaminobutyric acetyltransferase L-2,4diaminobutyric acid trans-aminase L-ectoine synthase Trehalose-6-P synthase Trehalose-6-P phosphatase Source Halomonas elongata Cellular role(s) Ectoyne Eschericia coli Trehalose Osmolyte and Compatible Solutes Gene Pro-DH HAL-3 Protein Source Cellular role(s) Proline Arabidopsi Proline dehydrogena s thaliana se FMN-binding Saccharo Na+/K+ protein myces homeostas cerevisae is Ion Transporters and Ion Homeostasis Gene Protein Source AtNHX-1 Vacuolar Na+/H+ antiporter Plasma membrane Na+/H+ antiporter Vacuolar H+pyrophosphatase K+/Na+ transport regulation Arabidopsis thaliana Arabidopsis thaliana AtSOS-1 AVP-1 HAL-1 Arabidopsis thaliana Saccharomyc es cerevisiae Cellular role(s) Na+ vacuolar sequestration Na+ extrusion Vacuolar acidification K+/Na+ homeostasis Redox Proteins Gene MnSOD Gly-1 Protein Superoxide dismutase Glyoxylase Source Saccharomyces cerevisiae Brassica juncea Cellular role(s) Reduction of O2 content S-Dlactoylglutathion e TPX-2 Peroxidase Nicotiana tabacum Change cell properties GST Glutathione Stransferase Glutathione peroxidase Nicotiana tabacum Nicotiana tabacum ROS scavenging GPX Transcription and signal transduction factors Gene Protein DREB-1A Transcription factor Cnb-1 Calcineurin OsCDPK-7 Protein kinase Source Cellular role(s) Arabidopsis Improved thaliana gene expression Saccharomyc Improved es cerevisiae Ca++ signaling Oryza sativa Improved gene expression Mischelineous Gene DnaK Apo-Inv Protein Source Cellular role(s) Heat shock A. Protein protein halophytica stabilization Apoplastic yeast- Saccharomyc Sucrose derived invertase es cerevisiae synthesis Perspective in Abiotic Stress Tolerance 1. Abiotic stress elicit multigenic responses within the plant cells. The tolerance to different abiotic stress is contributed by a range of different biochemical/physiological mechanism 2. Only a limited number of plant genes with a definite function have been identified, cloned and characterized 3. Changing levels of transcription factors can alter the levels of several genes at the same time 4. The transgenics raised far for enhancing tolerance to abiotic stress have been achieved through employing strong constitutive promoters, for driving expression of the transgenes. Promoters which are induced by different abiotic stress including high temperature stress, anaerobic stress, salt stress and water stress have poor strength of expression when compared to constitutively expression promoter