** AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** ALGAE, INCLUDING CYANOBACTERIA, ARE AN EMERGING CROP FOR GENERATING CARBON NEUTRAL RENEWABLE COMMODITIES LIKE BIOFUELS, FOODS, AND PHARMACEUTICALS. BUT WATER SUSTAINABILITY IS A PROMINENT CONCERN IN AGRICULTURAL ALGAL POND CULTIVATION BOTH ECOLOGICALLY AND FINANCIALLY, WITH WATER SEPARATION ALONE ACCOUNTING FOR 20-30% OF CURRENT ALGAL PRODUCTION COSTS. IMMOBILIZED BIOFILM CULTURES ADDRESS THIS CHALLENGE BY REDUCING SEPARATION COSTS AND WATER SUSPENSION NEEDS. BIOFILMS ARE STICKY AGGREGATES OF BACTERIAL CELLS THAT ARE OFTEN STUDIED IN THE CONTEXT OF BACTERIAL PATHOGENS, AS THEY FACILITATE BACTERIAL RESILIENCE TO HARSH ENVIRONMENTS LIKE HUMAN AND OTHER ANIMAL BODIES. CYANOBACTERIAL BIOFILMS ARE NOT PATHOGENIC BUT POSSESS DYNAMIC, STRESS-ADAPTIVE COMPOSITIONS - HOLDING CONSTITUENTS THAT PROTECT BOTH THE CYANOBACTERIA AND NEIGHBORING PLANTS AND SOILS. HOWEVER, LITTLE IS KNOWN ABOUT HOW THESE BIOFILMS, OR WHAT IN THEM, PROTECTS THE CYANOBACTERIA FROM HARSH ABIOTIC OR ENVIRONMENTAL CLIMATE-RELEVANT STRESSES, LIKE DROUGHT, TEMPERATURE, SALT, AND PH. THEREFORE, I AM GENETICALLY REPROGRAMMING OR ENGINEERING NATIVE BIOFILM MATRIX MODULATORS IN A MODEL INDUSTRIAL CYANOBACTERIUM TO DETERMINE MECHANISMS CONFERRING ABIOTIC STRESS RESILIENCE IN A STRAIN WITH INHERENTLY POOR BIOFILM FORMATION AND STRESS TOLERANCE. GENETIC MUTANT BIOFILM FORMATION AND VIABILITY UNDER ABIOTIC STRESS WILL BE COMPARED WITH UN-ENGINEERED OR WILD TYPE CYANOBACTERIAL CULTURES. INITIAL MUTANTS EXHIBIT ~10X INCREASED VIABILITY UNDER DROUGHT STRESS COMPARED TO WILD TYPE, PRESENTING COMPELLING PRELIMINARY EVIDENCE THAT ABIOTIC STRESS TOLERANCE CAN BE ENGINEERED INTO A NON-TOLERANT CYANOBACTERIUM. NEXT, MUTANTS WILL BE EXPOSED TO DIFFERENT STRESSES AND PROFILES OF THEIR BIOFILM CONTENTS AND GENE EXPRESSION WILL BE COMPARED UNDER IDEAL CONDITIONS VS STRESS TO ASCERTAIN WHAT BIOLOGICALLY CONTRIBUTES TO STRESS ADAPTATION.OVERALL, THIS FUNDAMENTAL RESEARCH AIMS TO GENERATE MORE STRESS-ROBUST INDUSTRIAL CYANOBACTERIAL STRAINS, REDUCING ECONOMIC AND ECOLOGICAL BURDENS IN THE ALGAL CULTIVATION INDUSTRY AND HELPING FACILITATE THE EXPANSION AND ACCESSIBILITY OF ALGAL CROPS. UNCOVERING BIOLOGICAL MECHANISMS OF STRESS-TOLERANCE IN CYANOBACTERIA IS TRANSLATABLE TOWARDS IMPROVEMENT OF CLIMATE RESILIENCE WITHIN THE AG SECTOR ACROSS SOIL AND CROP HEALTH. THE PROJECT WILL IMPACT THE PUBLIC COMMUNITY BY GENERATING OPEN-ACCESS PEER-REVIEWED JOURNAL ARTICLE(S), NUMEROUS PUBLIC RESEARCH PRESENTATIONS TO DIVERSE AUDIENCES, TRAINING AND MENTORSHIP FOR UNDERREPRESENTED AND EARLY-CAREER SCIENTISTS AND FACILITATING ENGAGING PUBLIC SCIENTIFIC OUTREACH AROUND TOPICS LIKE CLIMATE CHANGE AND ADAPTATION IN AGRICULTURE. THIS RESEARCH IS RELEVANT TO THE AFRI PLANT HEALTH AND PRODUCTION AND PLANT PRODUCTS PRIORITY BY TARGETING TUNABLE STRESS RESILIENCE PHENOTYPES IN MODEL CYANOBACTERIA WITH TRANSLATIONAL APPLICATIONS TOWARDS TERRESTRIAL CROP AND SOIL HEALTH IN THE FACE OF INCREASING CLIMATE INSTABILITY. THIS STUDY ALSO ADV,ANCES AFRI PRIORITIES IN BIOENERGY, NATURAL RESOURCES, AND ENVIRONMENT BY ENHANCING ENVIRONMENTAL STRESS TOLERANCE IN AN EMERGING BIOFUEL CROP AND IMPROVING WATER SUSTAINABILITY.