Genetic Engineering and Increasing Resistance of Plants to chilling by Creating Mutants Bacteria Lacking the Ice Nucleation

Abstract

Abstract. Ice nucleation active bacteria are important in damages which are caused by chilling and freezing in plants that are sensitive to cold. Every treatmentwhich leads to a reduction in the number of ice- causing nucleation bacteria or activity of bacteria ice-causing nucleation decreases the amount of frost damage in plants.One of these ways is a change in genetic structure of coding inaz protein so that, this protein isn’t synthesized in bacteria or is made imperfectly and therefore, ice nucleation formation and freezing is delayed in plant.The Purpose of the present study is recognition of INA isolates of Pseudomonas Putidabacteria and creating INA mutants. For this study, plant samples weregathered from almond trees which showed symptoms of freezing in spring. After separating and recognition of bacteria strains fromplant tissue the range of ice nucleation activity was determined by droplet freezing method.Using polymer chain reaction(PCR) the presence or absence of INA gene in bacteria isolates was studied and detected. Production of PCR was done and by Electrophoresis Erwiniaherbicola epiphyte strain Vector of colonized gene of inazed was used as positive control. For creating mutation inp.putida strain containing nucleation activity,electroporation method and for creating mutation in ice gene,Tn 5 transposonwas used.Mutations got resistant to kanamycin and were chosen with selective medium containing Kanamycin. Studyingthe effect of mutations on cucumber planting showed that some mutations didn’tshow freezing symptoms and planting remained normal, whereas some of isolations andcontrol providedpositive freezing evidence.