Potential of the Ice-minus Bacteria

Ice-minus bacteria is a nickname given to a variant of the common bacteria Pseudomonas syringae (P. syringae). This breed of P. syringae misses the power to develop a certain surface protein, generally detected on wild-type “ice-plus” P. syringae. The “ice-plus” protein (Ina protein, “Ice nucleation-active” protein) detected on the outer bacterial cell membrane acts as the nucleating cores for ice crystals. This facilitates ice formation, therefore the designation “ice-plus.” The ice-minus form of P. syringae is a mutation, missing the gene accountable for ice-nucleating surface protein output. This deficiency of surface protein provides a less complimentary surroundings for ice formation. Both strains of P. syringae occur naturally, but genetic engineering has provided the celluloid removal or alteration of specific genes, sanctioning the conception of the ice-minus strain.

The success of the agricultural world is to a great extent dependent on the weather. Cold weather circumstances are straightaway responsible for the appearance of frost on plants and first and foremost, crops. In the United States alone, it has been approximated that frost accounts for approximately $1 billion in crop damage each year. As P. syringae commonly inhabits plant surfaces, its ice nucleating nature motivates frost development, freezing the buds of the plant and destructing the occurring crop. The creation of an ice-minus strain of P. syringae to the surface of plants would incur competition between the strains. Should the ice-minus strain win out, the ice nucleate provided by P. syringae would no longer be present, lowering the level of frost evolution on plant surfaces at normal water freezing temperature (0oC). Even if the ice-minus strain does not win out, the amount of ice nucleate present from ice-plus P. syringae would be reduced due to competition. Reduced degrees of frost generation at normal water freezing temperature would transform into a lowered quantity of crops lost due to frost damage, rendering higher crop yields overall.

To consistently produce the ice-minus breed of P. syringae, its ice-forming gene must be detached, exaggerated, inactivated and re-introduced into P. syringae bacterium. The following steps are often utilized to isolate and generate ice-minus strains of P. syringae:

• Digest P. syringae’s DNA with restriction enzymes.

• Insert the individual DNA pieces into a plasmid. Pieces will insert randomly, allowing for different variations of recombinant DNA to be produced.

• Transform the bacterium Escherichia coli (E.coli) with the recombinant plasmid. The plasmid will be taken in by the bacteria, rendering it part of the organism’s DNA.

• Identify the ice-gene from the numerous newly developed E. coli recombinants. Recombinant E. coli with the ice-gene will possess the ice-nucleating phenotype, these will be “ice-plus.”

• With the ice nucleating recombinant identified, amplify the ice gene with techniques such as polymerase chain reactions (PCR).

• Create mutant clones of the ice gene through the introduction of mutagenic agents such as UV radiation to inactivate the ice gene, creating the “ice-minus” gene.

• Repeat previous steps (insert gene into plasmid, transform E. coli, identify recombinants) with the newly created mutant clones to identify the bacteria with the ice-minus gene. They will possess the desired ice-minus phenotype.

• Insert the ice-minus gene into normal, ice-plus P. syringae bacterium.

• Allow recombination to take place, rendering both ice-minus and ice-plus strains of P. syringae

Read more about Impact Of Pharmacogenomics On Public Healthcare Policy