β-1,3 glucan is the major constituent of the cell wall of fungus. β-1,3 glucanase acts as a cell wall degrading enzyme which degrades the cell wall of the fungus. The expression of the glucanase gene in transgenic plants has conferred increased resistance against fungal pathogens.
We have tried the same experiment at the callus stage, means we have not made the plants but the screening was done at calli stage.
We have inoculated the transformed calli with plant pathogenic fungus. But there was no inhibition of fungal mycelium observed at the callus stage. what could be the possible reasons?
Depend on the promoter you have used. If you use a tissue specific promoter then it will express only in that particular tissue, not in callus. Otherwise an ubiquitous promoter will probably express in callus also. To understand the phenomenon, you can take the example of selection marker, which is expressed in the callus, thereby helping in selection of the transformed explants. It proves that inserted genes can express in callus also.
Yes, depending on the promoter, like CaMV 35S can cause expression in all tissues. For the level of expression you can check RNA expression of the transgene with RT-PCR or protein with western blot in callus tissues.
A Ti plasmid is a circular piece of DNA found in almost all bacteria. This natural ability to alter the plant's genetic makeup was the foundation of plant transformation using Agrobacterium. ... modified T-DNA region where the genes that cause crown gall formation are removed and replaced with the genes of interest.
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