An Other Busy Week

On Monday, I worked on my abstract for submitting it on Estralla community college conference. We narrowed down my project to an examination of the "microbiology and genetics of agrobacterium." In my project, I characterize Agrobacterium by infecting different plants grow in laboratory. I already infected 15 Soy beans in three different ways and I have to wait three to four weeks for tumors growing on Soy beans.  Also, I genetically characterize Agrobacterium by two different method including RAPD (Random Amplification of Polymorphic DNA) and RFLP (Restriction Fragment Length Polymorphism) analysis. I want to look at morphological and metabolic properties of Agrobacterium by doing thing such as gram sting and growing in different type of media.

After finishing my abstract and submitting it for the esteralla Community college conference, I did gram staining on powder Agrobacterium that we cultured it in the TSA (Trypticase Soy Agar) media last week to be sure that I have a correct bacteria for infecting soy beans. The bacterium was gram negative. Then, I subcultured my Agrobacterium in the TSB (Tryptic Soy Broth) media.

When I checked my soy beans on Tuesday, I saw that 12 of them grew enough and they are ready for be infected and planted in to the pot that contains garden potting soil mix.

On Tuesday, I set up another gel for bacteria 5-8 because I couldn’t get a good result on Monday. Meanwhile, I started to transfer my soy beans from Vermiculite which is ideal for the germination of seeds to the garden potting soil mix but before that I had to infect my soy beans with Agrobacterium. I didn’t like to infect my Soy beans because I planted and grown them by myself but I didn’t have any choice, I had to do that.  Thus, I choose three soy beans that grown most and infected them in three different ways with a fresh Agrobacterium that I cultured in Brain Heart Infusion broth. For the first soybeans, I cut its stem with a razor blade a little bit then I injected Agrobacterium to that part with a syringe. For the second Soy beans, I injected Agrobacterium directly to the stem with a syringe.

 For the third one, I just mixed Agrobacterium with potting soil around the Soy bean’s stem and place it in the separate plant saucer. For the reason that Agrobacterium that I mixed with soil would go through water and I didn’t want other soy beans observe water that is polluted by Agrobacterium. I’ll monitor my three infected soy beans to see which way of infection would be more effective and accountable. 

Infected Soybeans after 7 days

Infected Soybeans after 15 days