The objective of this project was to study the diversity on maple tree leaves with location as a variable. This was focused on how locations could influence microbial diversity in the same species of maple tree. Samples from three locations (figure 1 and 2) were obtained and analyzed. The experiment was divided into two major parts: culture dependent and culture independent analyses.
In the culture dependent analyses, leaf swaps were plated on three types of media (LB, Nutrient agar and Mannitol salt) to see which type of plate would allow for better growth of the bacteria. Figure 3 shows a master plate with the bacteria that were isolated from the initial test plate. Master plates were also made for LB and Mannitol salt plates (data not shown), but only LB was as effective as nutrient agar at growing the bacteria, most likely because these are nutrient rich and complex media plates. The fact that the organism grew better in rich and complex media, indicates that the organisms were in majority heterotrophs. PCR was done on all the colonies in the master plates in several occasions, both with 16s and 18s primers, but when the samples were analyzed through gel electrophoresis, no bands appeared, therefore the PCR samples from the culture dependent organisms were not sequenced. Although sequences for these organisms were not obtained, the samples were Gram stained in order to observe the morphological and biochemical differences in the colonies. Figure 4 shows images from colonies obtained from a maple tree next to …show more content…
The phylogenetic tree was built solely based on bacteria, but from what we saw in the culture dependent studies, fungi have a big effect on microbial diversity. In addition, 16S and 18S rDNA analyses need to be performed on the culture dependent microbes to effectively characterize them, and further compare the differences in each of the
In the culture dependent analyses, leaf swaps were plated on three types of media (LB, Nutrient agar and Mannitol salt) to see which type of plate would allow for better growth of the bacteria. Figure 3 shows a master plate with the bacteria that were isolated from the initial test plate. Master plates were also made for LB and Mannitol salt plates (data not shown), but only LB was as effective as nutrient agar at growing the bacteria, most likely because these are nutrient rich and complex media plates. The fact that the organism grew better in rich and complex media, indicates that the organisms were in majority heterotrophs. PCR was done on all the colonies in the master plates in several occasions, both with 16s and 18s primers, but when the samples were analyzed through gel electrophoresis, no bands appeared, therefore the PCR samples from the culture dependent organisms were not sequenced. Although sequences for these organisms were not obtained, the samples were Gram stained in order to observe the morphological and biochemical differences in the colonies. Figure 4 shows images from colonies obtained from a maple tree next to …show more content…
The phylogenetic tree was built solely based on bacteria, but from what we saw in the culture dependent studies, fungi have a big effect on microbial diversity. In addition, 16S and 18S rDNA analyses need to be performed on the culture dependent microbes to effectively characterize them, and further compare the differences in each of the