I want to determine the colonization ability of some Bacillus strains on plant leaves and root (Rhizoplane, Rhizosphere and Endorhiza ) but problem is that when I isolate the bacteria the from leaves or roots there will be many different types of bacteria and enumeration of bacterial colonies on the basis of colony morphology is not an accurate method so I am searching for the marking method that can increase the accuracy during enumeration so please suggest some suitable marking methods. If anyone knows about any other method for evaluation of bacterial colonization abilities please guide me. Thanks
You can introduce some genes like GFP into the bacterium and inoculate the engineered bacterium for assessing colonization both qualitatively and quantitatively
Apart from going to the trouble of marking the strains, your Bacillus should be spore forming, and spores should survive heating, so you could eliminate a large proportion of your "background" organisms by heating your samples to say 80C for 10 min before dilution plating. But you might still need marking to distinguish between your strains and any other heat-resistant colonies. For marking, you could try using transposon Tn7 insertion and checking growth rates of insertion mutants to try and select an insertion mutant that is as "fit" as the parent (i.e. no decrease in growth rate in the lab). As for previous answer, best not to use these insertion mutants in the field.
If molecular tools are not available, generate spontaneous rifampicin resistant mutants of your strain. Compare the desired phenotypic traits (PGP traits and growth pattern) of mutant with wild type. If unchanged, go ahead with mutant for colonization studies.
Have a look at the following publication :
https://www.researchgate.net/publication/251571943_Endophytic_Lifestyle_of_Biocontrol_Strains_of_Pseudomonas_spp._in_Olive_Roots
Chapter Endophytic Lifestyle of Biocontrol Strains of Pseudomonas sp...
Hi Jamil
There are many methods to study on this subject, you can find some references on attachment 1
https://www.scielo.br/scielo.php?pid=S0100-06832006000200004&script=sci_abstract
5
https://microbiomejournal.biomedcentral.com/articles/10.1186/s40168-017-0241-2
6
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0094710
15
https://journals.tubitak.gov.tr/biology/issues/biy-17-41-1/biy-41-1-5-1510-16.pdf
17
https://journal.frontiersin.org/article/10.3389/fmicb.2016.01341/full
18
http://www.microbiologyresearch.org/docserver/fulltext/micro/137/1/mic-137-1-187.pdf?expires=1498090489&id=id&accname=guest&checksum=8EA6E35FAD97D22D8643C4CD4059CC4C
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