Gene expression of lipopeptide biosynthesis in Bacillus subtilis analyzed by RT-qPCR using the method 2-ΔΔ CT
DOI:
https://doi.org/10.29019/enfoqueute.1206Keywords:
Alternaria sp., Botrytis sp., plant-induced chemoresistance, ribonucleic acid, complementary DNA, Fengicins, Iturins, PhytopathogensAbstract
The genus Bacillus is a critical ally in combating phytopathogens such as Alternaria sp. and Botrytis sp., which cause losses to the agricultural sector in Ecuador. Bacillus subtilis generates important lipoproteins belonging to the fengicin and iturin families; therefore, this study analyzed the expression of genes involved in the synthesis of FEND and ITUDI lipoproteins. In addition, the antagonism of Bacillus subtilis against phytopathogenic microorganisms was analyzed through the PICR (percentage of radial growth). Using molecular techniques such as RT-qPCR, the expression levels of the aforementioned genes were quantified on three specific days: 1, 5 and 9 when Bacillus subtilis is in confrontation with phytopathogens. For this purpose, three treatments with the phytopathogens mentioned above were carried out. Total RNA was extracted, followed by retrotranscription, and finally, the cDNA was subjected to qPCR analysis to determine gene expression values. It was determined that the gene coding for phengicins is expressed seven times on the fifth day of treatment when Bacillus subtilis is in the presence of Alternaria sp. It was also determined that the ITUDI gene is expressed an average of 4.56 times more on the fifth day of treatment in the presence of Alternaria sp. Finally, the expression levels of FEND and ITUDI are expressed an average of 3.6 and 2.3 times, respectively, when it is in antagonism with Botrytis sp.
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