The synthesis of capsaicinoids occurs in the placenta of the fruits of pungent peppers. However, the mechanism of capsaicinoids' biosynthesis in pungent peppers under salinity stress conditions is unknown. The Habanero and Maras genotypes, the hottest peppers in the world, were chosen as plant material for this study, and they were grown under normal and salinity (5 dS m−1) conditions. The results showed that salinity stress harmed plant growth but increased the capsaicin content by 35.11% and 37.00%, as well as the dihydrocapsaicin content by 30.82% and 72.89% in the fruits of the Maras and Habanero genotypes, respectively, at 30 days after planting. The expression analysis of key genes in capsaicinoids biosynthesis revealed that the PAL1, pAMT, KAS, and PUN1 genes were overexpressed in the vegetative and reproductive organs of pungent peppers under normal conditions. However, under salinity stress, overexpression of PAL1, pAMT, and PUN1 genes was identified in the roots of both genotypes, which was accompanied by an increase in capsaicin and dihydrocapsaicin content. The findings showed that salinity stress caused an enhancement in the capsaicin and dihydrocapsaicin contents in the roots, leaves, and fruits of pungent peppers. Nonetheless, it was found that the production of capsaicinoids is generally not restricted to the fruits of pungent peppers.

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