A variety of environmental issues are affecting crops all across the world, but rising temperatures are posing the greatest threat. High temperature has been found to drastically inhibit seedling emergence and cause leaf necrosis at the seedling stage, which results in poor plant stand and significantly decreased yields. Polyamines (PAs) are positively charged, low-molecular-weight aliphatic nitrogenous bases present in all living organisms and are involved in various biological processes in plant growth and development, including senescence and response to different abiotic stresses. Putrescine (Put) functions as a master growth regulator that promotes optimal plant development and greater stress tolerance. Here, the current study aimed to elucidate how Put (1 mM) functions in reducing the negative impacts of high temperature on four varieties of Brassica juncea (RH-1707, RH-1708, RH-1566 and RH-1999-42). Exposure of plants to high temperature resulted in decrease in growth parameters, chlorophyll content and relative water content. Simultaneously, increases were found in antioxidant enzymes, electrolyte leakage, lipid peroxidation, hydrogen peroxide content and stomatal density. High temperature more significantly affected varieties RH-1707 and RH-1708, while RH-1566 and RH-1999-42 showed lesser effects. Exogenous application of Put mitigated the negative impacts of high temperature by enhancing growth, chlorophyll content, relative water content and antioxidant enzyme activities and, simultaneously, it reduces oxidative damage and stomatal density. This study specifies that varieties RH-1707 and RH-1708 are sensitive whereas RH-1566 and RH-1999-42 are tolerant of high temperature and provides an insight into the effectiveness of Put in mitigating the effects of high temperature to a significant extent in B. juncea seedlings.