Heliotropium thermophilum can survive at a soil temperature of 65 degrees C in natural and laboratory conditions, but the mechanism of survival at high soil temperatures is not completely known. The objective of this study was to determine whether changes in abscisic acid (ABA), osmolytes and heat shock factors (HSFs) levels have an effective role in the development of thermotolerance in H. thermophilum at high temperatures. Soil temperature at which the thermophilic plant could live was gradually increased in laboratory conditions and the effects of four different temperatures (20 +/- 5 degrees C: low, 40 +/- 5 degrees C: mild, 60 +/- 5 degrees C: medium, 80 +/- 5 degrees C: extreme heat) were observed for 15 days. The results showed that the content of thiobarbituric acid reactive substances (TBARS) did not significantly change in extreme heat, whereas the leaf water potential and stomatal conductivity decreased. ABA biosynthesis, accumulation of osmolyte compounds including proline and total soluble sugars, and the expression levels of heat shock transcription factor A4A (HSFA4A), heat shock transcription factor A3 (HSFA3), and heat shock factor (HSF4) genes significantly increased with increase of soil temperature from 20 +/- 5 degrees C to 80 +/- 5 degrees C. In conclusion, we observed that H. thermophilum is an extreme thermophile. This plant can adjust osmotic activity to effectively take water through the osmolytes accumulation, reducing water loss by ABA-mediated stomatal closing and survive at high soil temperatures by stimulating the increased transcription level of HSFs.