Gene expression analysis of heat stress tolerance during pollination in tomato pollen tubes

RNA-Seq analysis of the pistil tubes of 4 tomato varieties after 120 minutes of acute heat stress at 34 degrees Celcius.Anthers of flowers from each genotype were harvested into a 15mL falcon tube and resuspended in 5000uL of pollen germination media (PGM). 2000uL was removed and divided into two 1.5mL Eppendorf tubes per genotype and incubated in a 28C water bath for 30 minutes. One of the tubes was moved to a 34C water bath and both tubes were incubated for an additional 90 minutes. The samples were centrifuged, PGM removed, flash frozen in liquid nitrogen, and stored at -80 until extraction with the Qiagen RNeasy Plant Mini Kit.