To detect the expression of LOS5 gene, the leaf samples
were collected after 6°C treatment for 24 h. RNA samples
were extracted using trizol reagent following the
manufacturer’s instructions (Invitrogen Inc., USA).
SuperScriptTM II reverse transcriptase (Invitrogen Inc, USA)
was used for first-strand cDNA synthesis based on the
manufacturer’s protocol using LOS5 primers, and PCR conditions
are: 94°C for 2 min, 30 cycles of 94°C for 30 s, 60°C
for 90 s, 72°C for 90 s, and finally 72°C for 10 min