Identification of nitric oxide responsive genes in the floral buds of Litchi chinensis

Litchi (Litchi chinensis Sonn.) is an important tropical and subtropical evergreen woody fruit tree, and it has been shown that nitric oxide (NO) could promote litchi flowering. NO responsive genes of litchi (cv. Nuomici) primordia were identified through a suppression subtractive hybridization (SSH) library screen. We obtained 1 563 expressed sequences tags (ESTs) that were enriched in the NO treated inflorescence primordia. We then used a reverse Northern analysis to identify 728 true NO responsive ESTs, the sequences of which have been further analyzed. They represent 70 litchi unique genes that could be classified into 9 categories: 14 % of them were involved in transport facilitation, 7 % in transcription regulation, 9 % in stress response, 7 % in sugar metabolism, 9 % in secondary metabolism, 10 % in intracellular signalling, and 44 % in other metabolism, whereas 11 % were genes with unknown functions, and 7 % were genes with no hit found. Next, we performed a real-time quantitative polymerase chain reaction (RT-qPCR) to determine the expression of selected candidate genes during a time-course of NO treatment and of normal floral tissue development.