Cloning and expression of pineapple sucrosephosphate synthase gene during fruit development
A 1132-base pairs (bp) polymerase-chain-reaction product of sucrose-phosphate synthase (SPS) (EC 126.96.36.199) from pineapple (Ananas comosus cv. Comte de paris) fruit was cloned and nominated as Ac- SPS1. The sequence encodes a putative 377 amino acids protein containing two serine conserved features that had been found in other plant SPS genes: the presence of a 14-3-3 protein special binding domain and an activated site of osmosis stress, which can been activated by phosphorylation and dephosphorylation. The Neighbour-joining tree revealed that Ac-SPS1 belonged to the first kind of sucrose phosphate synthase gene. The results indicated that, the Ac-SPS1 expression was low in the earlier period of fruit growth, then, increasing from 20 days after anthesis and gradually a falling on 40 days, reached the peak with the highest value around 70 days. The SPS activity and sucrose content reached their maximum 80 days after anthesis. It proved that the accumulation of sucrose was correlated with SPS activity and mRNA content and it maximally occurred at 10 d after SPS mRNA and activity had reached its maxima. These results indicated that Ac-SPS1 gene played a key role in sucrose accumulation during the pineapple fruit development and transcriptional activation with increase in Ac- SPS1 expression might be important regulatory events of sugar during pineapple fruit maturation.
Key words: Pineapple fruit, sucrose phosphate synthase, gene cloning, expression.