Expression Analysis Of SPS Gene Families And Study On Alternative Splicing Of SPSⅢ From Saccharum Officinarum

Sucrose phosphate synthase(SPS) is one of key enzymes in sugar metabolism. The research mainly focused on expression of SPS gene families and alternative splicing analysis of SPSâ…¢Pre-mRNA from sugarcane.1. During the early, mid and late of accumulate sugar, relative expression of each of the five SPS gene families (SPSâ… ,SPSâ…¡,SPSâ…¢,SPSâ…£,SPSâ…¤) in the different tissues of sugarcane was determined by real-time PCR. Two sugarcane varieties were FN95-1702(high sugar) and ROC-5(low sugar). The different tissues of sugarcane were young leaves, the third function of leaves and four different parts of the cane. Real-time PCR results show that:During different time of accumulate sugar, between two sugarcane varieties, most of the SPS gene family members showed very significant difference (P<0.01), a few showed significant difference (P<0.05), very few of them showed no difference (P>0.05). In addition, during accumulate of sucrose, in the canes, relative expression of SPS gene families at a high level, and then gradually decreased. However, in the main photosynthetic tissues, relative expression of SPS gene families had remained high level.2. Designed primers according to EU278617 of NCBI, SPSâ…¢gene exon 8-11 fragment was cloned into pMD18-T vector and sequenced. The middle fragment deleted mutants of SPSâ…¢8-11 from sugarcane were induced by PCR-based site-directed mutagenesis, digested and sequenced.4 gene fragments were constructed into plant expression vectors. The recombinants were transformed into Arabidopsis thaliana by floral-dip method. Putative transformants were obtained respectively. It establishes the foundation for molecular mechanism of SPSâ…¢exon skipping.