Characterisation of the subunit genes of pyrophosphate-dependent phosphofructokinase from loquat (Eriobotrya japonica Lindl.)

摘要: Pyrophosphate-dependent phosphofructokinase (PFP) catalyses the reversible phosphorylation of fructose-6-phosphate to fructose-1,6-bisphosphate in glycolysis pathway. Two full-length complementary DNAs encoding putative PFP α- and β-subunits, named EjPFPa EjPFPb1, were isolated by reverse transcription polymerase chain reaction (RT-PCR) rapid amplification cDNA ends (RACE) from Eriobotrya japonica Lindl. (loquat). The other β-subunit gene was identified transcriptome data obtained Illumina sequencing, designated as EjPFPb2. They share up 88 % identity with plant α/β-subunits. EjPFPb clustered separately clades or respectively. both shown be mainly localised cell membrane using confocal microscopy GFP fusion proteins 35S:EjPFPa/b-GFP. Monitoring dynamic changes transcripts demonstrated that did not show coordinated expression during fruit development loquat. transcript levels leaves loquat seedlings significantly enhanced after 3 h treatment 0.5, 1.0 1.5 M fructose glucose, which indicates are modulated glucose vivo. Transgenic tobacco plants overexpressed visible phenotype changes, while overexpressing EjPFPb1 grew faster at cost reduced leaf size sucrose content. Meanwhile, content increased growing compared wild-type plants. results confirm genes two subunits have different transcriptional regulation systems play roles carbohydrate metabolism. Elevation can partly impact glycolytic carbon allocation