Allelic Variation in the Gene Encoding Pyrophosphate Dependent Phosphofructokinase (pfp) in Sugarcane

Materials and Methods The enzyme pyrophosphate dependent phosphofructokinasc (PFP), considered to play an important role in sucrosc Forwal-cl ~ ~ r i m c ~ BI was designed l.rom 3 rcgion of high metabolism i n plants, comprises two protein subunits encodhomology idcntil'icd by comparison ol. know11 PFP gcnc cd by separate genes. Part of the P-subunit gene was used as sequences f'rom castor bean and 1)ot;1t0 while rcvcrsc primer a model in investigating allelic variation in the polyploid B8 was designed from a sugat-cane PFP sequence (Sarita genetic environment of sugarcane. The gene l'ragnicnt was GI-oenewald; PhD thesis, University ol'Natal, 1998, and pcramplified by the polymerase chain reaction (PCR) from varsonal communication): ious total gcnomic DNA samples and a number ol' individual DNA molcculcs cloncd randomly From each source. B l (20bp) (5' to 3') ATI-GAT-TTC-ATI-CCI-GAG-GT Seclucnce analysis of clones has demonstrated that two strikB8 (2 1 bp) (5' to 3') CCA-TCA-GTG-ATG-TCA-ATG-CTG ingly different alleles arc prcscnt i n ancestral Snccl~ar~~rrrr The pl-cdictcd product ol'B IIB8 amplification is a rcgion of species as well as modern hybrid var~eties. the PFP gcnc rcprcscnting cxons 10. 1 1 and 12 and their l'l~~nking introns, approxirn,~[cly 1200bp i n size i n all.