PCR Identify The Waxy And Shrunken-2 Gene From F2 Population Of Maize

The waxy(wx) locus of maize encodes the Granule-Bound Starch Synthase I (GBSS I) which is expressed only in the endosperm,pollen and embryo sac.It catalyses the formation of amylose in the developing endosperm.The starch of wild-type maize is composed of about 25% amylase and 75% amylopectin whereas homozygous wx mutants produce amylopectin exclusively.Inserting of a controlling element in or near the waxy gene can reduce or eliminate expression of the waxy gene.For this reason,over 50 wx mutants alleles have been identified.Mutantion at waxy abolishes 5% to 95% of Granule-Bound Starch SynthaseI activity.This research select several pairs of special PCR primers from wild-type maize waxy gene.they can cover with the waxy gene.So they can identified the mutantion locus between the wild-type maize and the wx mutant mazie.According to the result,the upper primer P1(5 '-GGCGTACGAGGAGATGGTGAGGAA-3') and lower primer P2 (5 '-CGAAACAAA CAGGGCCAAAGATAG-3') near the 3' (4054-4760bp) can identify the variance between the wild-type maize and the wx mutant mazie. We identify the F2 population of maize by the primers.Identify 97 wild-type individual and 33 wx mutant individual in 130 F2 individual.The result is consist with the result of identify the seed by I2 /kI.According to the result of variance analysis of 19 waxy maize inbred lines by these primers.We find that R5-R19 have same mutant locus near 3'(4054-4760bp),Rl and R3 have same mutant locus near 5' (223-1864bp) ,R2 and R4 have same mutant locus near 5'(1696-2721bp). According to the result of formation of amylose analysis of parts of R5-R19 which have same mutant locus.We find that they have little different in formation of amylose,but have different in production.The shrunken-2(sh) locus of maize encodes the AGPPase. The wild-typen Shrunken-2 maize is turned to recessive shrunken-2 maize by inserting of controlling element.The mutantion decrease synthesis of amylose of endosperm and increase the quantity of sugar.We47select two pairs of primers from The wild-typen Shrunken-2 gene.The upper primer PI (5' -TGTATTCTTACCTCAGATGC-3') and lower primer P2(5'-TCATAGGAATATCAATAAG C-3') can identify the wild-typen and recessive Shrunken-2 gene.