Cloning and Sequence Analysis of Anthocyanin Biosynthesis Related Genes (PgANS, PgCHS and PgACT) in Pomegranate (Punica granatum L.)

Objective To obtain PgANS,PgCHS and PgACT genes in Kashgar pomegranate kernels and analyze their sequence.Method Xinjiang pomegranate in Kashgar was selected as the materials.The total RNA of pomegranate Kernels was extracted with Phenol-chloroform.The first cDNA was synthesized with TIANScript kit,and amplified to cDNA by RT-PCR using special primer.The pMD19-T vector linked with the cDNA fragment was transformed into Escherichia coli DH5α,and the positive clone was sequenced.Result The fragment of PgANS was 474bp,GenBank accession number was KT779433.Compared with Punica granatum L.which the accession numbers were KF841619,GU376749 on NCBI,the PgANS homology of nucleotide sequence was 99;.At the same time,the sequence analysis demonstrated that the deduced nucleotide sequence shared 85;,79; and 76; homology with that of Ipomoea batatas (GU598212),Morus alba (JF499385) and Fragariax ananassa (AY695817);the fragment of PgCHS was 140bp and the PgACT was 148bp.Compared with Punica granatum L.(JF747148,GU376750) on NCBI,the PgCHS homology of nucleotide sequence was 99;,and the PgACT was 93;.Conclusion The PgANS,PgCHS and PgACT genes have been obtained,which has laid the foundation for further studying the influence of storage conditions on the anthocyanins of Punica granatum L.kernels.