6-phosphogluconate dehydrogenase (6PDGH) gene cloning of kenaf (Hibiscus cannabinus L.) and its RNA interference vector construction.

[Objective] The 6-phosphogluconate dehydrogenase (6PDGH) gene of kenaf sterile line and maintainer line were cloned in present study and its RNAi vector was constructed in order to lay the foundation for further research of this gene function in anther development of kenaf(Hibiscus cannabinus L.). [Method] The assembling cDNA sequences of 6PDGH gene were obtained based on high-throughput sequencing data of kenaf anther transcriptome and bioinformatics method. The cDNA and total DNA of CMS P3A and maintainer line P3B anthers were used as template to clone the full-length of 6PGDH gene According to RNAi vector designing rule, the 389 bp gene-specific fragment of 6PGDH cDNA sequence was used as RNA interference fragment to insert into pKANNIBAL vector in forward and reverse direction, respectively, to construct the RNAi vector. [Result] The full-length of cloned 6PGDH gene in P3A and P3B anthers was 1751 bp in length, which contained a 1458 bp open reading frame(ORF) encoding 485 amino acids and had no intron. Only two bases' differences were found between base sequence of CMS and maintainer line. The amino acid sequence of 6PGDH gene shared 75.07%, 84.57%, 86.50%, 87.24%, 91.19% and 92.01% of homology with Arabidopsis thaliana (AY084486), Zea mays (AF061838), Glycine max (B007907), Medicago sativa (U18239), Populus trichocarpa (XM_002329211), Ricinus communis (XM_002530757). The RNAi vector of 6PGDH gene in kenaf was constructed successfully to obtain the interference expression vector pART27-pKANNIBAL-R1+2. [Conclusion] The full-length sequence of 6PGDH gene in kenaf was cloned successfully, and the constructed RNA interference vector pART27-pKANNIBAL-R1+2 could be used for studying 6PGDH gene function of kenaf. [ABSTRACT FROM AUTHOR]