Development Of Amplified Consensus Genetic Markers In Taxodiaceae And The Applying Of It

Amplified Consensus Genetic Markers (ACGM) were developed for Taxodiaceae species usingCryptomeria japonica ESTs data. Based on the ACGMs, several fragments amplificated by differentprimers were choose for sequencing and gene cloning. The main conclusions were as follows:1. After a comparison of Cryptomeria japonica and Arabidopsis ESTs to search for conservedsequence,237single e-PCR products were obtained. We randomly selected110candidate ACGMmarkers to test. In the110candidate ACGM markers tested,106yielded stable and clear PCR productsin C. japonica. We then tested the utility of these106primer pairs in10species, representing7generaof Taxodiaceae. The results showed that the number of specific amplification primer pairs among those10species varied from49to103（46.2~97.2%）. The106primer pairs （ACGM loci） were hightransferable to Cryptomeria fortunei Hooibrenk （97.2%） but were low in Metasequoiaglyptostroboides （46.2%）. The PCR bands per primer pair varied from1.06to1.15, which means thatmost of the ACGM primers can obtain single band within these10Taxodiaceae species. In summary,our study shows that ACGM is a technique applicable for marker development even in species withlimited sequence data.2. We selected25PCR products screening by4ACGMs for sequencing. The results of BLAST andalignment showed that those PCR fragment were predicted as the homology gene of the target genes.4. Based on the sequenced gene fragments, corresponding to their putative functions,12genes representMYB and CHS in different Taxodiaceae species were cloned. The structures and functions wereanalysis.