Development of PCR based markers in Terpene synthase genes for marker assisted selection of high resin yielders in Pinus roxburghii Sarg

Pine oleoresin is a complex mixture of volatile and nonvolatile terpenes and is exploited for commercial production in India. Pines have long reproductive cycle vegetative phase extending upto many years. Therefore, there is an urgent need to identify high resin pine yielders at an early stage. Due to the strong influence of genetic factors on resin yield in pine species, marker assisted selection may serve as a potential tool for early identification of the genotype of interest. This study was planned to identify high resin pine yielders at early stage to avoid exploitation of natural germplasm of chir pine. Protein sequences of different terpene synthases were downloaded from the NCBI database and were multiply aligned to identify conserved and variable regions across the sequences. Primers were designed based on the related coding sequences to target these regions using Primer 3 software to amplify and isolate related genomic loci in chir pine genotypes. These primers were characterized for specificity using Primer Blast and the presence of hairpin loop formation using OligoCalc software. DNA was extracted from different resin yielding pine genotypes and primers were run on them for molecular studies. With the help of bioinformatics tools, we were able to associate some markers with resin yield. This study holds a key promise for the conservation of pine germplasm in natural habitat.