Use of Nssr Markers for Determination of Clonal Identity and Genetic Structure in A Pinus Brutia Ten. Clonal Seed Orchard

Abstract

Determination of clonal identity of seed orchard clones is required for the forest tree breeding programs, mating system and pollen contamination studies. Clonal identity and genetic structure in a Pinus brutia Ten. clonal seed orchard were analyzed by 5 nSSR loci (ITPH4516, FRPP94, PEST2669, NZPR544 and EPI3). Needles from 5 ramets per clone (totally 30 clones) were sampled from the seed orchard for microsatellite analysis. Considering the studied ramets of the P. brutia seed orchard clones, all the studied nSSR loci revealed polymorphism. nSSR analysis clearly indicate that the possible multigenotype number of the seed orchard clones (30) was 13 with 7 of these being unique. 39 out of 150 ramets (26%) did not completely match with the genotypes of their supposedly related clones. In 9 out of the 30 clones, all 5 ramets matched each other and in 10 out of the 30 clones, 1 of the 5 ramets did not match the other 4 ramets. This indicates these ramets were mis-lanted or mislabeled during and/or before planting in the orchard. We conclude that genetic identification of orchard ramets seems to be necessary before beginning mating system, pollen contamination studies and forest tree breeding programs.