• Journal of Korean Society of Forest Science
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• v.95 no.4
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• pp.435-442
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• 2006

A total of 167 peculiar haplotypes confirmed from 28 cpSR variants that were observed in 19 populations of Japanese red pine in Korea through cpSSR marker analysis. Thirteen individuals that showed identical haplotype dispersed evenly in 10 populations, and the average number of effective haplotype within population was 13.37. Estimate of genetic diversity (He) was 0.987 on the basis of cpSSR haplotype variants that was equivalent to or higher than the estimates reported in other studies on some forest tree species. Estimation of genetic diversity (S.I.) on the basis of cpSSR variants composing each haplotype revealed the highest estimate of 1.109 for the population of Gangwon-Yeongwol and the lowest estimate of 0.411 for the population of Gyeongbuk Mungyeong with the average of 0.887. Most of observed cpSSR variants appeared to exist commonly in 19 populations (97.62%), and genetic differentiation of cpSSR variants among populations was turned out to be weak (${\Phi}_{ST}=0.024$). Relatively fast rate of mutation of cpSSR marker might be a major cause for such weak population differentiation. There was no identical haplotype shared between 39 population pairs of 173 pair-wise population pairs. Estimation of genetic distance among 19 populations on the basis of population pairs was also impossible, that might be resulted from restricted migration among 19 populations. Considering the observed distribution patterns of cpSSR variants in addition to the previous studies on I-SSR variants, informations on the present geographic location and genetic status of populations should be considered together for effective sustainable management of the genetic resources of Japanese red pine in Korea.

• Journal of Korean Society of Forest Science
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• v.94 no.3 s.160
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• pp.178-182
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• 2005

To identify the seedlings from controlled pollination between one paternal tree and three maternal trees of Japanese red pine, cpSSR markers of the paternally inherited haploid genome were analyzed in two year old 114 seedlings of full sib families. Individual specific DNA fingerprint like haplotypes of the parental trees were determined by PCR with three cpSSR primers. Haplotypes of the 114 seedlings were also identified by PCR with the same primers. On the basis of the comparison of cpDNA haplotypes of the 114 seedlings with those of the parental trees, 14 seedlings revealed to have distinguished haplotypes from those of the paternal tree. It was tentatively concluded that they were generated via pollination with the non-paternal trees. A seedling of Gangwon30 revealing non-paternal haplotype might have been generated via self pollination with the pollens of maternal tree through improper emasculation or contamination during artificial pollination. DNA fingerprint like cpSSR profiles observed in this study could be successfully applied to the various plant forensic analyses, such as identification of siblings of individual trees, asexually reproduced ramets of a specific clone, vegetatively propagated individuals via tissue culture, and pure full sib progenies.

• Journal of Korean Society of Forest Science
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• v.99 no.3
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• pp.344-352
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• 2010

To investigate the mating system of clones in the seed orchard of Japanese red pine, parameters of mating system, such as outcrossing rates, number of potential pollen contributors, and degree of pollen contamination, were estimated in the seed orchard of Japanese red pines on the basis of DNA data including 4 nSSR and 6 cpSSR markers. Estimates of outcrossing rates were ranged from 94.9 to 100% with an average of 98.9% on the basis of the analysis of cpSSR haplotypes. They were ranged from 90.3% to 100% with an average of 95.9% on the basis of the analysis of nSSR genotypes. However, cross checking of both DNA markers revealed that the seeds presumed to be products of self pollination were turned out to be generated by pollination between mother tree and other tree (i.e., 100% of cumulative outcrossing rate). Estimates of pollen contamination ranged from 43.6% (Gangwon-10) to 56.4% (Gangwon-12) with the average of 48.9%. On the basis of pooled cpSSR haplotype of each seed, maximum number of 21 pollen contributors were verified from the seeds reproduced by Kyungbuk-38. Minimum number of 13 pollen contributors were verified in Gangwon-10. Mean of 16.2 pollen contributors were verified from a total of 5 mother trees. In conclusion, considering pretty high outcrossing rates between clones within a seed orchard, it may be expected that a fairly good genetic potential of the seeds, produced in '77 plot of the seed orchard of Japanese red pines at Anmyeon island, may be guaranteed. Observed results from the analysis of mating system of Japanese red pines in a '77 plot of the seed orchard may also provide useful information for the establishment and management of the seed orchard of the progressive generation.

• Journal of Korean Society of Forest Science
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• v.104 no.4
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• pp.549-557
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• 2015

Novel cpSSR primers were developed based on the sequence information of the Pinus koraiensis chloroplast genome. A total of 30 cpSSR loci were detected in the chloroplast genome, and a total of 30 primer sets flanking those loci were designed. All primer sets were successfully amplified for chloroplast DNA in P. koraiensis. The cross-species transferability of the 30 primer sets was considerably high in P. pumila (100%) and P. paviflora (97%) belonging to the same Subgenus (Strobus) of P. koraiensis. Meanwhile, the transferability was relatively low (73%) in P. densiflora and P. sylvestris belonging to Subgenus Pinus. A total of 13 cpSSR loci out of the 30 loci were polymorphic in the Mt. Jumbong population of P. koraiensis. The mean of haploid diversity(H) was 0.512. The number of haplotypes(N) and the haplotype diversity($H_e$) were 25 and 0.992, respectively. Of the 25 haplotypes, 22 were unique in the analyzed population. The unique haplotypes differentiated 22 individuals (79%) from the total of 28 individuals. In conclusion, the novel cpSSR primers developed in this study would be applicable to other Pinus species, especially the subgenus Strobus, and provide a high level of polymorphism for the study of genetic variation of P. koraiensis.

• Korean Journal of Plant Resources
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• v.25 no.1
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• pp.63-71
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• 2012

To assess parameters of mating system in seed orchard, such as outcrossing rates, number of potential pollen contributors, and degree of pollen contamination, seeds, produced in '77 plot of the Japanese red pine (Pinus densiflora S et Z) seed orchard at Anmyeon island, were collected in 2007 and analysed by nSSR and cpSSR markers. Estimates of outcrossing rates ranged from 91.2 to 100% (mean 97.7%) on the basis of the analysis of cpSSR haplotypes and from 81.6 to 100% (mean 95.3%) on the basis of the analysis of nSSR genotypes. By cross checking of both DNA markers, seeds, presumed to be products of self pollination on the basis of single marker, were confirmed as outcrossed seeds, which resulted in cumulative outcrossing rates of 98.9%. On the basis of pooled cpSSR haplotype of each seed, the number of pollen contributors and paternal contribution rates were estimated as 14.8 and 0.512, respectively. In conclusion, considering pretty high level of outcrossing rates observed in a seed orchard, good genetic potential of the seeds, produced in '77 plot of the seed orchard of Japanese red pines at Anmyeon island, may be guaranteed. Investigated results from the analysis of mating system of Japanese red pines in a '77 plot of the seed orchard may also be expected to provide useful information for the management and establishment of the seed orchard of the progressive generation.

• Journal of Korean Society of Forest Science
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• v.104 no.4
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• pp.578-587
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• 2015

Nuclear SSR (nSSR) and chloroplast SSR (cpSSR) markers were analyzed to assess the parameters of mating system in seed orchard, such as outcrossing rates, the number of potential pollen contributors, paternal contribution rates, degree of pollen contamination, and biparental inbreeding ($t_m-t_s$). In 2006, 2007, seeds were collected from the seed orchard of Pinus densiflora, established in 1977 at Anmyeon island. Estimates of outcrossing rates ranged from 94.9 to 100% (mean 98.9%) in 2006 and from 91.2 to 100% (mean 97.7%) in 2007 on the basis of the analysis of cpSSR haplotypes and from 90.3 to 100% (mean 95.9%) in 2006 and from 81.6 to 100% (mean 95.3%) in 2007 on the basis of the analysis of nSSR genotypes. By cross checking of both DNA markers, mean cumulative outcrossing rates of 100% and 98.9% were estimated in each year. Mean contamination rates were estimated as 48.9% and 42.4%, respectively. On the basis of cpSSR haplotype observed in each seed, paternal contribution rates (the number of pollen contributors) were estimated as 0.458 (mean 16.2) in 2006 and 0.512 (mean 14.8) in 2007. In conclusion, considering pretty high level of outcrossing rates observed in a seed orchard, there might be little to be influenced by inbreeding depression for genetic potential of the seeds induced by selfing. Estimates of the mating system parameters obtained from the two reproductive years were not statistically different, which revealed stable genetic quality of seeds produced in different years. Observed results from this study may provide useful information for the management and establishment of the seed orchard of the progressive generation.