• Journal of Korean Society of Forest Science
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• v.90 no.4
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• pp.565-572
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• 2001

The spatial genetic structure of Needle fir(Abies holophylla Max.) seedlings on forest gap within a Needle fir forest at Mt. Odae in Korea was analyzed on the basis of ISSR(inter-simple sequence repeats) marker analysis. The gap size was $1,500m^2(50m{\times}30m)$, and we sampled 416 one- or two-year-old seedlings by 2m intervals. Some trees at the upper crown layer except Needle firs and all trees at the middle and lower crown layers were removed, and Needle firs at the upper crown layer showed very weak growth strength or to be withering to death. The results of spatial autocorrelation using 31 polymorphic ISSR markers revealed that it was genetically homogeneous within spatial distance of 15.6m and the randomness of genetic distribution was from 15.6m to 31.2m. The genetic patch size of seedlings in forest gap might be restricted by the density of mother trees, making allow for the average height of adult Needle firs, the seed dispersal area, and the average distance between adults. For the directionality of seedling distribution, we investigated the variography using 'genetic configuration' which was the value of configuration in Multidimensional Scaling by genetic distance. In directional variogram, the increment of spatial distance from East to West direction was inversely proportional to genetic homogeneity. We presumed that this anisotrophy of seedling distribution at this forest gap resulted from the directionality of seed dispersal rather than the difference of fecundity between mother trees or the microhabitat variation, taking the evenness of forest floor condition, a vast seed production and the random distribution of seedlings at the studied site into consideration.

• Journal of Forest and Environmental Science
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• v.38 no.3
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• pp.152-158
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• 2022

A cryopreservation is an essential tool for preservation of germplasm. In this study, the possibility for cryopreservation of embryogenic cells of Siberian ginseng (Eleutherococcus senticosus) in liquid nitrogen (-196℃) was evaluated. The effects of glycerol and dimethyl sulfoxide (DMSO) at different concentrations (5%, 10% and 20%) as cryoprotectants on regrowth of cryopreserved E. senticosus embryogenic cells were tested. There was significant effect of cryoprotectants on regrowth of embryogenic cells (p=0.0019). The highest and lowest fresh mass gain were achieved when embryogenic cells were frozen with 10% DMSO and 5% glycerol (138.2±5.9 and 61.3±14.6, respectively). The effect of the cryoprotectants on the frequency embryo germination was tested. There was no significant difference between glycerol and DMSO (p=0.846). Three different concentrations of cryoprotectants did not significantly affect the frequency embryo germination (p=0.534). Finally, the genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cells was tested by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. RAPD and ISSR analysises showed that there was no genetic variation among regenerants.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.19-28
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• 2019

An efficient shoot regeneration condition for pea cv. 'Sparkle' was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 mg/L BA and a combination of 2 mg/L BA and 1 mg/L TDZ) when cotyledonary node explants were cultured. Moreover, the pretreatment of explant in 200 mg/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. By histological study, cell division and proto-meristem were formed near the surface of the sub-epidermal and epidermal cell layers of cotyledonary node in earlier than 3 days after culture. The analysis of genetic stability of regenerants by using thirteen ISSR markers showed that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.08a
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• pp.71-71
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• 2020

Xanthoceras sorbifolia Bunge, the sole species in the genus Xanthoceras, is a flowering plant in the family Sapindaceae. It is an important tree species being a source of edible oil and biodiesel with a capacity as a pioneer of degraded and desert land. Seeds of X. sorbifolia were collected from two plantations and two superior trees in Inner Mongolia; and one plantation and one superior tree in Liaoning, China. An inter simple sequence repeat (ISSR) analysis showed genetic variation among four artificial populations in China: two in Inner Mongolia (IM), one in Liaoning (LN), and one in Shandong (SD). The average percentage of polymorphic loci was 81.25 % for these four populations. Based on an analysis of molecular variance, 23 % of the total genetic variation was found among populations, and 77 % within populations. Traits of seeds varied considerably between and among areas, for example two trees produced quite different seeds in several traits although they are adjacent to each other in the same farm. As much attention has not been paid to the traits of seeds, there should be a genetic test to understand this variation. It is necessary to obtain information on seed characteristics first and then provide basic information for further research on the selection of superior trees and provenances.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.121-126
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• 2008

Protoplasts from cabbage and radish were isolated and fused symmetrically by PEG treatment. The PEG treated mixture of high concentrated protoplasts produced lots of micro-calli after $2{\sim}3$ weeks. The microcalli developed to normal calli and shoots were regenerated from the calli. A total of 218 shoots were regenerated, but none of them contained the NWB-CMS specific DNA marker, indicating that the transfer of the radish NWB-CMS character into cabbage did not occur. However, ISSR analysis revealed that the cell fusion between protoplasts from radish and cabbage was occurred (3 out of 208 plantlet). The fused regenerants possessed the characteristics of source plants used for protoplast fusion. After vernalization, three regenerants were flowered with white petal color as seen in radish. Only three seeds were able to obtain from one regenerant by backcrossing with the cabbage pollen.

• Molecules and Cells
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• v.25 no.4
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• pp.467-478
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• 2008

Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

• Korean Journal of Plant Taxonomy
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• v.41 no.4
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• pp.299-314
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• 2011

The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.

• Plant Biotechnology Reports
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• v.5 no.1
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• pp.35-43
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• 2011

This communication describes for the first time an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans. Nodal segments collected from field-grown clumps and cultured on Murashige and Skoog (MS) medium supplemented with $4.4{\mu}M$ benzylaminopurine (BA) and $2.32{\mu}M$ kinetin (Kin) gelled with 0.2% gelrite yielded 80% aseptic cultures with 100% bud-break. The in vitro-formed shoots obtained after bud-break were successfully multiplied in MS liquid medium supplemented with $13.2{\mu}M$ BA, $2.32{\mu}M$ Kin, and $0.98{\mu}M$ indole-3-butyric acid (IBA). Sub-culturing of shoots every 3 weeks on fresh multiplication medium yielded a consistent proliferation rate of 3.5-fold. Shoot clusters containing three to five shoots were successfully rooted with 100% success on half-strength MS liquid medium supplemented with $9.8{\mu}M$ IBA, $2.85{\mu}M$ indole-3-acetic acid (IAA), $2.68{\mu}M$ naphthaleneacetic acid (NAA), and 3% sucrose. Plantlets grown in vitro were acclimatized and subsequently transferred to the field. Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.30-30
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• 2019

Pea (Pisum sativum) is one of important legume crops in the world. It is commonly used as a protein source for animal and human diet, and also used as a natural nitrogen source which is produced by a symbiotic bacterium in their root nodule and helpful for terrestrial ecosystem. The successful in vitro manipulation is depended on three main factors including physiology of plant donor, in vitro manipulation approach, and stress physiology during plant cultivation. Moreover, genotype is an important for plant manipulation; different genotype gives the different response to regeneration efficiency. An efficient condition of shoot induction for pea (Pisum sativum cv. 'Sparkle') was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 mg/L BA and a combination of 2 mg/L BA and 1 mg/L TDZ) with cotyledonary node explants culture. Moreover, the pretreatment of explant in 200 mg/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. The analysis of genetic stability of regenerants by using 13 ISSR markers presented that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

• Korean Journal of Plant Taxonomy
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• v.53 no.1
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• pp.14-24
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• 2023

Saussurea dorogostaiskii Palib. (Asteraceae) is a critically endangered medicinal plant in Mongolia and Russia. We studied the genetic variation of S. dorogostaiskii from three mountains of northern Mongolia. The genetic profile was assessed in 70 individuals from eight populations using five inter-simple sequence repeat markers, producing 53 loci with 96.4% polymorphism across all bands. Shannon's index (I) and Nei's gene diversity (H) value at the species level of S. dorogostaiskii are 0.25 and 0.17, respectively. An AMOVA showed high genetic variation among the populations (22% of populations and 32% of mountains), consistent with the high genetic differentiation (G_ST = 0.49) and low gene flow (N_m = 0.51) in S. dorogostaiskii populations. Eight populations were clustered into two groups, corresponding to their geographic locations. The low within-population genetic diversity and high genetic differentiation among S. dorogostaiskii populations factor into their endangered designation. This genetic analysis reveals that all populations are equally threatened, and community-based conservation is appropriate for these species.