• Korean journal of applied entomology
• /
• v.48 no.4
• /
• pp.547-551
• /
• 2009

Two closely related species, the soybean podworm, Matsumuraeses phaseoli, and the podborer, M. falcana, gives differential economic damages on crops. It is difficult to discriminate these potential sympatric species by morphological characters. The goal of this study was to develop a discriminating molecular marker based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). A partial genomic fragment (500 bp) of mitochondrial cytochrome oxidase I (mtCOI) was sequenced in both species, in which restriction site by Rsa I was selected as a dichotomous marker. PCR-RFLP in the mtCOI region clearly discriminated both species.

• Parasites, Hosts and Diseases
• /
• v.35 no.2
• /
• pp.127-134
• /
• 1997

Transmission electron microscopy of an ArGnthnmoebo isolate (KA/LS) from a contact lens case revealed bacterial endosymbionts within cytoplasm of the amoebae. The Acnnthamoebn isolate belonged to the morphological group ll. Based on the polymerase chain reaction (PCR) - restriction fragment leilgth polymorphism (RFLP) of 185 ribosomal RNA coding DNA (rDNA) , the isolate was identified as A. Iwnunensis. Strain typing by isoenzyme analysis using isorlectric focusing (IEF) and mitochondrial (Ent) DNA RFLP revealed that the isolate was closely related with KA/Ll , the most predominant type of isolates from contact lens storage casas, KA/E2, a clinical isolate, KA/W4, previou:fly reported to host endosymbionts. and L3a strains of A. Iwnunensis. The endosymbionts were similar to those of KA/W4 in a.jpects that they were randomly distributed in both trophozoites and cysts, and were rod-shaped bacteri3 measuring approximately 1.38 x 0.50 ㎛. But the number of endosymbionts per amoeba was significantly lower than that of KA/W4. They were neither limited by phagosomal membranes nor included in lacunae- like stnlcture.

• Mycobiology
• /
• v.33 no.2
• /
• pp.104-108
• /
• 2005

Genus Phellinus taxonomically belongs to Aphyllophorales and some species of this genus have been used as a medicinal ingredients and Indian folk medicines. Especially, P. linteus and morphological-related species are well-known medicinal fungi that have various biological activities such as humoral and cell-mediated, anti-mutagenic, and anti-cancer activities. However, little is known about the rapid detection for complex Phellinus species. Therefore, this study was carried out to develop specific primers for the rapid detection of P. linteus and other related species. Designing the species-specific primers was done based on internal transcribed spacer sequence data. Each primer set detected specifically P. linteus (PL2/PL5R) and P. baumii (PB1/PB4R). These primer sets could be useful for the rapid detection of specific-species among unidentified Phellinus species. Moreover, restriction fragment length polymorphism analysis of the ITS region with HaeIII was also useful for clarifying the relationship between each 5 Phellinus species.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.3
• /
• pp.167-173
• /
• 1997

The embryogenic callus (EC), from which somatic embryos could be induced, was compared with nonembryogenic callus(NE) to study the origin and features of totipotent cell in water dropwort (Oenanthe stolonifera DC). To induce and maintain of EC and the NE, meristematic stem and immature floret were inoculated in MS media supplemented with 1 mg/L 2,4-D, and with 2.5 mg/L NAA and 5mg/L BA, respectively, The EC was not induced from the NE even after subculturing in MS medium supplemented with 1 mg/L 2,4-D. Plantlets were not regenerated from the NE in hormone-free medium. In histochemical comparison of the EC with the NE by light microscopy, the EC had smaller cells in size, dense cytoplasm, and more starch granules of cells compared to the NE cells. The cell from the EC, as observed by transmission electron microscopy, had smaller vaculoes, well developed ribosomes, mitochondria, and endoplasmic reticulum, whereas the cells from the NE had larger vacuoles and underdeveloped organelles. In protein pattern from NE, EC and Somatic embryo (SE), as analyzed by SDS polyacrylamide gel electrophoresis, different proteins specific for tissue were observed: 17 and 28 KD for NE, 50, 52, 57, 66, 68 KD for EC and 20 KD for SE. DNA polymorphism was also observed between EC and NE as analyzed by RAPD (randomly amplified polymorphic DNA) method. The origin of totipotent stem cell and the relationship between irreversible genomic change arose in differentiation and the loss of totipotency in plant were discussed.

• Horticultural Science & Technology
• /
• v.18 no.2
• /
• pp.103-106
• /
• 2000

The genetic relationship of Bletilla striata native to Korea and Japan was investigated using random amplified polymorphic DNA (RAPD). The 156 reproducible DNA bands, consisted of 58 polymorphic and 98 monomorphic bands, were obtained by polymerase chain reaction (PCR) with selected 10 random primers. The 8 Bletilla lines have been classified into three groups according to the similarity coefficient obtained by RAPD analysis. The dendrogram showed overall correlationship between similarity coefficient of 0.48 and 0.84. The first group included A (Bletilla striata native to Korea), B (Bletilla striata variegated and native to Korea in Mokpo), C (Bletilla striata variegated and native to Korea), D and E (Bletilla striata native to Japan). In this group, it was showed that B and C had the most similar genetic relationship. The similarity coefficient between D and E was 0.77. D and E had a very close resemblance in plant height and flower color with A native to Korea, respectively. The second group included only G (dwarf Bletilla native to Japan) and had a different morphological character compared to other cultivars. The last group included F and H (dwarf and variegated Bletilla native to Japan) and they had a similarity of variegation.

• Plant Resources
• /
• v.2 no.1
• /
• pp.22-25
• /
• 1999

Molecular markers are useful to confirm the hybridity of F1 plant derived from cross of two homozygous parents with similar morphological traits. RAPD markers were used to test F1 hybrid plant obtained from cross of two homozygous soybean (Glycine max) parents. Fl plant for cross I was made from the mating of Hobbit87 (female) and L63-1889 (male) and Fl plant for cross II was obtained from the mating of H1053 (female) and L63-1889 (male). Selfing plant per each cross was also obtained. Among 20 Operon primers used, OPA04 and OPA09 show polymorphism between cross I and II parent. Band in size 1Kb of OPA04 and 2.1Kb of OPA09 primer was polymorphic band. This fragment identified Fl hybrid plant and selfing plant in cross I and II. Female parent Hobbit87 in cross I and H1053 in cross II has no this fragment (recessive allele). However, male parent L63-1889 and Fl hybrid plant in cross I and II has this size of polymorphic band (dominant allele). This indicated that Fl hybrid and selfing plants were detected by RAPD marker before phenotypic marker would be used to identify Fl hybridity. Amplification products of selfing plant for cross I and II were completely same to the those of female parent. When mature, flower color of Fl hybrid plant in cross I and II was purple and flower color of selfing plant in cross I and II was white. Purple flower is dominant trait. Fl hybridity was successfully detected at very early growth stage using RAPD marker. Therefore, RAPD marker can be used broadly to confirm Fl hybridity in many crops.

• Horticulture, Environment, and Biotechnology : HEB
• /
• v.59 no.6
• /
• pp.875-888
• /
• 2018

Tulip (Tulipa L.) is one of the most important ornamental geophytes in the world. Analysis of molecular variability of tulips is of great importance in conservation and parental lines selection in breeding programs. Of the 70 genic microsatellites, 15 highly polymorphic and reproducible markers were used to assess the genetic diversity, structure, and relationships among 280 individuals of 36 wild and cultivated tulip accessions from two countries: Iran and the Netherlands. The mean values of gene diversity and polymorphism information content were 0.69 and 0.66, respectively, which indicated the high discriminatory power of markers. The calculated genetic diversity parameters were found to be the highest in wild T. systola Stapf (Derak region). Bayesian model-based STRU CTU RE analysis detected five gene pools for 36 germplasms which corresponded with morphological observations and traditional classifications. Based on analysis of molecular variance, to conserve wild genetic resources in some geographical locations, sampling should be performed from distant locations to achieve high diversity. The unweighted pair group method with arithmetic mean dendrogram and principal component analysis plot indicated that among wild tulips, T. systola and T. micheliana Hoog exhibited the closest relationships with cultivated tulips. Thus, it can be assumed that wild tulips from Iran and perhaps other Middle East countries played a role in the origin of T. gesneriana, which is likely a tulip species hybrid of unclear origin. In conclusion, due to the high genetic variability of wild tulips, they can be used in tulip breeding programs as a source of useful alleles related to resistance against stresses.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2020.08a
• /
• pp.47-47
• /
• 2020

The Korean National Agrobiodiversity Center holds the more than 1300 accessions of proso millet, but a large portion of accessions are landrace of Korea that has very similar traits. To comprehend the maximum genetic diversity of this crop, a core collection with minimum number of accessions will facilitate easy access to genetic material. Here we assessed the genetic diversity and population structure in a germplasm collection of 830 accessions by employing EST-SSR markers and morphological traits. A total of 107 alleles were detected with an average allele number of 4.9 per locus among the 830 accessions based on 37 EST-SSR markers. The number of alleles per locus ranged from 2 to 7. Polymorphism information content and expected heterozygosity ranged from 0.06 to 0.68 (mean = 0.21) and 0.06 to 0.73 (mean = 0.23), respectively. The germplasm collection was separated into two groups based on population structure analysis, whereas principal coordinate analysis (PCoA) could not cluster accessions according to their geographic origin. Subsequently, a preliminarily developed core collection with a total of 141 accessions (17%) was selected from the whole set of germplasm by combining allelic variations of EST-SSR markers and eight different phenotypic traits. The core collection optimally represented the whole germplasm collection and displayed a similar level of PCoA value and genetic variation from the initial collection. The results obtained here provide a primary resource for further genetic analysis and establish a reference for further development of appropriate genetic breeding strategies.

• Horticultural Science & Technology
• /
• v.28 no.5
• /
• pp.828-835
• /
• 2010

Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.

• Horticultural Science & Technology
• /
• v.16 no.1
• /
• pp.21-24
• /
• 1998

RAPD markers were analyzed in order to detect the genetic variation and diversity of the fifty-two melon lines. SDS extraction method produced more and purer DNA than CTAB method. RAPD reaction conditions were optimized as follows ; 10ng template DNA, 270nM primer, $200{\mu}M$ each of dATP, dCTP, dGTP and dTTP, $0.3{\mu}unit$ dynazyme and 10x buffer brought to $15{\mu}l$ final volume with distilled water. The adequate annealing temperature was $39^{\circ}C$ and forty cycles of amplification produced the best RAPD band patterns. Among a total of 123 bands from 12 random primers, 25 polymorphic bands(20%) were selected as reliable markers. The average number of polymorphic bands per primer was 2.1 among the 52 lines. Intragroup genetic relationship based on the marker difference was closer than intergroup genetic relationship. The 52 lines could be grouped into two major group (Korean landraces and melon lines) and then melon group subdivided into two subgroups (net melon lines and no-net melon). This result corresponded to morphological grouping. Eight RAPD markers separated the Korean landraces and melon groups and four RAPD markers separated net melon and no-net melon groups.