• ALGAE
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• v.32 no.1
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• pp.15-28
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• 2017

Morphological, molecular and chromosomal studies were carried out on Tauya basicrassa, an endemic kelp species distributed on the northern continental coast of the Sea of Okhotsk in Russia. The sporophytes of T. basicrassa grow up to 3-6 m long, 1.8-2.2 m wide, and 6.5-7 kg wet weight. The thallus has a blade with very thick narrow basal portion and thinner and much broader upper portion, which usually splits into 3 bullated lobes. A dwarf laminariacean alga, which did not show any morphological similarity to the other species of the order Laminariales, was found from the same locality. The blade of this alga is thin and soft, reached 26-34 cm long and 6-6.5 cm wide and had 4 longitudinal rows of bullations that covered the entire blade. Molecular analysis showed that the dwarf alga has 100% sequence identity in plastid-encoded RuBisCo spacer, mitochondrial cytochrome c oxidase subunit 1 and nuclear-encoded rDNA genes with normal sporophytes of T. basicrassa, indicating that they are different life forms of the same species. Fluorescent DAPI staining showed that the nucleus in the normal sporophyte was 50-65% larger than those of the dwarf ones. Chromosome count using acetocarmine staining showed n = ca. 20 for the normal sporophytes of T. basicrassa and n = ca. 10 for the dwarf one. These results suggest that the dwarf thallus is a haploid parthenosporophyte of T. basicrassa, which developed in nature. This is the first evidence of parthenosporophytes of the laminariacean algae occurring naturally in the field.

• Korean Journal of Plant Taxonomy
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• v.33 no.4
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• pp.339-357
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• 2003

The general morphological characters and chromosome numbers about 7 taxa of Korean Allium sect. Sacculiferum were reviewed. From the results, authors elucidated the key character as well as the range of variations in each taxon, and considered the relationships among taxa in this section. Clear taxonomic treatments also carried out based on the type specimens and the original descriptions. Shape and growing pattern of leaves, size of inflorescences, shape of perianths and tepals, as well as chromosome numbers were key characters in identifying the taxa and in estimating their relationships. On the basis of these characters, A. thunbergii var. thunbergii, A. sacculiferum and A. deltoide-fistulosum were recognised as a distinct species respectively. Hal-la-bu-chu which had been misidentified as A. cyaneum (sect. Reticulato-bulbosa) in Korea was proved to be A. taquetii and belonged to the sect. Sacculi/erum. In addition, A. cyaneum var. deltoides, had been described as a variety of A. cyaneum, was recombined as a variety of A. thunbergii [A. thunbergii var. deltoides (S. Yu, W. Lee et S. Lee) H. J. Choi et B. U. Oh] with the new Korean name of 'Se-mo-san-bu-chu'. In conclusion, Korean sect. Sacculiferum was composed of five species and two varieties. A key to identify the taxa in this section was provided.

• Korean Journal of Plant Tissue Culture
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• v.25 no.2
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• pp.135-139
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• 1998

Reciprocal interspecific hybrids between N. glauca(2n=24) and N. langsdorffii(2n=18) were obtained by intercrossing. One hundred percent of F$_1$ seeds was produced from intercrossing of N. glauca $\times$ N. langsdorffii, whereas the frequency of F$_1$ hybrid seed formation from N. langsdorffii $\times$ N. glauca was very low. However, all the hybrid seeds were germinated well and then grown to normal plantlets. All the plants of F$_1$ hybrids have chromosome number of interspecific hybrids (2n=21). From observation of morphological characteristic, the structure of petrol, leaf, flower, and the morphology of pollen have characteristics of F1 hybrid. Spontaneous tumors (genetic tumor) were formed from each F$_1$ hybrid; the genetic tumor arose at the reproductive phase when the maternal type of F$_1$ hybrid came from N. glauca, while the genetic tumor arose only after reproductive phase when the maternal type of F$_1$ hybrid came from N. langsdorffii. The genetic tumor actively proliferated on hormone-free medium and produced numerous teratoma shoots. In addition, normal leaf or stem explants of F$_1$ hybrid produced calli on hormone-free medium after 15 days of culture, the calli produced new numerous teratoma shoots after 30 days. The frequency of teratoma shoot formation from rnterspecific hybrid was higher in the N. glauca $\times$ N. langsdorffii than in the N. langsdorffii $\times$ N. glauca. Root development from the teratoma shoots was hardly obtained. Teratoma shoots without roots in vitro can form genetic tumor at the vegetative growth phase after tissue culture.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.269-274
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• 2008

The purpose of this study was to determine the effects of Taxol pre-treatment to in vitro matured bovine oocytes, and sucrose and trehalose added to vitrification solution on spindle morphology and embryonic development following cryopreservation. Bovine oocytes were collected from ovaries and matured in tissue culture medium 199 (TCM 199) supplemented with 10% Fetal Bovine Serum (FBS), 0.05ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone and $1{\mu}g/ml$ estradiol for 22h in $39^{\circ}C$, 5% $CO_2$, TCM 199-HEPES containing 20% FBS was used as basic medium (BM) to prepare vitrification solution. Oocytes were pre-treated with $1\;{\mu}M$ Taxol in maturation medium for 15 min prior to vitrification. Oocytes were exposed to 1.6 M ethylene glycol (EG) and 1.3M dimethyl sulfoxide (DMSO) in BM and then were exposed to 3.2 M EG, 2.6 M DMSO and 0.5 M sucrose in BM or 3.2 M EG, 2.6 M DMSO and 0.5 M trehalose in BM. Oocytes with cumulus cells and oocytes without cumulus cells were considered as control 1 and control 2, respectively and held in TCM 199-HEPES at $39^{\circ}C$. Oocytes were frozen using modified solid surface vitrification and were stored in cryotubes in liquid nitrogen for more than 1 week. Frozen oocytes were thawed in TCM 199-HEPES containing 0.5 M, 0.25 M and 0.1 M sucrose in BM for 2 min, respectively or 0.5 M, 0.25 M and 0.1 M trehalose in BM for 2 min, respectively. Immunoflurorescence staining of oocytes was performed to assess spindle morphology and chromosome configuration of oocytes. The rates of cleavage and blastocyst were examined following in vitro fertilization. Normal spindle morphology rate of oocytes pre-treated with Taxol prior to vitrification was not higher than that of other vitrified groups. Taxol pre-treatment did not increase cleavage and blastocyst formation rates, although control groups showed significantly higher rates (p<0.05). Percentages of normal spindle and embryonic development were not significantly different among vitrified groups regardless of type of sugar. In conclusion, Taxol pre-treatment of oocytes before cryopreservation did not reduce the damage induced by vitrification and subsequently did not improve embryonic development following vitrification. Trehalose may be used as an alternative non-permeating cryoprotectant in vitrification solution.

• Horticultural Science & Technology
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• v.28 no.5
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• pp.810-817
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• 2010

We observed abnormal morphology of cotyledons occurring in seedlings derived from open-pollinated and cross-pollinated tetraploid grapes and selected aneuploids, especially hypo- and hyper-tetraploid in seedlings with abnormal morphology of cotyledons. Five types of morphologically abnormal cotyledons were observed. In open-pollination of four tetraploid grapes, the frequency of abnormal cotyledons was 1.6% (49 of 3029 seeds). Percentage of aneuploids in the seedlings of abnormal cotyledons was 20.4% (10 of 49 seedlings). Aneuploids in open-pollination consisted of three (4n = 4x-2), four (4n = 4x-1), and three (4n = 4x+1) seedlings. In cross-pollination of tetraploids, the frequency of abnormal cotyledons was 3.4% (59 of 1729 seeds). Percentage of aneuploids in the seedlings with abnormal cotyledons was 22.0% (13 of 59 seedlings). Aneuploids from cross-pollination of tetraploids consisted of two (4n = 4x-2), nine (4n = 4x-1), one (4n = 4x+1), and one (4n = 4x+3) seedlings. According to the results, although the abnormal cotyledon morphology of seedlings obtained from crossing between tetraploid grapes appeared at low rate (2.3%), aneuploid seedlings occurred at high rate (22.0%); therefore, it indicated that this selection strategy might be very efficient in the initial seedling stage.

• KSBB Journal
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• v.9 no.1
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• pp.16-25
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• 1994

Sporulation mutants of Bacillus subtilis were developed for overproduction of heterologous proteins. The strains spoOJ spoIIG, and spoOJ spoIIG double mutant were constructed from two pretense-delfted mutant (DB104). The vector containing aprE gene was integrated in the chromosome of each strain, then the morphology of each strain was observed by TEM (trasmission electron microscopy). The morphology of spoOJ mutant and spoIIG mutant coincides with the description of the previous reports, respectively. The sporulating cells of spoOJ SpoIIG double mutation resemble spoIIG mutant more similarly, but with a little rougher cell wall membrane. The spoOJ mutation in B. subtilis gives negative effect on aprE activity with only a decreased sporulation frequency. On the contrary spoIIG mutation increases the aprE activity twice with an undetectable sporulation frequency. In the case of spoOJ and spolIG, i. e. double mutation, the effect of spoOJ on aprE activity seems to be relieved and the double mutant shows more or less the same aprE activity compared to spoIIG mutant.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.492-499
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• 2009

A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.901-910
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• 2003

Using the G-, C-, and NOR-banding techniques, a karyotyping for Korean Native Pig was performed. Blood samples were collected from 50 male Korean Native Pigs that had been bred at the National Livestock Research Institute and then blood cells were prepared from in vitro cultures followed by karyotyping; G-, C-, and NOR-banding patterns of metaphase chromosomes were analyzed. The karyotype of Korean Native Pig is 38, XX or XY which consists of 5 pairs of submetacentric chromosomes(Group I), 2 pairs of acrocentric chromosomes with short p-arm(Group II), 5 pairs of medium metacentric chromosomes(Group III), 6 pairs of acrocentric chromosomes(Group IV) and metacentric X and Y sex chromosomes. On GTG-banding, the Korean Native Pig exhibited a typical and identical banding pattern in each homologous chromosomes. Overall chromosomal morphology and positions of typical landmarks of the Korean Native Pig were virtually identical to those of Committee for the Standardized Karyotype of the Domestic Pig(CSKDP). However, numbers of G-bands of the Korean Native Pig chromosomes were more than those of CSKDP. In chromosomes 1, 3, 5, 6, 7, 8, 13, 14, 15, 16, 17, 18 and X, the Korean Native Pig exhibited more separated bands as compared with CSKDP. In C-banding patterns, although the quantity of heterochromatin was variable in each chromosome, most of the Korean Native Pig chromosomes had heterochromatic C-bands on centromeres. However, the heterochromatic C-band was constantly observed on the whole Y chromosome. In AgNOR staining, the NORs were located at centromeres on the chromosomes 8 and 10. The number of NORs per metaphase ranged from 2 to 4 giving a mean value of 2.13. The number of NORs were distributed on all chromosome pair 10 but not on chromosome 8. The sizes of NORs were also differed between homologous chromosomes 8. Numbers of NORs of Korean Native Pig were significantly higher than those of Yorkshire. The pattern of pig NORs was polymorphic in breeds, individuals and cells, especially on chromosome 8.

• Fisheries and Aquatic Sciences
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• v.16 no.3
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• pp.177-185
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• 2013

The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

• Korean Journal Plant Pathology
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• v.10 no.1
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• pp.39-46
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• 1994

Pseudomonas fluorescens Biovar III strains S-2 antagonistic to Rhizoctonia solani was subjected to Tn5 mutagenesis by the transposon vector pGS9. Ampicillin and kanamycin resistant (Ampr, Kmr) transconjugants were recovered at a frequency of 1.3$\times$10-7 per initial recipient cell, when recipient cells were washed twice in TE buffer before conjugation. Of the ca. 3000 transconjugants, a frequency of noninhibitory (Inh-), nonfluorescent (Flu-) and auxotorphic (Pro-) mutants were 0.27%, 0.47% and 0.40%, respectively. In these mutants, all Inh- mutants showed the same colony morphology as wild type, whereas all Flu- and Pro- mutants inhibited the growth of R. solani. These mutants were also susceptible to chloramphenicol, indicating only the Tn5 element, except for parts of pGS9, was integrated into the recipient genome. In a Southern blot analysis, the Tn5 element inserted into one site on the chromosome for each of the chosen mutants. However, Tn5 insertion sites of Inh-, and Pro- mutants were differed in each other. These indicate that the genes essential for R. solani inhibition, fluorescent production and auxotrophic are chromosomally located, but not linked to each other.