Comparative Analysis of Tissue and Cell Cycle on the Far Eastern Catfish, Silurus asotus between Diploid and Triploid

Gil, Hyun Woo,Lee, Tae Ho,Han, Ho Jae,Park, In-Seok The Korean Society of Developmental Biology 2017 발생과 생식 Vol.21 No.2

The influence of triploidization on histological characteristics of retina, trunk kidney, liver and midgut tissue, and cell cycle of tail fin and gill tissue in far eastern catfish, Silurus asotus were analyzed. In the infertile triploid fish, the nucleus and/or cell size of secondary proximal tubule cells of trunk kidney, hepatocyte and midgut epithelium are much larger than those of the corresponding cells in the diploid fish (P<0.05). However, triploid tissue showed fewer number of outer nuclear layer in retina and nuclei in secondary proximal tubule of trunk kidney than those for diploid tissue. The mean percentages of the $G_l-$, the S- and the $G_2+M-phase$ fractions were 92.5%, 3.2% and 4.3% in tail fin tissue of diploid, and 93.4%, 2.6% and 4.0% in those of triploid, respectively. There were no significant differences in the percentages of each cell cycle fraction between diploid and triploid. The mean percentages of each phase fractions were 75.1%, 11.1% and 13.8% in gill tissue of diploid and 85.2%, 8.9% and 5.9% in those of triploid, respectively. The differences of cell cycle between tail fin tissue and gill tissue were statistically significant in diploid and triploid (P<0.05). Also, the differences between diploid and triploid were statistically significant in tail fin tissue and gill tissue (P<0.05). Cyclin D1 and cyclin E expressions were not significantly difference between gill tissue and tail fin tissue, and protein expressions of induced triploid were higher than those of diploid. Results from this study suggest that some characteristics in the triploid exhibiting larger cell and nucleus size with fewer number of cell than diploid can be used as an indicator in the identification of triploidization and ploidy level in far eastern catfish.

Comparative Analysis of Tissue and Cell Cycle on the Far Eastern Catfish, Silurus asotus between Diploid and Triploid

Hyun Woo Gil,Tae Ho Lee,Ho Jae Han,In-Seok Park 한국발생생물학회 2017 발생과 생식 Vol.21 No.2

The influence of triploidization on histological characteristics of retina, trunk kidney, liver and midgut tissue, and cell cycle of tail fin and gill tissue in far eastern catfish, Silurus asotus were analyzed. In the infertile triploid fish, the nucleus and/or cell size of secondary proximal tubule cells of trunk kidney, hepatocyte and midgut epithelium are much larger than those of the corresponding cells in the diploid fish (P<0.05). However, triploid tissue showed fewer number of outer nuclear layer in retina and nuclei in secondary proximal tubule of trunk kidney than those for diploid tissue. The mean percentages of the Gl-, the S- and the G2+M-phase fractions were 92.5%, 3.2% and 4.3% in tail fin tissue of diploid, and 93.4%, 2.6% and 4.0% in those of triploid, respectively. There were no significant differences in the percentages of each cell cycle fraction between diploid and triploid. The mean percentages of each phase fractions were 75.1%, 11.1% and 13.8% in gill tissue of diploid and 85.2%, 8.9% and 5.9% in those of triploid, respectively. The differences of cell cycle between tail fin tissue and gill tissue were statistically significant in diploid and triploid (P<0.05). Also, the differences between diploid and triploid were statistically significant in tail fin tissue and gill tissue (P<0.05). Cyclin D1 and cyclin E expressions were not significantly difference between gill tissue and tail fin tissue, and protein expressions of induced triploid were higher than those of diploid. Results from this study suggest that some characteristics in the triploid exhibiting larger cell and nucleus size with fewer number of cell than diploid can be used as an indicator in the identification of triploidization and ploidy level in far eastern catfish.

Evaluation of Cytotoxicity, Antimicrobial and Antioxidant Enzyme Activity of Diploid and Tetraploid Platycodon grandiflorum

부희옥,김영선,김학현,권수정,우선희 한국작물학회 2015 한국작물학회지 Vol.60 No.2

This experiment was conducted to obtain the have higher contents of pharmaceutical constituents as well as higher yield from colchicine induced diploid and tetraploid extracts of Platycodon grandiflorum. In order to determine the biological activity, this study was focused to evaluate the cytotoxicity, antimicrobial on the bronthus disease bacteria, antioxidant enzyme activity of diploid and tetraploid extracts in P. grandiflorum. The activities of antioxidant enzyme according to different solvent extracts were measured as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX). The cytotoxicity of methanol extracts of P. grandiflorum showed significant differences between tetraploid and diploid. That is, the cytotoxic effect against human cancer cell was higher in tetraploid than in diploid. At all extracts concentration, tetraploid samples showed high toxicity and the IC50 (concentration causing 50% cell death) value showed the highest on HCT-116 cell (105.91 μg/mL), and exhibited significant activity against the Hep 3B cell (140.67 μg/mL), SNU-1066 cell (154.01 μg/mL), Hela cell (158.37 μg/mL), SNU-601 cell (182.67 μg/mL), Calu-6 cell (190.42 μg/mL), MCF-7 cell (510.19 μg/mL). Antimicrobial activities of diploid P. grandiflorum were relatively low compared to tetraploid P. grandiflorum on most of the bacterial strains. In tetraploid P. grandiflorum, K. pneumoniae showed the clear zone formation (18~19 mm) of growth inhibition, followed by the clear zone formation of 13~15 mm on C. diphtheria and S. pyogenes. The antimicrobial activities in diploid P. grandiflorum were the highest on K. pneumonia (14~15 mm), and showed the clear zone formation of 11~12 mm on C. diphtheria and 12~13 mm on S. pyogenes. The antimicrobial activity is thought to look different depending on the bacterial strains and the polyploidy of P. grandiflorum. The root extract of P. grandiflorum had the highest (97.2%) SOD enzyme activity in ethyl acetate partition layer of tetraploid while water partition layer of diploid showed thelowest (48.6%) SOD enzyme activity. The activity of CAT showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all partition layers except butyl alcohol. The activities of APX and POD showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all fraction solvents except water layer. These results indicate that the tetraploid P. grandiflorum can be used as a source for developing cytotoxic agent and antimicrobials which can act against bronchus diseases bacterial strains

Evaluation of Cytotoxicity, Antimicrobial and Antioxidant Enzyme Activity of Diploid and Tetraploid Platycodon grandiflorum

Hee-Ock Boo,Young-Sun Kim,Hag-Hyun Kim,Soo-Jeong Kwon,Sun-Hee Woo 한국작물학회 2015 Korean journal of crop science Vol.60 No.2

This experiment was conducted to obtain the have higher contents of pharmaceutical constituents as well as higher yield from colchicine induced diploid and tetraploid extracts of Platycodon grandiflorum. In order to determine the biological activity, this study was focused to evaluate the cytotoxicity, antimicrobial on the bronthus disease bacteria, antioxidant enzyme activity of diploid and tetraploid extracts in P. grandiflorum. The activities of antioxidant enzyme according to different solvent extracts were measured as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX). The cytotoxicity of methanol extracts of P. grandiflorum showed significant differences between tetraploid and diploid. That is, the cytotoxic effect against human cancer cell was higher in tetraploid than in diploid. At all extracts concentration, tetraploid samples showed high toxicity and the IC50 (concentration causing 50% cell death) value showed the highest on HCT-116 cell (105.91 μg/mL), and exhibited significant activity against the Hep 3B cell (140.67 μg/mL), SNU-1066 cell (154.01 μg/mL), Hela cell (158.37 μg/mL), SNU-601 cell (182.67 μg/mL), Calu-6 cell (190.42 μg/mL), MCF-7 cell (510.19 μg/mL). Antimicrobial activities of diploid P. grandiflorum were relatively low compared to tetraploid P. grandiflorum on most of the bacterial strains. In tetraploid P. grandiflorum, K. pneumoniae showed the clear zone formation (18~19 mm) of growth inhibition, followed by the clear zone formation of 13~15 mm on C. diphtheria and S. pyogenes. The antimicrobial activities in diploid P. grandiflorum were the highest on K. pneumonia (14~15 mm), and showed the clear zone formation of 11~12 mm on C. diphtheria and 12~13 mm on S. pyogenes. The antimicrobial activity is thought to look different depending on the bacterial strains and the polyploidy of P. grandiflorum. The root extract of P. grandiflorum had the highest (97.2%) SOD enzyme activity in ethyl acetate partition layer of tetraploid while water partition layer of diploid showed the lowest (48.6%) SOD enzyme activity. The activity of CAT showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all partition layers except butyl alcohol. The activities of APX and POD showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all fraction solvents except water layer. These results indicate that the tetraploid P. grandiflorum can be used as a source for developing cytotoxic agent and antimicrobials which can act against bronchus diseases bacterial strains.

Polyploidy in Lilium lancifolium: Evidence of autotriploidy and no niche divergence between diploid and triploid cytotypes in their native ranges

Chung, M.Y.,Lopez-Pujol, J.,Chung, J.M.,Kim, K.J.,Park, S.J.,Chung, M.G. G. Fischer Verlag 2015 FLORA -JENA- Vol.213 No.-

Lilium lancifolium, the tiger lily, constitutes a polyploid complex with both diploids (reproduced by seeds and bulbils) and triploids (propagated exclusively via bulbils). An autopolyploid origin for the triploid forms has been previously suggested based on classical cytogenetics, chromosome mapping techniques, ecological data, and geographic distribution in their native range (Korea and the Japanese Tsushima Island). Using 13 allozyme loci, we comparatively assessed clonal structure and levels of genetic diversity in four diploid and 11 triploid populations in South Korea to test the autopolyploid origin of the triploid cytotype and to infer which seedling recruitment strategy is operating within the diploid populations. We also employed ecological niche modeling and multivariate analysis to determine whether triploids of L. lancifolium occupy different and broader niches to those of diploids in Korea and Tsushima Island. The diploids harbored higher levels of within-population genetic diversity than triploids, and allele profiles found in triploids were exactly subsets of those in diploids. Repeated seedling recruitment was inferred for the diploids, whereas all the studied triploid populations were monoclonal since there is no seedling (sexual) recruitment. Although we found no niche divergence between cytotypes of L. lancifolium, the triploids have a broader niche breadth. Genetic data further confirm the autotriploid origin of L. lancifolium, and the lack of a clear, strong evidence for niche divergence between cytotypes of L. lancifolium supports the view that ecological differentiation is not a pre-requisite for the establishment of new polyploid lineages.

Haematological Parameters and Respiratory Function in Diploid and Triploid Far Eastern Catfish, Silurus asotus

Dong Won Seol,Soo Yeon Im,Woo June Hur,Min Ouk Park,Dong Soo Kim,Jae Yoon Jo,In Seok Park 한국유전학회 2008 Genes & Genomics Vol.30 No.3

Haematological features pertaining to aerobic capability were compared between diploid and triploid specimens of the Far Eastern catfish, Silurus asotus. No significant differences between diploids and triploids were found for the haematocrit value, total haemoglobin, and mean corpuscular haemoglobin concentration, while the mean corpuscular volume, mean corpuscular haemoglobin, and plasma glucose concentration were significantly higher in triploids than in diploids, and the number of red blood cells was significantly lower in triploids than in diploids. The oxygen consumption rate did not differ significantly between diploid and triploid fish ( P>0.05). Nevertheless, the respiratory frequency was higher in triploids than in diploids ( P< 0.05). Triploids were characterized by a lower concentration of circulating blood cells, and aquaculture practice should consider the need for a lower surface to volume ratio.

Agro-morphological Characterization of Anther Derived Plants in Sweet Pepper (Capsicum annuum L. cv. Boogie)

Surendra Lal Shrestha,Binod Prasad Luitel,강원희 한국원예학회 2011 Horticulture, Environment, and Biotechnology Vol.52 No.2

This study was conducted in sweet pepper ‘Boogie’ to produce the haploids and diploids via anther culture, and to characterize their plant and fruit characters. Anthers were cultured on Dumas de Vaulx medium, (C-medium) supplemented with 0.1 mg・L-1 kinetin and 0.1 mg・L-1 2, 4-D, and 3% sucrose and 0.32% phytagel. Cultures were first incubated in dark at 35℃ for one week and then transferred at 25℃ under 16 h photoperiod for 40 days. Regenerated plantlets including haploids and diploids were evaluated for their plant and fruit characters at Hwacheon in spring, 2007. Results revealed that 39.6 % callus was formed in total anthers (6100) whereas only 1.78% plants were regenerated among them. Ploidy analysis showed that 78.1% regenerants were diploids. Haploid plants were characterized by short plant height, narrow leaf size, short internode length, small flower bud size, and small fruit with low volume than diploid plants. Average fruit volume measured in diploids was 62.7% higher than haploids. Variation in plant and fruit characters was observed within the diploid population and pepper breeder could utilize such variation in breeding program. Based on this characterization and compared to original cultivar, SP94, SP117, SP148, SP171 and SP174 are found superior diploid lines and recommended them to evaluate for variety improvement in sweet pepper.

배수성이 다른 만다린의 교잡에서 종자 형성과 3배체 후대 생성의 반응

박지영,오은의,김어진,정승용,송관정 한국육종학회 2024 한국육종학회지 Vol.56 No.1

Seedlessness is the most important characteristic of citrus and one of its breeding purposes. Triploids are the most reliable way to secure seedlessness and can be developed through crosses between tetraploids and diploids or between diploids and diploids. We evaluated the characteristics of seed formation and recovery frequency of triploids in interploid crosses in this study. Normal seed formation was the highest (85.0%) in crosses of 2x×2x, followed by crosses 4x×2x and 2x×4x. Partially developed seeds were obtained at the highest rate (35.3%) from crosses of 4x×2x with 0.7 seeds per fruit, followed by 2x×2x and 2x×4x with 0.2 and 0.1 seeds per fruit, respectively. The number of undeveloped seeds per fruit was 1.1, 1.0, and 0.3 from the 4x×2x, 2x×4x, and 2x×2x crosses, respectively, and the lowest frequency was recorded in the 2x×2x cross. Diploid plants were mostly obtained in the 2x×2x cross (98.8%), while 73.0% triploids and 27.0% tetraploids were obtained with 4x×2x, and 14.3% diploids, 57.1% triploids, and 28.6% tetraploids were achieved with the 2x×4x cross. Diploid plants were mostly obtained from normal seeds, while ad triploid and tetraploid plants were mostly obtained from partially developed and undeveloped seeds. The results indicated that crosses of 4x×2x were the most efficient and that in vitro culture should be applied to rescue plants from partially developed and undeveloped seeds in triploid breeding programs.

삼배체와 이배체 굴, Crassostrea gigas의 형태 및 생리학적 특성 비교

김수경,심나영,이원영,최민섭,최은희,임현정 한국패류학회 2013 The Korean Journal of Malacology Vol.29 No.3

The morphological relations and physiological characteristics of the triploid and diploid oysters (Crassostrea gigas) in Taean area, west coast of Korea, were investigated from May 2012 to April 2013. Mophometric analysis indicated that the triploid oysters have the same shell length to shell height ratio but higher shell depth to shell height ratio than diploids. Consistent with morphological characteristics, triploid oysters showed greater values of fatness, condition index and RNA/DNA ratio during the period of experiment. The DNA concentration in adductor muscle and mantle of triploid were either lower or equal to the nucleic acids of diploid. However, RNA/DNA ratio were significantly higher than diploid. It appears that RNA/DNA ratio could be a useful indicator of health condition of triploid and diploid oysters when taken in correlation with the morphological indices.

2배체 대장균의 제조와 그 특성

정혜임,임동빈,Jung, Hyeim,Lim, Dongbin 한국미생물학회 2016 미생물학회지 Vol.52 No.4

대장균에서 6개의 Leu 코돈중 가장 흔한 코돈은 CUG이다. 이 코돈을 인식하는 tRNA는 4개의 유전자에 의해 합성되는데, leuPQV와 leuT 2개의 locus로 나누어져 있다. 이 CUG를 인식하는 모든 tRNA가 결핍된 균주를 만들기 위해, 우선 leuPQV가 삭제된 균주(${\Delta}leuPQV$)와, leuT의 anticodon CAG를 GAG로 돌연변이시킨 균주[$Km^R$, $leuT^*$(GAG)]를 각각 만들었다. 이 두 돌연변이 유전자를 모으기 위해 ${\Delta}leuPQV$ 균주를 recipient로, $leuT^*$(GAG) 균주를 donor로하는 transduction을 수행한 결과, 콜로니 크기가 큰 것과 작은 것 두 종류의 transductant를 얻었다. PCR 후 염기서열 분석 결과 큰 콜로니는 예측한 recombinant로 판명됐으나, 작은 콜로니는 donor와 recipient 염색체 간의 상호교환재조합(reciprocal recombination)으로는 설명이 되지 않는, 돌연변이 유전자[$leuT^*$(GAG)]와 야생형 유전자(leuT(CAG)]를 모두 가진 균주로 밝혀졌다. 이 heterozygous diploid는 광학현미경으로 관찰시 세포의 형태와 크기에서 특이점이 발견되지 않았으나, 영양배지에서 야생형에 비해 생장이 한참 느리면서, 선형생장곡선(linear growth curve)이라는 예측하지 못한 생장특성을 보였다. 이 2배체 균주는 선택배지에서는 항상 작은 균일한 콜로니를 형성하였으나, 배지에 선택항생제 없을 경우, $leuT^*$(GAG) 유전자형 세포와 leuT(CAG) 유전자형 세포로 분리가 일어났다. 우리의 결과를 종합해볼 때, 이 2배체 균주는, $leuT^*$(GAG)와 leuT(CAG) 부분만 2배체로 갖는 부분이배체(merodiploid)라기 보다는, $leuT^*$(GAG)와 leuT(CAG)가 서로 다른 염색체에 있는 완전이배체라는 모델을 지지했다. 우리는 이러한 2배체가 어떻게 생성되었으며, 어떻게 분리되는지, 또 이 균주는 왜 선형생장곡선을 보이는지 등에 대한 모델을 토론하였다. Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.