• Title/Summary/Keyword: Tetraploid

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Novel approaches for generating and manipulating diploid strains of Chlamydomonas reinhardtii

  • Kariyawasam, Thamali;Joo, Sunjoo;Goodenough, Ursula;Lee, Jae-Hyeok
    • ALGAE
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    • v.34 no.1
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    • pp.35-43
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    • 2019
  • Genetic study of haploid organisms offers the advantage that mutant phenotypes are directly displayed, but has the disadvantage that strains carrying lethal mutations are not readily maintained. We describe an approach for generating and performing genetic analysis of diploid strains of Chlamydomonas reinhardtii, which is normally haploid. First protocol utilizes self-mating diploid strains that will facilitate the genetic analysis of recessive lethal mutations by offering a convenient way to produce homozygous diploids in a single mating. Second protocol is designed to reduce the chance of contamination and the accumulation of spontaneous mutations for long-term storage of mutant strains. Third protocol for inducing the meiotic program is also included to produce haploid mutant strains following tetraploid genetic analysis. We discuss implication of self-fertile strains for the future of Chlamydomonas research.

Production and development of porcine tetraploid parthenogenetic embryos

  • Lin, Tao;Lee, Jae Eun;Shin, Hyeon Yeong;Lee, Joo Bin;Kim, So Yeon;Jin, Dong Il
    • Journal of Animal Science and Technology
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    • v.61 no.4
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    • pp.225-233
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    • 2019
  • The aim of this study was to produce porcine tetraploid (4N) parthenogenetic embryos using various methods and evaluate their developmental potential. In method 1 (M1), porcine 4N parthenogenetic embryos were obtained by inhibiting extrusion of both first (PB1) and second (PB2) polar bodies; in methods 2 (M2) and 3 (M3), 4N parthenogenetic embryos were obtained by electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 or PB1 extrusion, respectively. We found no differences in the rates of cleavage or blastocyst formation or the proportion of 4N embryos among M1, M2, and M3 groups. The different methods also did not influence apoptosis rates (number of TUNEL-positive cells/number of total cells) or expression levels of apoptosis-related BAX and BCL2L1 genes. However, total cell and EdU (5-ethynyl-2'-deoxyuridine)-positive cell numbers in 4N parthenogenetic blastocysts derived from M1 were higher (p < 0.05) than those for M2 and M3 groups. Our results suggest that, although porcine 4N parthenogenetic embryos could be produced by a variety of methods, inhibition of PB1 and PB2 extrusion (M1) is superior to electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 (M2) or PB1 (M3) extrusion.

In vitro Long Term Conservation of Potato Germplasms (감자 유전자원의 기내 장기보존 방법)

  • Yi, Jung-Yoon;Cho, Hyun-Mook;Park, Kuen-Woo
    • Horticultural Science & Technology
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    • v.16 no.4
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    • pp.508-510
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    • 1998
  • This study was carried out to establish the in vitro culture system of potato germplasms for minimizing the occurrence of variation and maximizing the culture period. We used osmoticum such as sorbitol or mannitol with sucrose in the absence of plant growth regulators. The growth of potato germplasms in the medium containing osmoticum was increased when the growth temperature was lowered. After six months storage in low temperature, plant heights of tetraploid was somewhat higher than those of diploid with the exception of stn-16 and the difference due to media was not observed. But after twelve months storage, survival rates of plants cultured in LSM 1(sucrose and sorbitol) was higher than those of plants cultured in LSM 2(sucrose and mannitol). The survival rate of stn-16, diploid wild species, was approximately 75% and it was considerably high. In Atlantic, tetraploid cultivated variety, every individual was survived.

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Sperm Cryopreservation of Tetraploid Pacific Oysters Crassostrea gigas (4배체 참굴(Crassostrea gigas)의 정자 동결보존)

  • Park, Mi Seon;Min, Byung Hwa;Lim, Hyun-Jeong;Hur, Young Baek;Do, Yong Hyun;Myeong, Jeong-In
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.6
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    • pp.853-859
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    • 2014
  • The goal of this study was to evaluate the effects of cryoprotectants [dimethyl sulfoxide (DMSO), methanol, polyethylene glycol, and propylene glycol], cryoprotectant concentrations (10% and 20%), equilibration time (3, 10, and 30 min), cooling rate ($3^{\circ}$, $5^{\circ}$, $7^{\circ}$, and $10^{\circ}/min$), and straw size (0.25 and 0.5 mL) for sperm cryopreservation of tetraploid Pacific oysters. There was a significant difference among the four cryoprotectants, with 10% DMSO yielding the highest post-thaw survival and activity index of sperm. A significant relationship was observed between the cryoprotectant and its concentration. The sperm with equilibration times of 30 min yielded higher post-thaw survival and activity indices than those with 3 and 10 min equilibration times. The sperm cooled at a rate of $5^{\circ}/min$ yielded the highest post-thaw survival and activity index, and the results were significantly different from those observed for cooling at $7^{\circ}$ and $10^{\circ}/min$. Post-thaw survival and activity indices of sperm using a 0.25-mL straw were significantly higher than those using a 0.5-mL straw.

Genetic Diversity among Korean Bermudagrass (Cynodon spp.) Ecotypes Characterized by Morphological, Cytological and Molecular Approaches

  • Kang, Si-Yong;Lee, Geung-Joo;Lim, Ki Byung;Lee, Hye Jung;Park, In Sook;Chung, Sung Jin;Kim, Jin-Baek;Kim, Dong Sub;Rhee, Hye Kyung
    • Molecules and Cells
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    • v.25 no.2
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    • pp.163-171
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    • 2008
  • The genus Cynodon comprises ten species. The objective of this study was to evaluate the genetic diversity of Korean bermudagrasses at the morphological, cytological and molecular levels. Morphological parameters, the nuclear DNA content and ploidy levels were observed in 43 bermudagrass ecotypes. AFLP markers were evaluated to define the genetic diversity, and chromosome counts were made to confirm the inferred cytotypes. Nuclear DNA contents were in the ranges 1.42-1.56, 1.94-2.19, 2.54, and 2.77-2.85 pg/2C for the triploid, tetraploid, pentaploid, and hexaploid accessions, respectively. The inferred cytotypes were triploid (2n = 3x = 27), tetraploid (2n = 4x = 36), pentaploid (2n = 5x = 45), and hexaploid (2n = 6x = 54), but the majority of the collections were tetraploid (81%). Mitotic chromosome counts verified the corresponding ploidy levels. The fast growing fine-textured ecotypes had lower ploidy levels, while the pentaploids and hexaploids were coarse types. The genetic similarity ranged from 0.42 to 0.94 with an average of 0.64. UPGMA cluster analysis and principle coordinate analysis separated the ecotypes into 6 distinct groups. The genetic similarity suggests natural hybridization between the different cytotypes, which could be useful resources for future breeding and genetic studies.

AFLP analysis to assess genomic stability in Solanum regenerants derived from wild and cultivated species

  • Aversano, Riccardo;Di Dato, Francesco;Di Matteo, Antonio;Frusciante, Luigi;Carputo, Domenico
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.265-271
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    • 2011
  • The cultivated potato as well as its tuber-bearing relatives are considered model plants for cell and tissue culture, and therefore for exploiting the genetic variation induced by in vitro culture. The association between molecular stability and tissue culture in different genetic backgrounds and ploidy levels has already been explored. However, it still remains to be ascertained whether somaclonal variation differs between callus-derived chromosome-doubled and undoubled regenerants. Our research aimed at investigating, through amplified fragment length polymorphism (AFLP) markers, the genetic changes in marker-banding patterns of diploid and tetraploid regenerants obtained from one clone each of Solanum bulbocastanum Dunal and S. cardiophyllum Lindl (both 2n = 2x = 24) and tetraploids from cultivated S. tuberosum L. (2n = 4x = 48). Pairwise comparisons between the banding patterns of regenerants and parents allowed detecting considerable changes associated to in vitro culture both at diploid and tetraploid level. The percentages of polymorphic bands between diploid and tetraploid regenerants were, respectively, 57 and 69% in S. bulbocastanum and 58 and 63% in S. cardiophyllum. On average, the frequencies of lost parental fragments in regenerants were significantly higher than novel bands both in S. bulbocastanum (48 vs. 22%) and S. tuberosum (36 vs. 18%) regenerants. By contrast, in S. cardiophyllum, a similar incidence of the two events was detected (32 vs. 29%). Our results revealed that structural changes after tissue culture process strongly affected the genome of the species studied, but diploid and tetraploids regenerated plants responded equally.

Determination of Chimera Types and Ploidy Level of Sports from 'Campbell Early' Grape (Vitis labruscana) (포도 '캠벨얼리' 품종에서 발생한 아조변이체의 배수성 및 키메라 형태 검정)

  • Noh, Jung-Ho;Park, Kyo-Sun;Yun, Hae-Keun;Do, Gyung-Ran;Hur, Youn-Young;Kim, Seung-Hui;Lee, Han-Chan;Ryou, Myung-Sang;Park, Seo-Jun;Jung, Sung-Min
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.996-1002
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    • 2010
  • Flow cytometry (FCM) was used to measure the ploidy level of three different sports from 'Campbell Early' ($Vitis$ $labruscana$) grape. Results of the study showed different ploidy levels. FCM analysis for 'Campbell Early' grape which contains 2C DNA diploid cells showed single peak around 35-40 while 'Kyoho' grape with 4C DNA tetraploid cells had a different level of 70-80. However, analysis of the sports displayed a histogram with 2 peaks containing both 2C and 4C nuclei. There was no difference in histograms of 2C DNA flesh and pericarp; on the other hand, 4C DNA flesh type of sports had a different histogram from that of the 2C DNA pericarp. Chromosome numbers of diploid ('Campbell Early'), tetraploid ('Kyoho'), and three sports were counted under the microscope. 'Campbell Early' and 'Kyoho' have 38 and 76 chromosomes, respectively. Three different sports are mixoploids with mixtures of diploid and tetraploid cells. Microscopic observations of shoot apical meristems in sports from 'Campbell Early' grape were carried out to determine the type of plant chimera. 'Campbell Early' grape (diploid) and 'Kyoho' grape (tetraploid) showed that both had 2 tunica layers covering corpus cells, while the three different sports had tunica layers showing mostly oblique division. Most cells from 'Kyoho' grape were larger than 'Campbell Early' grape. Cells from L-2 and L-3 layers of the three sports were similar to 'Kyoho' grape in size, although all cells in L-1 surface layer were uniform in size like 'Campbell Early' grape. Results of FCM analysis indicated that both normal and polyploid cells could be intermixed in sports and could become mixoploidy consisting of diploid and tetraploid. All sports used in the tests were periclinal chimera plants with two distinct L-1 and L-2 cell layers. The result of this study suggests that all three sports which originated from 'Campbell Early' grape might be 2-4-4 type chimera formation.

Proteomic Responses of Diploid and Tetraploid Roots in Platycodon grandiflorum (2배체와 4배체 도라지의 단백질 발현양상 비교 분석)

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Moon, Young-Ja;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.394-400
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    • 2015
  • The roots of Platycodon grandiflorum species either dried or fresh, are used as an ingredient in salads and traditional cuisine in Korea. To interpret the root proteins, a systematical and targeting analysis were carried out from diploid and tetraploid roots. Two dimensional gels stained with CBB, a total of 39 differential expressed proteins were identified from the diploid root under in vivo condition using image analysis by Progenesis Same Spot software. Out of total differential expressed spots, 39 differential expressed protein spots (${\geq}\;1.5$-fold) were analyzed using LTQ-FTICR mass spectrometry. Except two proteins, the rest of the identified proteins were confirmed as down-regulated such as Isocitrate dehydrogenase, Proteasome subunit alpha type-2-B. However, the most of the identified proteins from the explants were mainly associated with the oxidoreductase activity, nucleic acid binding, transferase activity and catalytic activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

In Vitro Culture of Immature Embryo Obtained by Crossing between Tetraploid Grape 'Fujiminori' and Triploid 'Summer Black' (포도 4배체 '후지미노리'와 3배체 '썸머블랙'의 교배로 얻은 미숙배의 기내배양)

  • Koh, Jae Chul;Oh, Ju Eun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.352-358
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    • 2013
  • For the germination and differentiation of immature embryos obtained by artificial crossing between tetraploid grape 'Fujiminori' (Vitis vinifera ${\times}$ V. labruscana) and triploid 'Summer Black' (V. labruscana ${\times}$ V. vinifera), were incubated in vitro using MS medium supplemented with $GA_3$ or coconut water (CW) at various concentrations. The percentage of embryo formation of 'Fujiminori' ${\times}$ 'Summer Black' was 64.3%. Embryo germination percentage was higher than 95% in all the $GA_3$ treatments at the concentrations of 0.01, 0.05, 0.25, and $1.25mg{\cdot}L^{-1}$. However, only 15.8-31.6% of the germinated embryos successfully developed into normal plantlets. At higher concentration of $GA_3$, the plantlets developed infirm hypocotyls with over elongated and less enlarged structure. Among the treatments of CW at the concentrations of 5, 10, 15, and 20% (v/v), 10% and 15% were more effective and plantlet achievement percentage were 68.4 and 66.7%, respectively. The addition of 10% CW was most effective to obtain plantlets with optimal shoot length, node and root numbers. 15% CW was suitable to obtain plantlets with longer roots. Accordingly, the embryo culture using the MS medium supplemented with 10-15% CW was observed to be more efficient for germinating and growing the immature embryos produced from artificial crossing between tetraploid grape 'Fujiminori' and triploid 'Summer Black'.