• Title/Summary/Keyword: Somaclones

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Chromosomal Variation in Female and Male Somaclones of Rumex acetosa L. (수영 (Rumex acetosa L.) 암.수 체세포클론에서 염색체 변이)

  • 김수영;이미경;김동순;방재욱
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.113-116
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    • 2001
  • Chromosomal variation was investigated in the female and male somaclones regenerated from the leaf segment culture of Rumex acetosa L. Difference in phenotype depending on the sexuality was not observed. In female somaclones, 21 among 25 somaclones carried the same chromosome complements (2n=14) with wildtype and others were tetraploids (2n=28), Considerable chromosomal variation was found in male somaclones. Only 4 among 20 somaclones carried normal chromosome number (2n=15) and 13 somaclones were aneuploids.

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Genetic Analysis of Somaclones Derived from Different Cytotype Plants of Scilla scilloides Complex using RAPD (PCR-RAPD 기법을 이용한 세포 유형이 다른 무릇 (Scilla scilloides Complex) 체세포클론의 유전적 분석)

  • 오정순;방재욱
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.235-240
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    • 1999
  • The usefulness of randomly amplified polymorphic DNA (RAPD) was evaluated to access the genetic variation in somaclones derived from different cytotype plants of Scilla scilloides Complex, AA (2n=16), BB (2n=18) and AABB (2n=34). Three arbitrary decamer primers were successfully used to amplify genomic DNA from the somaclones. DNA polymorphism was observed between cytotypes. The total number of bands in AA, BB and AABB somaclones were 110, 116 and 103, and marker bands examined were 15, 19 and 26, respectively. The diversity of types using PCR in AA, AABB and BB somaclones were 39.2%, 72.3% and 45.7%, respectively. RAPD band patterns suggest that type AA is more stable than type BB and AABB. The frequencies of specific band in AA, BB or AABB somaclones were 0.9%, 4.3% and 4.9%, respectively. The applicability and reliability of RAPD markers for evaluating the somaclones are discussed.

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Metabolism Characteristics of Acifluorfen Tolerant in Somaclones of Eastern Black Nightshade (Solanum ptycanthum Dun.)

  • Yu, Chang-Yeon;Li, Cheng Hao;Lee, Dong-Wook;Lim, Jung-Dae;Kim, Jae-Kwang
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.126-130
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    • 2005
  • Acifluorfen tolerance charateristics determined the involvement of absorption, translocation, and metabolism in acifluorfen tolerance. Less than 6% of the applied $^{14}C-acifluorfen$ was absorbed. There were no differences in acifluorfen absorption between susceptible and tolerant somaclones. More $^{14}C-acifluorfen$ was translocated in the susceptible than the tolerant somaclones. The susceptible somaclone did not metabolize acifluorfen while some somaclones (i.e.,EBN-3A) metabolized $^{14}C-acifluorfen$. Nomenclature: Acifluorfen, 5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitrobenzoic acid; eastern black night shade, Solanum ptycanthum Dun.,$#^3$ SOLPT.

Saikosaponin Contents in Somaclones Derived from Different Aged Calli of Bupleurum falcatum L. (배양 기간이 다른 캘러스로부터 재분화된 시호 (Bupleurum falcatum L.) 체세포클론의 사이코사포닌 함량 변이)

  • Kim, Eun-Kyung;Lee, Mi-Kyung;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.4
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    • pp.323-327
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    • 1998
  • The contents of saikosaponin a, c and d in the root of somaclones and wild plants of B. falcatum L. were determined. The saikosaponin contents of wild plants collected from Chongson-gun, Kangwon Province and Yongdok-gun, Kyongbuk Province were higher than those of the somaclones derived from 3-month and 2-year aged calli. The mean values of saikosaponin content in 2 year-old somaclones derived from the callus cultured three months were higher than those in 1 year-old somaclones from 2-year aged callus. Variation of saikosaponin content depending on the culture periods was not found. Four plants among 1-year old somaclones showed saikosaponin content twice as much as wild plants had. This result implies the possible use of somaclonal variation for crop improvement.

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In Vitro Regeneration of Lycium chinense Miller and Detection of Silent Somaclones with RAPD Polymorphisms

  • Ahn, In-Suk;Park, Young-Goo;Shin, Dong-Ill;Sul, Ill-Whan
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.157-163
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    • 2004
  • An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

Elimination of Grapevine leafroll associated virus-3, Grapevine rupestris stem pitting associated virus and Grapevine virus A from a Tunisian Cultivar by Somatic Embryogenesis and Characterization of the Somaclones Using Ampelographic Descriptors

  • Bouamama-Gzara, Badra;Selmi, Ilhem;Chebil, Samir;Melki, Imene;Mliki, Ahmed;Ghorbel, Abdelwahed;Carra, Angela;Carimi, Francesco;Mahfoudhi, Naima
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.561-571
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    • 2017
  • Prospecting of local grapevine (Vitis vinifera L.) germplasm revealed that Tunisia possesses a rich patrimony which presents diversified organoleptic characteristics. However, viral diseases seriously affect all local grapevine cultivars which risk a complete extinction. Sanitation programs need to be established to preserve and exploit, as a gene pool, the Tunisian vineyards areas. The presence of the Grapevine leafroll associated virus-3 (GLRaV-3), Grapevine stem pitting associated virus (GRSPaV) and Grapevine virus A (GVA), were confirmed in a Tunisian grapevine cultivar using serological and molecular analyses. The association between GRSPaV and GVA viruses induces more rugose wood symptoms and damages. For this reason the cleansing of the infected cultivar is highly advisable. Direct and recurrent somatic embryos of cv. 'Hencha' were successfully induced from filament, when cultured on $Ch{\acute{e}}e$and Pool (1987). based-medium, enriched with $2mg1^{-1}$ of 2,4-dichlorophenoxyacetic acid and $2.5mg1^{-1}$ of Thidiazuron, after 36 weeks of culture. After six months of acclimatization, RT-PCR carried on 50 somaplants confirmed the absence of GVA, GRSPaV as well as GLRaV-3 viruses in all somaplants. Ampelographic analysis, based on eight OIV descriptors, was carried out on two years acclimated somaplants, compared to the mother plant. Results demonstrated that the shape and contours of 46 somaclones leaves are identical to mother plant leaves and four phenotypically off-type plants were observed. The healthy state of 100% 'Hencha' somaclones and the high percentage of phenotypically true-to-type plants demonstrate that somatic embryogenesis is a promising technique to adopt for grapevine viruses elimination.

In vitro Selection of Acifluorfen-tolerant Solanum ptycanthum and Phenotypic Variation in Regenerated Plants

  • Yu, Chang-Yeon;Lim, Jeong-Dae;Kim, Myong-Jo;Kang, Won-Hee;Hyun, Tae-Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.263-268
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    • 2002
  • Acifluorfen-tolerant callus lines of Solanum ptycanthum were isolated by stepwise selection. Growth of the unselected line was completely inhibited at 0.5 uM. while some selected lines grew at 8 uM acifluorfen. Twenty-two of twenty-five acifluorfen-tolerant callus lines regenerated shoots. Many of the regenerated somaclones were variants, differing in leaf shape, leaf color, number of flower parts, flower color, and fertility. The acifluorfen tolerant S. ptycanthum callus lines differed.

Identification of DNA Variations Using AFLP and SSR Markers in Soybean Somaclonal Variants

  • Lee, Suk-Ha;Jung, Hyun-Soo;Kyujung Van;Kim, Moon-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.69-72
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    • 2004
  • Somaclonal variation, defined as phenotypic and genetic variations among regenerated plants from a parental plant, could be caused by changes in chromosome structure, single gene mutation, cytoplasm genetic mutation, insertion of transposable elements, and DNA methylation during plant regeneration. The objective of this study was to evaluate DNA variations among somaclonal variants from the cotyledonary node culture in soybean. A total of 61 soybean somaclones including seven $\textrm{R}_1$ lines and seven $\textrm{R}_2$ lines from Iksannamulkong as well as 27 $\textrm{R}_1$ lines and 20 $\textrm{R}_2$ lines from Jinju 1 were regenerated by organogenesis from the soybean cotyledonary node culture system. Field evaluation revealed no phenotypic difference in major agronomic traits between somaclonal variants and their wild types. AFLP and SSR analyses were performed to detect variations at the DNA level among somaclonal variants of two varieties. Based on AFLP analysis using 36 primer sets, 17 of 892 bands were polymorphic between Iksannamulkong and its somaclonal variants and 11 of 887 bands were polymorphic between Jinju 1 and its somaclonal variants, indicating the presence of DNA sequence change during plant regeneration. Using 36 SSR markers, two polymorphic SSR markers were detected between Iksannamulkong and its somaclonal variants. Sequence comparison amplified with the primers flanking Satt545 showed four additional stretches of ATT repeat in the variant. This suggests that variation at the DNA level between somaclonal variants and their wild types could provide basis for inducing mutation via plant regeneration and broadening crop genetic diversity.

Detection of DNA Instability Induced from Tissue Culture and Irradiation in Oryza sativa L. by RAPD Analysis

  • Kim, Dong-Sub;Lee, In-Sok;Hyun, Do-Yoon;Jang, Cheol-Seong;Song, Hi-Sup;Seo, Yong-Weon;Lee, Young-Il
    • Journal of Plant Biotechnology
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    • v.5 no.1
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    • pp.25-31
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    • 2003
  • As a consequence of tissue culture of rice, RAPD analysis was peformed to determine whether extended culture periods as undifferentiated calli affected the subsequent genetic constancy, and whether any resulting DNA rearrangements could be detected between sibling plants produced from the same callus. Somaclonal variation was induced at the initial stage of tissue culture and it increased with the length of culture maintenance. Of the 192 total bands, the number of polymorphic bands was 22 (11.5%), 33 (17.2%), and 49 (25.5%) in the callus of 1,3, and 6 months culture, respectively. A significantly higher level of genotypic polymorphisms between regenerants from two different somaclones was also detected, although all the regenerants were derived from a single genotype. In comparison of DNA polymorphisms between regenerants from non-irradiated and from irradiated calli, a scope of variation spectrum by gamma ray irradiation was larger than that by tissue culture. Consideration must be given to this genomic variation where attempts are to be made to use desirable somaclonal variants for plant breeding purpose and in genetic engineering program.

Genetic polymorphism analysis of somatic embryo-derived plantlets of Cymbopogon flexuosus through RAPD assay

  • Bhattacharya, S.;Dey, T.;Bandopadhyay, T.K.;Ghosh, P.D.
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.245-252
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    • 2008
  • The genetic status of somatic embryo-derived plantlets of Cymbopogon flexuosus was examined by randomly amplified polymorphic DNA (RAPD) analysis. Auxins such as 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1-4 mg/l) were used in Murashige and Skoog (MS) medium for induction of calli from rhizomatous explants of Cymbopogon flexuosus. Optimum calli were induced on MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (2, 4-D) (3.5 mg/l) alone or in combination with $N^6-benzyladenine$ (2 mg/l). Somatic embryogenesis was achieved from long term calli when cultured on MS medium containing 2, 4-dichlorophenoxyacetic acid (2, 4-D) (2 mg/l) along with $N^6-benzyladenine$ (BA) (1-2 mg/l). Regeneration was achieved when freshly induced embryogenic calli were sub-cultured on MS medium supplemented with $N^6-benzyladenine$ (3 mg/l) alone. Long-term cultured embryos showed profuse minute rooting on regeneration medium supplemented with N6 -benzyladenine (3 mg/l). Microshoots were rooted in the presence of indole-butyric acid (IBA) (2 mg/l). DNA samples from the mother plant and 18 randomly selected regenerated plants from a single callus were subjected to RAPD analysis with 6 arbitrary decamer primers for the selection of putative somaclones. A total of 64 band positions were scored, out of which 19 RAPD bands were polymorphic. From genetic similarity coefficient based on RAPD band data sharing, it was found that the majority of the clones were almost identical or more than 92% similar to the mother plant, except CL2 and CL9 (66%) which showed highest degree of genetic change with CL2 and CL9 showing presence of two non-parental bands each.