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Journal ArticleDOI

Cell selection as a possible reason for the specificity of somaclonal variation in pea

01 Dec 1995-Plant Breeding (Blackwell Publishing Ltd)-Vol. 114, Iss: 6, pp 520-524
TL;DR: These experiments suggest that mutations affecting developmental characters may be expressed in tissue culture and may cause increased adaptation in mutant cells to the in vitro conditions, which may lead to a higher proportion of quantitative mutations among the regenerated plants.
Abstract: A study of regenerants obtained from long-term callus cultures of different pea (Pisum sativum L.) genotypes revealed specificity of in vitro mutagenesis. The specificity was displayed in the preferential generation of the somaclonal variations affecting quantitative and developmental characters. About 60% of regenerated lines obtained from the cultivar Ranny Zeleny carried mutations in the Lf and Sn loci, which control initiation of flowering. An in vitro study of isogenic lines differing at the Lf and Sn loci illustrates the growth advantages of mutant genotypes in tissue culture. These experiments suggest that mutations affecting developmental characters (e.g. mutations in loci which control flowering behaviour) may be expressed in tissue culture and may cause increased adaptation in mutant cells to the in vitro conditions. Rapid propagation of mutant cells during in vitro culture may lead to a higher proportion of quantitative mutations among the regenerated plants.
Citations
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Journal ArticleDOI
TL;DR: Assessing the stability of repetitive sequences, such as simple sequence repeats (SSR) and retrotransposons, both comprising a large part of genome suggests multiple shoot culture of pea maintained over a long period may be considered as a true to type multiplication method of the original genotype.
Abstract: In vitro clonal propagation of plants should generate identical copies of the selected genotype. However, associated stress might result in a breakdown of control mechanisms and consequent instability of the genome. We have used several molecular methods to assess the genetic stability of long-term propagated (24 years) multiple shoot in vitro culture of pea (Pisum sativum L.). We focused on assessing the stability of repetitive sequences, such as simple sequence repeats (SSR) and retrotransposons, both comprising a large part of genome. No differences were found when seedlings (Co-2004) or original seed (Co-1982) controls and long-term or newly established in vitro (one subculture cycle) samples were investigated by the SSR, inter-repeats (ISSR) or inter-retrotransposon amplified polymorphism (IRAP) method. However, the more global amplified fragment length polymorphism (AFLP) and particularly the methylation sensitive MSAP methods detected 11 and 18% polymorphism among samples, respectively. Interestingly, investigation of the global cytosine methylation status by HPCE measurement revealed no statistically significant differences. Some evidence of retrotransposon re-arrangement was observed by sequence-specific amplification polymorphism. This occurred mostly in the abundant Ty3-gypsy type Cyclop element and to a smaller extent in the Ogre element. Alternatively, no polymorphism was detected among the PDR-1 element of the Ty1-copia type retrotransposon. Based on these results, multiple shoot culture of pea maintained over a long period may be considered as a true to type multiplication method of the original genotype.

122 citations


Cites background from "Cell selection as a possible reason..."

  • ...The phenomenon of somaclonal variation was documented in pea, where several authors reported morphologically deviant regenerants (Gostimskii et al. 1995; Ezhova et al. 1995; Stejskal and Griga 1992; Griga 2000a, b; Griga et al. 1995)....

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01 Jan 2005

46 citations

Journal ArticleDOI
TL;DR: The main approaches have been considered to studying the genetic control of plant cell totipotency in an in vitro culture and the activity of genes that determine and maintain the meristematic state of cells, level of hormones in the cells, and sensitivity to hormones.
Abstract: The main approaches have been considered to studying the genetic control of plant cell totipotency in an in vitro culture. The capacity of cultured plants for callusogenesis, organ formation, and somatic embryogenesis depends on the activity of genes that determine and maintain the meristematic state of cells, level of hormones in the cells, and sensitivity to hormones, as well as on the activity other genes that control different stages of plant morphogenesis.

20 citations


Cites background from "Cell selection as a possible reason..."

  • ...In the tissue culture, both genes affected the growth of calluses and frequency of shoot regeneration: Sn and Lf determined the sensitivity of cells to illumination and exogenous auxin, respectively (Ezhova et al., 1995)....

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  • ...Thus, it was shown that the genes Lf and Sn controlling the time of lowering in the pea affected the growth and shoot formation in vitro (Ezhova and Kovalenko, 1992; Ezhova et al., 1995)....

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  • ...affected the growth and shoot formation in vitro (Ezhova and Kovalenko, 1992; Ezhova et al., 1995)....

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Journal ArticleDOI
TL;DR: The results presented herein demonstrate the utility of long-term pea cultures as model systems for studying the influence oflong-term cultivation on genetic stability, and suggest ways of circumventing the rooting problems associated with such cultures.
Abstract: Forty-seven endogenous cytokinin metabolites were determined in samples of long-term in vitro multiple shoot cultures of pea (Pisum sativum L.). Significant differences were found in the levels of all cytokinin groups except cis-zeatin and some of its derivatives. Old cultures (30 or 10 years old) contained larger total and active cytokinin pools than freshly prepared cultures. Long-term shoot cultures maintained on cytokinin-supplemented media initially exhibit gradually increasing concentrations of endogenous cytokinins that subsequently stabilise at a high level, which can make it more difficult to induce rooting later on. This is consistent with previous findings concerning the growth and development of plantlets derived from long-term cultures and the generally accepted role of cytokinins during root development. The results presented herein demonstrate the utility of long-term pea cultures as model systems for studying the influence of long-term cultivation on genetic stability, and suggest ways of circumventing the rooting problems associated with such cultures.

16 citations


Cites background from "Cell selection as a possible reason..."

  • ...In addition, several authors have reported that also long term in vitro pea cultures can give rise to morphologically deviant regenerants (Gostimskii et al. 1995; Ezhova et al. 1995)....

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Journal ArticleDOI
TL;DR: The presence of the same molecular markers in the somaclones and in different non-somaclonal maize variants suggests that in some cases, the same mechanisms determine both in vitro and in vivo variability and that cell culture enhances the rate of heritable genomic changes that naturally occur in living organisms.

9 citations

References
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Journal ArticleDOI
TL;DR: This review provides a glimpse at the specific kinds of genetic changes encountered among regenerated plants and their progeny, including cytosine methylation alterations of the genome.
Abstract: Plants regenerated from relatively undifferentiated callus cultures possess a vast array of genetic changes. Such variations can result in useful agricultural and horticultural products. For other purposes, however, variations in traits other than those of interest may be undesirable--for example, using cultured cells for genetic engineering. Any steps made toward understanding the basis of tissue culture-induced genetic variation should be helpful in developing a more stable and manipulatable somatic cell system. This review provides a glimpse at the specific kinds of genetic changes encountered among regenerated plants and their progeny. Included among these variations are cytosine methylation alterations of the genome. The repeat-induced point mutation (RIP) phenomenon, reported for filamentous fungi, is invoked to provide a framework to consider the origin of variation in plant tissue cultures.

587 citations

Journal ArticleDOI
TL;DR: Callus formation and shoot production occurred within 4 to 6 weeks in defined media containing 0.2 to 5.0 μM benzyladenine and 1 μM naphthaleneacetic acid while most callus produced one or more shoots at high frequency, root formation did not occur regularly.
Abstract: Shoot formation was observed in callus from apical cells of pea (Pisum sativum L. cv. Century). Shoot apices from 4-day-old plants were macerated and the resulting cell masses grown on agar media. The callus formation and shoot production occurred within 4 to 6 weeks in defined media containing 0.2 to 5.0 μM benzyladenine and 1 μM naphthaleneacetic acid. While most callus produced one or more shoots at high frequency, root formation did not occur regularly. Plants obtained by these procedures were grown to maturity producing flowers and pods.

114 citations

Journal ArticleDOI
TL;DR: For several mutant types that could be scored unambiguously, somaclonal variation was compared to variation induced by treatment of seeds with ethyl methane sulphonate (EMS), and the results showed that the mutant frequencies were higher after EMS treatment than those generated through tissue culture.
Abstract: Plants were regenerated from leaf, cotyledon, and hypocotyl explants of tomato cv Moneymaker. Various phenotypic alterations were observed among regenerated plants (R1), but were not transmitted to the progenies, except for ploidy variation. Variation in ploidy level, mainly tetraploidy, occurred in R1 plants and their R2 progenies, and the frequency of polyploid plants depended on the explant source. More than 50% of the regenerants derived from hypocotyl explants were found to be polyploid. A correlation was observed between the percentage of polyploid cells present in the explant material in vivo and the frequency of polyploid plants. Several monogenic mutations were recovered in the R2, four of which were shown to be allelic to known, recessive, single-gene mutants. No significant effect of explant source or duration of tissue culture period on mutant frequency or spectrum was found. For several mutant types that could be scored unambiguously, somaclonal variation was compared to variation induced by treatment of seeds with ethyl methane sulphonate (EMS). The results showed that the mutant frequencies were higher after EMS treatment than those generated through tissue culture. With respect to the mutant spectrum, no clear differences were observed between the spectra obtained after EMS treatment and those after tissue culture. However, tissue culture gave rise to polyploid plants, whereas no ploidy variants occurred after EMS treatment.

107 citations

Journal ArticleDOI
TL;DR: Whole plant regeneration via somatic embryogenesis was obtained in pea using explants from immature embryos or shoot apex segments and plantlets obtained from both zygotic embryos and shoot apices were transferred to soil and were grown to maturity.
Abstract: Whole plant regeneration via somatic embryogenesis was obtained in pea (Pisum sativum L.) using explants from immature embryos or shoot apex segments. The induction of somatic embryos required picloram or 2,4-D. Germination of fully-developed embryos was accomplished by subculture on medium with only cytokinin and then on medium supplemented with cytokinins in combination with a reduced auxin concentration. Plantlets obtained from both zygotic embryos and shoot apices were transferred to soil and were grown to maturity. Nine plants were examined cytologically, revealing three tetraploids (2n=4x=28) and six diploids (2n=2x=14).

86 citations

Journal ArticleDOI
TL;DR: A comparison was made of the type and frequency of mutational events found in the progeny of tomato plants regenerated after one passage in vitro with those induced by chemical mutagenesis with ethyl methane sulphonate, indicating that the two sources of variability differ in their effect.
Abstract: A comparison was made of the type and frequency of mutational events found in the progeny of tomato plants regenerated after one passage in vitro with those induced by chemical mutagenesis with ethyl methane sulphonate. Several mutants were recovered in the progeny of regenerated and mutagenized plants of two cultivars of tomato. They can be grouped into the following categories: seedling lethality, male sterility, resistance to Verticillium, short stature, change in number of lateral shoots or in leaf shape. The results indicate that the two sources of variability differ in their effect, changing the spectrum and frequency of the mutants as well as, at least in some cases, their pattern of segregation.

82 citations