In Vitro Micropropagation of Freesia hybrida and the Assessment of Genetic and Epigenetic Stability in Regenerated Plantlets

Gao, Xiang; Da-yuan, Yang; Cao, Donghui; Ao, Man; Sui, Xin; Wang, Qinmei; Kimatu, Josphert N.; Wang, Li

doi:10.1007/s00344-009-9133-4

Cited by 75 publications

(45 citation statements)

References 59 publications

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“…In our conditions, the number of methylation changes per plant was small and the resulting total polymorphism was \1 % independently of the age of the cell culture, indicating little disruption to the epigenetic Values are means of 3-4 independent measurements ± SD status. Similarly, low levels of methylation variation were found in freesia somatic seedlings derived from short-term cultures (Gao et al 2010) and pea long-term multiple shoot cultures (Smýkal et al 2007). Many studies with methylation-sensitive molecular markers that reported a high level of epigenetic variation reported no effect on phenotype or morpho-agronomic traits (Bednarek et al 2007;Li et al 2007;Schellenbaum et al 2008;Fiuk et al 2010).…”

Section: Discussionmentioning

confidence: 94%

Assessment of genetic and epigenetic changes during cell culture ageing and relations with somaclonal variation in Coffea arabica

Landey

Cenci²,

Guyot

et al. 2015

Plant Cell Tiss Organ Cult

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Long-term cell cultures were used in coffee to study the cytological, genetic and epigenetic changes occurring during cell culture ageing. The objective was to identify the mechanisms associated with somaclonal variation (SV). Three embryogenic cell lines were established in Coffea arabica (2n = 4x = 44) and somatic seedlings were regenerated after 4, 11 and 27 months. Phenotyping and AFLP, MSAP, SSAP molecular markers were performed on 199 and 124 plants, respectively. SV were only observed from the 11 and 27-month-old cultures, affecting 30 and 94 % of regenerated plants, respectively. Chromosome counts performed on 15 plants showed that normal plants systematically displayed normal chromosome numbers and that, conversely, aneuploidy (monosomy) was systematically found in variants. The allopolyploid structure of C. arabica allowed aneuploid cells to survive and regenerate viable plants. No polymorphic fragments were observed between the AFLP and SSAP electrophoretic profiles of mother plants and those of the in vitro progeny. Methylation polymorphism was low and ranged between 0.087 and 0.149 % irrespective of the culture age. The number of methylation changes per plant-normal or variant-was limited and ranged from 0 (55-80 % of the plants) to 4. The three cell lines showed similar SV rate increases during cell culture ageing and produced plants with similar molecular patterns indicating a non random process. The results showed that cell culture ageing is highly mutagenic in coffee and chromosomal rearrangements are directly linked to SV. Conversely, the analysis of methylation and transposable elements changes did not reveal any relation between the epigenetic patterns and SV.

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Section: Discussionmentioning

confidence: 94%

Assessment of genetic and epigenetic changes during cell culture ageing and relations with somaclonal variation in Coffea arabica

Landey

Cenci²,

Guyot

et al. 2015

Plant Cell Tiss Organ Cult

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“…As the methylation-sensitive isoschizomers MspI and HpaII differ in their sensitivity to cytosine methylation (Lei et al, 2006;Schulz et al, 2013;Yaish et al, 2014), the comparison of the differential digestion pattern generated by each enzyme allows us to identify the methylation status of the DNA. The MSAP technique has been considered efficient in the detection and characterization of somaclonal variants in several crops (Lei et al, 2006;Gao et al, 2010;Bobadilla Landey et al, 2013). Despite the economic importance of sugarcane and also the impact of tissue culture technique to this crop, reports on the methylation status of sugarcane plants derived from tissue culture have not been published yet.…”

Section: Introductionmentioning

confidence: 99%

DNA methylation in sugarcane somaclonal variants assessed through methylation-sensitive amplified polymorphism

Francischini

Kemper

Costa³

et al. 2017

Genet. Mol. Res.

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ABSTRACT. Micropropagation is an important tool for large-scale multiplication of plant superior genotypes. However, somaclonal variation is one of the drawbacks of this process. Changes in DNA methylation have been widely reported as one of the main causes of somaclonal variations in plants. In order to investigate the occurrence of changes in the methylation pattern of sugarcane somaclonal variants, the MSAP (methylation-sensitive amplified polymorphism) technique was applied to micro-propagated plantlets sampled at the third subculture phase. The mother plant, in vitro normal plantlets, and in vitro abnormal plantlets (somaclonal variants) of four sugarcane clones were screened against 16 MSAP selective primers for EcoRI/MspI and EcoRI/HpaII restriction enzymes. A total of 1005 and 1200 MSAPderived markers with polymorphism percentages of 28.36 and 40.67 were obtained for EcoRI/HpaII and EcoRI/MspI restriction enzyme combinations, respectively. The genetic similarity between the mother plant and the somaclonal variants ranged from 0.877 to 0.911 (EcoRI/ MspI) and from 0.928 to 0.955 (EcoRI/HpaII). Most of the MASPs among mother plant and micro-propagated plantlets were derived from EcoRI/MspI restriction enzymes suggesting alteration due to gain or loss of internal cytosine methylation. A higher rate of loss of methylation (hypomethylation) than gain of methylation (hypermethylation) was observed in the abnormal in vitro sugarcane plantlets. Although changes in the methylation pattern were also observed in the in vitro normal plantlets, they were lower than those observed for the in vitro abnormal plantlets. The MASP technique proved to be a promising tool to early assessment of genetic fidelity of micro-propagated sugarcane plants.

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“…En los explantes de Hedysarum theinum también con regeneración adventicia no se afectó al genotipo (Erst et al, 2015) y en Nothapodytes foetida, tampoco se encontró variación en los regenerantes obtenidos de manera adventicia (Chandrika et al, 2010). En varios trabajos se ha reportado que son varios los factores que influyen en la variación genética entre ellos la vía regenerativa de manera indirecta, sin embargo no es regla el hecho que los cultivos de regeneración directa estables tienen menor riesgo de variación somaclonal y los que pasan por una fase de callo presentan mayor probabilidad de variación (Tang, 2001;Gao et al, 2010;Bairu et al, 2011). Esto se comprueba en este trabajo con los brotes regenerados en un medio conteniendo dicha combinación (Fig.…”

Section: Vía De Regeneración Sub-cultivosunclassified

Conservación de plantas regeneradas in vitro y análisis de la variación somaclonal de Cinchona officinalis, Linneo

González¹

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In Vitro Micropropagation of Freesia hybrida and the Assessment of Genetic and Epigenetic Stability in Regenerated Plantlets

Cited by 75 publications

References 59 publications

Assessment of genetic and epigenetic changes during cell culture ageing and relations with somaclonal variation in Coffea arabica

Assessment of genetic and epigenetic changes during cell culture ageing and relations with somaclonal variation in Coffea arabica

DNA methylation in sugarcane somaclonal variants assessed through methylation-sensitive amplified polymorphism

Conservación de plantas regeneradas in vitro y análisis de la variación somaclonal de Cinchona officinalis, Linneo

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