Cremart: A new chemical for efficient induction of micronuclei in cells and protoplasts for partial genome transfer

Ramulu, K. Sree; Dijkhuis, P.; Famelaer, I.; Cardi, Teodoro; Verhoeven, H. A.

doi:10.1007/bf00231625

Cited by 20 publications

(15 citation statements)

References 15 publications

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“…Although other spindle toxins, amiprophos-methyl and butamiphos, had been used for inducing efficient micronucleation in suspension-cultured cells of Nicotiana plumbaginifolia 11 , Solanum tuberosum 12,13,24 and Helianthus giganteus 2 , their efficiency on the induction of micronucleation in Hemerocallis hybrida was limited 21 .…”

Section: Micronucleationmentioning

confidence: 99%

Preparation of Microprotoplasts for Partial Genome Transfer via Microprotoplast Fusion in Liliaceous Ornamental Plants

Saito

Nakano

2002

JARQ

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We aimed to produce intergeneric hybrid plants with only one or a few alien chromosomes via microprotoplast fusion for genetic improvement and chromosome studies in Liliaceous ornamental plants. In order to apply this technique, it is essential to establish an efficient system for mass-preparation of microprotoplasts. We have established 2 different systems for isolating microprotoplasts, one from partially synchronized cell suspension cultures of Hemerocallis hybrida and the other from developing microspores of Lilium longiflorum. Here, the induction of micronucleated cells, isolation of microprotoplasts, and enrichment of smaller microprotoplasts containing one or a few chromosomes are described for both systems.

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Section: Micronucleationmentioning

confidence: 99%

Preparation of Microprotoplasts for Partial Genome Transfer via Microprotoplast Fusion in Liliaceous Ornamental Plants

Saito

Nakano

2002

JARQ

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“…According to the expected observations these should be small and contain 1 to a few chromosomes (Ramulu et al 1994, data above). An effective isolation procedure had, therefore, to be developed based on the previous observations with FCM and CLSM.…”

Section: Monitoring Of the Process Of Micro-nucleation And Mpp Isolatmentioning

confidence: 75%

“…For induction of micronuclei, the suspension cells were subsequently treated for 24 h with APM (amiprophos-methyl; Tokunol M; 0-methyl-0-0-(4-methyl-6-nitrophenyl)-N-isopropyl-phosphoro thiamidate from Bayer Mobay Corporation, Agricultural Chemicals Division, Kansas City, MO., USA) or CA (Cremart; Butamiphos; Q-ethyl-Q-(3-methyl-6-nitro-phenyl) N-sec-butylphosphorothioamidate from Sumitome Chemical Comp., Osaka, Japan) at 15-30 mM or 2.25-36 mM respectively. These concentrations were chosen on the basis of our previous work with Nicotiana plumbaginifolia and potato (Ramulu et al 1993(Ramulu et al , 1994.…”

Section: Synchronization and Induction Of Micronucleimentioning

confidence: 99%

A study of the process of synchronisation and micronucleation in Beta vulgaris and the monitoring of an isolation procedure for micro-nuclei and micro-protoplasts by confocal laser scanning microscopy and flow cytometry

Famelaer

Verhoeven²,

Dijkhuis³

et al. 2007

Plant Cell Tiss Organ Cult

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The process of synchronization and micro-nuclei induction in a suspension culture of Beta vulgaris, was induced by the sequential treatment with the DNA-synthesis inhibitor aphidicolin (30 mM, 24 h) and the spindle-toxin amiprophos-methyl (32 mM, 24 h). Mitotic arrest of divisions, spreading of G2-metaphase chromosomes, re-grouping of chromosomes, formation of a nuclear cell wall around single and re-grouped chromosomes and restitution of nuclei with a doubled DNA content was observed. The process of micro-nucleation was induced much less efficiently in Beta vulgaris than in Nicotiana plumbaginifolia. Cytological observations and monitoring of the process with flow cytometry and confocal laser scanning microscopy, was essential to follow up the course of events and to monitor the development of an efficient procedure for microprotoplast isolation. Micro-nucleated protoplasts were fractioned by iso-osmotic Percoll gradient centrifugation to obtain heterogeneous micro-protoplast populations with cytoplasts and micro-protoplasts of different size. An enriched fraction with small subdiploid micro-protoplasts was obtained with the equivalent DNA content of 1-4 chromosomes, as revealed by confocal laser scanning microscopy and flow cytometry. Sub-diploid micro-protoplasts with DNA amounts equivalent to 1-4 chromosomes were predominantly observed in the size classes of 1.8-10.2 mm at a frequency of 34.2-34.5%. The DNA measurements of micro-nuclei and micro-protoplasts, confirmed the hypothesis that the process of micronucleation followed the same course of cellular events as observed in N. plumbaginifolia. The correlation between DNA content and size of micro-nuclei and micro-protoplasts was not linear and affected by the degree of DNA condensation, total amount of DNA, and the presence of cytoplasm.

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“…Helianthus Schnabl et al (1999) Vinca Catharanthus Kostenyuk et al (1991) used for chromosomal fragmentation (Ramulu et al, 1994;Forsberg et al, 1998b). Treatment of the donor protoplasts with bromodeoxyuridine prior to UV and gamma irradiation could promote formation of highly asymmetric hybrids (Trick and Bates, 1996).…”

Section: Viciamentioning

confidence: 97%

Intergeneric somatic hybridization and its application to crop genetic improvement

Liu

Deng

2005

Plant Cell Tiss Organ Cult

112

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Related or distant species of cultivated crops are a large pool of many desirable genes. Gene transfer from these species through conventional breeding is difficult owing to post-and pre-zygotic sexual incompatibilities. Somatic hybridization via protoplast fusion is a possible alternative for gene transfer from these species to cultivated crops. Since the early days of somatic hybridization many intergeneric somatic hybrids have been developed through symmetric fusion, asymmetric fusion and microfusion. Somatic hybrids are mainly selected by using markers such as specific media or fusion parents with special features, biochemical mutants, antibiotic resistance and complementation strategy. The hybridity of the regenerants is determined based on morphological, cytological and molecular analysis. The inheritance patterns of nuclear and cytoplasmic genomes in the somatic hybrids are diverse. Nuclear DNA from both fusion parents co-exists congruously in some hybrids with translocation and rearrangement of chromosomes, but spontaneous elimination of chromosomes from either or both fusion parents has been observed very often. In asymmetric fusion, chromosome elimination is an important issue that is a complicated process influenced by many factors, such as irradiation dose, phylogenetic relatedness, ploidy level of fusion parent and regenerants. As for chloroplast genome, uniparental segregation is mainly detected, though co-existence is also reported in some cases. The mitochondrial genome, in contrast to chloroplast, undergoes recombination and very frequent rearrangements. Somatic cell fusion has potential applications for crop genetic improvement by overcoming sexual incompatibility or reproductive barriers, and by realizing novel combinations of nuclear and/or cytoplasmic genomes.

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Cremart: A new chemical for efficient induction of micronuclei in cells and protoplasts for partial genome transfer

Cited by 20 publications

References 15 publications

Preparation of Microprotoplasts for Partial Genome Transfer via Microprotoplast Fusion in Liliaceous Ornamental Plants

Preparation of Microprotoplasts for Partial Genome Transfer via Microprotoplast Fusion in Liliaceous Ornamental Plants

A study of the process of synchronisation and micronucleation in Beta vulgaris and the monitoring of an isolation procedure for micro-nuclei and micro-protoplasts by confocal laser scanning microscopy and flow cytometry

Intergeneric somatic hybridization and its application to crop genetic improvement

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