Characterization and molecular analysis of transgenic plants obtained by microprotoplast fusion in sunflower

Binsfeld, P. C.; Wingender, Ruth; Schnabl, Heide

doi:10.1007/s001220051604

Cited by 36 publications

(57 citation statements)

References 26 publications

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“…About 90% of the CIPCtreated microsporocytes formed more than 4 nuclei, and meiocytes with 7 or 8 nuclei were most frequently obtained. Until now, amiprophos-methyl had mainly been used for inducing micronucleation in microsporocytes of Solanum tuberosum 5 , and in suspension-cultured cells of several Solanaceous species 14,17 and Helianthus giganteus 2 . On the other hand, micronucleation of suspension-cultured cells of Hemerocallis hybrida was efficiently induced by propyzamide 21,22 .…”

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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“…Stimulation of micronucleation by CB had already been reported for human cells: although it had been considered to be difficult to obtain micronucleated human cells by sole treatment with colcemid, micronucleation could be induced by the addition of CB to the cultures during the colcemid treatment (Fournier 1982). As in the case of human cells, CB also stimulated micronucleation of cultured cells of Solanum tuberosum and Nicotiana plumbaginifolia (Ramulu et al 1990(Ramulu et al , 1993, and Helianthus giganteus (Binsfeld et al 2000) during spindle toxin treatment. In these previous studies, CB was added to the cultures when the MI reached a peak (Ramulu et al 1990, 1993, Binsfeld et al 2000.…”

Section: Discussionmentioning

confidence: 83%

“…cell fusion between microprotoplasts and somatic cell protoplasts, can be used for partial genome transfer in higher plants , Ramulu et al 1992, 1995, 1999. Several asymmetric hybrid plants have so far been produced by this technique in the Solanaceous species (Ramulu et al 1996a,b) and in the genus Helianthus (Binsfeld et al 2000).…”

Section: Introductionmentioning

confidence: 99%

“…For applying this technique, it is essential to develop efficient systems for inducing micronucleated cells as well as for isolating microprotoplasts Ramulu 1991, Ramulu et al 1993). To date, microprotoplasts have been successfully prepared from micronucleated suspension cells of Nicotiana plumbaginifolia, Solanum tuberosum, S. brevidens, Beta vulgaris (Ramulu et al 1992) and Helianthus giganteus (Binsfeld et al 2000).…”

Section: Introductionmentioning

confidence: 99%

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Isolation and Characterization of Microprotoplasts from Propyzamide-treated Cell Suspension Cultures of Hemerocallis hybrida.

Saito

Nakano

2002

Breed. Sci.

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To produce intergeneric hybrid plants with one or a few chromosomes via microprotoplast fusion in the Liliaceous ornamentals, we developed a system for isolating microprotoplasts from micronucleated cells in suspension cultures of Hemerocallis hybrida cv. Stella d'Oro (2n = 2x = 22). Micronucleation was efficiently induced with a micronucleus index up to 19.1 % by the following sequential treatment of suspension cultures: initially with 2 mM hydroxyurea for 24 h and then with 8 µM propyzamide for 60 h with the addition of 20 µM cytochalasin-B to the cultures 20 h after the initiation of the propyzamide treatment. Following the enzyme treatment of the micronucleated cells and ultra-centrifugation with a continuous iso-osmotic gradient of Percoll solution, microprotoplasts were isolated, each of which had a small nucleus surrounded by a thin rim of cytoplasm. Microprotoplasts less than 10 µm in diameter were obtained with yield of 2.9 × 10 4 cells per 1 ml packed cell volume of suspension cells through sequential filtration with nylon sieves with decreasing pore sizes (50, 20 and 10 µm). The DNA content of the microprotoplasts was less than the 2C level as indicated by flow cytometric analysis, and the relative fluorescence intensity in some of their nuclei corresponded to one or a few chromosomes.

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“…Molecular markers are widely used to estimate the genetic variations, as molecular markers are self-determining for confounding results of environment factors. SSR and RAPD markers were used to observe the genetic distance between sunflower genotypes (Binsfeld et al, 2000;Popov et al, 2002) and identification of hybrids and getting information of variation for future breeding (Saftić-Panković, 2007).…”

Section: Discussionmentioning

confidence: 99%

Assessment of RAPD Markers to Analyse the Genetic Diversity among Sunflower (Helianthus annuus L.) Genotypes

Raza

Shaukat

Ali

et al. 2018

Turkish JAF Sci.Tech.

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Genetic diversity estimation among different species is an important tool for genetic improvement to maximize the yield, desirable quality, wider adaptation, pest and insect resistance that ultimately boosting traditional plant breeding methods. The most efficient way of diversity estimation is application of molecular markers. In this study, twenty random amplified polymorphic DNA (RAPD) primers were utilized to estimate the genetic diversity between ten sunflower genotypes. Overall 227 bands were amplified by 20 primers with an average of 11.35 bands per primer. RAPD data showed 86.34% polymorophic bands and 13.65% of monomorophic bands. Genetic similarity was ranged from 50.22% to 87.22%. The lowest similarity (50.22%) was observed between FH-352 and FH-359 and the maximum similarity 87.22% was observed between A-23 and G-46. Polymorphic information content (PIC) values were varying from 0.05 to 0.12 with a mean of 0.09. Cluster analysis based on RAPD results displayed two major distinct groups 1 and 2. Group-2 contains FH-352 which was the most diverse genotype, while group-1 consists of few sub groups with all other genotypes. Ample diversity was found in all the genotypes. Present study reveals novel information about sunflower genome which can be used in future studies for sunflower improvement.

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Characterization and molecular analysis of transgenic plants obtained by microprotoplast fusion in sunflower

Cited by 36 publications

References 26 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

Isolation and Characterization of Microprotoplasts from Propyzamide-treated Cell Suspension Cultures of Hemerocallis hybrida.

Assessment of RAPD Markers to Analyse the Genetic Diversity among Sunflower (Helianthus annuus L.) Genotypes

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