Comparison of UV light, X-ray and restriction enzyme treatment as tools in production of asymmetric somatic hybrids between Brassica napus and Arabidopsis thaliana

Forsberg, Johanna; Lagercrantz, Ulf; Glimelius, Kristina

doi:10.1007/s001220050854

Cited by 29 publications

(8 citation statements)

References 26 publications

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“…The irradiation dose is one important factor. Chromosome elimination was enhanced with the increase of irradiation dose by analyzing the viability, division percentage, and plating efficiency of UV-irradiated protoplasts in our study, the same as in previous reports [6,30,31] . Chromosome elimination is a complex process and the exact mechanism needs comprehensive investigation.…”

Section: Discussionsupporting

confidence: 66%

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Regeneration and identification of interspecific asymmetric somatic hybrids obtained by donor-recipient fusion in cotton

Yang

Zhang

et al. 2009

Chin. Sci. Bull.

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et al. Regeneration and identification of interspecific asymmetric somatic hybrids obtained by donor-recipient fusion in cotton.Asymmetric hybrids between Gossypium hirsutum (YZ-1) and G. davidsonii were obtained by donor-recipient fusion. YZ-1 was considered the recipient and was pretreated with iodoacetamide (IOA), while G. davidsonii was considered the donor and was irradiated with ultraviolet (UV) before fusion. YZ-1 protoplasts stopped growth when treated with 0.5 mmol/L IOA for 20 min, and G. davidsonii protoplasts stopped growth when irradiated with 38.7 J/cm 2 UV for 30 s. Asymmetric somatic hybrids were obtained by electrofusion between the separately treated protoplasts of the 2 species. The regenerated plants were identified by morphological, cytological, and molecular analysis. Most regenerated plants derived from fused protoplasts displayed new morphology; some were intermediate between the two parents and a few displayed recipient-like morphology. Chromosome numbers in these somatic hybrids mostly ranged from 40 to 73. The hybridity was confirmed by random amplified polymorphic DNA and simple sequence repeat analysis. Organelle DNA inheritance of the YZ-1 and G. davidsonii somatic hybrid was investigated by cleaved amplified polymorphism sequence and chloroplast simple sequence repeat analysis, which indicated that recombination and rearrangements might have occurred in some regions of mitochondrial and chloroplastic DNA. This is the first report of completely asymmetric hybrid production via donor-recipient fusion between G. hirsutum and G. davidsonii, which is a novel case in hybrid production following the symmetric fusion and asymmetric fusion based on UV irradiation in cotton.cotton, donor-recipient fusion, IOA, protoplast, UV Conventional breeding programs have made great progress in cotton improvement, but it has become increasingly difficult to make further advances due to sexual incompatibility between some cultivars and wild cotton, especially if the favorable gene is only present in the wild species. An available method for bypassing sexual-crossing barriers is protoplast fusion, which permits transferring desirable agronomically relevant traits from wild cotton to cultivars. This technique has been exploited to enhance resistance and improve quality [1,2] and has produced novel hybrids for incorporation into plant breeding programs [3,4] . In recent years, symmetric and asymmetric protoplast fusion between wild and cultivated species has been successfully achieved in cotton [1,2,5,6] . However, symmetric protoplast fusion transfers not only the desired genes, but also all of the unfavorable ones [3,7] . In asymmetric protoplast fusion, only a part of the wild genome is transferred to a recipient protoplast, reducing the number of undesirable traits incorporated into the recipient genome, but selecting the regenerants is very difficult. So, the donor-recipient fusion with the do-

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Section: Discussionsupporting

confidence: 66%

“…From the FCM analysis (Figure 1(a) and (b)), IOA inactivated cytoplasts and caused the nuclear chromosomes to fragment synchronously. As for nuclei inactivation, UV has been frequently used to break and fragment chromosomes of donors for production of asymmetric hybrids [6,7,15,16,30] . The irradiation dose is one important factor.…”

Section: Discussionmentioning

confidence: 99%

Regeneration and identification of interspecific asymmetric somatic hybrids obtained by donor-recipient fusion in cotton

Yang

Zhang

et al. 2009

Chin. Sci. Bull.

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“…In order to transfer only a limited part of a genome of one species (donor) to the genome of another species (recipient), the technique of asymmetric somatic hybridization has been developed (Dudits et al 1980). Many studies show that asymmetric somatic hybrids often have a higher regeneration and rooting capacity as well as increased fertility compared to symmetric hybrids (Bates et al 1987;Fahleson et al 1994;Skarzhinskaya et al 1996;Forsberg et al 1998b). …”

Section: Introductionmentioning

confidence: 99%

Development of rapeseed with high erucic acid content by asymmetric somatic hybridization between Brassica napus and Crambe abyssinica

2003

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PEG-induced asymmetric somatic hybridization between Brassica napus and Crambe abyssinica was carried out. C. abyssinica is an annual cruciferous oil crop with a high content of erucic acid in the seed oil valuable for technical purposes. UV-irradiated mesophyll protoplasts of C. abyssinica cv 'Carmen' and cv 'Galactica' were fused with hypocotyl protoplasts of different genotypes of B. napus cv 'Maplus' and breeding line '11502'. Shoot regeneration frequency varied between 6.1% and 20.8% among the different doses of UV-irradiation, ranging from 0.05 J/cm(2) to 0.30 J/cm(2). In total, 124 shoots were regenerated, of which 20 asymmetric somatic hybrids were obtained and verified by nuclear DNA content and AFLP analysis. AFLP data showed that some of the characteristic bands from C. abyssinica were present in the hybrids. Cytological analysis of these hybrids showed that 9 out of 20 asymmetric hybrids had 38 chromosomes, the others contained 40-78 chromosomes, having additional chromosomes between 2 and 40 beyond the 38 expected for B. napus. The investigation into the fertility of asymmetric somatic hybrids indicated that the fertility increased with increasing UV-doses ranging from 0.05 J/cm(2) to 0.15 J/cm(2). All of the hybrids were cultured to full maturity, and could be fertilized and set seeds after self-pollination or backcrosses with B. napus. An analysis of fatty acid composition in the seeds was conducted and found to contain significantly greater amounts of erucic acid than B. napus. This study indicates that UV-irradiation could be used as a tool to produce asymmetric somatic hybrids and to promote the fertility of the hybrids.

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“…Mesophyll protoplasts were used in protoplast fusion of plants Arabidopsis thaliana and Brassica napus (FORSBERG et al 1998a), where UV-and X-radiation were used for the production of asymmetric somatic hybrids. Comparing asymmetric hybrids to symmetric hybrids, the asymmetric hybrids were a little taller and had larger leaves.…”

Section: Intergeneric Hybridization With the Genus Brassicamentioning

confidence: 99%

Protoplast cultures and protoplast fusion focused on Brassicaceae: A review

Navrátilová¹

2004

Hort. Sci. (Prague)

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ABSTRACT:The subjects of this article are protoplast isolations and protoplast fusions, in particular their history, a review of factors influencing the protoplasts isolation and fusion, selection of hybrid plants and utilization of somatic hybrids in plant breeding. Somatic hybridization through protoplast fusion can overcome sexual incompatibility among plant species or genera; transfer genes of resistance to diseases (viral, bacterial, fungal), pests, herbicides and others stress factors; obtain cybrid plants; transfer cytoplasmic male sterility or incease content of secondary metabolites in hybrid plants. The article is focussed mainly on the family Brassicaceae because among representatives are significant crops for the human population. Various successful combination of intraspecific, interspecific and intergeneric protoplast fusion were reported between representatives of the family Brassicaceae with the genus Brassica which belonged to the first agricultural crops used for the isolation of protoplast.

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Comparison of UV light, X-ray and restriction enzyme treatment as tools in production of asymmetric somatic hybrids between Brassica napus and Arabidopsis thaliana

Cited by 29 publications

References 26 publications

Regeneration and identification of interspecific asymmetric somatic hybrids obtained by donor-recipient fusion in cotton

Regeneration and identification of interspecific asymmetric somatic hybrids obtained by donor-recipient fusion in cotton

Development of rapeseed with high erucic acid content by asymmetric somatic hybridization between Brassica napus and Crambe abyssinica

Protoplast cultures and protoplast fusion focused on Brassicaceae: A review

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