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Obtaining carrot (Daucus carota L.) plants in isolated microspore cultures

100%

Gorecka K., Kowalska U., Krzyzanowska D., Kiszczak W.

vol. 51

issue 2

141-147

Microspores were cultured on the modified B5 liquid medium containing 2.4D (0.1 mg L?1), NAA (0.1 mg L?1), L-glutamine (500 mg L?1), L-serine (100 mg L?1), and sucrose (100 g L?1). The developmental stages of microspores and divisions were observed. Initially, the formation of binuclear and multicellular structures was noticed. Plants regenerated in the cultures in which the tetrad stage of microsporogenesis had predominated. Embryoids were still forming 24 weeks after the cultures were set up. Six weeks after the transfer of androgenetic embryos onto the B5 regeneration medium, they were converted into complete plants. Out of 90 androgenetic plants planted in a growth chamber, 42 plants adapted to the new conditions. All of those plants proved to be diploids in cytometric analysis.

Obtaining homozygous carrot plants with the aid of androgenesis in anther cultures

88%

Gorecka K., Krzyzanowska D., Kiszczak W., Kowalska U., Gorecki R.

Biotechnologia

2008

issue 2

142-152

Experiments with anther cultures of 22 carrot cultivars were carried out to study the effect of various factors on the effectiveness of embryogenesis in these cultures. The factors included: the stage of microsporogenesis, genotype, training of donor plants and their growth conditions. A modified B5 medium (Gamborg, et al. 1968) containing 500 mg L-1 glutamine, 100 mg L-1 serine, 0.1 mg L-1 of 2,4D, 0.1 mg L-1 NAA, 100 g L-1 sucrose and 6.5 g L-1 agar were used to induce androgenesis. Regeneration was carried out on MS media and B5 with reduced concentration of sucrose at 20 g L-1 without aminoacids and hormones or with small amount of hormones. Substrates that were a mixture of various components, such as peat, sand, mineral wool and charcoal, were used for adaptation. Ploidy of the obtained plants was determined by cytometry method. Homozygosity of the plants was established using two isoenzymatic systems: PGI ? phosphoglucose isomerase, and AAT ? aspartate aminotransferase. Anatomical studies of embryogenesis during anther cultures were also carried out to confirm the androgenetic origin of embryos. It was found that the uninucleate stage was the most suitable time to stimulate microspores to produce embryos, and that bud length was a good external indicator of the stage of microsporogenesis. The studied cultivars differed in their ability to undergo androgenesis in vitro. It was shown that it was not necessary to remove all shoots and umbels except the main one. Generally, the embryos were obtained regardless of the way the donor plants were trained, even when the plants were not trained at all. The donor plants grown in a greenhouse produced more embryos than the plants grown in the field. On MS and B5 media without hormones, used to regenerate plants from embryos, secondary embryogenesis was found to take place followed by a conversion of embryos to complete plants, which subsequently resulted in better adaptation (more than 80% of plants became adapted). Cytometric studies revealed that more than 90% of the obtained androgenetic plants had a doubled chromosome complement. By analyzing the AAT and PGI isoenzymes, it was found that the obtained carrot androgenetic plants were homozygotes. Anatomical studies confirmed that embryos were formed from microspores.

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1 Biotechnologia

1 Journal of Applied Genetics

1 2010

1 2008

2 Gorecka K.

2 Kiszczak W.

2 Kowalska U.

2 Krzyzanowska D.

1 Gorecki R.

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Obtaining carrot (Daucus carota L.) plants in isolated microspore cultures

Obtaining homozygous carrot plants with the aid of androgenesis in anther cultures