In vitro culture of unfertilized ovules in carrot (Daucus carota L.)

Kiełkowska, Agnieszka; Adamus, Adela

doi:10.1007/s11240-010-9735-3

Cited by 30 publications

(24 citation statements)

References 39 publications

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“…In cucumber, TDZ is successfully used for gynogenic induction and regeneration of embryos (Gémes-Juhász et al 2002;Diao et al 2009). Kielkowska and Adamus (2010) reported that medium supplement with indole-3-acetic acid (IAA) promoted embryo development (0.63%), while supplementation with 2,4-D (2,4-Dichlorophenoxyacetic acid) and 6-BA promoted callus development (1.34%) in culture of carrot unfertilized ovule. Addition of polyamine (spermidine and putrescine) in induction and regeneration media resulted in improved gynogenic embryos and haploid plantlets (Martínez et al 2000;Ebrahimi and Zamani 2009).…”

Section: Culture Mediummentioning

confidence: 99%

In vitro haploid and dihaploid production via unfertilized ovule culture

Chen

Cui

Malik

et al. 2010

Plant Cell Tiss Organ Cult

104

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Haploids and doubled haploids are very important in plant breeding, enabling the time needed to produce homozygous lines to be shortened compared with conventional breeding. In the present review, emphasis is given to haploid induction through unfertilized ovule/ovary culture. Attention is given to induction of haploid plants from female gametophyte culture through analysis of factors in the processes of gynogenesis, including genotype selection, stage of ovule development, pretreatment, and culture media containing nutritional components and phytohormones. The gynogenetic approach may be of great value in discovering novel genetic recombinations. Application of double haploids in genetics and plant breeding is also highlighted. This review also identifies some existing knowledge gaps where work may increase the efficiency of this process in different plant species.

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Section: Culture Mediummentioning

confidence: 99%

In vitro haploid and dihaploid production via unfertilized ovule culture

Chen

Cui

Malik

et al. 2010

Plant Cell Tiss Organ Cult

104

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“…It was first envisioned as a technique to accelerate the breeding process and to use for practical and basic research in many crops (Dunwell 2010;Ferrie and Caswell 2010;Germanà 2011). While efficient protocols for microspore embryogenesis induction have been developed to obtain haploid and DH plants in several crops (Maluszynski et al 2003;Dunwell 2010;Ferrie and Caswell 2010), limited progress has been reported using anther cultures (Dohya et al 1997;Tyukavin et al 1999;Adamus and Michalik 2003;Górecka et al 2005;Smýka-lová et al 2009;), isolated microspore cultures (IMC; Dohya et al 1997;Górecka et al 2010;, or unfertilized ovule cultures (Kiełkowska and Adamus 2010) of Apiaceae species.…”

Section: Introductionmentioning

confidence: 99%

Microspore embryogenesis and production of haploid and doubled haploid plants in carrot (Daucus carota L.)

Zhuang

et al. 2012

Plant Cell Tiss Organ Cult

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Protocols were developed for the generation of haploid and doubled haploid plants from isolated microspores of carrot (Daucus carota L.). Forty-seven carrot accessions, including six inbred lines, 11 cultivars, 20 F 1 s, two BC 1 F 1 s, four F 2 s, one F 3 , and three F 4 s, were screened to evaluate the genotype influence on isolated microspore embryogenesis over 4 years. Twenty-eight accessions responded by producing embryos and/or calli. A cytological analysis showed that two modes of carrot microspore embryogenesis exist: an indirect route via calli (C mode), and a direct route via embryos (E mode). Eleven accessions were in the C mode, and 17 were in both modes. The highest production rates were in 10Y25 (a European Nantes cultivar) with 27 calli and 307 embryos, and 100Q6 (a semi-Nantes F 1 hybrid) with 176 calli and 114 embryos. The time period to produce embryos or calli differed significantly between 2 and 6 months. Cold and heat pretreatment generally had a negative impact on the induction of microspore embryogenesis, but a short pretreatment showed a positive influence on some accessions. Twentyeight lines regenerated plants from the primary individual embryos or calli of three accessions were established to analyze the ploidy level. The percentage of spontaneous diploidization showed very wide differences among the accessions and lines. Differences in leaf color intensity, leaf size, and leaf dissection were found among haploid, doubled haploid, and triploid plants.

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“…Ovules were isolated from enlarged ovaries under sterile conditions and placed in 60-mm Petri dishes containing 5 ml medium. Explants were cultured on one of the following media: H — according to Kiełkowska and Adamus ( 2010 ) with 0.06 μM indole-3-acetic acid (IAA); O — 1/2-strength Murashige and Skoog (MS) (Murashige and Skoog 1962 ) with 100 mM MnSO 4 , 4.16 mM Edamin K (Sigma Aldrich Ltd., Poznań, Poland),) and 30 g l −1 sucrose; SHI — full-strength MS with B5 vitamins (Gamborg et al 1968 ) and 0.57 μM IAA, 1.44 μM gibberellic acid (GA 3 ), 4.6 μM kinetin, and 40 g l −1 sucrose; SHT — SHI medium with 9.08 μM thidiazuron (TDZ); HT — H medium with 9.08 μM TDZ; HP — H medium with 1 mM putrescine; or SHP — SHI medium with 1 mM putrescine. All media contained 7 g l −1 Lab-agar (Biocorp, Warszawa, Poland) and were adjusted to pH 5.8 before autoclaving.…”

Section: Methodsmentioning

confidence: 99%

“…The ploidy level of regenerants was determined by flow cytometry as described by Kiełkowska and Adamus ( 2010 ). All diploid plants were then screened for PGI isozyme variants and DNA polymorphism at chs2 and ipi3 loci according to the protocols used for evaluation of the ovule donor plants.…”

Section: Methodsmentioning

confidence: 99%

“…(Eenink 1974 ), Pisum sativum (Virk and Gupta 1984 ), and Gossypium hirsutum (Sacks 2008 , Kantartzi and Roupakias 2009 ). Parthenogenesis has also been described for carrots (Kiełkowska and Adamus 2010 ) after stimulation of ovule development by pollination by other Apiaceae species like parsley or celery. Haploid and diploid plants were obtained from unfertilized ovules cultured in vitro .…”

Section: Introductionmentioning

confidence: 94%

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An improved protocol for carrot haploid and doubled haploid plant production using induced parthenogenesis and ovule excision in vitro

Kiełkowska

Adamus

Barański

2014

In Vitro Cell.Dev.Biol.-Plant

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In this work, we describe an improved protocol for induced parthenogenesis and ovule culture of carrot (Daucus carota L.). The effects of pollination with parsley pollen and/or 2,4-dichlorophenoxyacetic acid (2,4-D) treatment on the stimulation of parthenogenesis were studied using heterozygous donor plants of 30 varieties and breeding populations of carrots. Isolated ovules, cultured in vitro, enlarged and developed embryos or calli. The application of 2,4-D on pollinated flowers stimulated callus development but did not increase the frequency of embryo development from ovules and, thus, was not useful for increasing the frequency of haploid plant recovery. The efficiency of embryo development was accession-dependent and varied from 0 to 24.29%. In optimized conditions, most accessions responded by embryo development exclusively. The highest frequency of embryo development was observed from ovules excised from ovaries 20–22 d after pollination with parsley pollen. Among several media used for ovule culture, 1/2-strength Murashige and Skoog medium with 0.06 μM indole-3-acetic acid (IAA) was the best. It allowed the production of embryos at a similar frequency as on the media supplemented with kinetin, gibberellic acid, putrescine, or thidiazuron, but restricted callus development. Most plants obtained were haploids and diploids derived from parthenogenesis, as evidenced by homozygosity at three independent loci based on isozyme and PCR analyses. In total, considering haploids and embryo-derived homozygous diploids together, 72.6% of regenerated plants were of gametic origin.

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In vitro culture of unfertilized ovules in carrot (Daucus carota L.)

Cited by 30 publications

References 39 publications

In vitro haploid and dihaploid production via unfertilized ovule culture

In vitro haploid and dihaploid production via unfertilized ovule culture

Microspore embryogenesis and production of haploid and doubled haploid plants in carrot (Daucus carota L.)

An improved protocol for carrot haploid and doubled haploid plant production using induced parthenogenesis and ovule excision in vitro

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