Enhanced regeneration of haploid plantlets from microspores of Brassica napus L. using bleomycin, PCIB, and phytohormones

Ahmadi, Behzad; Alizadeh, Khalil; Silva, Jaime A. Teixeira da

doi:10.1007/s11240-012-0119-8

Cited by 24 publications

(12 citation statements)

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“…The conditions leading to the induction and development of microspore-derived embryos vary depending on the species. Many factors influence microspore embryogenesis, including genotype, stage of microspore development, donor plant growth conditions, media composition, and culture conditions (Maluszynski et al 2003;Dunwell 2010;Ferrie and Caswell 2010;Ahmadi et al 2012). Matsubara et al (1995) firstly obtained small calli from a few carrot-isolated microspores.…”

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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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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“…It is known that treatment of colchicine instead of heat shock or with heat shock stimulates embryogenesis (Zhao et al 1996 ;Zhou et al 2002 ;Agarwal et al 2006 ). Furthermore, bleomycin (0.1-0.2 μg/ml), a glycopeptide antibiotic, for 20-30 min pretreatment increased embryo yields in B. napus , (Zeng et al 2010 ;Ahmadi et al 2012 ). Some mutagens such as γ-rays irradiation, N-Nitroso-N-Methyl Urethane (NMU) and ethyl methane sulphonate (EMS) were reported to promote microspore embryogenesis in B. napus , B. oleracea and B. juncea (Jeung and Lee 1997 ;Pechan and Keller 1989 ;Ferrie et al 2008 ).…”

Section: Stress and Pretreatmentmentioning

confidence: 99%

“…The anti-auxin, p -chlorophenoxyisobutyric acid (PCIB) improved microspore embryogenesis by 2-6 fold in B. juncea , B. rapa and B. napus (Agarwal et al 2006 ;Zhang et al 2011 ;Ahmadi et al 2012 ). The methods of treatment were different among these studies; one was pretreatment (1 day) of PCIB (Agarwal et al 2006 ;Ahmadi et al 2012 ), and another was treatment of PCIB during culture period . Brassinosteroids (BRs) such as 24-epibrassinolide (EBR) and brassinolide (BL) increased the production of microspore embryogenesis in B. napus and B. juncea , however, they had no effects in B. carinata , B. nigra and B. rapa (Ferrie et al 2005 ).…”

Section: Culture Mediummentioning

confidence: 99%

“…In contrast, Ferrie et al ( 2005 ) observed that brassinosteroids did not affect the plant regeneration from embryos. Although the exogenous plant growth regulators are not essential in the plant regeneration from microsporederived embryos, 0.05-0.1 mg/l GA 3 , 2.0 mg/l BA or 0.1 mg/l BA + 0.2 mg/l IAA were reported to enhance the regeneration in B. napus (Ahmadi et al 2012 ).…”

Section: Plant Regeneration and Dh Productionmentioning

confidence: 99%

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Microspore Culture and Doubled Haploid Technology

Takahata

Takahashi

Tsuwamoto³

2013

Biotechnology of Crucifers

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Haploids and doubled haploids (DHs) produced by in vitro culture of gametophytic cells, especially male gametophyto, are of great importance to plant breeding and basic science. The routine protocol of isolated microspore culture of Brassica has been established, and used worldwide. However, recently, new protocols dealing with multiple samples have been developed. Although various factors infl uencing microspore embryogenesis, plant regeneration and diplodization have been examined, attempts to enhance these effi ciencies have continued. With the advancement of molecular genetics, many approaches from both forward-and reverse-genetics for understanding the mechanism of microspore embryogenesis have been attempted. This chapter reviews the recent progress of microspore embryogenesis of Brassica in relation to culture protocol and factors affecting microspore embryogenesis, and recent achievements in elucidating the mechanism of microspore embryogenesis.

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“…If traditionally, plant breeders usually achieve homozygosity of the cross products through self-fertilization in 6-8 years, by microspore culture, homozygous plants are obtained within a year because such lines are obtained in a single generation In vitro, while several generations of inbreeding are required using traditional means. Even with 6 years of self pollination in a conventional plant breeding program, the breeder can only achieve about 98% homozygosity, compared to 100% with doubled haploidy techniques (Ahmadi, 2012). In order for doubled haploidy to be effective in a breeding program, an efficient microspore culture protocol is required.…”

Section: Introductionmentioning

confidence: 99%

The Influence of pH on Microspore Embryogenesis of White Cabbage (<i>Brassica oleracea</i> L.)

Cristea

2013

Not Sci Biol

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In vitro microspore culture is one of the top techniques utilised now-a-days for the obtaining of double haploid plants in many plant species, including Brassica. The pH of the medium is a critical factor for the success of In vitro microspore culture as it influences the invertase enzyme activity, translated at cellular level through an acceleration or reduction of sucrose cleavage. The results published until now shows rather contradictory findings, as the response of microspores have been proved to be highly depending on genotypes, most of them being focused on Brassica napus. Thus, in the present study, the effect of different NLN liquid medium pH, ranging between 5.0 to 7.0 were tested in order to establish the most suitable pH for the expression of embryogenic competences of microspores cultivated on medium In vitro and ultimately for the obtaining of microspore-derived embryos. Among the 11 values of pH tested, the best results were obtained on variants with pH 5.8 and 6.0, both in what concern the maintaining of microspores viability and the number of microspore-derived embryos. The findings of the present study provide a strong base for the establishment of an efficient protocol for the In vitro culture of microspore at Brassica oleracea L. genotypes with Romanian origin.

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Enhanced regeneration of haploid plantlets from microspores of Brassica napus L. using bleomycin, PCIB, and phytohormones

Cited by 24 publications

References 43 publications

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

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

Microspore Culture and Doubled Haploid Technology

The Influence of pH on Microspore Embryogenesis of White Cabbage (<i>Brassica oleracea</i> L.)

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