Haploids and doubled haploids in Brassica spp. for genetic and genomic research

Ferrie, A. M. R.; Möllers, Christian

doi:10.1007/s11240-010-9831-4

Cited by 101 publications

(63 citation statements)

References 82 publications

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“…Variation and mutagenesis of plantlets were also appreciated by geneticists and breeders (Dunwell 2010;Ferrie and Möllers 2010). We obtained several plantlets that were sensitive to bolt even without any cold treatment in culture or soon after transplanting (Fig.…”

Section: Discussionmentioning

confidence: 91%

“…Selection and production of inbred lines is time consuming and difficult to achieve. Even with 6 years of self-pollination by conventional breeding methods, only about 98 % homozygosity could be achieved (Germanà 2006;Ferrie and Möllers 2010). Moreover, the vigor of early-generation carrot inbred lines selected from open-pollinated cultivars decreases dramatically due to inbreeding depression (Peterson and Simon 1986).…”

Section: Introductionmentioning

confidence: 99%

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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: Discussionmentioning

confidence: 91%

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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“…Traits are fixed and homozygous true breeding lines are produced within one generation, which can help reduce the time needed to develop new varieties (Thomas et al 2003;Ferrie and Caswell 2011;Ferrie and Möllers 2011). For the same reasons, in vitro microspore culture has become an important tool for mutagenesis (Barro et al 2001(Barro et al , 2002Sonntag and Rudloff 2004;Liu et al 2005;Ferrie et al 2008) and in vitro selection (Albrecht et al 1995;Möllers et al 2000;Liu et al 2005).…”

Section: Introductionmentioning

confidence: 99%

“…In rapeseed, F 1 derived segregating DH populations have been extensively used for map construction and the genetic analysis of important traits (Ferreira et al 1994;Parkin et al 1995;Sharpe et al 1995;Uzunova et al 1995;Zhao et al 2005;Delourme et al 2006;Qiu et al 2006;Radoev et al 2008). In these mapping studies distorted marker segregations have been observed regularly (summarized in Ferrie and Möllers 2011). This is often explained by the assumption that the selection of favourable alleles at loci controlling microspore embryogenesis leads to skewed segregations at linked marker loci (Foisset et al 1993;Cloutier et al 1995;Foisset and Delourme 1996).…”

Section: Introductionmentioning

confidence: 99%

Identification and evaluation of intervarietal substitution lines of rapeseed (Brassica napus L.) with donor segments affecting the diploidization rate of isolated microspores

et al. 2016

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In Brassica species microspore derived doubled haploid lines are an important tool in breeding and research. A limiting step in the production of doubled haploid lines is the diploidization of the microspores. Strong differences have been observed in diploidization rates between different genotypes but little is know about the genetic control of these differences. To identify genomic regions that carry genetic factors controlling the diploidization rate of isolated microspores of rapeseed, marker segregations were compared between segregating populations of diploid and haploid microspore derived embryos and a BC 1 from a cross between 'Express 617' and 'RS239'. After map construction ten intervarietal substitution lines from the same cross were selected with donor segments covering five genomic regions that showed a pattern of skewed marker segregations across the three populations indicative of the segregation of genes controlling the diploidization rates. The diploidization rates of microspores of the ten lines ranged from 23.9 to 58.7 % while the recurrent parent 'Express 617' showed a rate of 52.5 %. For three lines the diploidization rates were significantly lower (P = 0.05) than the rate of 'Express 617'. By comparing donor segments between the significant and the non-significant lines, seven genomic regions that cover just between 4.17 and 6.16 % of the rapeseed genome were identified that may contain genetic factors controlling diploidization rates in rapeseed. In addition, one marker was found that has a high probability to be linked to such a factor. The significant lines represent an ideal material for further in depth studies of this trait.

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“…The biotechnology of producing pure breeding lines, the doubled haploids, via anthers and isolated microspore cultures is recently widely applicable in genetic studies and breeding cole crops (1)(2)(3). Due to this approach, the in vitro generated plants have different ploidy, and, together with doubled haploids, also haploid, tetraploid and mixoploid forms can occur (4).…”

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confidence: 99%

A RELATIONSHIP BETWEEN PLOIDY LEVEL AND THE NUMBER OF CHLOROPLASTS IN STOMATAL GUARD CELLS IN DIPLOID AND AMPHIDIPLOID Brassica SPECIES

Монахос¹,

Нгуен²,

Bezbozhnaya³

et al. 2014

S-h. biol.

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A b s t r a c t Doubled haploid lines production through isolated anthers and isolated microspore cultures has been used widely for genetic studies and plant breeding of Brassica crops. The ploidy level of microspore derived plants varies, and normally haploid, diploid and mixoploid plants could be obtained in vitro. The determination of ploidy level is essential in doubled haploid pure line production. The determination of ploidy level by counting the number of chloroplast in stomatal guard cells (NCSGC) is less time consuming, laborious and expensive comparing to chromosome counting in root tip cells or mother pollen cells and flow cytometry methods. Several studies have been reported concerning relationship between ploidy level and number of chloroplast in stomatal guard cells of Brassica rapa, B. napus and B. oleracea species, however a small number of genotypes had been analyzed. In our study, the NCSGCs of haploid (n) and diploid (2n) Chinese cabbage (B. rapa ssp. pekinensis), winter oilseed rape (B. napus var. napus) and haploid, di-and tetraploid white cabbage (B. oleracea var. capitata) microspore derived plants were estimated, and also the influence of plant growth temperature (6±2 °Ñ è 24±2 °Ñ) and development stage (vegetative or generative) was investigated. High correlation between the ploidy level of microspore-derived plants and NCSGC is found for white cabbage (Brassica oleracea, r = 0.94), Chinese cabbage (B. rapa, r = 0.90) and oilseed rape (B. napus, r = 0.94). The chloroplast average number in stomatal guard cells was very similar among the same ploidy genotypes of Chinese cabbage as well as rapeseed, while the variation of chloroplast number in diploid and tetraploid white cabbage plants was significant. In a range of early, middle and late maturing diploid white cabbage inbred lines there was established the tendency to form more chloroplasts in the early lines (Pl, Sus and others) and less in the late lines (AM2, Sa1, Ges2 and others), with the difference up to 1.7 times, that is comparable to the difference between haploid and diploid plants of Chinese cabbage or rapeseed. The chloroplast number in stomatal guard cells is 4.2-7.8 and 7.9-13.6 for Chinese cabbage (B. rapa) haploids and diploids, respectively, 7.5-12.4 and 14.1-20.3 for rapeseed (B. napus) amphihaploids and amphidiploid, respectively, and 7.7-9.9, 11.7-17.9 and 18.0-26.5 for white cabbage (B. oleracea) haploids, diploids and tetraploids, respectively. No significant influence of vegetative or generative stage of plant development or growth temperature on NCSGC was found.

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Haploids and doubled haploids in Brassica spp. for genetic and genomic research

Cited by 101 publications

References 82 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.)

Identification and evaluation of intervarietal substitution lines of rapeseed (Brassica napus L.) with donor segments affecting the diploidization rate of isolated microspores

A RELATIONSHIP BETWEEN PLOIDY LEVEL AND THE NUMBER OF CHLOROPLASTS IN STOMATAL GUARD CELLS IN DIPLOID AND AMPHIDIPLOID Brassica SPECIES

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