An improved protocol for androgenesis in cauliflowers (<i>Brassica oleracea</i> var. <i>botrytis</i>)

Stipic, M.; Campion, B.

doi:10.1111/j.1439-0523.1997.tb02170.x

Cited by 13 publications

(4 citation statements)

References 20 publications

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“…Most of the cabbage and cauliflower regenerants were also completely sterile with abortive anthers or self-sterile despite normal morphology of flowers. This problem was described previously by STIPIC and CAMPION (1997). Despite the high number of diploid regenerants, the frequency of fertile regenerants was very low in their experiments with cauliflower.…”

Section: Resultsmentioning

confidence: 70%

Production and utilization of doubled haploids in Brassica oleracea vegetables

Klíma¹,

Vyvadilová²,

Kučera³

2004

Hortic. Sci.

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Development of doubled haploid lines by means of in vitro microspore culture has been increasingly used for homozygous line production in breeding programmes of Brassica crops due to a possibility of significant time reduction of cultivar development. Whereas this method is routinely used in oilseed rape (Brassica napus), it is still difficult to apply microspore culture techniques to practical breeding of some vegetable brassicas. The main difficulties are the very low embryo yield and insufficient regenerative ability of microspore-derived plants in many of Brassica oleracea genotypes (CARLOS, DIAS 1999;RUDOLF et al. 1999). Very often abnormal embryos occur which need to be subcultured many times to induce normal shoots (KUGINUKI et al. 1999). Direct and rapid plant regeneration is very important for eliminating cytogenetic abnormalities and improving the efficiency of doubled haploid system. Our previous studies (VYVADILOVÁ et al. 1998a(VYVADILOVÁ et al. ,b, 2001 aimed to increase the efficiency of the microspore culture technique, especially by investigating factors affecting pollen embryogenesis and testing the embryogenic responsibility in a broad range of genotypes from Brassica oleracea collection and some breeding materials. The previously optimized microspore culture procedure was used in this study, which was directed at increasing the efficiency of whole plant production. Obtained doubled haploid lines will be evaluated for agronomic and quality traits with an emphasis on fungal disease and virus resistance and promising lines will be used in breeding programmes. MATERIAL AND METHODSBrassica oleracea genotypes including several botanical varieties such as cabbage, cauliflower and kohlrabi used for experiments were selected according to the previous results regarding the embryogenic responsibility. Eight cultivars and landraces of head cabbage (convar. capitata L.) var. alba DC. and var. rubra DC.; four kohlrabi (var. gongylodes L.) open-pollinated cultivars and one self-pollinated line, one commercial F1 hybrid and three experimental hybrids of cauliflower (var. botrytis L.) were used for experiments. After vernalization for 4 months at 4°C in a cold room plants of cabbage and kohlrabi were grown in the growth chamber under controlled environmental conditions with a 16 h photoperiod and day/night temperature 18/12°C. Cauliflower plants were grown in isolation cages in the openair conditions. The microspore culture technique was modified to make it applicable to a wider spectrum of genotypes (VYVADILOVÁ et al. 2001). Green cotyledonary embryos about 5 mm in size were subcultured (for 5 to 7 days) on differentiation medium with benzylaminopurine, indolyl acetic acid and 2% sucrose, solidified by 0.8% agar (Table 1). Afterwards two thirds of both cotyledons were cut off and embryos were transferred onto regeneration medium without phytohormones, with 1% sucrose and 1% agar (Table 2) More than 400 regenerants of R 1 generation were derived in kohlrabi, cabbage and cauliflower by means of different mod...

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Section: Resultsmentioning

confidence: 70%

Production and utilization of doubled haploids in Brassica oleracea vegetables

Klíma¹,

Vyvadilová²,

Kučera³

2004

Hortic. Sci.

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“…Previous anther culture studies in cauliflower reported 41% (Ockendon 1988), 26% (Wang et al 1999) and 79% spontaneous diploids (Stipic and Campion 1997).…”

Section: Ploidy Analysismentioning

confidence: 89%

Effect of incubation temperature and sucrose concentration on callus induction and plant regeneration in cauliflower (Brassica oleracea var. botrytis)

Singh,

Devi,

Sarao

2021

VegSci

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Androgenic and plant regeneration potential from anther culture were investigated in cauliflower genotypes under different incubation temperatures, sucrose concentrations and regeneration media compositions. Anthers were cultured on MS medium supplemented with 1.5 mg/L 2,4-D and 1.0 mg/L NAA growth hormone with four different treatments of sucrose i.e. 3%, 5%, 10% and 13%. Maximum androgenesis has been observed with 10% sucrose concentration. Anthers of all the varieties were incubated under three different temperatures i.e. 25°C, 32°C and 35°C. Among these incubation temperatures, higher androgenesis was observed at 32°C incubation temperature. Callus were cultured on five shoot regeneration media compositions, among which maximum regeneration has been observed on MS media supplemented with 5.0 mg/L BAP and 1.0 mg/L NAA followed by MS media supplemented with 6.0 mg/L BAP and 1.0 mg/L NAA. Maximum root regeneration has been observed in ½ MS medium supplemented with 1.0 mg/ L IBA growth hormone followed by ½ MS medium supplemented with 0.4 mg/L IBA. In cytological examination, 20.3% of the regenerated plants were found to be haploid while 29.8% were spontaneous doubled haploids.

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“…The production of DH plants from microspores is an important technique used for plant breeding and basic research (Ferrie and Caswell, 2011). Isolated microspore cultures have been successfully used for B. oleracea crops, including cabbage (Rudolf et al, 1999), broccoli (Dias, 2001), and cauliflower (Stipic and Campion, 1997). Mejza et al (1993) regenerated wheat plants from isolated microspores using an ovary co-culture.…”

Section: Discussionmentioning

confidence: 99%

Rapid Introgression of the Fusarium Wilt Resistance Gene into an Elite Cabbage Line through the Combined Application of a Microspore Culture, Genome Background Analysis, and Disease Resistance-Specific Marker Assisted Foreground Selection

et al. 2017

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Cabbage is an economically important vegetable worldwide. Cabbage Fusarium Wilt (CFW) is a destructive disease that results in considerable yield and quality losses in cole crops. The use of CFW-resistant varieties is the most effective strategy to mitigate the effects of CFW. 01-20 is an elite cabbage line with desirable traits and a high combining ability, but it is highly susceptible to CFW. To rapidly transfer a CFW resistance gene into 01-20 plants, we used microspore cultures to develop 230 doubled haploid (DH) lines from a cross between 01-20 (highly susceptible) and 96-100 (highly resistant). One of the generated DH lines (i.e., D134) was highly resistant to CFW and exhibited a phenotypic performance that was similar to that of line 01-20. Therefore, D134 was applied as the resistance donor parent. We generated 24 insertion–deletion markers using whole genome resequencing data for lines 01-20 and 96-100 to analyze the genomic backgrounds of backcross (BC) progenies. Based on the CFW resistance gene FOC1, a simple sequence repeat (SSR) marker (i.e., Frg13) was developed for foreground selections. We screened 240 BC1 individuals and 280 BC2 individuals with these markers and assessed their phenotypic performance. The proportion of recurrent parent genome (PRPG) of the best individuals in BC1 and BC2 were 95.8 and 99.1%. Finally, a best individual designated as YR01-20 was identified from 80 BC2F1 individuals, with homozygous FOC1 allele and genomic background and phenotype almost the same as those of 01-20. Our results may provide a rapid and efficient way of improving elite lines through the combined application of microspore culture, whole-genome background analysis, and disease resistance-specific marker selection. Additionally, the cabbage lines developed in this study represent elite materials useful for the breeding of new CFW-resistant cabbage varieties.

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An improved protocol for androgenesis in cauliflowers (Brassica oleracea var. botrytis)

Cited by 13 publications

References 20 publications

Production and utilization of doubled haploids in Brassica oleracea vegetables

Production and utilization of doubled haploids in Brassica oleracea vegetables

Effect of incubation temperature and sucrose concentration on callus induction and plant regeneration in cauliflower (Brassica oleracea var. botrytis)

Rapid Introgression of the Fusarium Wilt Resistance Gene into an Elite Cabbage Line through the Combined Application of a Microspore Culture, Genome Background Analysis, and Disease Resistance-Specific Marker Assisted Foreground Selection

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