Regeneration of plants from mesophyll protoplasts ofBrassica oleracea

Bidney, Dennis; Shepard, James F.; Kaleikau, Edward K.

doi:10.1007/bf01281788

Cited by 39 publications

(21 citation statements)

References 3 publications

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“…Generation of intergeneric somatic hybrids is no exception, TORIYAMA et al (1987), FAHLESON et al (1994) and NAVRÁTILOVÁ et al (1997) have done a great deal of work in this respect. The regeneration frequencies being genotype dependent vary between species, genotypes and source materials (BIDNEY et al 1983;GLIMELIUS 1984;VAMLING & GLIMELIUS 1990). Plant regeneration has been reported from hypocotyl and leaf protoplasts of B. oleracea var.…”

mentioning

confidence: 99%

A Simple Procedure for Mesophyll Protoplast Culture and Plant Regeneration in Brassica oleracea L. and Brassica napus L.

Kaur¹,

Vyvadilová²,

Klíma³

et al. 2006

CZECH J GENET PLANT

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An improved protocol for Brassica protoplast culture and plant regeneration was developed. Isolated protoplasts from four-weeks-old in vitro shoot tip culture of Brassica oleracea var. botrytis cv. Siria F1 and Brassica napus doubled haploid of breeding line OP-1 were cultured at a density of 9.8-11.2 × 10 4 protoplasts/ml in darkness at 25°C in a modified medium containing 2% glucose, 0.25 mg/l 2,4-D, 1 mg/l BAP and 1 mg/l NAA. The first divisions of protoplasts were observed on the third day of culture in B. oleracea and on the fourth day in B. napus. The protoplast cultures were diluted with low osmotic medium on 7 th and 11 th day. The frequency of dividing cells was about 80% in B. oleracea and 50% in B. napus. After one month, the microcalli of approximately 0.5-1 mm in size were transferred into an induction medium with various combinations of growth regulators. Minimum duration of enzyme treatment time and extended dark period in the initial phase of culture increased the survival rate of protoplasts. Organogenesis started when the calli enlarged in size on an induction medium (1 mg/l NAA, 0.02 mg/l GA 3 , 1 mg/l 2iP) with 2% sucrose and 0.8% agar. Regeneration frequency of calli was found to be 69-75% in B. oleracea and 2-3% in B. napus. Well-developed shoots were transferred for rooting to a half-strength MS medium without growth regulators. More than 100 B. oleracea regenerants were transferred into soil, and they produced normal heads and set seeds. This very simple procedure is efficient and suitable mainly for B. oleracea var. botrytis and represents a background for fusion experiments.

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confidence: 99%

A Simple Procedure for Mesophyll Protoplast Culture and Plant Regeneration in Brassica oleracea L. and Brassica napus L.

Kaur¹,

Vyvadilová²,

Klíma³

et al. 2006

CZECH J GENET PLANT

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“…The response of cultured protoplasts to the basic components (macronutrients, micronutrients and vitamins) of the following culture media was examined: Kao [10], K3 [18], A [11], B5 [7], S1 [22], NN [19] and BSK [4]. The basic components were supplemented with 0.38M glucose, NAA (1.0 mg/L), 2,4-D (0.2 mg/L), and BAP (0.5 mg/L), except for medium BSK, whose glucose concentration was reduced to 0.23 M, so that the osmolarity would fall within the range of the other media.…”

Section: Basic Mediummentioning

confidence: 99%

“…These were plated on several regeneration media under bright fluorescent light (70-100/~Em -2 s-t), 16 hour photoperiod and a continuous 25 °C temperature. The regeneration media tested contained Kao [10], Murashige and Skoog [17], B5 [7], K3 [18] or C1 [4] basal components with sucrose (3, 10, 20 or 30 g/L) and several growth regulator ratios of high cytokinin to low auxin.…”

Section: Growth Regulator Combinationsmentioning

confidence: 99%

“…Westar and CMS Westar regenerated shoots at a frequency of 5-20% (% shoots produced from plated calli) on K3 [18] or Murashige and Skoog [17] basal media supplemented with 3 g/L sucrose, 18.2g/L mannitol, 2mg/L zeatin riboside, 0.1 mg/L NAA and 7g/L agarose, pH 5.6. CMS Varuna shoots were recovered at a very low frequency (less than 1%) on C-1 [4] medium supplemented with 10 g/L sucrose, 18.2 g/L mannitol, 2 mg/L zeatin riboside, 0.1 mg/L NAA and 7 g/L agarose, pH 5.6. Cutlass calli have not regenerated shoots yet.…”

Section: Plant Regenerationmentioning

confidence: 99%

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Study of factors affecting the culture of Brassica napus L. and B. juncea Coss. mesophyll protoplasts

Kao

Séguin-Swartz

1987

Plant Cell Tiss Organ Cult

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Various factors affecting the culture of Brassica napus and B. juncea mesophyll protoplasts were examined in order to develop suitable culture media for these species. The basic components (salts and vitamins) of culture media K3 and Kao best supported cell division and colony development in protoplast culture of both species. The addition of casamino acids to Kao's medium resulted in colony browning in B. napus genotypes. B. napus protoplasts grew well with glucose as the osmotic stabilizer, whereas B. juncea protoplasts responded better to sucrose. High NAA and low 2,4-D combinations were effective in stimulating colony growth. Colony development was rapid for a range of genotypes cultured with these recommendations in these media and plant regeneration was obtained from protoplast-derived calli in both species.

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“…Also the use of an agarose support system has been reported for Brassica napus cultures [10] and in other species the use of agarose is a standard technique [11]. Ficoll treatment and agarose have been used in a recent study on plant regeneration from stem cortex protoplasts isolated from B. napus cv.…”

Section: Introductionmentioning

confidence: 99%

Effects of a Ficoll-agarose system on early division and development of mesophyll protoplasts of winter oilseed rape (Brassica napus L.)

Millam

Burns

Hocking

et al. 1988

Plant Cell Tiss Organ Cult

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A method is described for regenerating callus from mesophyll protoplasts of a winter variety of Brassica napus. The method combines the use of Ficoll in an initial liquid medium, enhancing early protoplast division and cell colony formation, with a transfer to an agarose system after 10 days culture to give rapid microcaUi formation. Further transfers resulted in callus regeneration and the initiation of organogenesis.

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Regeneration of plants from mesophyll protoplasts ofBrassica oleracea

Cited by 39 publications

References 3 publications

A Simple Procedure for Mesophyll Protoplast Culture and Plant Regeneration in Brassica oleracea L. and Brassica napus L.

A Simple Procedure for Mesophyll Protoplast Culture and Plant Regeneration in Brassica oleracea L. and Brassica napus L.

Study of factors affecting the culture of Brassica napus L. and B. juncea Coss. mesophyll protoplasts

Effects of a Ficoll-agarose system on early division and development of mesophyll protoplasts of winter oilseed rape (Brassica napus L.)

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