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

Kaur, Navtej; Vyvadilová, M.; Klíma, Miroslav; Bechynĕ, M

doi:10.17221/3649-cjgpb

Cited by 24 publications

(12 citation statements)

References 32 publications

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“…The timing of the first mitotic division is generally expected between 24 and 48 h of culture (Glimelius 1984 ; Mukhopadhyay et al 1991 ), but a shift of the first mitosis to days 3 or 4 of culture, as observed in our study, was also reported for B. oleracea (Fu et al 1985 ; Robertson and Earle 1986 ; Kaur et al 2006 ). Division frequencies in B. oleracea varied as a consequence of the source of tissue, culture system, culture maintenance, and genotype.…”

Section: Discussionsupporting

confidence: 83%

“…The use of some types of tissues (such as roots and cortex) leads to a higher risk of contamination, while others (such as hypocotyls and cotyledons) required the use of large amount of seeds to obtain satisfactory yields, and others (such as suspension cultures) are difficult to maintain for Brassica species (Simmonds et al 1991 ). In B. oleracea , protoplasts have previously been isolated mainly from leaves (Fu et al 1985 ; Robertson and Earle 1986 ; Kaur et al 2006 ) and hypocotyls (Glimelius 1984 ; Mukhopadhyay et al 1991 ; Chen et al 2004 ). In this study, both of these sources, taken from sterile conditions, were utilized and protoplasts were readily isolated for all tested accessions.…”

Section: Discussionmentioning

confidence: 99%

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An alginate-layer technique for culture of Brassica oleracea L. protoplasts

Kiełkowska

Adamus

2012

In Vitro Cell.Dev.Biol.-Plant

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Ten accessions belonging to the Brassica oleracea subspecies alba and rubra, and to B. oleracea var. sabauda were used in this study. Protoplasts were isolated from leaves and hypocotyls of in vitro grown plants. The influence of selected factors on the yield, viability, and mitotic activity of protoplasts immobilized in calcium alginate layers was investigated. The efficiency of protoplast isolation from hypocotyls was lower (0.7 ± 0.1 × 106 ml−1) than for protoplasts isolated from leaf mesophyll tissue (2 ± 0.1 × 106 ml−1). High (70–90%) viabilities of immobilized protoplasts were recorded, independent of the explant sources. The highest proportion of protoplasts undergoing divisions was noted for cv. Reball F1, both from mesophyll (29.8 ± 2.2%) and hypocotyl (17.5 ± 0.3%) tissues. Developed colonies of callus tissue were subjected to regeneration and as a result plants from six accessions were obtained.

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Section: Discussionsupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

An alginate-layer technique for culture of Brassica oleracea L. protoplasts

Kiełkowska

Adamus

2012

In Vitro Cell.Dev.Biol.-Plant

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“…Before and after the cold treatment, the uppermost fully developed leaf was harvested from each plant and used to assess proline and dehydrin contents. Protoplast-derived calli were prepared following Kaur et al (2006). When the calli had reached a diameter of 5 -8 mm, they were divided into two groups of 20 calli.…”

Section: ⎯⎯⎯⎯mentioning

confidence: 99%

Dehydrin and proline content in Brassica napus and B. carinata under cold stress at two irradiances

et al. 2012

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The accumulation of cold-induced dehydrin and proline was related to the frost tolerance (FT) in several Brassica species or cultivars. A dehydrin of molecular mass 47 kDa was detected in the leaves of an Ethiopian mustard (B. carinata) and a pair of dehydrins of similar molecular mass in the three (two winter, one spring) oilseed rape (B. napus) cultivars, when plants were maintained at 4 °C for one-month under two different irradiances. More dehydrin was accumulated in oilseed rape than in Ethiopian mustard under the high irradiance. A significant correlation was observed between leaf dehydrin content and FT, and no relationship between proline content and FT or between the proline and dehydrin contents. Protoplast-derived callus cells behaved differently from leaves sampled from intact plants, as they did not accumulate dehydrin and proline in response to cold stress.Additional key words: Ethiopian mustard, frost tolerance, oilseed rape, protein marker, protoplast-derived callus. ⎯⎯⎯⎯

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“…Light-stimulated H + extrusion was also reported by Petzold and Dahse (1988) from leaf discs of Vicia faba L. The H + extrusion may accelerate the browning process. Kaur et al (2006) believed that under illumination, the cultures turn brown, thus affecting the division efficiency of protoplasts of Brassica oleracea L. and Brassica napus L. It is possible that the accelerated browning quickly kills cells of leaf explants of 'Fire Flash' and provides little opportunity for cells to respond to the induction. Although more leaf explants produced callus in the dark culture, the induced calluses still died after culture under lighted conditions.…”

Section: Discussionmentioning

confidence: 99%

Regeneration of Chlorophytum amaniense ‘Fire Flash’ Through Indirect Shoot Organogenesis

Chen¹,

Liu²,

Chen³

et al. 2011

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Chlorophytum amaniense Engl. ‘Fire Flash’ is a popular exotic ornamental foliage plant as a result of its unique coral-colored midribs and petioles and tolerance to interior low light levels. Currently, demand for propagative materials exceeds the availability of seeds. This study was intended to develop an in vitro culture method for rapid propagation of this cultivar. Leaf and sprouted seed explants were cultured on a Murashige and Skoog basal medium supplemented with different cytokinins with 1.1 μM α-naphthalene acetic acid (NAA) or 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Leaf explants showed poor responses in callus production and no adventitious shoots were obtained. Callus formation frequencies from sprouted seeds were 71% and 85% when induced by 9.8 μM N6-(2-isopentyl) adenine (2iP) with 1.1 μM NAA and 9.1 μM N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with 1.1 μM NAA, respectively. Adventitious shoots occurred after the induced calluses were subcultured on the same concentrations of TDZ or 2iP with NAA. Shoot formation frequencies from calluses cultured on TDZ with NAA and 2iP with NAA were 92% and 85%, and the corresponding mean shoot numbers were 37 and 31 per piece of callus (1 cm3), respectively. Adventitious shoots rooted at 100% after transferring to the basal medium containing 4.4 μM 6-benzylaminopurine (BA) with 2.7 μM NAA. Plantlets, after transplanting to a soilless substrate were easily acclimatized in a shaded greenhouse under a photosynthetic photon flux (PPF) density of 200 μmol·m−2·s−1. Regenerated plants grew vigorously without undesirable basal branching or distorted leaves. This newly established regeneration method can provide the foliage plant industry with a means for rapidly propagating ‘Fire Flash’ liners in a year-round fashion.

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

Cited by 24 publications

References 32 publications

An alginate-layer technique for culture of Brassica oleracea L. protoplasts

An alginate-layer technique for culture of Brassica oleracea L. protoplasts

Dehydrin and proline content in Brassica napus and B. carinata under cold stress at two irradiances

Regeneration of Chlorophytum amaniense ‘Fire Flash’ Through Indirect Shoot Organogenesis

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