Plant regeneration from mesophyll protoplast culture of cabbage (Brassica oleracea var ‘capitata’)

1985 ◽  
Vol 71 (3) ◽  
pp. 495-499 ◽  
Author(s):  
You-ying Fu ◽  
Shi-rong Jia ◽  
Yun Lin
Keyword(s):  
Plant Regeneration ◽  
Brassica Oleracea ◽  
Protoplast Culture ◽  
Mesophyll Protoplast

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

2011 ◽  
Vol 42 (No. 3) ◽  
pp. 103-110 ◽  
Author(s):  
N.D. Kaur ◽  
M. Vyvadilová ◽  
M. Klíma ◽  
M. Bechyně
Keyword(s):  
Brassica Napus ◽  
Plant Regeneration ◽  
Brassica Oleracea ◽  
Growth Regulators ◽  
Protoplast Culture ◽  
Treatment Time ◽  
Simple Procedure ◽  
Enzyme Treatment ◽  
Induction Medium ◽  
Brassica Napus L

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&ndash;11.2 &times; 10<sup>4 </sup>protoplasts/ml in darkness at 25&deg;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<sup>th</sup> and 11<sup>th</sup> 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&ndash;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<sub>3</sub>, 1 mg/l 2iP) with 2% sucrose and 0.8% agar. Regeneration frequency of calli was found to be 69&ndash;75% in B. oleracea and 2&ndash;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. &nbsp;

Isolation, Culture and Plant Regeneration From Protoplasts of Nicotiana debneyi

1980 ◽  
Vol 7 (6) ◽  
pp. 635 ◽  
Author(s):  
WR Scowcroft ◽  
PJ Larkin
Keyword(s):  
Acetic Acid ◽  
Plant Regeneration ◽  
Indole Acetic Acid ◽  
High Efficiency ◽  
Protoplast Culture ◽  
Callus Cultures ◽  
Dna Uptake ◽  
Mesophyll Protoplasts ◽  
Nicotiana Debneyi ◽  
Protoplast Division

Mesophyll protoplasts of two genetically distinct genotypes of N. debneyi were cultured with sustained division following a plating efficiency in excess of 50%. Fully fertile mature plants were regenerated from callus cultures derived from protoplasts. Shoots were induced in medium containing 1 mg/l 6-benzylaminopurine and 0.5 mg/I indole acetic acid. The repeatably high efficiency of protoplast culture was used to evaluate the quantitative effects of two drugs, kanamycin and trimethoprim, which effectively inhibited colony formation at concentrations of 100 and 50 �g/ml, respectively. An enhancer of DNA uptake, poly-L-ornithine, had virtually no effect on sustained protoplast division at a concentration of 7.5 �g/ml or less.

Protoplast culture and plant regeneration from Brassica carinata Braun

1987 ◽  
Vol 6 (1) ◽  
pp. 67-69 ◽  
Author(s):  
Phan V. Chuong ◽  
K. P. Pauls ◽  
W. D. Beversdorf
Keyword(s):  
Plant Regeneration ◽  
Protoplast Culture ◽  
Brassica Carinata ◽  
Brassica Carinata Braun

scholarly journals Production and utilization of doubled haploids in Brassica oleracea vegetables

2011 ◽  
Vol 31 (No. 4) ◽  
pp. 119-123 ◽  
Author(s):  
M. Klíma ◽  
M. Vyvadilová ◽  
V. Kučera
Keyword(s):  
Plant Regeneration ◽  
Brassica Oleracea ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Doubled Haploid Line ◽  
Culture Technique ◽  
F1 Hybrids ◽  
Line Production ◽  
Mature Embryos ◽  
Whole Plants

A possibility to increase the efficiency of plant regeneration from microspore-derived embryos of selected botanical varieties of Brassica oleracea was investigated from 2001 to 2004. More than 400 regenerants of R<sub>1 </sub>generation were derived in kohlrabi, cabbage and cauliflower by means of different modifications of microspore culture technique. Distinct genotype differences in embryogenic responsibility and regenerative ability of microspore embryos to whole plants were detected. The highest frequency of embryogenesis and subsequent regeneration of plants were achieved in cauliflower cultivar Siria F1, kohlrabi line P7 and some experimental F1 hybrids of cauliflower. The best production of embryos was obtained when donor plants were grown in the growth chamber under controlled light and temperature conditions. The regeneration of plantlets was considerably improved by repeated subculture of cotyledonary embryos on media with various combinations of phytohormones and excision of the cotyledons from mature embryos. The percentage of plant regeneration from subcultured embryos in kohlrabi ranged from 11.11 to 63.64%, in cauliflower from 23.53 to 46.19% and in cabbage from 5.88 to 52.00%. The utilization of regenerants for doubled haploid line production is often complicated by male sterility also in plants with the normal diploid chromosome number. &nbsp; &nbsp;

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