Protoplast isolation and plant regeneration of different genotypes of Petunia and Calibrachoa

2009 ◽  
Vol 99 (1) ◽  
pp. 27-34 ◽  
Author(s):  
L. Meyer ◽  
M. Serek ◽  
T. Winkelmann
Keyword(s):  
Plant Regeneration ◽  
Protoplast Isolation

scholarly journals Plant regeneration from protoplast of Brazilian citrus cultivars

2000 ◽  
Vol 35 (4) ◽  
pp. 727-732 ◽  
Author(s):  
FERNANDA JANUZZI MENDES DA GLORIA ◽  
FRANCISCO DE ASSIS ALVES MOURÃO FILHO ◽  
BEATRIZ MADALENA JANUZZI MENDES
Keyword(s):  
Plant Regeneration ◽  
Adventitious Shoots ◽  
Cell Suspension Cultures ◽  
Protoplast Isolation ◽  
Direct Organogenesis ◽  
Malt Extract ◽  
Embryogenic Cell Suspension ◽  
Embryogenic Cell ◽  
Leaf Mesophyll

A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M), CaCl2 (24.5 mM), NaH2PO4 (0.92 mM), MES (6.15 mM), cellulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) and pectolyase Y-23 (Seishin, 0.2%). Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 muM) and malt extract (500 mg/L). Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L), BAP (1.32 muM), NAA (1.07 muM) and coconut water (10 mL/L). Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development.

scholarly journals Isolation and culture of leaf protoplasts from Tunisian grapes

OENO One ◽  
2003 ◽  
Vol 37 (3) ◽  
pp. 145 ◽  
Author(s):  
Ahmed Mliki ◽  
Rahma Jardak ◽  
Götz M. Reustle ◽  
Abdelwahed Ghorbel
Keyword(s):  
Cell Division ◽  
Plant Regeneration ◽  
Aspergillus Niger ◽  
Shoot Culture ◽  
Protoplast Isolation ◽  
Penicillium Funiculosum ◽  
Experimental Conditions ◽  
Grape Varieties ◽  
Leaf Protoplasts

<p style="text-align: justify;">Experimental conditions for leaf protoplast isolation and culture were optimised for <em>in vitro</em> plants deriving from shoot culture of two Tunisian grape varieties, Sakasly and Muscat d’Alexandrie (<em>Vitis vinifera</em> L.). The best yields were obtained from leaves of 4 to 5 weeks old <em>in vitro</em> plants, digested for 13 hours under 25 rpm agitation with an enzymatic mixture containing 0.25 % cellulase of <em>Aspergillus niger</em>, 0.25 % cellulase of <em>Penicillium funiculosum</em>, 0.5 % cellulysin of <em>Trichoderma viridae</em>, and 0.2 % macerozyme R-10 of <em>Rhizopus</em> sp. More than 50 % of the purified protoplasts had a diameter of 30-40 μm and were rich in chloroplasts. Best aptitude for cell division was found in protoplasts immobilised in sodium alginate layers at a density of 0.5x10<sup>6</sup> cell/ml, cultivated in CPW-13 medium containing 4 mg/l of NOA and 0.88 mg/l of TDZ. The variety Muscat d’Alexandrie gave better yield whereas Sakasly showed better cell division rates. Formation of micro and macrocallus have been obtained, but the oxidation of the medium has to be solved in order to promote plant regeneration.</p>

scholarly journals Protoplast isolation and plant regeneration in two cultivated coriander varieties, Co-1 and RS

2018 ◽  
Vol 99 (4) ◽  
pp. 345-355
Author(s):  
Muzamil Ali ◽  
Abdul Mujib ◽  
Nadia Zafar ◽  
Dipti Tonk
Keyword(s):  
Plant Regeneration ◽  
Protoplast Isolation

Protoplast Isolation, Culture, and Plant Regeneration from Passiflora

2003 ◽  
pp. 169-182 ◽  
Author(s):  
Paul Antiony ◽  
Wagner Otoni ◽  
J. Brian Power ◽  
Kenneth C. Lowe ◽  
Michael R. Davey
Keyword(s):  
Plant Regeneration ◽  
Protoplast Isolation

Cotyledon-derived diploid and haploid protoplast culture and diploid plant regeneration in Brassica napus cv. ' Topas '

1998 ◽  
Vol 76 (3) ◽  
pp. 530-541
Author(s):  
M Sun ◽  
H Kieft ◽  
AAM van Lammeren
Keyword(s):  
Cell Division ◽  
Brassica Napus ◽  
Plant Regeneration ◽  
Protoplast Culture ◽  
Shoot Formation ◽  
Protoplast Isolation ◽  
Brassica Napus L ◽  
Successful Isolation ◽  
Young Leaves ◽  
Haploid Embryos

The present paper describes a simple and reliable protocol for the successful isolation, purification, culture, and regeneration of diploid cotyledon-derived protoplasts of Brassica napus L. cv. 'Topas'. Various protoplast isolation media, nutrient media, subculture procedures, and protoplast sources were tested under two culture temperatures. Protoplast viability, cell wall regeneration, and cell division were monitored. Single cotyledon-derived protoplasts formed calli in liquid protoplast medium, and when these were subcultured on solid proliferation medium and solid regeneration medium of appropriate composition, plants regenerated either by shoot formation or embryogenesis. Continuous culture at 32°C instead of 25°C favoured the initiation of cell division and cell proliferation but prevented regeneration, although calli maintained regeneration capacity. Viable haploid protoplasts were isolated from cotyledons of heat-shock-induced, microspore-derived haploid embryos and from young leaves of secondary embryos that were formed on microspore-derived embryos. Cell divisions were triggered in the two types of haploid protoplast cultures, and microcalli were formed at high frequencies. Differences between haploid and diploid protoplast cultures are discussed.Key words: cotyledon protoplast culture, haploid culture, plant regeneration.

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