Plant Regeneration From Leaf-Derived Calli of Gentians and Their Protoplast Culture

Jomori, Hiroshi; Takahata, Y.; Kaizuma, Norihiko

doi:10.17660/actahortic.1995.392.9

Cited by 14 publications

(19 citation statements)

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“…Browning of the protoplast cultures of G. cruciata and G. septemfida, which was observed within 2 weeks from culture initiation, is thought to be caused by oxidation of phenolic compounds released from plant cells into the culture medium (Zhu et al 1997). This phenomenon was also described by Takahata and Jomori (1989); Jomori et al (1995); Kunitake et al (1995) and Nakano et al (1995), and indicates that it is one of the major problems in protoplast cultures of Gentianaceae. In the case of G. cruciata and G. septemfida, weekly medium refreshment was insufficient to prevent cell browning.…”

Section: Discussionmentioning

confidence: 94%

Comparison of the morphogenic potential of five Gentiana species in leaf mesophyll protoplast culture and ploidy stability of regenerated calli and plants

Tomiczak

Śliwińska

Rybczyński

2016

Plant Cell Tiss Organ Cult

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The morphogenic potential of five Gentiana species of medicinal and ornamental value, i.e.G. cruciata, G. kurroo, G. lutea, G. septemfida, and G. tibetica, was compared using agarose-bead leaf mesophyll protoplast culture. Modified Murashige and Skoog medium containing 2.0 mg l -1 1-naphthaleneacetic acid and 0.1 mg l -1 thidiazuron ensured the highest cell division frequency during both protoplast and protoplast-derived cell culture or callus formation. Indirect plant regeneration mainly via the somatic embryogenesis pathway was achieved for G. kurroo and G. tibetica with the greatest efficiency occurring with MS medium supplemented with 1.0 mg l -1 kinetin, 0.5 mg l -1 gibberellic acid, and 80 mg l -1 adenine sulfate. A considerable percentage of autopolyploid and aneuploid cells was detected in callus lines obtained from protoplasts using flow cytometry. The presence of polyploid cells in calli resulted in the regeneration of 85 % polyploid plants of G. kurroo and 14 % polyploids of G. tibetica. Chromosome counting and stomatal characteristic confirmed the ploidy variation among regenerants.

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Section: Discussionmentioning

confidence: 94%

Comparison of the morphogenic potential of five Gentiana species in leaf mesophyll protoplast culture and ploidy stability of regenerated calli and plants

Tomiczak

Śliwińska

Rybczyński

2016

Plant Cell Tiss Organ Cult

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“…In the next decade, studies concentrated mainly on Japanese ornamental gentian species and cultivars such as G. scabra , G. triflora Pall., and their hybrids (Takahata and Jomori 1989;Jomori et al 1995;Nakano et al 1995) as well as on lisianthus, Eustoma grandiflorum (Griseb.) Schinners (O'Brien and Lindsay 1993; Kunitake et al 1995).…”

Section: Protoplast Culture Of Gentiansmentioning

confidence: 99%

“…As demonstrated earlier, the concentration of ammonium salts in MS medium is too high for protoplast survival and mitotic division (Bajaj 1989). Consequently, modification has been made to MS macronutrient composition, mainly the (Takahata and Jomori 1989;Jomori et al 1995) or its complete withdrawal and replacement with glutamine (Fiuk and Rybczyński 2007;Tomiczak et al 2015). Other media used successfully for gentian protoplast culture include nutrient-rich KM8P medium developed by Kao and Michayluk (1975) for cells and protoplasts cultured at a very low densities (Meng et al 1996), V-KM medium reported by Bokelmann and Roest (1983) for potato protoplasts (O'Brien and Lindsay 1993), and B5 medium of Gamborg et al (1968), as used by Takahata and Jomori (1989) and Nakano et al (1995).…”

Section: Protoplast Culturementioning

confidence: 99%

“…Various approaches of protoplast culture, based on liquid or semisolid media and their combination, have been developed (Davey et al 2005a). The first Gentiana leaf mesophyll protoplast cultures were carried out in simple liquid systems (Takahata and Jomori 1989;Jomori et al 1995), resulting in a low plating efficiency, for example, 0.1 % as reported by Takahata and Jomori (1989) for G. scabra. Taking into consideration the many benefits of embedding protoplasts in semisolid media (Dons and Colijn-Hooymans 1989), Nakano et al (1995) tested 3 different gelling agents for cultures of G. triflora and G. triflora × G. scabra, with 0.2 % gellan gum giving the highest percentage (25.6 %) of divisions of protoplast-derived cells.…”

Section: Protoplast Culturementioning

confidence: 99%

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Protoplast Culture and Somatic Cell Hybridization of Gentians

Tomiczak

Mikuła

Rybczyński

2015

The Gentianaceae - Volume 2: Biotechnology and Applications

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During the last three decades, less than fifteen papers have described the results of scientific investigations in the field of gentian protoplast technology and somatic hybridization. Despite rather limited research already done on this subject, several important goals have been achieved. Protoplast-to-plant systems have been developed either for leading ornamental species or for specific medicinal plants. Two major protoplast sources were evaluated in gentians, namely differentiated leaf mesophyll cells and undifferentiated callus/cell suspensions. Plant regeneration proceeded by the two different pathways of shoot organogenesis or somatic embryogenesis. Some examples of somaclonal variation at the ploidy level were demonstrated within the pool of protoplast-derived regenerants. Totipotency exhibited by gentian protoplasts was exploited to create three different somatic hybrid combinations: intergeneric Swertia mussotii (+) Bupleurum scorzonerifolium, and interspecific Gentiana kurroo (+) G. cruciata and G. cruciata (+) G. tibetica. IntroductionPlant cell totipotency as postulated by Schleiden and Schwann in the middle of the nineteenth century is the basis of modern plant biotechnology (Vasil 2008). Based on the ability of a living single plant cell to dedifferentiate and to convert into other cell types, it is possible to obtain a completely new plant from a cell and even from its protoplast. Consequently, plant regeneration from single protoplasts underlies the genetic manipulation technologies of somatic hybridization and direct genetic transformation by DNA uptake. However, the prerequisite for practical application of these technologies is the development of efficient and reproducible protoplast-to-plant systems for the species of interest.

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“…The following major studies of gentians in vitro culture were performed: 1) description of regeneration potential via organogenesis (Jomori et al 1995, Hosokawa et al 1996, MomEilovi6 et al 1997b, 2) recognition and description of somatic embryogenesis (Mikuta et al 1996b, Mikuta and Rybczyfiski 2001, Mikuta et al 2001, 3) protoplast culture and somatic hybridization to create and improve horticultural value of new hybrid genomes (Nakano et al 1995, Meng et al 1996, 4) secondary metabolite content in regenerants, callus tissue, cell suspension and postcultural media (Yamada et aL 1991), 5) genome manipulation by the transformation using Agrobacterium tumefaciens, A. rhizogenes and particle bombardment to increase horticultural traits and secondary metabolites (MomEilovi6 et al 1997a, Hosokawa et al 1997, 2000, Vinterhalter et al 1999.…”

Section: Introductionmentioning

confidence: 99%

Somatic embryogenesis of Gentiana genus III. Characterization of three-year-old embryogenic suspensions of G. pannonica originated from various seedling explants

2002

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Experiments were carried out with three-year-old embryogenie suspension culture of Gentiana pannonica Scop. The initial explant for the suspension determinated both the embryogenic character and embryo production. Cultures were initiated by culture of hypocotyl, cotyledon and root explants on MS (Mnrashige and Skoog 1962) medium supplemented with 1.0 mg.l-Kinetin and 0.5 mg.l 2,4-D, later transferred and main-1 tained in liquid MS medium with 1.0 mg-l-Dicamba, 0.1 mg-l-I NAA, 2.0 mg-1-1 BAP and 80.0 mg-l-r AS. Regeneration medium included 0.0-1.0 mg.1-1 GA3 + 0.0 -2.0 mg-1-1 Kin. + 0.0-160 mg.1-1 AS.In these culture conditions, the effect of the explant was found to be the most important factor. The curve of growth, growth coefficient and % of participation of various size aggregates differed in the studied suspensions. Flow cytometry revealed various DNA content in nuclei from praembryogenic mass depending on the explant origin. To complete embryogenesis the medium was changed from liquid to solidified in the presence of the same plant growth regulators combination required. The most embryogenic culture appeared hypocotyl-derived and it yielded the highest number of somatic embryos. The suspension culture originating from root proliferated the highest number of embryogenic cell clusters but did not produce embryos for fraction 120-450 ~tm. One hundred mg of suspension of the fraction that was larger than 450 gm yielded 309, 175, 123 embryos for the following suspensions: root-, cotyledon-, hypocotyl-derived, respectively. Almost 50 % of non-deformed fully developed embryos from all studied suspensions passed conversion into germling stage and finally plants were regenerated.

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Plant Regeneration From Leaf-Derived Calli of Gentians and Their Protoplast Culture

Cited by 14 publications

References 0 publications

Comparison of the morphogenic potential of five Gentiana species in leaf mesophyll protoplast culture and ploidy stability of regenerated calli and plants

Comparison of the morphogenic potential of five Gentiana species in leaf mesophyll protoplast culture and ploidy stability of regenerated calli and plants

Protoplast Culture and Somatic Cell Hybridization of Gentians

Somatic embryogenesis of Gentiana genus III. Characterization of three-year-old embryogenic suspensions of G. pannonica originated from various seedling explants

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