Plant regeneration from protoplasts of Gentiana macrophylla Pall. using agar-pool culture

Hu, Xuemin; Yin, Yongguang; He, Tengbing

doi:10.1007/s11240-014-0705-z

Cited by 9 publications

(12 citation statements)

References 20 publications

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“…In aPL culture, the division frequency reached 39.6%, and the colony formation frequency was 16.9% ( Table 2). The result is similar to the one obtained in G. macrophylla [16]. Hu et al [16] assumed that these differences might be related to the cell browning.…”

Section: Protoplast Culturesupporting

confidence: 85%

“…The result is similar to the one obtained in G. macrophylla [16]. Hu et al [16] assumed that these differences might be related to the cell browning. We found that the cells or cell-groups near the Petri dish walls were more likely to turn brown in liquid and sLD culture systems.…”

Section: Protoplast Culturesupporting

confidence: 85%

“…Agar-pool culture (aPL). Preparation was based on the method described by Hu et al [16]. Briefly, 1.5 mL melted solid medium P1 (Table 1) was poured into a Petri dish.…”

Section: Protoplast Culturementioning

confidence: 99%

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Plant regeneration from protoplasts of Gentiana straminea Maxim

Shi

Yang

2016

Open Life Sciences

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A protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N6-benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

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Section: Protoplast Culturesupporting

confidence: 85%

Section: Protoplast Culturesupporting

confidence: 85%

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Plant regeneration from protoplasts of Gentiana straminea Maxim

Shi

Yang

2016

Open Life Sciences

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“…The physical and chemical conditions used for protoplast release allowed us to obtain yields of G. kurroo protoplasts similar to those of G. decumbens (Tomiczak et al 2015) and G. crassicaulis (Meng et al 1996), whereas the yields of protoplasts for other species was comparable to those that of G. acaulis , G. triflora, and G. triflora 9 G. scabra (Nakano et al 1995). A large number of factors related to plant material used, including plant genotype, age, and preculture conditions, influence the yield of released protoplast (Wright 1985;Grèzes et al 1994;Hu et al 2015). Our study indicates that yield of protoplasts can be also affected by protoplast size, thereby the cell size of source material.…”

Section: Discussionmentioning

confidence: 96%

“…So far, plants of the genus Gentiana have been regenerated only from green leaf mesophyll protoplasts of G. scabra (Takahata and Jomori 1989), G. triflora and G. scabra x G. triflora (Nakano et al 1995), from callus protoplasts of G. crassicaulis (Meng et al 1996) and from cell suspension protoplasts of G. kurroo (Fiuk and Rybczyński 2007;Wójcik and Rybczyński 2015) and G. macrophylla (Hu et al 2015). Although leaves are the most widely accessible plant material for protoplast isolation, cultures derived from leaf mesophyll protoplasts of gentians are usually associated with low frequency of morphogenetic processes.…”

Section: Introductionmentioning

confidence: 99%

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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