Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose

Kang, Hyun Hee; Naing, Aung Htay; Kim, Chang Kil

doi:10.3390/biology9080228

Cited by 22 publications

(16 citation statements)

References 31 publications

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“…The nature of the explant tissue and the thickness of the cell wall play an important role in high-efficiency protoplast isolation, which is a critical stage in the process of seedling regeneration or somatic hybridization. However, protoplasts were successfully isolated and cultured in different ornamentals, such as Dendrobium [ 70 ], lily [ 71 ], rose [ 72 ], chrysanthemum [ 73 ], petunia [ 74 ], carnation [ 75 ], coneflower [ 76 ], geraniums [ 77 , 78 ], Persian silk tree [ 79 ], etc. Pre-plasmolyzing the explant tissue with osmotic stabilizers, such as mannitol and sorbitol, before enzyme treatment is effective for protoplast isolation in most plant tissue [ 80 ].…”

Section: Standard Techniques Involved In Plantlet Generation In Vitromentioning

confidence: 99%

Tissue Culture in Ornamentals: Cultivation Factors, Propagation Techniques, and Its Application

Mumivand

Akter

et al. 2022

Plants

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Ornamentals come in a variety of shapes, sizes, and colors to suit a wide range of climates, landscapes, and gardening needs. Compared to demand, a shortage of plant materials and diversity force the search for solutions for their constant acquisition and improvement to increase their commercial value, respectively. In vitro cultures are a suitable solution to meet expectations using callus culture, somatic embryogenesis, protoplast culture, and the organogenesis of protocorm-like bodies; many of these techniques are commercially practiced. Factors such as culture media, explants, carbohydrates, plant growth regulators, and light are associated with the success of in vitro propagation. Techniques, especially embryo rescue and somatic hybridization, are widely used to improve ornamentals. The development of synthetic seed allows season-independent seed production and preservation in the long term. Despite the advantages of propagation and the improvement of ornamentals, many barriers still need to be resolved. In contrast to propagation and crop developmental studies, there is also a high scope for molecular studies, especially epigenetic changes caused by plant tissue culture of ornamentals. In this review, we have accumulated and discussed an overall update on cultivation factors, propagation techniques in ornamental plant tissue culture, in vitro plant improvement techniques, and future perspectives.

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Section: Standard Techniques Involved In Plantlet Generation In Vitromentioning

confidence: 99%

Tissue Culture in Ornamentals: Cultivation Factors, Propagation Techniques, and Its Application

Mumivand

Akter

et al. 2022

Plants

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“…The protoplast plating density can range from single cells up to a few million protoplasts per milliliter, but typically range from 5 × 10 4 –1 × 10 6 protoplasts/ml ( Table 2 ). In a comparison of plating densities of petunia ( Petunia hybrida ) leaf protoplast culture, 1 × 10 6 protoplasts/ml produced a significantly higher division frequency and number of calli than 5 × 10 4 protoplasts/ml ( Kang et al, 2020 ). However, the microcolony viability decreased with the plating density increasing to 1.5 × 10 6 protoplasts/ml, potentially due to high phenolics accumulation.…”

Section: Protoplast Culturementioning

confidence: 99%

Protoplast Regeneration and Its Use in New Plant Breeding Technologies

Reed

Bargmann

2021

Front. Genome Ed.

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The development of gene-editing technology holds tremendous potential for accelerating crop trait improvement to help us address the need to feed a growing global population. However, the delivery and access of gene-editing tools to the host genome and subsequent recovery of successfully edited plants form significant bottlenecks in the application of new plant breeding technologies. Moreover, the methods most suited to achieve a desired outcome vary substantially, depending on species' genotype and the targeted genetic changes. Hence, it is of importance to develop and improve multiple strategies for delivery and regeneration in order to be able to approach each application from various angles. The use of transient transformation and regeneration of plant protoplasts is one such strategy that carries unique advantages and challenges. Here, we will discuss the use of protoplast regeneration in the application of new plant breeding technologies and review pertinent literature on successful protoplast regeneration.

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“…Plant somatic cell fusion through cell wall separation (enzymolysis) and passivation includes a series of processes, and every step is likely to damage cells [ 10 , 11 ]. The influence of chemical factors on the regeneration ability of heterozygous cells is often the result of environmental conditions and gene interaction.…”

Section: Introductionmentioning

confidence: 99%

Transcriptome Analysis of Sugarcane Young Leaves and Protoplasts after Enzymatic Digestion

Zhang

Wang

Han

et al. 2022

Life

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Sugarcane somatic cell hybridization can break through the barrier of genetic incompatibility between distantly related species in traditional breeding. However, the molecular mechanisms of sugarcane protoplast regeneration and the conditions for protoplast preparation remain largely unknown. In this study, young sugarcane (ROC22) leaves were enzymatically digested, and the viability of protoplasts reached more than 90% after enzymatic digestion (Enzymatic combination: 2% cellulase + 0.5% pectinase + 0.1% dissociative enzyme + 0.3% hemicellulase, pH = 5.8). Transcriptome sequencing was performed on young sugarcane leaves and protoplasts after enzymatic digestion to analyze the differences in gene expression in somatic cells before and after enzymatic digestion. A total of 117,411 unigenes and 43,460 differentially expressed genes were obtained, of which 21,123 were up-regulated and 22,337 down-regulated. The GO terms for the 43,460 differentially expressed genes (DEGs) were classified into three main categories: biological process, cellular component and molecular function, which related to developmental process, growth, cell proliferation, transcription regulator activity, signal transducer activity, antioxidant activity, oxidative stress, kinase activity, cell cycle, cell differentiation, plant hormone signal transduction, and so on. After enzymatic digestion of young sugarcane leaves, the expressions of GAUT, CESA, PSK, CyclinB, CyclinA, CyclinD3 and cdc2 genes associated with plant regeneration were significantly down-regulated to 65%, 47%, 2%, 18.60%, 21.32%, 52% and 45% of young leaves, respectively. After enzymatic digestion, Aux/IAA expression was up-regulated compared with young leaves, and Aux/IAA expression was 3.53 times higher than that of young leaves. Compared with young leaves, these key genes were significantly changed after enzymatic digestion. These results indicate that the process of somatic enzymatic digestion process may affect the regeneration of heterozygous cells to a certain extent.

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Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose

Cited by 22 publications

References 31 publications

Tissue Culture in Ornamentals: Cultivation Factors, Propagation Techniques, and Its Application

Tissue Culture in Ornamentals: Cultivation Factors, Propagation Techniques, and Its Application

Protoplast Regeneration and Its Use in New Plant Breeding Technologies

Transcriptome Analysis of Sugarcane Young Leaves and Protoplasts after Enzymatic Digestion

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