Applications of In Vitro Techniques in Plant Breeding

Ichwan, Budiyati; Tapingkae, Tanya; Taji, Acram

doi:10.1007/978-3-319-22521-0_10

Cited by 18 publications

(17 citation statements)

References 169 publications

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“…Protoplast-based transient expression system (PTES) has been proven to be a powerful experimental tool in molecular biology. It is widely used for protein localization, protein-protein interactions, identification of the gene function and gene regulation [14,34,44,[46][47][48][49]. Transient protoplast transfection has been used due to its advantages in high screening efficiency with gene silencing and genome editing technologies, prior to the development of transgenic plants [43,[50][51][52].…”

Section: Introductionmentioning

confidence: 99%

Highly Efficient Protoplast Isolation and Transient Expression System for Functional Characterization of Flowering Related Genes in Cymbidium Orchids

Ren

Gao

et al. 2020

IJMS

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Protoplast systems have been proven powerful tools in modern plant biology. However, successful preparation of abundant viable protoplasts remains a challenge for Cymbidium orchids. Herein, we established an efficient protoplast isolation protocol from orchid petals through optimization of enzymatic conditions. It requires optimal D-mannitol concentration (0.5 M), enzyme concentration (1.2 % (w/v) cellulose and 0.6 % (w/v) macerozyme) and digestion time (6 h). With this protocol, the highest yield (3.50 × 107/g fresh weight of orchid tissue) and viability (94.21%) of protoplasts were obtained from flower petals of Cymbidium. In addition, we achieved high transfection efficiency (80%) through the optimization of factors affecting polyethylene glycol (PEG)-mediated protoplast transfection including incubation time, final PEG4000 concentration and plasmid DNA amount. This highly efficient protoplast-based transient expression system (PTES) was further used for protein subcellular localization, bimolecular fluorescence complementation (BiFC) assay and gene regulation studies of flowering related genes in Cymbidium orchids. Taken together, our protoplast isolation and transfection protocol is highly efficient, stable and time-saving. It can be used for gene function and molecular analyses in orchids and other economically important monocot crops.

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Section: Introductionmentioning

confidence: 99%

Highly Efficient Protoplast Isolation and Transient Expression System for Functional Characterization of Flowering Related Genes in Cymbidium Orchids

Ren

Gao

et al. 2020

IJMS

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“…The presence of auxin, either alone or in combination with a low concentration of cytokinin is a critical factor for the proliferation of root primordia on cultured explants (Zulkarnain, 2009). This is probably due to their participation in the regulation of cell cycling, division, and differentiation (Tapingkae et al, 2012, Zulkarnain et al, 2015.…”

Section: Resultsmentioning

confidence: 99%

Plantlets Regeneration from Crown Bud Slicing of Pineapple (Ananas comosus)

2018

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Pineapple propagation by lateral shoots, suckers or crowns is often confronted with limited number of regenerated seedlings and high diversity in flowering and fruit formation. In order to solve this problem, this study offer an alternative method by using tissue culture techniques. This study aimed to determine the effect of growth regulators on plantlet regeneration from bud slicing of pineapple cv. Tangkit. Four levels of 2.4-D (0.0, 0.001, 0.01 and 0.1 ppm) in combination with BA (0.0, 0.1, 1.0 and 10.0 ppm) were tested on solid MS medium. Cultures were incubated in total darkness for a week followed by transfer to 16-hour photoperiod. Results showed that explants treated with 2,4-D and/or BA succeeded in regenerating adventitious shoots. Average leaf number did not differ significantly among treatments (P = 0.60). Highest leaf number (2.99 ± 0.23) was obtained on medium with 0.01 ppm 2,4-D without BA, followed by 0.1 ppm 2,4-D without BA (2.85 ± 0.33). Meanwhile, roots were only formed on medium with 0.1 ppm 2.4-D without BA (4.2 ± 0.37 per shoot). Thus, complete plantlets were regenerated only on medium supplemented with 0.1 ppm 2,4-D without BA. The growth of plantlets was relatively uniform, and plantlet acclimatization succeeded 100% on Jiffy pots. The finding of optimum concentration of 2.4-D and BA in this study is important to develop standard protocol for in vitro propagation of pineapple cv. Tangkit. Thus, the benefit of producing seeds in large quantities and relatively uniform in growth is made possible through tissue culture technique.

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“…The link between flowering in rose and ethylene was discussed, and two main hypotheses could be formulated with actual experiment, the hypothesis of inhibition of flowering induction by ethylene in rose and the importance of ethylene for flower petal development. In vitro flowering also has physiological interest, helping to understand the mechanism of rose flowering, but could also be used for breeding purposes, using early in vitro flowering for techniques such as in vitro pollination and fertilization (Zulkarnain et al, 2015).…”

Section: Agno 3 Influenced Induction and Development Of In Vitro Flowmentioning

confidence: 99%

AgNO3 improved micropropagation and stimulate in vitro flowering of rose (Rosa x hybrida) cv. Sena

Matos

Oliveira

et al. 2021

Ornam. Hortic.

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Rose is one of the most important cut flower in the world. Rose micropropagation was used for production of clonal and disease-free plantlets and to breeding purposes. However, many important rose cultivars showed physiological disorders as early-leaf senescence and very low multiplication rate under in vitro conditions. Our hypothesis is that these symptoms were associated with high sensibility of these cultivars to ethylene accumulation on in vitro environment. The rose cv. Sena was in vitro cultivated under different concentrations of AgNO3 and two light sources, LED and fluorescent lamps, as a way to investigate in vitro similar symptoms to ethylene accumulation. AgNO3 at 1.0-2.0 mg L-1 solved the main in vitro physiological disorders observed in this rose cultivar. Also, AgNO3 stimulated induction of 50% of rose shoots to in vitro flowering at 2.0 mg L-1. Higher concentrations also resulted in flowering induction, but with imperfect flower development.

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Applications of In Vitro Techniques in Plant Breeding

Cited by 18 publications

References 169 publications

Highly Efficient Protoplast Isolation and Transient Expression System for Functional Characterization of Flowering Related Genes in Cymbidium Orchids

Highly Efficient Protoplast Isolation and Transient Expression System for Functional Characterization of Flowering Related Genes in Cymbidium Orchids

Plantlets Regeneration from Crown Bud Slicing of Pineapple (Ananas comosus)

AgNO3 improved micropropagation and stimulate in vitro flowering of rose (Rosa x hybrida) cv. Sena

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