Plant regeneration from adventitious roots of spinach (Spinacia oleracea L.) grown from protoplasts

Komai, Fuminori; Masuda, Kiyoshi; Harada, Takashi; Okuse, Ichiro

doi:10.1016/s0168-9452(96)04463-9

Cited by 7 publications

(6 citation statements)

References 21 publications

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“…These morphological differences were probably due to the lower light intensity and higher humidity under in vitro conditions. Similar results were reported by Komai et al (1996), who obtained 1.46 x 10 7 protoplasts/g leaves when spinach (Spinacia oleracea L.) seedlings cultured in vitro were treated from 4 to 10 hours with 2% cellulase R-10 and 0.5% macerozyme R-10. Hu et al (1999) also used the enzymatic solution of 2% cellulase R-10, 1.0% macerozyme, 0.5% driselase, between 15 and 20 hours, to isolate protoplasts from in vitro leaves and hypocotyl of several Brassica species.…”

Section: Hybridssupporting

confidence: 88%

Protoplast production from napier grass and pearl millet triploid hybrids

et al. 2010

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The objective of this work was to obtain protoplasts from napier grass and pearl millet triploid hybrids as a basis for future studies on chromosomal duplication. Explants were taken from mesophyll of in vitro-and in vivo-cultured plants or from calli of two triploid hybrids (H1 and H2), which were treated with enzymatic solutions containing different concentrations of cellulase R-10 (0.5, 1.0, 1.5 and 2.0%) with an additional 0.2% macerozyme and 0.1% driselase or 1.0% pectolyase Y-23 and 0.5% hemicellulase. Enzymatic digestion was monitored once every hour for five hours. Protoplasts were obtained from in vitro and in vivo leaflets of both triploid hybrids, and in vitro leaflets were the best explant sources. The quantity of produced protoplasts varied according to the hybrid, the enzymatic solution and the treatment time.Index terms: Pennisetum purpureum, Pennisetum glaucum, In vitro cultivation, Interspecific hybrids, Forage. RESUMOObjetivou-se, neste trabalho, a obtenção de protoplastos de híbridos triplóides entre o capim-elefante e o milheto como base para futuros trabalhos de duplicação cromossômica. Foram utilizados explantes de mesofilo de plantas cultivadas in vitro e in vivo, ou de calos de dois híbridos triplóides (H1 e H2), os quais foram tratados com soluções enzimáticas em diferentes concentrações da enzima celulase R-10 (0,5; 1,0; 1,5 e 2,0%), acrescidas de 0,2% macerozyme e 0,1% driselase ou 1,0% pectolyase Y-23 e 0,5% hemicelulase. A digestão enzimática foi monitorada a cada hora durante 5 horas. Obtiveram-se protoplastos a partir de folhas in vitro e in vivo dos dois híbridos triplóides, sendo as folhas in vitro as melhores fontes de explante. A quantidade de protoplastos variou em função do híbrido, da solução enzimática e do tempo de tratamento. Termos para indexação:Pennisetum purpureum, Pennisetum glaucum, cultivo in vitro, híbridos interespecíficos, forrageiras.

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Section: Hybridssupporting

confidence: 88%

Protoplast production from napier grass and pearl millet triploid hybrids

et al. 2010

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“…Yet, only a few adventitious buds were obtained that could not be grown into whole plant due to lack of root induction. Since then, spinach is supposed to be a recalcitrant plant. , Later, numerous research articles describing protocols of spinach in vitro regeneration were published, and almost all articles suggested a genotype-dependent response of spinach, which has appeared as a major bottleneck affecting its regeneration capacity. ,,,,− The purpose of this study was to establish an efficient and reproducible in vitro regeneration system for the cultivar “Local Palak”. The explants taken from in vitro germinated plantlets were cultured onto various regeneration media containing a variety of plant hormones.…”

Section: Discussionmentioning

confidence: 99%

“…NAA has a significant positive effect on in vitro regeneration. However, it was found that gibberellic acid (GA 3 ), in combination with auxins like NAA, IAA, and 2,4-D was very effective in inducing embryogenic calli spinach. , The effect of GA 3 has been emphasized in many studies for somatic embryogenesis as well as organogenesis. ,,, GA 3 is famous for encouraging the development of shoot primordium in plants, but its effects depend directly on the cultivar used, ,, the explant selected, concentration of GA 3 added to media, amount and types of other plant growth regulators, and the photoperiod and light intensity. The present study revealed the decisive role of GA 3 concentration on direct regeneration of spinach from petiole explants as described in Section .…”

Section: Discussionmentioning

confidence: 99%

In Vitro Plant Regeneration from Petioles of Spinach (Spinacia oleracea L.)

Hussain,

Ali,

Mustafa

et al. 2023

ACS Agric. Sci. Technol.

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Spinach (Spinacia oleracea L.) is an economically important leafy vegetable crop with an annual global production of 32 million tons. However, it is vulnerable to chewing as well as sucking insects. In vitro plant regeneration can significantly contribute to efforts toward crop improvement. This study is aimed at developing a highly efficient protocol for direct in vitro regeneration of S. oleracea from five different explants (cotyledons, leaves, petioles, hypocotyls, and root segments) and five different regeneration media. Results intimated that only petioles exhibited a response for direct regeneration; hence, they were used in further experiments. More than 35% of the petioles regenerated directly into shoots, which were later separated and cultured onto a rooting medium. All other explants showed variable responses; however, none of them could regenerate in vitro. As far as the role of plant growth regulators is concerned, gibberellic acid (GA 3 ) appeared to be the most imperative one for direct in vitro regeneration and bringing about maximum regeneration in Murashige and Skoog (MS) medium augmented with 2 mg L −1 gibberellic acid (GA 3 ), 0.4 mg L −1 naphthalene acetic acid (NAA), and 1 mg L −1 benzyl amino purine (BAP). The resultant plants were acclimatized in the greenhouse and resulted in healthy, fleshy leaves. For the first time, the protocol describes efficient direct in vitro regeneration from green tissues of S. oleracea. As this is an efficient in vitro regeneration system, it may be a step forward to engineer/edit the genome of this green vegetable for valuable traits including agronomic traits improvement, biofortification, and biopharmaceutical production in the future.

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“…It took almost twenty years to achieve the next success, when a plethora of reports appeared. Briefly, the system of micropropagation from shoot primordia was established [73], while de novo shoot organogenesis was induced from hypocotyls [74][75][76][77][78][79][80], cotyledons [78,[80][81][82], roots [78,83,84], leaves [68,85,86], mature dry seeds [87], and leaf protoplasts [88][89][90][91]. In addition, spinach calli are suitable for the establishment and maintenance of cell suspension cultures [92].…”

Section: Introductionmentioning

confidence: 99%

Somatic Embryogenesis in Spinach—A Review

Zdravković-Korać,

Belić,

Ćalić

et al. 2023

Horticulturae

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A spinach-supplemented diet exerts numerous health benefits, but high levels of oxalic acid and nitrate can cause medical problems, so their levels should be reduced, while the levels of vitamins and phytochemicals could be further increased by breeding. Conventional spinach breeding is limited by the very complex sex determination. However, these limitations could be circumvented in synergy with a biotechnological approach. Accordingly, tissue culture techniques allow rapid and efficient clonal propagation of selected valuable genotypes, and somatic embryogenesis has been recognized as a superior process for clonal propagation because somatic embryos resemble zygotic embryos and therefore can spontaneously develop into complete plants. Since spinach has been considered recalcitrant to in vitro regeneration for decades, a deeper insight into the mechanisms underlying somatic embryogenesis is important for a better understanding and further improvement of the efficiency of this process. In this review, a comprehensive overview of the major factors affecting somatic embryogenesis in spinach is presented and discussed, with particular emphasis on the synergistic effects of α-naphthaleneacetic acid, gibberellic acid, light, and the intrinsic predisposition of individual seedlings to somatic embryogenesis, as well as the expression of genes encoding key enzymes involved in the maintenance of gibberellin homeostasis and the levels of endogenous gibberellins.

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Plant regeneration from adventitious roots of spinach (Spinacia oleracea L.) grown from protoplasts

Cited by 7 publications

References 21 publications

Protoplast production from napier grass and pearl millet triploid hybrids

Protoplast production from napier grass and pearl millet triploid hybrids

In Vitro Plant Regeneration from Petioles of Spinach (Spinacia oleracea L.)

Somatic Embryogenesis in Spinach—A Review

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