Mass cloning of Rose and Mussaenda, popular garden plants, via somatic embryogenesis

Protocols were developed for propagation of Rosa hybrida cv. Landora and Mussaenda erythrophylla cv. Rosea via somatic embryogenesis by manipulating growth regulators and culture conditions. Calli were induced from young leaf explants of Rosa hybrida cv. Landora and Mussaenda erythrophylla cv. Rosea on Murashige, Skoog medium supplemented with 6-benzylaminopurine or kinetin along with indole-3-acetic acid or 2,4-dichloroacetic acid within four weeks of culture. The calli were subcultured either in the same medium or in a modified medium for induction of embryogenic callus. Embryogenic calli in rose were developed on Murashige, Skoog medium supplemented with 0.5–1.0 mg/l 6-benzylaminopurine, 2.0 mg/l 2,4-dichloroacetic acid, and 400–800 mg/l l-proline or l-glutamine. The results showed that stimulation of auxin-induced somatic embryogenesis by proline has a great impact on development of somatic embryos and secondary somatic embryogenesis in rose. In Mussaenda, embryogenic calli were developed on Murashige, Skoog medium supplemented with 0.5–1.0 mg/l 6-benzylaminopurine, 2.0–3.0 mg/l indole-3-acetic acid, and 10 mg/l ascorbic acid. Somatic embryos were isolated and transferred to half-strength Murashige, Skoog medium supplemented with 0.25–0.5 mg/l 6-benzylaminopurine + 0.1 mg/l gibberelic acid + 5.0 mg/l adenine sulfate and 2% sucrose for maturation and germination. About 70% somatic embryos of Mussaenda germinated. The rose somatic embryos, however, did not germinate. The somatic embryos of rose, when incubated in the dark at 4°C for two weeks and transferred to 1/2 strength Murashige, Skoog medium supplemented with 0.5 mg/l 6-benzylaminopurine, 0.25 mg/l gibberelic acid, and 2% sucrose, showed 60% germination. The seedlings showed a distinct shoot development but the radicles were blunt without well-defined root system. The shoots were harvested and cultured in the multiplication medium containing Murashige, Skoog medium supplemented with 1.0 mg/l 6-benzylaminopurine and 0.1 mg/l indole-3-acetic acid for four weeks and then subcultured in the same medium for further multiplication. The somatic embryos of Mussaenda erythrophylla cv. Rosea germinated into normal plantlets with distinct shoot and well-developed root system. The somatic embryo-derived plantlets grew normally and flowered within two months of transfer to the field.

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Somatic Embryogenesis and Plant Regeneration via Solid-liquid Alternating Culture in Elite Genotypes of Upland Cotton

10.21203/rs.3.rs-67726/v1 ◽

2020 ◽

Author(s):

Zhengjie Liu ◽

Xinwang Wang ◽

Jinping Hua

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Plant Regeneration ◽

Somatic Embryos ◽

Upland Cotton ◽

Culture System ◽

Embryonic Callus ◽

Solid Liquid ◽

Indole 3 Acetic Acid ◽

Glandless Cotton

Abstract Background: Cotton is one of the most genotype-dependent plants for regeneration, in order to expand cotton regeneration genotypes and establish efficient regeneration system platform, Jiwu 2031 (glandless cotton), ND 58 and CAU 102, were selected for studying the highly efficient somatic embryos formation and regeneration via solid-liquid alternating culture system.Results: In present research, the MSB medium (MS salts adding B5 vitamins) containing 0.571 µM indole-3-acetic acid (IAA), 0.465 µM kinetin (KT) and 0.904 µM 2, 4-dichlorophenoxyacetic acid (2, 4-D) was effective for callus initiation of fourteen Upland cotton with ‘Corker 201’ as the control. ‘Jiwu 2031’, ‘ND 58’ and ‘CAU 102’ could form somatic embryos and regenerate fertile plants in a solid MSB medium containing 10.31 mM NH4NO3, and supplemented with 0.027 mM glycin, 2.460 µM indole-3-acetic acid (IBA), 0.930 µM KT, 3.784 mM asparagine and 6.843 mM glutamine. Under the solid-liquid alternating culture system, the non-embryonic callus was induced to form embryonic callus and the identical status somatic embryos were obtained in 42 days, and the period for plants regeneration was shortened to 90~120 days, with the higher proportion of normal plant regeneration.Conclusions: The solid-liquid alternating culture method could increase the rate of embryogenesis and shorten the period of plants regeneration of Upland cotton. This work provides the evidence that the glandless cotton is beneficial for somatic embryogenesis (SE) and plant regeneration.

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SOMATIC EMBRYOGENESIS OF MUSSAENDA `QUEEN SIRIKIT'

HortScience ◽

10.21273/hortsci.29.4.251f ◽

1994 ◽

Vol 29 (4) ◽

pp. 251f-251 ◽

Cited By ~ 1

Author(s):

Christopher S. Cramer ◽

Mark P. Bridgen

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Somatic Embryo ◽

Somatic Embryos ◽

Basal Medium ◽

Leaf Number ◽

Callus Production ◽

Indole 3 Acetic Acid

Disinfected midrib sections of Mussaenda `Queen Sirikit' ≈3 to 4 mm in size were cultured on a basal medium of Murashige and Skoog salts and vitamins, 87.7 mm sucrose, and 5 g Sigma agar/liter supplemented with several concentrations of indole-3-acetic acid (IAA) (0, 5.0, 10.0, 20.0 μm) and 6-benzylaminopurine (BAP) (0, 0.5, 1.0, 2.5, 5.0, 10.0, 25.0, 50.0 μm). Cultures were subculture onto the same treatment after 5 weeks and observed weekly for 15 weeks for the presence of somatic embryos. As somatic embryos were produced, they were subculture onto basal medium supplemented with 0.5, 1.0, 2.5, or 25.0 μm BAP. Callus was first observed at 2 weeks in cultures grown on basal medium supplemented with 5.0–20.0 μm IAA and 0–50.0 μm BAP. Somatic embryos were observed at 8 weeks on basal medium supplemented with 5.0–10.0 μm IAA and 2.5–5.0 μm BAP. Callus cultured on 0–10 μm IAA and 5.0–10.0 μm BAP produced the greatest number of somatic embryos by 15 weeks. Somatic embryos subculture to basal medium supplemented with 25.0 μm BAP proliferated shoots, while eliminating BAP from the medium resulted in root and callus production. Shoots and entire plants were removed from in vitro conditions and successful] y acclimated to greenhouse conditions. Somatic embryo-derived plants flowered sporadically 25 to 35 weeks after removal from in vitro conditions. Variations in sepal number and leaf number per node were observed at 1% to 5%.

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Somatic Embryogenesis from Roots of Camellia japonica Plantlets Cultured in Vitro

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.116.4.753 ◽

1991 ◽

Vol 116 (4) ◽

pp. 753-757 ◽

Cited By ~ 24

Author(s):

Ana M. Vieitez ◽

Carmen San-José ◽

F. Javier Vieitez ◽

Antonio Ballester

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Dicarboxylic Acid ◽

Butyric Acid ◽

Somatic Embryos ◽

Basal Medium ◽

Camellia Japonica ◽

Secondary Embryos ◽

Indole 3 Acetic Acid

Somatic embryos were induced on the roots of Camellia japonica L. plantlets regenerated from an in vitro clone of juvenile origin. The embryos appeared to differentiate from epidermic cells and to be connected with the root via a few parenchymatous cells. Somatic embryogenesis occurred on basal medium and with or without various combinations of zeatin, BA, and IBA. Secondary embryos were induced on cotyledons and/or hypocotyl regions of somatic embryos. Two morphological types of somatic embryos were developed, seed-like and bud-like types, and their formation was influenced by the presence of BA in the medium. Embryogenic capacity has been maintained for more than 24 months by subculturing secondary embryos at 7- to 8-week intervals. The best gibberellin/auxin combination for inducing the germination of isolated somatic embryos was GA at 5 mg·liter-1 G A3 and IAA at 1 mg·liter-1. P1antlets were successfully established in planting medium and have continued to grow in a greenhouse. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine (BA); (1α, 2β, 4aα, 4bβ, 10β)-2,4a,7-trihydroxy-l-methyl-8-methylenegibb-3-ene-1,10-dicarboxylic acid l,4a-lactone (GA); 1 H -indole-3-acetic acid (IAA); 1 H- indole-3-butyric acid (IBA); 2-methyl-4-(1 H- purine-6-ylamino)-2-buten-l-ol (zeatin).

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The Effects of Exogenous Auxins on Endogenous Indole-3-Acetic Acid Metabolism (The Implications for Carrot Somatic Embryogenesis)

PLANT PHYSIOLOGY ◽

10.1104/pp.112.2.549 ◽

1996 ◽

Vol 112 (2) ◽

pp. 549-558 ◽

Cited By ~ 67

Author(s):

D. M. Ribnicky ◽

N. Ilic ◽

J. D. Cohen ◽

T. J. Cooke

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Acid Metabolism ◽

Indole 3 Acetic Acid

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Can Ethylene Inhibitors Enhance the Success of Olive Somatic Embryogenesis?

Plants ◽

10.3390/plants11020168 ◽

2022 ◽

Vol 11 (2) ◽

pp. 168

Author(s):

Muhammad Ajmal Bashir ◽

Cristian Silvestri ◽

Amelia Salimonti ◽

Eddo Rugini ◽

Valerio Cristofori ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Salicylic Acid ◽

Silver Nitrate ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Cobalt Chloride ◽

Zygotic Embryos ◽

Ethylene Inhibitors ◽

Embryogenic Calli ◽

Stimulatory Action

An efficient in vitro morphogenesis, specifically through somatic embryogenesis, is considered to be a crucial step for the application of modern biotechnological tools for genetic improvement in olive (Olea europaea L.). The effects of different ethylene inhibitors, i.e., cobalt chloride (CoCl2), salicylic acid (SA), and silver nitrate (AgNO3), were reported in the cyclic somatic embryogenesis of olive. Embryogenic callus derived from the olive immature zygotic embryos of the cultivar Leccino, was transferred to the expression ECO medium, supplemented with the ethylene inhibitors at 20 and 40 µM concentrations. Among these, the maximum number of somatic embryos (18.6) was obtained in media containing silver nitrate (40 µM), followed by cobalt chloride (12.2 somatic embryos @ 40 µM) and salicylic acid (40 µM), which produced 8.5 somatic embryos. These compounds interfered on callus traits: white friable embryogenic calli were formed in a medium supplemented with 40 µM cobalt chloride and salicylic acid; in addition, a yellow-compact embryogenic callus appeared at 20 µM of all the tested ethylene inhibitors. The resulting stimulatory action of silver nitrate among all the tested ethylene inhibitors on somatic embryogenesis, clearly demonstrates that our approach can efficiently contribute to the improvement of the current SE protocols for olive.

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Effect of auxin and phenobarbital on the ultrastructure and digitoxin content in Digitalis purpurea tissue culture

Canadian Journal of Botany ◽

10.1139/b96-047 ◽

1996 ◽

Vol 74 (3) ◽

pp. 378-382 ◽

Cited By ~ 5

Author(s):

Mercedes Bonfill ◽

Javier Palazón ◽

Rosa M. Cusidó ◽

M. Teresa Piñol ◽

Carmen Morales

Keyword(s):

Acetic Acid ◽

Volume Fraction ◽

Volume Ratio ◽

Naphthaleneacetic Acid ◽

Acetic Acid Medium ◽

Digitalis Purpurea ◽

Murashige Skoog ◽

Mitochondrial Volume ◽

Callus Tissues ◽

Indole 3 Acetic Acid

Callus derived from Digitalis purpurea hypocotils were grown during a 6-week period on solid Murashige–Skoog medium supplemented with 1 mg/L 6-benzylaminopurine, 0.01 mg/L gibberellic acid and 0.1 mg/L indole-3-acetic acid or α-naphthaleneacetic acid, with or without phenobarbital (40 mg/L). The presence of phenobarbital in the culture medium caused a reduction of the vacuole/cytoplasm ratio. At the same time, the chloroplastic volume fraction decreased in callus tissue cells grown in media supplemented with phenobarbital, while the mitochondrial volume ratio increased. Digitoxin content was enhanced in callus tissues, especially in those grown on indole-3-acetic acid medium supplemented with phenobarbital. The relationship between ultrastructure of D. purpurea callus and digitoxin content is discussed. Keywords: Digitalis purpurea tissue cultures, digitoxin, phenobarbital, mitochondria, chloroplast.

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Immuno-cytochemical localization of indole-3-acetic acid during induction of somatic embryogenesis in cultured sunflower embryos

Planta ◽

10.1007/s00425-002-0791-8 ◽

2002 ◽

Vol 215 (4) ◽

pp. 577-583 ◽

Cited By ~ 51

Author(s):

Cl�ment Thomas ◽

Roberte Bronner ◽

Jean Molinier ◽

Els Prinsen ◽

Harry van Onckelen ◽

...

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Cytochemical Localization ◽

Indole 3 Acetic Acid

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Efficient Somatic Embryogenesis and Plant Regeneration from Immature Embryos of Tapiscia sinensis Oliv., an Endemic and Endangered Species in China

HortScience ◽

10.21273/hortsci.49.12.1558 ◽

2014 ◽

Vol 49 (12) ◽

pp. 1558-1562 ◽

Cited By ~ 4

Author(s):

Yuyu Wang ◽

Faju Chen ◽

Yubing Wang ◽

Xiaoling Li ◽

Hongwei Liang

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Activated Charcoal ◽

Somatic Embryos ◽

Basal Medium ◽

Immature Embryos ◽

Zygotic Embryos ◽

Naphthaleneacetic Acid ◽

Embryogenic Calli ◽

Induction Rate

High-frequency somatic embryogenesis and plant regeneration were achieved from immature cotyledonary-stage embryos in the endangered plant, Tapiscia sinensis Oliv. Plant growth regulators with different concentrations and combinations on embryogenesis capacity were studied. The optimal explants for in vitro somatic embryogenesis were immature embryos in T. sinensis. A high callus induction rate of 100% was achieved on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg·Ll−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5% (w/v) activated charcoal. Alternatively, a high induction rate (96.16%) of somatic embryogenesis was obtained on MS basal medium supplemented with the combination of 0.05 mg·L−1 α-naphthaleneacetic acid (NAA) and 0.2 mg·L−1 6-benzylaminopurine (6-BA), and somatic embryos proliferated fastest on the mentioned medium supplemented with 0.5% (w/v) activated charcoal and 3% (w/v) sucrose, inoculation of explants proliferating 21 times in the 23-day subculture. Of the 100 plantlets transferred to field after the acclimation, 95 (95%) survived. Based on the histocytological observations, the development of somatic embryos was similar to that of zygotic embryos. There were two accumulation peaks of starch grains in the embryogenic calli and in the globular-stage embryos, both closely related to the energy supply, and the embryoids were of multicelluar origin.

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Somatic Embryos Derived from Cotyledons of Cucumber

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.115.4.691 ◽

1990 ◽

Vol 115 (4) ◽

pp. 691-696 ◽

Cited By ~ 12

Author(s):

Rebecca M. Cade ◽

Todd C. Wehner ◽

Frank A. Blazich

Keyword(s):

Tissue Culture ◽

Acetic Acid ◽

Somatic Embryogenesis ◽

Somatic Embryos ◽

Culture Media ◽

Naphthaleneacetic Acid ◽

Histological Evidence ◽

Tissue Culture Media ◽

Maturation Medium ◽

Dichlorophenoxy Acetic Acid

Two studies were conducted to test the effects of various tissue culture media on somatic embryogenesis from cotyledon tissue of cucumber (Cucumis sativus L.). The two best media for embryo initiation were Murashige and Skoog (MS) salts and vitamins containing either 1 or 2 mg 2,4-D/liter and 0.5 mg kinetin/liter. In the second study, embryos developed more normally. More plantlets developed when tissue was removed from the initiation medium after 3 weeks and transferred to MS containing 1 mg NAA/liter and 0.5 mg kinetin/liter for 3 weeks, rather than leaving the embryos on a medium containing 2,4-D. Histological evidence indicated that the embryos were multicellular in origin. Charcoal in the maturation medium inhibited embryo development. Chemical names used: (2,4-dichlorophenoxy) -acetic acid (2,4-D); N-(2-furanylmethyl)-lH-purine-6-amine (kinetin); 1-naphthaleneacetic acid (NAA).

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