Somatic Embryogenesis in Muscadine Grape

Low frequency of in vitro regeneration has hampered the adoption of genetic engineering technique for improving the quality of muscadine grape. This study is to develop a straightforward method for high-frequency regeneration of muscadine grapes in vitro. Leaves, petioles, and immature ovules of muscadine grapes were cultured on various media. Embryogenic callus, somatic embryos were formed after 9 weeks inoculated on embryo rescue (ER) medium. The somatic embryos were isolated and subcultured on fresh medium to promote enlargement and increase the number of uniformly sized somatic embryos. Of the medium tested (MS, NN, and ER), the ER medium was the best for somatic embryo growth and multiplication. The somatic embryogenic lines were maintained by transferring the embryos to the fresh ER medium every 4 weeks. Germination was achieved by transferring these embryos to woody plant medium or NN medium. The frequency of somatic embryogenesis of embryo germination appeared to be genotype dependent. The establishment of the somatic embryogenesis system in this study should be a step forward in directly transferring a foreign gene into muscadine grape.

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In vitro regeneration and somatic embryogenesis in citrus

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v24i2.23558 ◽

2015 ◽

Vol 24 (2) ◽

pp. 247-262 ◽

Cited By ~ 1

Author(s):

El Sawy A Mohamed ◽

Amina Gomaa ◽

Nancy Danial

Keyword(s):

Somatic Embryogenesis ◽

Genetic Stability ◽

Somatic Embryos ◽

Rapd Analysis ◽

In Vitro Regeneration ◽

Rt Pcr ◽

Mother Plants

Better results were obtained when stigma explants of variegated lemon and citron were used. After ten months, somatic embryos developed into plantlets at a frequency ranged from 13.3 for lime to 66.7% for lemon. Virus presence was tested by ELISA and RT?PCR. The results indicated that the plantlets regenerated through somatic embryogenesis are CTV?free. RAPD analysis was used to asses the genetic stability of plantlets as compared to the mother plants. The results indicated that most plantlets belong to the respective mother plants and the polymorphism percentage was genotype and explant?dependant.Plant Tissue Cult. & Biotech. 24(2): 247-262, 2014 (December

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THIDIAZURON: A POTENT GROWTH REGULATOR FOR INDUCING HIGH-FREQUENCY ORGANOGENESIS AND SOMATIC EMBRYOGENESIS IN VITRO

HortScience ◽

10.21273/hortsci.27.6.693a ◽

1992 ◽

Vol 27 (6) ◽

pp. 693a-693

Author(s):

K.A. Malik ◽

Christena Visser ◽

praveen K. saxena

Keyword(s):

Somatic Embryogenesis ◽

High Frequency ◽

Somatic Embryos ◽

Direct Evidence ◽

Shoot Organogenesis ◽

In Vitro Regeneration ◽

Endogenous Phytohormones ◽

Potent Growth

In vitro regeneration by shoot organogenesis and-or somatic embryogenesis is accomplished by culturing the explants on a nutrient medium supplemented with phytohormones. Auxins in general, and 2,4-D in particular, have been shown to induce somatic embryogenesis whereas shoot regeneration is stimulated by cytokinins. In studying the morphoregulatory role of thidiazuron (TDZ) - a substituted urea with cytokinin-like activity - we found that it induces a high frequency of both organogenesis and somatic embryogenesis depending upon the plant species. For instance, whole seedlings of peanut developed somatic embryos and those of bean and pea produced shoots in response to culture on TDZ (1-40 μM)-supplemented media. In cultured explants of geranium, the use of TDZ (0.2-1 μM) effectively replaced the requirement of 2,4-D or BAP and IAA for obtaining somatic embryos. The frequency of regeneration was two to ten times higher than that achieved with auxin-cytokinin combinations. While no direct evidence is currently available to establish a relationship between TDZ and endogenous phytohormones, our results suggest that it may act by establishing endogenously the auxin:cytokinin ratio permissive of induction and expression of morphogenically competent cells.

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Plant Regeneration in Buckwheat (Fagopyrum esculentum Moench.) via Somatic Embryogenesis and Induction of Meristemoids in Abnormal Embryos

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v29i1.41977 ◽

2019 ◽

Vol 29 (1) ◽

pp. 33-47 ◽

Cited By ~ 1

Author(s):

Ribha Saraswat ◽

Mithilesh Kumar

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Shoot Organogenesis ◽

In Vitro Regeneration ◽

Hypocotyl Explants ◽

Regenerated Plants ◽

Half Strength ◽

Fagopyrum Esculentum Moench ◽

Full Utilization

An efficient in vitro regeneration protocol is reported for common buckwheat. A combination of 0.5 mg/l 2,4-D and 0.2 mg/l BAP with sucrose showed highest induction of somatic embryogenesis from cotyledon and hypocotyl explants. More than 35% of normal somatic embryos matured on MS. MS with 2% sucrose were found best for germination and conversion of somatic embryos to plantlets. In tissue culture, abnormal somatic embryos usually occur. In this report, abnormal embryos are also used to induce shoot organogenesis, adding to the number of final regenerants and ensuring full utilization of regenerative propagules. A treatment of 0.2 mg/l BAP induced meristemoids in 60% of underdeveloped embryos and a combination of 0.5 mg/l BAP and 0.5 mg/l AgNO3 led browning and senescence-free progression of shoot buds to well developed shoots, which were subsequently rooted in half strength MS containing 2% sucrose and 0.25 mg/l IBA. The regenerated plants survived acclimatization, flowered and set seeds. Plant Tissue Cult. & Biotech. 29(1): 33-47, 2019 (June)

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Somatic Embryogenesis and Plant Regeneration from Commercial Soybean Cultivars

Plants ◽

10.3390/plants9010038 ◽

2019 ◽

Vol 9 (1) ◽

pp. 38 ◽

Cited By ~ 3

Author(s):

Ghulam Raza ◽

Mohan B. Singh ◽

Prem L. Bhalla

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Somatic Embryo ◽

Somatic Embryos ◽

In Vitro Regeneration ◽

Crop Improvement ◽

Soybean Cultivars ◽

Immature Cotyledons ◽

Soybean Genotypes

The efficient regeneration of plants from commercial genotypes is a pre-requisite for successful genetic transformation, to apply modern crop improvement techniques such as CRISPR-based genome editing. Plant regeneration through the somatic embryogenesis pathway offers an advantage over the organogenesis approach, avoiding the risk of developing chimeras. Plant genotype, explant type, and media compositions play an essential role in the in-vitro regeneration of plants. This study aimed to characterize the commercially grown Australian soybean genotypes for their potential to induce somatic embryos, embryo proliferation, maturation, germination, and plant regeneration. Overall, nine soybean cultivars belonging to different maturity groups were evaluated. Immature cotyledon ranging from 2–4 and 4–6 mm in size were used as explants for somatic embryogenesis induction. Maximum somatic embryo induction frequency (86%) was observed from 4–6 mm immature cotyledons of the cv. Jack (MG III), followed by 66%, 26%, 21%, and 6% in cultivars Williams (MG III), Snowy (MG III), MoonB1 (MG V), and PNR791 (MG V), respectively. On the other hand, cv. Snowy showed maximum somatic-embryo-inducing potential (67%) in 2–4 mm immature cotyledons followed by Williams, Jack, MoonB1, and PNR791. Somatic embryos from Jack, Williams, and Snowy cultivars were further tested for embryo proliferation, maturation, and germination. Maximum proliferation and maturation were observed in cv. Jack, followed by Snowy and Williams. However, cv. Snowy showed a significantly higher conversion of cotyledonary stage embryos to plantlets (85%), than both Jack and Williams cultivars (53% each). In conclusion, this study outlined a protocol for somatic embryogenesis and plant regeneration from three soybean cultivars. Our findings suggest commercial cv. Snowy could be a good candidate for developing transgenic plants through somatic embryogenesis.

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Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.4990 ◽

1970 ◽

Vol 19 (1) ◽

pp. 89-99

Author(s):

K. Choudhary ◽

M. Singh ◽

M. S. Rathore ◽

N. S. Shekhawat

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Somatic Embryos ◽

Basal Medium ◽

Long Term Study ◽

Continuous Application ◽

First Time ◽

Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l of 2,4-D and 0.5 mg/l of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

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A temporary immersion system improves in vitro regeneration of peach palm through secondary somatic embryogenesis

Annals of Botany ◽

10.1093/aob/mcr033 ◽

2011 ◽

Vol 108 (8) ◽

pp. 1463-1475 ◽

Cited By ~ 56

Author(s):

D. A. Steinmacher ◽

M. P. Guerra ◽

K. Saare-Surminski ◽

R. Lieberei

Keyword(s):

Somatic Embryogenesis ◽

In Vitro Regeneration ◽

Secondary Somatic Embryogenesis ◽

Temporary Immersion System ◽

Temporary Immersion ◽

Peach Palm

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Agrobacterium-mediated transformation of the wild orchid Cattleya maxima Lindl

Universitas Scientiarum ◽

10.11144/javeriana.sc23-1.amto ◽

2018 ◽

Vol 23 (1) ◽

pp. 89

Author(s):

Augusta Yadira Cueva-Agila ◽

Rino Cella

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Attractive Alternative ◽

Gene Encoding ◽

Anatomical Structures ◽

Large Numbers ◽

Hygromycin Resistance Gene ◽

Growing Conditions ◽

Somatic Embryogenesis Receptor Kinase

Protocorms are unique anatomical structures; they are akin to rhizoids and are formed by young orchid seedlings under physiological conditions. Explanted orchid tissues produce similar structures called protocorm-like bodies (PLBs) when exposed to appropriate in vitro growing conditions. Both the propagative nature of PLBs and the easiness by which they can be generated, make these structures an attractive alternative to seed-mediated production for growing large numbers of plants. To increase somatic embryogenesis and optimize the procedure, PLBs of Cattleya maxima were transformed using the Agrobacterium tumefaciens method. The T-DNA carried a Hygromycin-resistance gene, a visible marker (GFP5-GUSA) and a rice gene encoding the Somatic Embryogenesis Receptor Kinase, deemed to be important for somatic embryogenesis. Treated PLBs generated somatic embryos developing Hygromycin-resistant plantlets. The insertion of T-DNA was confirmed by PCR, and GFP expression was observed using a fluorescent stereomicroscope. Transformed Cattleya maxima PLBs were more efficient in forming somatic embryos (60-80%) than untransformed controls (45-57%), and this contrast was maximized in hormone-free, Murashige and Skoog (MS) medium (80% of the transformed plants compared to 57% of the untransformed ones). This finding supports the notion that SERK plays an important role in Orchid embryogenesis.

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EFFECT OF DIFFERENT SOURCES OF PLANT GROWTH REGULATOR ON THE INDUCTION AND DEVELOPMENT OF MANGOSTEEN SOMATIC EMBRYOS

Indonesian Journal of Agricultural Science ◽

10.21082/ijas.v17n1.2016.p9-16 ◽

2017 ◽

Vol 17 (1) ◽

pp. 9

Author(s):

Yosi Zendra Joni ◽

Riry Prihatini ◽

Darda Efendi ◽

Ika Roostika

Keyword(s):

Somatic Embryogenesis ◽

Plant Growth ◽

Embryogenic Callus ◽

Plant Growth Regulator ◽

Somatic Embryos ◽

Casein Hydrolysate ◽

Malt Extract ◽

Mass Propagation ◽

Embryogenic Callus Induction

Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.

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