scholarly journals Somatic Embryogenesis and Plant Regeneration in Viola canescens Wall. Ex. Roxb.: An Endangered Himalayan Herb

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 761
Author(s):  
Arun Kumar Khajuria ◽  
Christophe Hano ◽  
Narendra Singh Bisht
Keyword(s):  
Somatic Embryogenesis ◽  
Culture Medium ◽  
Casein Hydrolysate ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Mature Embryos ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Root Axis ◽  
Somatic Embryo Induction

Viola canescens Wall. ex. Roxb. is an important but threatened medicinal herb found at 1500–2400 m above mean sea level in the Himalayas. Overexploitation and habitat preference have put the plant under serious threat. Thus, the present study was undertaken to develop an efficient protocol for in vitro propagation via somatic embryogenesis. The results revealed that plant can be regenerated successfully through somatic embryogenesis using leaf derived calli. Regular subculturing of calli on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D)/indole-3-butyric acid (IBA)/kinetin (Kn) and varying combinations of 2,4-D+Kn induced somatic embryogenesis. The maximum average number of somatic embryos (SE) (19.15 ± 2.66) was induced on the medium with 0.15 + 0.05 mg L−1 of 2,4-D and Kn, respectively, and this medium was used as a control. To enhance somatic embryo induction, the control MS medium was supplemented with l-glutamine (200–400 mg L−1) and casein hydrolysate (1–4%). The maximum average number of SE (27.66 ± 2.67) and average mature SE (13.16 ± 3.48) were recorded on the medium having 2 % l-glutamine and 50 mg L−1 casein hydrolysate. The induced SE were asynchronous, so, to foster their maturation, the culture medium (free from growth regulators) was supplemented with abscisic acid (ABA) and silver nitrate (AgNO3). The maximum average number (35.96 ± 3.68) of mature SE was noticed on MS medium supplemented with 1.5 mg L−1 ABA. Mature embryos had two well-developed cotyledons and an elongated hypocotyl root axis. The development of SE into plantlets was significant for embryos matured on the medium with AgNO3 and ABA, with 86.67% and 83.33% conversion on the medium with 0.20 mg L−1 6-benzylaminopurine (BAP). The plantlets thus produced acclimatized in a growth chamber before being transferred to the field, which showed 89.89% survival. The plants were morphologically similar to the mother plant with successful flowering.

Embryo induction and plant regeneration of Callerya speciosa (Fabaceae) through anther culture

2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li
Keyword(s):  
Anther Culture ◽  
Culture Medium ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Anther Wall ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.

scholarly journals Impact of Osmotica and Plant Growth Regulators on Somatic Embryogenesis of Date Palm

2019 ◽  
Vol 7 (3) ◽  
pp. 296-303
Author(s):  
Mouaad Amine Mazri ◽  
Ilham Belkoura ◽  
Reda Meziani ◽  
Hajar Es-Saoudy ◽  
Fahd Rachad ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Date Palm ◽  
Dichlorophenoxyacetic Acid ◽  
Adventitious Bud ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Mature Embryos ◽  
Maturation Medium ◽  
Semi Solid ◽  
2,4 Dichlorophenoxyacetic Acid

An efficient somatic embryogenesis system is reported for date palm cv. Al-Fayda, a genotype resistant to the bayoud disease. Callus induction was achieved from adventitious bud explants cultured for 6 months on semi-solid Murashige and Skoog (MS) medium containing 4.5 μM 6-(dimethylallylamino) purine (2iP) and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or picloram. The highest somatic embryogenesis frequency (89%) was obtained on MS medium supplemented with 225 μM 2,4-D. Subsequently, embryogenic cultures were transferred to agitated liquid MS medium (maturation medium) containing various concentrations of mannitol, polyethylene glycol (PEG) or sorbitol. The highest rate of somatic embryo maturation (71.4 mature embryos per 100 mg callus) was achieved on the medium supplemented with 40 g l-1 PEG. Mature somatic embryos were then transferred to MS medium supplemented with gibberellic acid (GA3) or 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) at various concentrations. The highest frequency of germination and conversion (26%) was obtained on the medium containing 5 μM NAA and 5 μM BAP. The developed plants were then transferred to ex vitro conditions, where a survival rate of 77.02% was observed. The regeneration protocol established in the present investigation will be used for mass propagation of date palm cv. Al-Fayda.

EFFICIENT CALLUS INDUCTION AND PLANT REGENERATION VIA SOMATIC EMBRYOGENESIS FROM IMMATURE LEAF-DERIVED PROTOPLASTS OF GROUNDNUT (ARACHIS HYPOGAEA L.)

1996 ◽  
Vol 44 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Perumal Venkatachalam ◽  
Narayanasamypillai Jayabalan
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Arachis Hypogaea ◽  
Callus Cultures ◽  
Alternative Methods ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
High Yields ◽  
2,4 Dichlorophenoxyacetic Acid

High yields of protoplasts were obtained from immature leaves of aseptically grown plants of Arachis hypogaea using an enzyme solution containing cellulase 2.0% (w/v) and Macerozyme 1.0% (w/v) in 0.6 M mannitol. Isolated protoplasts were cultured in Kao's medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and mini calli in 4 weeks. After 4 weeks, protoplast colonies were transferred to the Murashige and Skoog (MS) medium supplemented with a-naphthalene acetic acid (NAA) and BAP. Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones differentiated roots on the same medium. Alternative methods for plant regeneration from protoplast derived callus cultures were tried through somatic embryogenesis. Protoplast-derived calli treated with 2,4-D and BAP formed somatic embryos. Somatic embryogenesis began in the proembryo stage and proceeded from globular to dicotyledonary stage. Embryos were then transferred onto hormone-free MS medium for germination. Five to ten percent of these embryoids germinated and grew to plantlets. Regenerated plants were transferred to plastic cups and grown to maturity.

scholarly journals Somatic Embryogenesis and Organogenesis in Magnolia dealbata Zucc. (Magnoliaceae), an Endangered, Endemic Mexican Species

HortScience ◽  
2006 ◽  
Vol 41 (5) ◽  
pp. 1325-1329 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Ángel Jiménez-Rodríguez ◽  
Victor M. Chávez-Avila
Keyword(s):  
Somatic Embryogenesis ◽  
Basal Medium ◽  
Direct Somatic Embryogenesis ◽  
Direct Organogenesis ◽  
Limiting Factor ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Magnolia Dealbata

Plants of Magnolia dealbata were regenerated from zygotic embryos through somatic embryogenesis and direct organogenesis. Medium and incubation conditions were determinating factors for the development of morphogenetic responses. Photoperiodic exposure was a limiting factor in the general development of the explants, and incubation in darkness allowed their development. The highest formation of shoots per responding explant were obtained on woody plant (WP) medium supplemented with 13.3 μM or 22.2 μM 6-benzylaminopurine (BA) in combination with 2.26 μM or in absence of 2,4-dichlorophenoxyacetic acid (2,4-D) from which 2.5 shoots per explant were induced. Subcultures on WP medium, supplemented with polyvinylpyrrolidone (PUP) 40,000 1 g·L–1) avoided necrosis of explants. Somatic embryos were formed in 85% of explants cultivated on WP medium with 2,4-D (2.3 μM or 4.5 μM); 20% induced indirect embryogenesis and 65% formed direct somatic embryogenesis. The plants were transferred to soil to acclimatize under greenhouse conditions, achieving 90% survival. Somatic embryo conversion to plantlets was obtained with subculture on WP basal medium without growth regulators. In vitro culture can play a key role in the propagation and conservation of this endangered species.

In Vitro Shoot Regeneration of NAA-Pulse Treated Plumular Leaf Explants of Cowpea

2010 ◽  
Vol 2 (2) ◽  
pp. 60-63 ◽  
Author(s):  
Muhammad AASIM
Keyword(s):  
Somatic Embryogenesis ◽  
Shoot Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Grain Legume ◽  
Ms Medium ◽  
Pulse Treatment ◽  
Mature Embryos ◽  
Legume Crop

Cowpea (Vigna unguiculata L.) is an economically important grain legume crop and is an important source of dietary protein in many of the developing countries. The present study reports the effect of pulse treatment duration, concentration of NAA and presence of NAA in the culture medium on shoot regeneration from plumular leaf explant of Turkish cowpea cv. ‘Akkiz’ and ‘Karagoz’. Pulse treatment of mature embryos with 20 mg l-1 NAA for 1 and 3 weeks followed by culturing of plumular leaf explant on MS medium containing 0.25, 0.50 and 1.0 BAP with 1.0, 2.0 and 4.0 mg l-1 NAA promoted somatic embryogenesis in both cultivars. Longer duration of pulse treatment was deleterious resulting in browning and consequently death of the embryos on explants. Pulse treatment with 20 mg l-1 NAA for one week was less deleterious and developed two plantlets after the explants were transferred to MS0 medium after 6 weeks through somatic embryogenesis in cv. ‘Akkiz’. Pulse treatment with 10 mg l-1 NAA for 1 week showed 33.33-50.00% and 25.00-50.00% shoot regeneration frequency in cv. ‘Akkiz’ and ‘Karagoz’ respectively on MS medium containing 0.25-1.00 mg l-1 BAP. Maximum number of 2.50 shoots each per explant were recorded in cv. ‘Akkiz’ and ‘Karagoz’ on MS medium containing 1.00 and 0.50 mg l-1 BAP respectively. Contrarily, maximum shoot length of 8.98 cm of cv. ‘Akkiz’ and 9.42 cm of cv. ‘Karagoz’ was recorded on MS medium containing 0.50 mg l-1 BAP and 1.00 mg l-1 BAP respectively. Regenerated shoots were rooted on MS medium containing 0.5 mg l-1 IBA and and acclimatized in growth room at room temperature where they produced viable seeds.

SOMATIC EMBRYOGENESIS IN CELL SUSPENSION CULTURE OF COWPEA (VIGNA UNGUICULATA (L.) WALP)

1995 ◽  
Vol 43 (4) ◽  
pp. 385-390 ◽  
Author(s):  
S. Kulothungan ◽  
A. Ganapathi ◽  
A. Shajahan ◽  
K. Kathiravan
Keyword(s):  
Somatic Embryogenesis ◽  
Liquid Medium ◽  
Vigna Unguiculata ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Seedling Leaf ◽  
Cotyledonary Stage

Embryogenic callus was induced from seedling leaf explants of cowpea (Vigna unguiculata (L.) Walp. cv. C152 on Murashige and Skoog (MS) medium containing 2.0 mg 1−1 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of somatic embryogenesis was noticed when this callus was transferred to MS liquid medium supplemented with 2 mg 1−1 2,4-D. Further studies on ontogeny of somatic embryos showed that the cells destined to become somatic embryos divided into spherical or filamentous proembryos. Subsequent divisions in the proembryo led to globular, heart, torpedo-shaped, and cotyledonary-stage somatic embryos. Tiny plantlets were obtained by transferring the cotyledonary-stage somatic embryos to MS liquid medium containing 0.5 mg 1−1 2,4-D.

Influence of auxins on somatic embryogenesis and alkaloid accumulation in Leucojum aestivum callus

2013 ◽  
Vol 8 (6) ◽  
pp. 591-599 ◽  
Author(s):  
Agata Ptak ◽  
Anna Tahchy ◽  
Edyta Skrzypek ◽  
Tomasz Wójtowicz ◽  
Dominique Laurain-Mattar
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Symptomatic Treatment ◽  
Dichlorophenoxyacetic Acid ◽  
Leucojum Aestivum ◽  
Anisic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Response

AbstractIn vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).

scholarly journals Somatic Embryogenesis and Massive Shoot Regeneration from Immature Embryo Explants of Tef

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Likyelesh Gugsa ◽  
Jochen Kumlehn
Keyword(s):  
Culture Medium ◽  
Important Determinant ◽  
Immature Embryo ◽  
Eragrostis Tef ◽  
Embryogenic Tissue ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Horn Of Africa ◽  
2,4 Dichlorophenoxyacetic Acid

Tef (Eragrostis tef) provides a major source of human nutrition in the Horn of Africa, but biotechnology has had little impact on its improvement to date. Here, we report the elaboration of an in vitro regeneration protocol, based on the use of immature zygotic embryos as explant. Explant size was an important determinant of in vitro regeneration efficiency, as was the formulation of the culture medium. Optimal results were obtained by culturing 0.2–0.35 mm embryo explants on a medium containing KBP minerals, 9.2–13.8 μM 2,4-dichlorophenoxyacetic acid, 6 mM glutamine, and 0.5% Phytagel. Although this protocol was effective for both the improved cultivar “DZ-01-196” and the landrace “Fesho”, the former produced consistently more embryogenic tissue and a higher number of regenerants. An average of more than 2,800 shoots could be obtained from each “DZ-01-196” explant after 12 weeks of in vitro culture. These shoots readily formed roots, and plantlets transferred to soil were able to develop into morphologically normal, fertile plants. This regeneration and multiplication system should allow for the application of a range of biotechnological methods to tef.

scholarly journals Secondary Somatic Embryogenesis in Centaurium erythraea Rafn

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 199
Author(s):  
Milica D. Bogdanović ◽  
Katarina B. Ćuković ◽  
Angelina R. Subotić ◽  
Milan B. Dragićević ◽  
Ana D. Simonović ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Histological Analysis ◽  
Developmental Process ◽  
Developmental Pathways ◽  
Dichlorophenoxyacetic Acid ◽  
Embryogenic Cells ◽  
Endangered Medicinal Plant ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Centaurium Erythraea

Somatic embryogenesis (SE) is a developmental process during which plant somatic cells, under suitable conditions, produce embryogenic cells that develop into somatic embryos (se). SE is the most important method for plant propagation in vitro, having both fundamental and applicative significance. SE can be induced from different tissues and organs, but when se are used as explants, the process is recognized as secondary or cyclic SE. We induced secondary SE in Centaurium erythraea by application of 2,4-dichlorophenoxyacetic acid (2,4-D) and N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU). A medium containing 0.1 mgL−1 2,4-D and 0.25 mgL−1 CPPU was optimal in terms of the number of primary SE explants forming se, the number of well-developed se per explant, and morphological appearance of the obtained se. These concentrations allowed SE to progress through three cycles, whereas at higher concentrations of 0.2 mgL−1 2,4-D and 0.5 mgL−1 CPPU, only two cycles were achieved. Histological analysis revealed that secondary se are formed both directly and indirectly. Secondary SE readily germinated and converted into plantlets. Induction of cyclic SE contributes to the conservation efforts of this endangered medicinal plant and expands the spectrum of in vitro developmental pathways described in centaury—an emerging model in developmental biology.

scholarly journals Shoot Proliferation, Callus Induction And Plant Regeneration In Tripsacum Laxum Nash

2021 ◽  
Author(s):  
Yuping Xiong ◽  
Jinhui Pang ◽  
Kunlin Wu ◽  
Jaime A. Teixeira Silva ◽  
Xinhua Zhang ◽  
...  
Keyword(s):  
Peat Soil ◽  
Shoot Proliferation ◽  
Multiple Shoots ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Rooting Medium ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract The peduncles of Tripsacum laxum Nash were used as explants to induce axillary shoots. Multiple shoots were proliferated on Murashige and Skoog (MS) medium to establish, for the first time, efficient shoot proliferation and plant in vitro regeneration systems. Optimal shoot proliferation medium was MS with 3.0 mg/L 6-benzyladenine (BA) and 0.2 mg/L α-naphthaleneacetic acid (NAA), resulting in a shoot proliferation coefficient of 11.0 within 45 d. Optimal rooting medium was MS with 0.1 mg/L NAA and/or 0.1 mg/L indole-3-butyric acid (IBA), inducing 100% root formation from shoots within 30 d. When young roots, leaf sheaths and shoot bases were used as explants, MS medium with 1.0 mg/L thidiazuron (TDZ) and 0.2 mg/L BA induced most shoots, with the least callus. Shoot bases induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), while leaf sheaths induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L BA. Rooted plantlets showed 99.3% survival when transplanted into a substrate of vermiculite: peat soil (1:3, v/v).

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