scholarly journals Phytochemical Analysis and Establishment of Embryogenic Cell Suspension and Agrobacterium-mediated Transformation for Farmer Preferred Cultivars of West African Plantain (Musa spp.)

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 789 ◽  
Author(s):  
Temitope Jekayinoluwa ◽  
Jaindra Nath Tripathi ◽  
George Obiero ◽  
Edward Muge ◽  
Leena Tripathi
Keyword(s):  
Cell Suspension ◽  
Culture Media ◽  
Crop Improvement ◽  
Genotypic Variation ◽  
Cell Suspensions ◽  
Phytochemical Analysis ◽  
Embryogenic Calli ◽  
Embryogenic Cell ◽  
Agrobacterium Mediated Transformation

Banana and plantain are among the foremost staple food crops providing food and livelihood to over 500 million people in tropical countries. Despite the importance, their production is hampered due to several biotic and abiotic stresses. Plant tissue culture techniques such as somatic embryogenesis and genetic transformation offer a valuable tool for genetic improvement. Identification and quantification of phytochemicals found in banana and plantain are essential in optimizing in vitro activities for crop improvement. Total antioxidants, phenolics, flavonoids, and tannins were quantified in various explants obtained from the field, as well as in vitro plants of banana and plantain cultivars. The result showed genotypic variation in the phytochemicals of selected cultivars. The embryogenic cell suspensions were developed for three farmer-preferred plantain cultivars, Agbagba, Obino l’Ewai, and Orishele, using different MS and B5-based culture media. Both culture media supported the development of friable embryogenic calli (FEC), while MS culture media supported the proliferation of fine cell suspension in liquid culture media. The percentage of FEC generated for Agbagba, Obino l’Ewai, and Orishele were 22 ± 24%, 13 ± 28%, and 9 ± 16%, respectively. Cell suspensions produced from FECs were successfully transformed by Agrobacterium-mediated transformation with reporter gene constructs and regenerated into whole plants.

scholarly journals Efficient plant regeneration from embryogenic cell suspension culture of two deepwater rice (Oryza sativa L.) Varieties

2013 ◽  
pp. 91-103
Author(s):  
L Khaleda ◽  
M Al-Forkan
Keyword(s):  
Plant Regeneration ◽  
Cell Suspension ◽  
Culture Media ◽  
Oryza Sativa L ◽  
Cell Suspension Cultures ◽  
Cell Suspensions ◽  
Deepwater Rice ◽  
Embryogenic Calli ◽  
Rice Varieties ◽  
Embryogenic Cell

Cell suspension cultures were initiated from 28-30 d old scutellar-derived embryogenic calli of two deepwater rice varieties HAJA-1 and HAJA-8 and maintained on R2 liquid medium containing 2 mg l-1 2,4-D. The formation of fine embryogenic cell suspensions and subsequent plant regeneration were dependent on plant genotype and culture media. Variety HAJA-8 was found less viable than the other variety HAJA-1. In general, it was observed that variety HAJA-1 showed better performance compared to variety HAJA-8 to R2 medium for the initiation of embryogenic cell lines and the rate of the cell growth was higher in this variety. Plant regeneration was not obtained from cell suspensions of both varieties when cells were cultured on regeneration media semi-solidified with 0.4% (w/v) agarose. Plant regeneration from cell suspensions of the both varieties were obtained when the agrose concentration of the regeneration media was increased from 0.4 to 1% (w/v). The highest plant regeneration frequencies were obtained from cell suspensions of the varieties HAJA-1 and HAJA-8 (48%) and (42%), respectively, pre-treated with water for 5 h and cultured on MS medium supplemented with 2 mg l-1 BAP+1 mg l-1 Proline. Washing cell suspensions with water for a prolonged period of time (15 h and more) inhibited plant regeneration. DOI: http://dx.doi.org/10.3329/cujbs.v5i1.13374 The Chittagong Univ. J. B. Sci.,Vol. 5(1 &2):91-103, 2010

scholarly journals Initiation of coconut cell suspension culture from shoot meristem derived embryogenic calli: A preliminary study

2016 ◽  
Vol 8 (1) ◽  
Author(s):  
U. Bhavyashree ◽  
K. Lakshmi Jayaraj ◽  
K. S. Muralikrishna ◽  
K. K. Sajini ◽  
M. K. Rajesh ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Fluorescent Microscopy ◽  
Cell Aggregation ◽  
Shoot Meristem ◽  
Malt Extract ◽  
Embryogenic Calli ◽  
Embryogenic Cell

<p>An attempt was made to establish highly competent embryogenic cell suspension culture in coconut, a species recalcitrant to in vitro culture. Embryogenic calli were initiated from shoot meristem explants of coconut. Y3 medium supplemented with 2.4-D (4.5 μM) and glutamine (34.2 μM) was found to be the best medium to initiate cell suspension. Growth evaluation was done by packed cell volume (PCV) and it was found that maximum growth volume of 9.9% was reached at 200 days of culture initiation. About 52% of viable cells were detected through fluorescent microscopy. Cell aggregation was noticed in Y3 medium supplemented with glutamine (34.2 μM), malt extract (100mg/l), biotin (40.9 μM) and kinetin (9.3 μM), but further progress could not be achieved. It was also observed that embryogenic calli were not of a friable type, but were associated with densely aggregated cells. Because of its hard nature, we were unsuccessful to obtain high quality cell suspension.</p>

scholarly journals Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Somatic Embryos ◽  
Culture Media ◽  
Ms Medium ◽  
Regenerated Plants ◽  
Male Flowers

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

scholarly journals Identificación, establecimiento in vitro y análisis fitoquímico preliminar de especies silvestres de ñame (Dioscorea spp.) empleadas con fines medicinales

2015 ◽  
Vol 17 (1) ◽  
pp. 9-17 ◽  
Author(s):  
Víctor Andrés Ramos Duarte ◽  
Silvia Lizette Bustamante, R. ◽  
Javier Rincón Velandia ◽  
Maritza Adelina Rojas Cardozo ◽  
Lauren Raz ◽  
...  
Keyword(s):  
Culture Media ◽  
Phytochemical Analysis ◽  
Extraction Solvent ◽  
Palabras Clave ◽  
Edible Species ◽  
Medicinal Use ◽  
Advanced Analysis ◽  
Layer Chromatography ◽  
Wild Yam

<strong><strong>Título en ingles: Identification, <em>in vitro</em> establishment and preliminary phytochemical analysis of wild yam (<em>Dioscorea spp</em>.) used for medicinal purposes</strong></strong><p><strong><strong><strong>Título corto: Identificación, establecimiento <em>in vitro </em>y análisis fitoquímico preliminar de especies silvestres de ñame</strong></strong></strong></p><p><strong>Resumen:  </strong>Tubérculos del género<em> Dioscorea </em>comercializados con fines medicinales, fueron recolectados con el propósito de lograr su establecimiento a condiciones <em>in vitro. </em>Previamente se lograron identificar taxonómicamente las especies y por medio de análisis fitoquímicos demostrar su potencial farmacéutico. El material recolectado fue identificado como <em>Dioscorea coriacea, D. lehmannii, D. meridensis, D. polygonoides </em>y una especie comestible<em> D. trifida</em>. Tubérculos recolectados de centros de acopio y traídos de campo fueron lavados, desinfectados, asperjados con Ácido Giberélico (AG<sub>3</sub>) y sembrados en sustrato BM-2®, en invernadero a 18°C día y 10°C noche. Los tubérculos completos o por secciones fueron almacenados en bolsas herméticas a temperatura ambiente. Posteriormente se desinfectó material vegetal de las especies <em>D. coriacea, D. lehmannii, D. meridensis y D polygonoides, </em>seleccionando explantes de brotes sanos (<em>D. coriacea / </em>laboratorio) para su establecimiento. Se evaluaron tres medios de cultivo para establecimiento, el que presentó los mejores resultados fue Medio Murashige &amp; Skoog (1962) suplementado con BAP 1 mL/L, AG<sub>3</sub> 1 mL/L y Putrescina 2 mL/L. Para la extracción y análisis de metabolitos secundarios se utilizaron tubérculos de <em>D. coriacea, D. lehmannii</em> y <em>D. polygonoides</em>, empleando como solvente de extracción metanol. Se  encontró mayor concentración de extracto vegetal en <em>D. coriacea </em>(54%), y mediante cromatografía en capa delgada (CCD), se confirmó la presencia de saponinas, que resultó mayor en comparación con <em>D. polygonoides</em> especie reconocida por su alto contenido de saponinas. Estos resultados permitirán realizar análisis más avanzados de los compuestos presentes y plantear su propagación masiva en condiciones <em>in vitro</em>. </p><p><strong>Palabras clave:</strong> diosgenina, micropropagación, ñame silvestre, cultivo de tejidos vegetales, saponinas, fitoquímica.</p><p><strong>Abstract:  </strong>Wild tubers of the genus<em> Dioscorea</em> sold for medicinal use were collected for the purpose of achieving its establishment under <em>in vitro</em> conditions. First we taxonomically identified the species and through phytochemical analysis demonstrated pharmaceutical potential. The material collected was identified as <em>Dioscorea coriacea</em>, <em>D. lehmannii</em>, <em>D. meridensis</em>, <em>D. polygonoides</em> and the edible species <em>D. trifida</em>. Tubers collected from wholesale distributors and from the field were washed, disinfected, sprayed with Gibberellic Acid (GA<sub>3</sub>) and planted in substrate BM-2®, in a greenhouse at 18 ° C during the day and 10 ° C overnight. Whole tubers or sections thereof were stored in sealed bags at room temperature. Subsequently plant material of the species <em>D. coriacea, D. lehmannii, D. </em>meridensis<em> </em>and<em> D. polygonoides</em> was disinfected and healthy buds (<em>D. coriacea</em> / laboratory) were selected for <em>in vitro</em> establishment. Three different culture media were evaluated for establishment; that which presented the best results was the Murashige &amp; Skoog (1962) medium, supplemented with BAP 1 mL / L, GA<sub>3</sub> 1 mL / L and Putrescin 2 mL / L. For the collection and analysis of secondary metabolites, tubers of <em>D. coriacea</em>, <em>D. lehmannii</em> and <em>D. polygonoides</em> were used, using methanol as the extraction solvent. The highest concentration of plant extract, 54%, was found in<em> D. coriacea</em>, a higher value than that of <em>D. polygonoides, </em>which had been reported previously; the presence of saponins was confirmed by thin layer chromatography (TLC). These results will enable more advanced analysis of the present compounds and enhance their mass propagation under <em>in vitro</em> conditions.</p><p><strong>Key words:</strong> diosgenin, micropropagation, wild yam, tissue culture, saponins, phitochemistry.</p><p><strong>Recibido: </strong>agosto 20 de 2014<strong>      Aprobado: </strong>abril 20 de 2015</p>

scholarly journals In vitro Development of Cauliflower Synthetic Seeds and Development of Plantlets In vivo

2014 ◽  
Vol 24 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Zahida Qamar ◽  
Md. Belal Hossain ◽  
Idrees A. Nasir ◽  
Bushra Tabassum ◽  
Tayyab Husnain
Keyword(s):  
Cell Suspension ◽  
Somatic Embryos ◽  
Cell Suspension Cultures ◽  
Synthetic Seeds ◽  
Embryogenic Calli ◽  
Globular Cells ◽  
Vitro Development ◽  
Direct Use

Synthetic seeds of cauliflower cv. Chillout were developed by encapsulating mature somatic embryos in neutral gel media. Somatic embryos were obtained by optimizing callus and cell suspension cultures of cauliflower. Friable, yellowish embryogenic calli were obtained on MS supplemented with 2 mg/l  2,4-D and 0.5 mg/l BAP using hypocotyl as explants, while calli were regenerated in media consisting of 5 mg/l BAP, 2 mg/l Kn and 6 mg/l GA3. Somatic embryo-genesis was induced in cell suspension culture where auxins were removed in successive steps triggering  conversion of globular cells into the heart, torpedo stage (71%) and finally into cotyledonary/somatic embryos (28%). The mature somatic embryos were encapsulated by mixing mature cell suspension with sodium alginate and calcium chloride mixture (1 : 4). Developed synthetic seeds germinated into complete plantlets when placed in neutral gel media.  Germination efficiency of synthetic seeds decreased to about 50 per cent after 12 weeks of storage at 4ºC followed by a rapid decrease to zero per cent after 16 weeks. It was also observed that cauliflower plantlets from synthetic seeds survived successfully when transferred to soil demonstrating  that cauliflower synthetic seeds is a promising step towards their  in vivo direct use. Plant Tissue Cult. & Biotech. 24(1): 27-36, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19193

Evaluation of the Enhanced Regeneration System for in vitro regeneration in barley

2006 ◽  
Vol 86 (1) ◽  
pp. 63-69
Author(s):  
Seedhabadee Ganeshan ◽  
Brian J Weir ◽  
Monica Båga ◽  
Brian G Rossnagel ◽  
Ravindra N Chibbar
Keyword(s):  
Hordeum Vulgare ◽  
Embryogenic Callus ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Cold Treatment ◽  
Regeneration System ◽  
Embryogenic Calli ◽  
Hordeum Vulgare L ◽  
Best Response

A simple two-step model for evaluation of in vitro regeneration protocols is proposed based on callus induction and regeneration from immature scutella of two Canadian barley (Hordeum vulgare L.) genotypes, AC Metcalfe and SB92559 using the Enhanced Regeneration System (ERS). The number of explants producing embryogenic callus, the number of plants per embryogenic callus and the number of plants per explant were considered. Tissue culture parameters included three combinations of growth regulators, two carbon sources in culture media, and three cold treatment regimes of spikes prior to scutella isolation. Culture medium containing 5 µM 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 µM benzyl adenine (BA) induced the highest percent of embryogenic calli and the highest number of shoots per embryogenic callus from AC Metcalfe. Medium containing 3.75 µM 2,4-D and 0.75 µM BA gave the best response for SB92559. Both genotypes produced more shoots on maltose than on sucrose medium. A 2-d treatment of spikes at 4°C resulted in best response for SB92559. Regeneration response from AC Metcalfe scutella from spikes was unaffected by being subjected to 2, 4 or 6 d of cold. Conditions resulting in best responses from both genotypes were tested on four commercial barley varieties. However, these lines showed inferior regeneration compared to SB92559 and AC Metcalfe. Key words: Hordeum vulgare, scutella, embryogenic callus, shoot production

Agrobacterium?-mediated transformation of embryogenic cell suspensions of the banana cultivar Rasthali (AAB)

2001 ◽  
Vol 20 (2) ◽  
pp. 157-162 ◽  
Author(s):  
T. R. Ganapathi ◽  
N. S. Higgs ◽  
P. J. Balint-Kurti ◽  
C. J. Arntzen ◽  
G. D. May ◽  
...  
Keyword(s):  
Cell Suspensions ◽  
Embryogenic Cell ◽  
Agrobacterium Mediated Transformation ◽  
Embryogenic Cell Suspensions ◽  
Banana Cultivar

scholarly journals INDUCTION OF A HEMOLYSIN RESPONSE IN VITRO

1970 ◽  
Vol 132 (6) ◽  
pp. 1267-1278 ◽  
Author(s):  
Klaus-Ulrich Hartmann
Keyword(s):  
Bone Marrow ◽  
Immune Response ◽  
T Cells ◽  
Cell Suspension ◽  
Cell Population ◽  
Cell Suspensions ◽  
Spleen Cells ◽  
Bone Marrow Derived Cells ◽  
Thymus Cells

The immune response to foreign erythrocytes was studied in vitro. Two subpopulations of cells were prepared. One was a population of bone marrow-derived spleen cells, taken from thymectomized, irradiated, and bone marrow-reconstituted mice; there was evidence that most of the precursors of the PFC had been present in this cell population, but few PFC developed in cultures of these cells alone in the presence of immunogenic erythrocytes. Another cell suspension was made from spleens of mice which had been irradiated and injected with thymus cells and erythrocytes; these cells were called educated T cells. The two cell suspensions together allow the formation of PFC in the presence of the erythrocytes which were used to educate the T cells, but not in the presence of noncross-reacting erythrocytes. If bone marrow-derived cells and T cells were kept in culture together with two different species of erythrocytes, and if one of the erythrocytes had been used to educate the T cells, then PFC against each of the erythrocytes could be detected.

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