scholarly journals Maturation and Conversion of Somatic Embryos of Three Genetically Diverse Rose Cultivars

HortScience ◽  
2002 ◽  
Vol 37 (6) ◽  
pp. 973-977 ◽  
Author(s):  
Javier Castillón ◽  
Kathryn Kamo
Keyword(s):  
Polyethylene Glycol ◽  
Carbon Source ◽  
Liquid Medium ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Callus Cultures ◽  
Embryo Maturation ◽  
Somatic Embryo Maturation ◽  
Globular Stage ◽  
Cotyledonary Stage

Embryogenic callus cultures of three genetically diverse cultivars of rose (Rosa hybrida L.), the floribunda `Trumpeter', the multiflora `Dr. Huey', and the hybrid tea `Tineké', were used to study the effect of various carbohydrates and osmotically active compounds on somatic embryo maturation and conversion. Cotyledonary-stage embryos were produced by dispersing callus in liquid medium followed by filtration to isolate globular-stage embryos. Quantitative experiments were conducted to determine maturation and conversion of the three rose cultivars in response to medium with sucrose, glucose, fructose, or maltose as the primary carbon source and also in response to various concentrations of either myo-inositol, polyethylene glycol, or mannitol in combination with 3% sucrose. Conversion of 27% was achieved for `Trumpeter' embryos following their maturation on 3% fructose. `Dr. Huey' embryos required maturation on medium containing 3% sucrose supplemented with either 2.5% or 5% mannitol for 36% and 61% conversion, respectively. Maturation of `Tineké' embryos on either 3% sucrose, 3% glucose, or 3% fructose resulted in a maximum 12% conversion.

scholarly journals Bioproduction of Diosgenin in Callus Cultures of Balanites aegyptiaca: Effect of Growth Regulators, Explants and Somatic Embryogenesis

2006 ◽  
Vol 1 (3) ◽  
pp. 1934578X0600100
Author(s):  
Bishnu P. Chapagain ◽  
Vinod Saharan ◽  
Dan Pelah ◽  
Ram C. Yadav ◽  
Zeev Wiesman
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Basal Medium ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Balanites Aegyptiaca ◽  
Basal Media

This study describes the effects of plant growth regulators, explants, and somatic embryogenesis on in vitro production of the steroidal sapogenin, diosgenin, in callus cultures of the Balanites aegyptiaca (L.) Del.(desert date). Root, shoot, hypocotyl, and epicotyl callus culture of B. aegyptiaca, were raised on MS basal media supplemented with various combinations of either 2,4-D and NAA alone, or with BAP. The diosgenin content (on a dry weight basis) was found to be highest when calli were cultured in MS basal medium supplemented with 1.0 mg l−1 2,4-D alone and/or in combination with 0.5 mg l−1 BAP. However, the callus growth was highest in media supplemented with 2.5 or 3.0 mg l−1 2,4-D. MS basal media supplemented with 2,4-D 2.5 mg l−1 alone and in combination with 0.5 mg l−1 BAP induced pre-embryogenic callus formation on root cultures. When these pre-embryogenic callus cultures were used to establish cell suspension cultures, two growth densities were obtained in embryogenic suspension cultures, inducing clusters of somatic embryos at various stages of development. The maximum number of somatic embryos were obtained at the fifth week on the medium supplemented with 1.0 mg l−1 2,4-D. However, the diosgenin content in these somatic cells was found to be lower compared to the explant calluses. This study revealed that production of diosgenin in callus cultures of B. aegyptiaca is possible, but the amount is significantly affected by the growth regulators, type of explants, and somatic embryogenesis.

scholarly journals Effects of Carbon Source and Explant Type on Somatic Embryogenesis of Four Cacao Genotypes

HortScience ◽  
2006 ◽  
Vol 41 (3) ◽  
pp. 753-758 ◽  
Author(s):  
Abdoulaye Traore ◽  
Mark J. Guiltinan
Keyword(s):  
Somatic Embryogenesis ◽  
Carbon Source ◽  
Somatic Embryos ◽  
Slow Growth ◽  
Carbon Sources ◽  
The Other ◽  
Root Production ◽  
Explant Type ◽  
Embryo Maturation ◽  
Genotypic Effect

The effects of five carbon sources (glucose, fructose, maltose, sorbitol, and sucrose) and two explant types (petals and staminodes) on cacao somatic embryogenesis was studied. No growth was observed on both types of explants cultured on sorbitol containing media and slow growth was obtained on media supplemented with maltose. Depending on the genotype, the percentage of explants producing one or more embryos ranged from 6% to 99%, 18% to 98%, and 3% to 82% on media containing glucose, fructose and sucrose respectively. Explants cultured continuously on maltose or sorbitol-containing media failed to produce embryos. Staminode explants produced 3 to 10 times more somatic embryos than petals. A strong genotypic effect on somatic embryogenesis was observed. Staminode explants of the Forastero clones Laranja and PSUSca 6 produced 2 to 30 times more somatic embryos than the Trinitarios UF 613 and ICS 16. During embryo maturation and conversion, no significant differences were observed among glucose, fructose, maltose, or sucrose for embryo weight, total shoot and root production. However, we found that all plantlets produced on glucose had shoots with normal cacao leaves while the other carbon sources sometimes produced plantlets with cotyledon-like leaves.

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.

scholarly journals Development of an Efficient Protocol to Obtain Transgenic Coffee, Coffea arabica L., Expressing the Cry10Aa Toxin of Bacillus thuringiensis

2019 ◽  
Vol 20 (21) ◽  
pp. 5334 ◽  
Author(s):  
Eliana Valencia-Lozano ◽  
José L. Cabrera-Ponce ◽  
Miguel A. Gómez-Lim ◽  
Jorge E. Ibarra
Keyword(s):  
Mass Spectrometry ◽  
Bacillus Thuringiensis ◽  
Somatic Embryo ◽  
Somatic Embryos ◽  
Leaf Tissue ◽  
Stable Transformation ◽  
Embryo Maturation ◽  
Embryogenic Cell ◽  
Somatic Embryo Maturation ◽  
Coffee Plants

This report presents an efficient protocol of the stable genetic transformation of coffee plants expressing the Cry10Aa protein of Bacillus thuringiensis. Embryogenic cell lines with a high potential of propagation, somatic embryo maturation, and germination were used. Gene expression analysis of cytokinin signaling, homedomains, auxin responsive factor, and the master regulators of somatic embryogenesis genes involved in somatic embryo maturation were evaluated. Plasmid pMDC85 containing the cry10Aa gene was introduced into a Typica cultivar of C. arabica L. by biobalistic transformation. Transformation efficiency of 16.7% was achieved, according to the number of embryogenic aggregates and transgenic lines developed. Stable transformation was proven by hygromycin-resistant embryogenic lines, green fluorescent protein (GFP) expression, quantitative analyses of Cry10Aa by mass spectrometry, Western blot, ELISA, and Southern blot analyses. Cry10Aa showed variable expression levels in somatic embryos and the leaf tissue of transgenic plants, ranging from 76% to 90% of coverage of the protein by mass spectrometry and from 3.25 to 13.88 μg/g fresh tissue, with ELISA. qPCR-based 2−ΔΔCt trials revealed high transcription levels of cry10Aa in somatic embryos and leaf tissue. This is the first report about the stable transformation and expression of the Cry10Aa protein in coffee plants with the potential for controlling the coffee berry borer.

Soybean somatic embryo maturation: composition, respiration and water relations

1991 ◽  
Vol 1 (4) ◽  
pp. 251-262 ◽  
Author(s):  
J. Slawinska ◽  
R. L. Obendorf
Keyword(s):  
Somatic Embryo ◽  
Somatic Embryos ◽  
Soybean Seed ◽  
Dry Weight ◽  
Zygotic Embryos ◽  
Naphthaleneacetic Acid ◽  
Embryo Maturation ◽  
Fresh Weight ◽  
Somatic Embryo Maturation ◽  
2,4 Dichlorophenoxyacetic Acid

AbstractEmbryogenesis was induced on cotyledons of immature zygotic embryos of soybean (Glycine max (L.) Merrill) placed on solid medium containing 62.5 mm glutamine, soybean seed growth medium salts and vitamins, and 40 mg I−1 2,4-dichlorophenoxyacetic acid (2,4-D) plus 175 mm maltose, or 8 mg I−1 α-naphthaleneacetic acid (NAA) plus 88 mM sucrose. Somatic embryo development was continued in liquid medium containing 0.16 mg I−1 indole-3-butyric acid and 2.64 mg I−1 abscisic acid, glutamine and salts as above, and 88–438 mM sucrose in progressively increasing steps. Germination was on solid half-strength Murashige-Skoog medium. During maturation, somatic embryos mimicked zygotic embryos in colour, protein concentration, water and solute potentials, and respiration. Protein and lipid accumulated to 329 and 86 g kg−1 dry weight in somatic embryos. Fatty acid composition was similar to that of axes of mature seeds. Before desiccation, the water and solute potentials of maturing somatic embryos declined to −1.13 and −1.99 MPa while turgor increased to 0.86 MPa. Concomitantly, a 60% reduction in activity of the cytochrome oxidase pathway of respiration occurred with somatic embryo maturation at 600 g water kg−1 fresh weight. Although small (about 8 mg per embryo), 60% of the somatic embryos formed roots and shoots during germination after maturation without drying and 30% germinated after drying to 60 g water kg−1 fresh weight. In the greenhouse, somatic plantlets grew to mature plants with seeds.

scholarly journals CPPU IN THE MEDIUM FOR SEED GERMINATION PROMOTES EMBRYOGENESIS FROM SEEDLING EXPLANTS IN COMMON BEAN.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 617f-617
Author(s):  
Mohamed F. Mohamed ◽  
Paul E. Read ◽  
Dermot P. Coyne
Keyword(s):  
Somatic Embryogenesis ◽  
Seed Germination ◽  
Liquid Medium ◽  
Common Bean ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Bipolar Structure ◽  
Seedling Explants ◽  
Explant Cultures ◽  
Cotyledonary Stage

Few studies on embryogenesis in common bean (Phaseolus vulgaris L.) have been reported and only the early stages of somatic embryogenesis were observed. Dry seeds from two common bean lines were germinated in darkness on L-6 medium containing 4% sucrose, 0.2 g casein hydrolysate /liter and 2.0 g phytagel /liter. The medium for seed germination was supplemented with 0, 2, 4 or 6μM forchlorfenuron (CPPU). Explants from cotyledonary leaves, petioles, hypocotyls and shoot apices were prepared from 14 day-old seedlings. Callus was derived from explant cultures incubated in darkness at 26C on the medium containing 4 μM 2,4-D and 1 μM Kinetin. The callus was transferred after 4 weeks into 125 ml Erlenmeyer flasks containing 50 ml liquid medium and placed on a gyrotary shaker (120 rpm) under cool-white light (12 μmol.m-2.s-1). The liquid medium was used with 2, 4 or 6 μM of 2,4-D alone or with zeatin supplements at relative concentrations of 0.25 and 0.5. Up to 200 somatic embryos from 40 to 50 mg callus inoculations were induced after 4 to 5 weeks. Callus derived from seedlings grown on CPPU-containing medium gave more repetitive somatic embryos. Cotyledonary stage embryos with clear bipolar structure were observed only from callus derived from seedlings grown on CPPU when transferred to suspension cultures containing 2,4-D and zeatin. All somatic embryos differentiated strong roots and some developed leaf-like structures on conversion medium.

scholarly journals Perkembangan kalus embriogenik sagu (Metroxylon sagu Rottb.) pada tiga sistem kultur in vitro Development of embryogenic callus of sago (Metroxylon sagu Rottb.) on three systems of in vitro culture

2016 ◽  
Vol 76 (1) ◽  
Author(s):  
Pauline D KASI ◽  
. SUMARYONO
Keyword(s):  
In Vitro Culture ◽  
Liquid Medium ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Temporary Immersion System ◽  
Embryogenic Cells ◽  
Temporary Immersion ◽  
Friable Embryogenic Callus ◽  
Metroxylon Sagu

Summary Embryogenic callus of sago (Metroxylon sagu Rottb.) has been grown on three systems of in vitro culture i.e. agar-solidified medium, liquid medium, and temporary immersion system (TIS) medium to observe and compare the development of embryogenic callus over one passage of six weeks.  A-half gram of embryogenic callus was cultured on a modified MS medium containing 10 mg/L   2,4-D and 0.1 mg/L kinetin. For histological studies, embryogenic callus was fixed in FAA and embedded in paraplast wax. Serial sections were stained with safranin 1% and observed microscopically. By the end of culture period, the development of embryogenic callus in TIS medium was relatively better than those of the other two media.  Fresh weight of callus in liquid medium and TIS increased by 6.5-fold, while on agar-solidified medium increased by 5.4-fold in six weeks.  About 40% of callus in liquid medium and TIS and 20% of callus on agar solidified medium have changed into somatic embryos at globular stage. Histology structure of embryogenic callus of the three systems of in vitro culture shows different pattern. On agar-solidified medium, secondary callus and friable embryogenic callus that consist of meristematic cells were formed. In contrast, more embryogenic cells were formed in liquid medium and TIS to support maturation process to somatic embryos. Therefore, temporary immersion system and liquid medium are recommended for maturation of embryogenic callus, whereas agar-solidified medium is for proliferation of embryogenic callus of sago.  Ringkasan Kalus embriogenik sagu (Metroxylon sagu Rottb.) telah ditumbuhkan pada tiga sistem kultur in vitro yaitu medium padat, medium cair, dan medium dengan sistem perendaman sesaat (SPS) untuk mempelajari dan mem-bandingkan perkembangan dari kalus embrio-genik selama periode enam minggu. Setengah gram kalus embriogenik dikulturkan pada medium MS modifikasi yang mengandung  2,4-D 10 mg/L dan kinetin 0,1 mg/L.  Untuk studi histologi, kalus embriogenik difiksasi dengan FAA dan embedding menggunakan lilin paraplast. Irisan diwarnai dengan safranin 1% dan diamati menggunakan mikroskop. Pada akhir periode kultur, pertumbuhan kalus pada medium dengan SPS lebih baik dibandingkan dengan medium cair dan padat. Bobot basah kalus pada  medium cair dan SPS meningkat 6,5 kali sedangkan pada medium padat meningkat 5,4 kali dalam waktu enam minggu. Sebanyak 40% kalus pada medium cair dan SPS serta 20% kalus pada medium padat berubah menjadi embrio somatik fase globuler. Struktur histologi kalus embriogenik pada ketiga jenis sistem kultur in vitro menunjukkan pola yang berbeda. Pada medium padat terjadi pembentukan kalus sekunder dan kalus embriogenik remah yang terdiri atas sel-sel meristematik. Sebaliknya pada medium cair dan SPS pembentukan sel embriogenik lebih banyak yang menunjang proses pendewasaan menjadi embrio somatik. Oleh karena itu, medium cair dan SPS direkomendasikan untuk pendewasaan kalus embriogenik, sedangkan medium padat untuk proliferasi kalus embriogenik sagu. 

scholarly journals Somatic embryogenesis and plant regeneration in elite clones of Theobroma cacao

2008 ◽  
Vol 43 (10) ◽  
pp. 1433-1436 ◽  
Author(s):  
Thiago Édson Ribeiro da Silva ◽  
Luciana Cardoso Cidade ◽  
Fátima Cerqueira Alvim ◽  
Júlio Cézar de Mattos Cascardo ◽  
Marcio Gilberto Cardoso Costa
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Carbon Source ◽  
Embryogenic Callus ◽  
Theobroma Cacao ◽  
Somatic Embryos ◽  
Embryogenic Calli ◽  
Embryogenic Potential ◽  
Petri Dishes ◽  
Theobroma Cacao L

The objective of this work was to evaluated a procedure for somatic embryogenesis and regeneration of cacao (Theobroma cacao L.) elite clones. Petal explants from cacao clones TSH 565 and TSH 1188 were cultured on PCG and SCG-2 media, for calli growth. Somatic embryos were formed on the surface of embryogenic calli after transfer to embryo development (ED) medium. Clone TSH 565 showed a higher embryogenic potential than TSH 1188. The best combination of carbon source for embryo induction in ED medium was genotype-specific. Embryogenic callus formations increased in micropore tape-sealed Petri dishes, irrespective of cacao genotype. Mature somatic embryos were successfully converted into plantlets.

scholarly journals Perkembangan kalus embriogenik sagu (Metroxylon sagu Rottb.) pada tiga sistem kultur in vitro Development of embryogenic callus of sago (Metroxylon sagu Rottb.) on three systems of in vitro culture

2016 ◽  
Vol 76 (1) ◽  
Author(s):  
Pauline D KASI ◽  
. SUMARYONO
Keyword(s):  
In Vitro Culture ◽  
Liquid Medium ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Temporary Immersion System ◽  
Embryogenic Cells ◽  
Temporary Immersion ◽  
Friable Embryogenic Callus ◽  
Metroxylon Sagu

Summary Embryogenic callus of sago (Metroxylon sagu Rottb.) has been grown on three systems of in vitro culture i.e. agar-solidified medium, liquid medium, and temporary immersion system (TIS) medium to observe and compare the development of embryogenic callus over one passage of six weeks.  A-half gram of embryogenic callus was cultured on a modified MS medium containing 10 mg/L   2,4-D and 0.1 mg/L kinetin. For histological studies, embryogenic callus was fixed in FAA and embedded in paraplast wax. Serial sections were stained with safranin 1% and observed microscopically. By the end of culture period, the development of embryogenic callus in TIS medium was relatively better than those of the other two media.  Fresh weight of callus in liquid medium and TIS increased by 6.5-fold, while on agar-solidified medium increased by 5.4-fold in six weeks.  About 40% of callus in liquid medium and TIS and 20% of callus on agar solidified medium have changed into somatic embryos at globular stage. Histology structure of embryogenic callus of the three systems of in vitro culture shows different pattern. On agar-solidified medium, secondary callus and friable embryogenic callus that consist of meristematic cells were formed. In contrast, more embryogenic cells were formed in liquid medium and TIS to support maturation process to somatic embryos. Therefore, temporary immersion system and liquid medium are recommended for maturation of embryogenic callus, whereas agar-solidified medium is for proliferation of embryogenic callus of sago.  Ringkasan Kalus embriogenik sagu (Metroxylon sagu Rottb.) telah ditumbuhkan pada tiga sistem kultur in vitro yaitu medium padat, medium cair, dan medium dengan sistem perendaman sesaat (SPS) untuk mempelajari dan mem-bandingkan perkembangan dari kalus embrio-genik selama periode enam minggu. Setengah gram kalus embriogenik dikulturkan pada medium MS modifikasi yang mengandung  2,4-D 10 mg/L dan kinetin 0,1 mg/L.  Untuk studi histologi, kalus embriogenik difiksasi dengan FAA dan embedding menggunakan lilin paraplast. Irisan diwarnai dengan safranin 1% dan diamati menggunakan mikroskop. Pada akhir periode kultur, pertumbuhan kalus pada medium dengan SPS lebih baik dibandingkan dengan medium cair dan padat. Bobot basah kalus pada  medium cair dan SPS meningkat 6,5 kali sedangkan pada medium padat meningkat 5,4 kali dalam waktu enam minggu. Sebanyak 40% kalus pada medium cair dan SPS serta 20% kalus pada medium padat berubah menjadi embrio somatik fase globuler. Struktur histologi kalus embriogenik pada ketiga jenis sistem kultur in vitro menunjukkan pola yang berbeda. Pada medium padat terjadi pembentukan kalus sekunder dan kalus embriogenik remah yang terdiri atas sel-sel meristematik. Sebaliknya pada medium cair dan SPS pembentukan sel embriogenik lebih banyak yang menunjang proses pendewasaan menjadi embrio somatik. Oleh karena itu, medium cair dan SPS direkomendasikan untuk pendewasaan kalus embriogenik, sedangkan medium padat untuk proliferasi kalus embriogenik sagu. 

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