scholarly journals Somatic Embryogenesis and Plant Regeneration from Immature Persimmon (Diospyros kaki Thunb.) Embryos

HortScience ◽  
2008 ◽  
Vol 43 (1) ◽  
pp. 211-214 ◽  
Author(s):  
Akira Sugiura ◽  
Yoshiko Matsuda-Habu ◽  
Mei Gao ◽  
Tomoya Esumi ◽  
Ryutaro Tao
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Cultured Cells ◽  
Callus Formation ◽  
Formation Rate ◽  
Adventitious Bud ◽  
Diospyros Kaki ◽  
Embryogenic Calli ◽  
Globular Embryos ◽  
2,4 Dichlorophenoxyacetic Acid

In persimmon, plant regeneration from cultured cells usually takes place through adventitious bud formation. If somatic embryogenesis were possible, the efficiency of mass propagation and genetic engineering would be greatly improved. We attempted to induce somatic embryogenesis from immature embryos and plant regeneration from the induced embryos. Hypocotyls and cotyledons from immature ‘Fuyu’ and ‘Jiro’ seeds were cultured in the dark in Murashige and Skoog medium solidified with gellan gum and supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) at various concentrations. Callus formation started at ≈2 weeks of culture, and the callus formation rate was highest at 3 or 10 μm combinations of 2,4-D and BA. The initially formed calli gradually became brown or black from which white embryogenic calli (EC) appeared secondarily. After ≈8 weeks of culture, globular embryos were formed from these EC, and the formation proceeded until 20 weeks of culture. Formation of globular embryos was higher with ‘Fuyu’ than ‘Jiro’, especially with hypocotyls. When EC with globular embryos were transferred to fresh medium with no plant growth regulators, ≈70% developed to the torpedo-type embryo stage in 6 weeks. The torpedo-type embryos thus formed were germinated and rooted in agar medium with or without zeatin in several weeks without entering dormancy. After germination and rooting, the plantlets were transferred to the same medium and acclimatized for another 4 weeks. As the embryos germinated and rooted simultaneously, the plantlets were easy to grow in pots without transplanting shock. This is the first report on plant regeneration through somatic embryogenesis of persimmon.

Study on Tissue Culture of Sweet Broad Pea

2014 ◽  
Vol 644-650 ◽  
pp. 5407-5410
Author(s):  
Hui Fang Chi
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Culture Medium ◽  
Callus Formation ◽  
Formation Rate ◽  
Adventitious Bud ◽  
Experimental Basis ◽  
Research Results

s. The cotyledons, Internodes, leaves and stems of sweet broad pea were studied on tissue culture. Research results show that: The ability of different explants for callus formation and adventitious bud differentiation in different culture medium is different. The callus formation rate and sprouting rate of Internodes is significantly higher than other explants, which is a ideal material for tissue culture. The callus formation rate of Internodes was 100% in MS +BA1.0 mg/L+NAA 1.0 mg/L and MS+ 2, 4-D 0.5 mg/L; The bud differentiation is best at the medium of MS+ 6-BA 2 mg/L, which reached 86.7%; the rooting rate was 83.3% at the medium of MS+ NAA 3mg/L. The study provides a experimental basis for further study on the plant regeneration in the sweet broad pea.

scholarly journals Plant regeneration from protoplasts of Gentiana straminea Maxim

2016 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Guomin Shi ◽  
Lina Yang ◽  
Tao He
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Protoplast Culture ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Embryogenic Calli ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Solid Liquid

AbstractA protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N6-benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

scholarly journals Induction of somatic embryogenesis in two cultivars of anthurium analysed by scanning electron microscopy

2019 ◽  
Vol 13 ◽  
pp. 1
Author(s):  
Priscila Bezerra Dos Santos Melo ◽  
Ana Cristina Portugal Pinto de Carvalho ◽  
Cândida Hermínia Campos de Magalhães Bertini ◽  
Celli Rodrigues Muniz ◽  
Adroaldo Guimarães Rossetti
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Somatic Embryogenesis ◽  
Callus Formation ◽  
Nodal Segments ◽  
Embryogenic Calli ◽  
Evaluation Period ◽  
Mean Values ◽  
Embryogenic Callus Formation ◽  
Scanning Electron

Somatic embryogenesis is an advantageous tool in the commercial production of micropropagated anthurium plantlets. As such, the aim of this study was to establish a protocol for the induction of somatic embryogenesis in Jureia and Luau cultivars. Defoliated nodal segments, 1.0 cm in length and containing one bud, were used as explants. The experimental design was completely randomised, in a 2 x 3 x 5 factorial scheme (cultivar: Jureia and Luau x auxin: 2,4-D, NAA and Picloram x concentration: 0, 2.5, 5.0, 7.5, and 10.0 μM), with 30 treatments in a scheme of plots split over time (15, 30, 45, 60, 75 and 90 days). The anatomy and percentage of embryogenic callus formation were analysed. The structures formed, analysed by scanning electron microscopy, corresponded to embryogenic calli. The Luau cultivar was superior in forming embryogenic calli. For the two cultivars, among the auxins under study, NAA demonstrated a greater induction potential for somatic embryogenesis, with the concentration of 7.5 μM giving the highest mean values. The 90-day evaluation period showed the maximum formation of embryogenic calli; however, mean values were fairly similar to the 75-day evaluation period. To induce embryogenic calli, therefore, it is suggested that the nodal segments be inoculated into a culture medium with added NAA growth regulator at a concentration of 7.5 μM, and that the explants remain in this medium for 75 days after inoculation.

scholarly journals Obtaining calli and regenerated plants in anther cultures of pea

2014 ◽  
Vol 50 (No. 2) ◽  
pp. 123-129 ◽  
Author(s):  
S. Bobkov
Keyword(s):  
Developmental Stages ◽  
Callus Formation ◽  
Naphthaleneacetic Acid ◽  
Embryogenic Calli ◽  
Nutrient Media ◽  
Shoot Morphogenesis ◽  
Regenerated Plants ◽  
Wide Range ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

Pea (Pisum sativum L.) is a species for which there is no efficient method for the recovery of haploid plants yet. This research investigated the influence of various genotypes, nutrient media, and stress treatments on callus formation, embryogenesis and plant regeneration in anther cultures of pea. A wide range of pea genotypes and nutrient media was studied. Morphogenic calli were initiated on media supplemented with &alpha;-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and 2,4-dichlorophenoxyacetic acid (2,4-D) without application of stress treatments. Embryogenic calli and embryos were regenerated on media with low sucrose content in the presence of 2,4-D or indole-3-butyric acid (IBA) after cold stress (4&deg;C) of isolated buds, alone or in combination with in&nbsp;vitro treatment of isolated anthers at higher temperatures (35&ndash;38&deg;C). The efficiency of regeneration via shoot morphogenesis on different nutrient media and the peculiarities of regeneration from embryogenic calli were investigated. Green embryogenic calli initiated on 2,4-D were able to develop through shoot morphogenesis on a medium supplemented with BA and NAA. This process led to regeneration of hypertrophic embryos at various developmental stages. The origin of regenerated plants (i.e. from microspores or somatic anther cells) was estimated using marker alleles determining morphological traits. Almost all R<sub>0</sub> regenerants derived from morphogenic calli originated from anther somatic cells.

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

HortScience ◽  
2014 ◽  
Vol 49 (12) ◽  
pp. 1558-1562 ◽  
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.

scholarly journals REGENERATION OF Pimpinella pruatjan THROUGH SOMATIC EMBRYOGENESIS

2016 ◽  
Vol 8 (2) ◽  
pp. 60
Author(s):  
I. Roostika ◽  
R. Purnamaningsih ◽  
I. Darwati ◽  
I. Mariska
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Callus Formation ◽  
Natural Habitat ◽  
Casein Hydrolysate ◽  
Shoot Multiplication ◽  
Dry Weight ◽  
Embryogenic Calli ◽  
The Media ◽  
Medicinal Properties

Pruatjan (Pimpinella pruatjan Molk.) is an Indonesian endangered plant which has various medicinal properties such as aphrodisiac, diuretic, and tonic. The plant is commonly harvested from its natural habitat, therefore it becomes endangered. Regeneration of pruatjan through organogenesis has been studied, but its shoot multiplication was very low (5 shoots per explant). The study aimed to investigate the best regeneration technique of pruatjan through somatic embryogenesis. This research was conducted at the tissue culture laboratory, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development in 2004-2005. Callus formation of pruatjan was induced from the petioles and leaves in Driver and Kuniyaki’s (DKW) based medium containing 2,4-D combined with picloram at the level of 0.5, 1.0, 1.5, and 1.5 ppm. Embryogenic calli were then transferred into embryo development medium in two ways. First, they were directly transferred into media containing IBA/NAA at the level of 0.5, 1.0, and 1.5 ppm. Second, they were indirectly transferred into media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate prior to the IBA/ NAA media. Parameters evaluated were fresh weight, dry weight, time initiation of embryogenic callus formation, and total number of embryos. The result showed that calli of pruatjan were successfully induced from the petioles and leaves. The best calli were induced from the leaves in the DKW medium containing 2.0 ppm 2,4-D and 0.5 ppm picloram. Embryo development of the calli was best if they were first grown in the media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate then transferred to the IBA/NAA media. The total number of somatic embryos was counted up to 103 on the medium containing 1.5 ppm IBA. This study indicated that pruatjan somatic embryogenesis regeneration required three different media, i.e. for callus induction, development and maturation, and for germination.

scholarly journals Plant Regeneration through Somatic Embryogenesis from Leaf Sheath Derived Callus of Sugarcane (Saccharum officinarum L.) var. Isd -16

2011 ◽  
Vol 21 (2) ◽  
pp. 143-149 ◽  
Author(s):  
Mohashweta Roy ◽  
M. Hossain ◽  
A. Biswas ◽  
M. K. Biswas ◽  
R. Islam
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Saccharum Officinarum ◽  
Leaf Sheath ◽  
Coconut Water ◽  
Friable Callus ◽  
Light Yellow ◽  
Half Strength

Leaf sheath explants of an indigenous variety Isd-16 of sugarcane (Saccharum officinarum L.) produced light yellow friable callus after culturing on to MS with 2,4-D (2 - 4 mg/l) and NAA (3 - 5 mg/l) singly. Callus formation was the maximum on MS + 3 mg/l 2,4-D. Callus underwent embryogenesis producing huge number of somatic embryos when subcultured on MS with 15 - 30 mg/l             L-proline, 3 mg/l 2,4-D + 5 - 10% coconut water (v/v) and 3 mg/l 2,4-D + 10% CW  (v/v) + 300 - 500 mg/l CH. L-proline significantly enhanced somatic embryo-genesis and 25 mg/l L-proline in MS was the best culture medium formulation. Most of the somatic embryos germinated and developed plantlets after 1 - 2 weeks of incubation in proline-supplimented medium. On the other hand, maturation and germination of embryos were achieved on half-strength MS with or without 0.25 - 1.0 mg/l L-proline, and 5% coconut water (v/v). Somatic embryos derived plantlets were then successfully transferred to natural condition through successive phages of acclimation.   Key words: Plant regeneration, Leaf sheath, Somatic embryogenesis, Sugarcane   D. O. I. 10.3329/ptcb.v21i2.10237   Plant Tissue Cult. & Biotech. 21(2): 143-149, 2011 (December)

scholarly journals Plant regeneration through somatic embryogenesis from calli derived from leaf bases of Laurus nobilis L. (Lauraceae)

2015 ◽  
Vol 24 (2) ◽  
pp. 213-221 ◽  
Author(s):  
Ahmad H Al Gabbiesh ◽  
M Ghabeish ◽  
I H ◽  
M Kleinwächter ◽  
D Selmar
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Root Formation ◽  
Callus Formation ◽  
Germplasm Conservation ◽  
Laurus Nobilis ◽  
The Media ◽  
Regenerated Plantlets ◽  
Culture Technology

Somatic embryogenesis was induced in embryo culture on half MS medium supplemented with NAA (8 mg/l) as the sole plant growth regulator after incubation of the media in the refrigerator at 4°C for two weeks to promote callus induction and somatic embryogenesis in Laurus nobilis. Both embryogenetic calli and somatic embryos were induced in the above selected medium. Embryo growth and development were stimulated by separation of embryos successfully from embryo clusters and transferred onto fresh half MS. Among the selected explants, only leaf bases were found to respond actively to plant regeneration, especially in inducing callus formation and in sustaining faster callus growth. Root formation of regenerated plantlets tended to decrease with time on regeneration media. Overall, 75% of the plantlets derived from the callus survived in the greenhouse; and they all grew to phenotypically normal plants. This procedure will enable the use of regeneration tissue culture technology for germplasm conservation of L. nobilis, a plant of high medicinal and commercial value.Plant Tissue Cult. & Biotech. 24(2): 213-221, 2014 (December)

scholarly journals Somatic embryogenesis from leaf transverse thin cell layer derived-callus of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.)

2016 ◽  
Vol 14 (1) ◽  
pp. 63-73
Author(s):  
Vu Thi Hien ◽  
Nguyen Phuc Huy ◽  
Bui Van The Vinh ◽  
Hoang Xuan Chien ◽  
Hoang Thanh Tung ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Layer ◽  
Culture Media ◽  
Callus Formation ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Transverse Thin Cell Layer ◽  
Cell Layers ◽  
2,4 Dichlorophenoxyacetic Acid

No report on plant regeneration via somatic embryogenesis of P. vietnamensis has been previously published. In the present study, somatic embryogenesis via callus formation from cultures of leaf transverse thin cell layers (tTCLs) of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) was investigated. α-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and thidiazuron (TDZ) were added separately and in combination into the culture media. Explant necrosis or low callogenesis rates were observed when 1-mm wide leaf tTCLs were cultured on media with TDZ, BA, 2,4-D or NAA. On the other hand, calli were successfully induced from the tTCL explants cultured on medium supplemented with either 2,4-D and BA or 2,4-D and TDZ. Callogenesis was observed under both light and dark conditions. The highest callogenesis rate (100%) was obtained on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg l-1 2,4-D in combination with 0.1 mg l-1 TDZ in darkness after eight weeks of culture. White calli were cut into small pieces (1.0 x 1.0 cm dimension) and placed on MS media containing 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and TDZ at various concentrations (0.01; 0.1; 0.2; and 0.5 mg l-1), and the best callus proliferation was recorded on medium containing 1.0 mg l-1 2,4-D and 0.2 mg l-1 TDZ. Somatic embryogenesis, with a success rate of 53.3% and 35 embryos per explant, was achieved when calli were subcultured onto MS medium supplemented with 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and 0.2 mg l-1 TDZ.

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