scholarly journals Somatic embryogenesis in the leaf tissue of Hedyotis diffusa Willd.

2018 ◽  
Vol 1 (6) ◽  
pp. 76-85
Author(s):  
Anh Thi Kim Nguyen ◽  
Tham Thi Thu Hoang ◽  
Hoang Ngo Phan
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Morphological Changes ◽  
Leaf Tissue ◽  
Naphthalene Acetic Acid ◽  
Mature Embryos ◽  
Embryo Formation ◽  
Hedyotis Diffusa ◽  
Leaf Segments

Hedyotis diffusa is a valuable medicinal herb belong to the Rubiaceae family. It is widely used in the treatment of various types of cancer and other diseases related to leukemia. Besides, the plant usually contains triterpenoids (oleanolic acid, ursolic acid) and flavonoids which have a range of pharmacological activities of antiinflammatory, antibacterial, hypoglycemic, antifree radicals, reducing blood lipids and anticancer. Leaf segments of 3 weeks old H. diffusa were cultured on half strength Murashige and Skoog (½MS) medium supplemented with different concentrations of indole acetic acid (IAA; 0.1, 0.2 and 0.4 mg/L) or α-naphthalene acetic acid (NAA; 0.1, 0.2 and 0.4 mg/L) and benzyladenine (BA) at a concentration of 1mg/L. In this study, the highest percentage of somatic embryogenesis was obtained using 0.1 mg/L IAA and 1 mg/L BA, and 0.2 mg/L IAA and 1 mg/L BA. The somatic embryos develop through a series of morphological stages: globular type, heart, torpedo and mature embryos. Besides, the highest number of shoots per explant was achieved in the same media. The middle and the distal end segments were found the most suitable for somatic embryogenesis. Morphological changes and the role of endogenous hormones in somatic embryo formation were analyzed. The position of the leaf segments of the same leaf, respiration rate, and endogenous hormone and somatic embryo formation were also discussed.

Plant Regeneration from Cultured Medicago truncatula with Particular Reference to Abscisic Acid and Light Treatments

1998 ◽  
Vol 46 (1) ◽  
pp. 151 ◽  
Author(s):  
K. E. Nolan ◽  
R. J. Rose
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Abscisic Acid ◽  
Plant Regeneration ◽  
Medicago Truncatula ◽  
Callus Induction ◽  
Naphthalene Acetic Acid ◽  
Marked Inhibition ◽  
Embryo Formation ◽  
Auxin Concentration

Medicago truncatula (Jemalong 2HA) can be regenerated by somatic embryogenesis utilising 1-naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP). There is a requirement for both NAA and BAP for callus induction and embryo formation. There is no requirement for a drop in auxin concentration to induce embryos. Abscisic acid (ABA) when present with NAA and BAP during embryo formation at a concentration of 1 µM, increases the number of embryos per callus. The ABA treatment stimulates embryo numbers in both light and darkness. The conversion efficiency of embryo to plant is unchanged irrespective of the presence of ABA during embryo formation, indicating that ABA does not improve the regeneration of the embryos once formed. Importantly, the presence of light in the embryo formation period causes a marked inhibition of embryo conversion.

scholarly journals Plant Regeneration Via Organogenesis and Somatic Embryogenesis in Verbascum Sinuatum L.

2014 ◽  
Vol 56 (1) ◽  
pp. 97-103 ◽  
Author(s):  
Roya Karamian ◽  
Fatemeh Ghasemlou
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Casein Hydrolysate ◽  
Shoot Multiplication ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Shoot Buds ◽  
Traditional Medicines ◽  
Mature Embryos

Abstract The genus Verbascum L. belongs to the family Scrophulariaceae and its members are used as medicinal herbs in traditional medicines worldwide. In this study we achieved plant regeneration in Verbascum sinuatum L. via organogenesis and somatic embryogenesis by culture of mature embryos. Embryogenic and nonembryogenic calli were induced from mature embryos on Murashige and Skoog (MS) medium supplemented with different concentrations of benzyl adenine (BA) and a-naphthalene acetic acid (NAA) (but not for 1.5 and 3 mg l−1 NAA). For multiplication of somatic embryoids and differentiation of shoot buds, yellow and friable embryonic calli were transferred to MS medium containing 30 g/l sucrose, 0.5 mg l−1 charcoal and 0.1 or 1 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) or to MS medium containing 60 g l−1 sucrose, 50 mg l−1 casein hydrolysate (CH), 0.5 mg l−1 kinetin (Kin), 5 mg l−1 2,4-D and 0.5 mg l−1 charcoal. Shoot multiplication and plantlet regeneration were achieved by transferring shoot buds to MS medium supplemented with 1 mg l−1 BA or Kin.

scholarly journals Pengaruh Sitokinin, Jenis Eksplan, dan Genotipe terhadap Embriogenesis Somatik Kakao

2016 ◽  
Vol 3 (2) ◽  
pp. 71
Author(s):  
Nur Ajijah ◽  
RR. Sri Hartati
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Theobroma Cacao ◽  
Somatic Embryos ◽  
Interaction Effects ◽  
Embryo Formation ◽  
Primary Somatic Embryogenesis ◽  
Primary Somatic Embryos

<p><em>Information on the effect of cytokinins on cacao (</em>Theobroma cacao<em> L.) primary somatic embryogenesis and its interaction with explant types and genotypes is not yet known. This study aimed to evaluate the effect of cytokinins and its interaction with explant types and genotypes on cacao somatic embryogenesis. The study was conducted at tissue culture laboratory of IAARD, Bogor from April until December 2012 and October 2014 until February 2016. Three types of cytokinins i.e. kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M), thidiazuron (0.01, 0.02, and 0.04 </em><em>μ</em><em>M) and benzylaminopurine (0.55, 1.11, and 2.22 </em><em>μ</em><em>M) in combination with 9 </em><em>μ</em><em>M 2,4-D were tested for their effectiveness in inducing somatic embryogenesis from petals and staminoid explants of Cimanggu 1 genotype. Furthermore, three levels of kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M</em><em>) also in combination with 9 </em><em>μ</em><em>M 2,4-D were evaluated for their influences on the somatic embryogenesis from petals and staminoid explants of three cacao genotypes i.e. Sulawesi 02, ICCRI 04 and Cimanggu 3. The result demonstrated that 2.32 </em><em>μ</em><em>M kinetin and staminoids explant were more effective to induce cacao somatic embryogenesis of Cimanggu 1 genotype (7%, 0.23 embryos/explant). Additionally, there were interaction effects between the level of kinetin with explant types and genotype on the percentage of explants forming embryo at 12 weeks after culture. The highest percentage of somatic embryo formation was shown by ICCRI 04 genotype with the use of petals explant and a kinetin level of 1.16 </em><em>μ</em><em>M (31.85%), but not significantly different from the level of kinetin 2.23 </em><em>μ</em><em>M (25.55%). The formation of primary somatic embryos of cacao is largely determined by the type and level of cytokinins, type of explant, and genotype.</em></p>

scholarly journals Effect of the combination of NAA, kinetin and sucrose on the induction of somatic embryogenesis in lulo (Solanum quitoense Lam.)

2014 ◽  
Vol 32 (2) ◽  
pp. 170-179 ◽  
Author(s):  
Hernando Criollo ◽  
Margarita Perea ◽  
Mariano Toribio ◽  
Johanna Muñoz
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Plant Propagation ◽  
Cotyledonary Leaf

Lulo is a species of great importance to the fruticulture of Colombia, but has significant phytosanitary problems that require an aggressive breeding program oriented toward the production of genotypes with tolerance to phytopathogens. These programs need to establish highly efficient mass plant propagation protocols, such as somatic embryogenesis. This study focused on research on the somatic embryogenesis of lulo using kinetin, naphthalene acetic acid-NAA (Plant Growth Regulators, PGRs), and different sucrose concentrations in a MS medium. Two lulo varieties, Solanum quitoense var. septentrionale and S. quitoense var. quitoense, and two explant types (hypocotyl and cotyledon) were used, incubated in dark conditions at 25±2°C. The highest production percentage of the embryos was obtained when 50 mM of NAA were added to the medium with sucrose (50.0 and 263.1 mM) for the two explant types used. In lulo with spines, the highest percentage of embryonic structures (50%) was observed with cotyledonary leaf explants and 50 mM of NAA ; while in the spineless lulo, the embryonic structures were observed in the same type of explant with 50 mM of NAA + 263.1 mM of sucrose (32%).

scholarly journals Evaluation of Endogenous Phytocomponents and Amino Acids Associated with Direct Somatic Embryogenesis of Coffee (Coffea arabica L.)

2018 ◽  
Vol 8 ◽  
pp. 1293-1308
Author(s):  
D. K. Isutsa ◽  
R. N. Mayoli ◽  
A. B. Nyende ◽  
C. M. Mweu
Keyword(s):  
Amino Acids ◽  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Embryo Development ◽  
Direct Somatic Embryogenesis ◽  
Embryogenic Cultures ◽  
Mature Embryos ◽  
The Status ◽  
Somatic Embryo Development ◽  
Set Up

Coffee is one of the most important crops cultivated in the world for use in beverages and confectionaries. Embryogenesis is a complex process that begins with a single cell and ends with the formation of mature embryos. Somatic embryo development involves accumulation of complex metabolites and storage reserves. This present experiment identified and quantified endogenous phytocomponents and amino acids present during somatic embryogenesis of ‘Ruiru 11’. Laboratory experiments for this study were set up in the Coffee Research Institute, Kenya at Ruiru. Third leaf pair explants were excised from 8-month-old greenhouse-grown mother plants sterilized and cultured in half strength Murashige and Skoog basal salts augmented with Thidiazuron. Once embryos had developed, the cultures were analysed for phytocomponents using GCMS and HPLC. The results showed that palmitoleic and stearic acids were highest (23.3 µg/g and 69.9 µg/g respectively) in brown embryogenic cultures. Cis 7,8 epoxy-2-methyl octadecane was highest (253 µg/g) in green embryogenic cultures. (Z)-3-Tetradecene was highest (25 µg/g) in brown non-embryogenic cultures. Z, Z-3,13- Octadecedien-1-ol and (Z)-7-Hexadecenal were highest (32.1 µg/g and 70.2 µg/g respectively) in green embryogenic cultures. Alanine content was highest (4.4 µg/g) in embryos of brown cultures. Amino acids, fatty acids and their derivatives are potential biomarkers for embryogenesis. Other phytocomponents should be identified and their role in coffee somatic embryogenesis determined. Further studies regarding the status of the phytocomponents identified in the present study, especially in particular stages of embryo development are needed to propose treatments to improve coffee somatic embryo development.

scholarly journals SOMATIC EMBRYOGENESIS OF MUSSAENDA `QUEEN SIRIKIT'

HortScience ◽  
1994 ◽  
Vol 29 (4) ◽  
pp. 251f-251 ◽  
Author(s):  
Christopher S. Cramer ◽  
Mark P. Bridgen
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Leaf Number ◽  
Callus Production ◽  
Indole 3 Acetic Acid

Disinfected midrib sections of Mussaenda `Queen Sirikit' ≈3 to 4 mm in size were cultured on a basal medium of Murashige and Skoog salts and vitamins, 87.7 mm sucrose, and 5 g Sigma agar/liter supplemented with several concentrations of indole-3-acetic acid (IAA) (0, 5.0, 10.0, 20.0 μm) and 6-benzylaminopurine (BAP) (0, 0.5, 1.0, 2.5, 5.0, 10.0, 25.0, 50.0 μm). Cultures were subculture onto the same treatment after 5 weeks and observed weekly for 15 weeks for the presence of somatic embryos. As somatic embryos were produced, they were subculture onto basal medium supplemented with 0.5, 1.0, 2.5, or 25.0 μm BAP. Callus was first observed at 2 weeks in cultures grown on basal medium supplemented with 5.0–20.0 μm IAA and 0–50.0 μm BAP. Somatic embryos were observed at 8 weeks on basal medium supplemented with 5.0–10.0 μm IAA and 2.5–5.0 μm BAP. Callus cultured on 0–10 μm IAA and 5.0–10.0 μm BAP produced the greatest number of somatic embryos by 15 weeks. Somatic embryos subculture to basal medium supplemented with 25.0 μm BAP proliferated shoots, while eliminating BAP from the medium resulted in root and callus production. Shoots and entire plants were removed from in vitro conditions and successful] y acclimated to greenhouse conditions. Somatic embryo-derived plants flowered sporadically 25 to 35 weeks after removal from in vitro conditions. Variations in sepal number and leaf number per node were observed at 1% to 5%.

scholarly journals Somatic embryogenesis and plant regeneration in purple food yam (Dioscorea alata L.)

2008 ◽  
pp. 22-33 ◽  
Author(s):  
Marilyn Belarmino ◽  
Jocelyn Gonzales
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Adventitious Shoots ◽  
Pantothenic Acid ◽  
Naphthalene Acetic Acid ◽  
Dioscorea Alata ◽  
Ms Medium ◽  
Leaf Petiole

A study was conducted to establish a reliable procedure for somatic embryogenesis and plant regeneration from callus cultures of purple food yam (Dioscorea alata L.). The procedure involved three steps; (1) culture of nodal stem segments from greenhousegrown plants to generate in vitro plantlets; (2) induction of callus from the leaf, petiole and nodal stem tissues; and (3) initiation of somatic embryo from callus. Results showed that the agar-solidified Murashige and Skoog (MS) medium containing 30 gl-1 sugar, 0.1 gl-1 α-cysteine , 10 mgl-1 calcium pantothenic acid, 2.0 mgl-1 asparagine, 2.0 mgl-1 arginine, 80.0 mgl-1 adenine sulfate (AdSO4) and 0.1 mgl-1 naphthalene acetic acid (NAA) effectively broke dormancy of lateral buds of nodal stem cultures from both ‘VU-2’ and ‘Kinampay‘ varieties. Production of multiple adventitious shoots occurred after transfer of in vitro nodal pieces to the same medium added with 1.0 mgl-1 benzylamino purine (BAP) or, MSA medium. Callus was effectively induced from the vegetative tissues in MS medium added with 1.0 mgl-1 2,4-Dichlorophenoxy acetic acid (2,4-D) or, with picloram. Among the three types of explants, the nodal stem was the most suitable which produced purplish nodular embryogenic callus. A higher percentage of nodal stem-derived calli produced globular embryos in MS medium containing 1.0 mgl-1 2,4-D and 0.5 mgl-1 BAP, or in 1.0 mgl-1 picloram and 0.5 mgl-1 BAP than, in the plant growth regulator-free medium (control). The maturation of embryos was facilitated by one-month culture in MS medium containing 0.1 mgl-1 ABA and 100 mgl-1 glutamine. This step improved the germination of somatic embryos in one-half strength PGR-free MS medium containing 100 mgl-1 glutamine (regeneration medium). All somatic embryoderived plantlets were morphologically normal and established well in soil.

scholarly journals Micropropagation of Tulip via Somatic Embryogenesis

Agronomy ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1857
Author(s):  
Małgorzata Podwyszyńska ◽  
Agnieszka Marasek-Ciolakowska
Keyword(s):  
Somatic Embryogenesis ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Vascular Bundles ◽  
Embryonic Cells ◽  
Embryogenic Calli ◽  
Pyridine Carboxylic Acid ◽  
Embryo Formation ◽  
Pyridine Carboxylic ◽  
2,4 Dichlorophenoxyacetic Acid

An effective method of tulip regeneration via somatic embryogenesis (SE) was developed. Explants, flower stem slices excised from cooled bulbs were incubated in darkness on MS modified media containing auxins alone (2,4-dichlorophenoxyacetic acid—2,4-D, 1-naphthalene acetic acid—NAA and 4-amino-3,5,6-trichloro-2-pyridine carboxylic acid—picloram) or combined with thidiazuron (TDZ) at 0.1 and 0.5 mg L−1. Yellowish-white callus with a granular structure was developed in the presence of all auxins on the cut surface from the tissues of the vascular bundles. From this, lines of embryogenic calli were derived. The addition of TDZ to the medium with auxins significantly stimulated somatic embryo formation. Cyclic and the most intensive proliferation of embryogenic callus as well as embryo formation was obtained in the presence of 2,4-D at 0.1 mg L−1 combined with TDZ at 0.5 mg L−1. Addition of proline enhanced either callus proliferation rate or frequency of embryo formation. The best quality embryos with cotyledons longer than 10 mm able to form bulbs were recorded when TDZ was replaced with 6-benzylaminopurine (BAP) at the concentration of 0.1 mg L−1. Histomorphology showed that the development of somatic embryos could have either external or internal origins. Embryos of external origin were initiated by cell division on the edge of embryogenic calli. Embryos of internal origin resulted from the division of parenchyma cells inside the tissue. Embryonic cells were characterized by their small volume, regular shape, dense cytoplasm and large nuclei. The globular embryos were covered by a distinct layer of periderm. Then, the embryos developed into structures having leaf-shaped cotyledons with a procambial strand and a sideward-orientated meristem of the vegetative apex (stolon). Cotyledon embryos did not show vascular connections with the parent tissue, and they did not develop embryonic roots.

scholarly journals Efficient and reproducible somatic embryogenesis and micro propagation in tomato via novel structures -Rhizoid Tubers

2018 ◽  
Author(s):  
Wajeeha Saeed ◽  
Saadia Naseem ◽  
Daniyal Gohar ◽  
Zahid Ali
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Naphthalene Acetic Acid ◽  
Light Conditions ◽  
Indirect Organogenesis ◽  
Novel Structure ◽  
Micro Propagation ◽  
Novel Structures ◽  
Indole 3 Acetic Acid

AbstractAn improved and highly reproducible system for invitro regeneration via somatic embryogenesis (S.E), applicable to several varieties of tomato (cv. Riogrande, cv. Roma grande, hybrid 17905 and model cv. M82) has been developed. First, we developed a conventional indirect organogenesis for all four varieties used in this study. The cotyledons and hypocotyls of 6-day-old tomato were used as explants (1-2 cm) for callus induction (CI) on different callus induction media (CIM) T0 – T12 (6-Benzylaminopurine BAP, NAA Naphthalene acetic acid, ZEA Zeatin, IAA Indole-3-acetic acid, KIN Kinetin). Maximum CI response was seen on CIMT6 (0.5 mg/L NAA, 1 mg/L BAP) and CIMT7 (2 mg/L IAA, 2 mg/L NAA, 2 mg/L BAP, 4mg/L KIN) in a period of 2 weeks for commercial varieties cvs. Riogrande and Roma. However, cv. M82 responded after 4 weeks to a combination of treatments [CIMT6 (0.5 mg/L NAA + 1 mg/L BAP) and CIMT8 (2 mg/L IAA + 2 mg/L NAA + 2 mg/L BAP + 4 mg/L ZEA)] for the production of calli. The Riogrande, being the most responsive commercial variety, was selected for invitro morphogenesis via S.E. During S.E. young cotyledons and hypocotyls explants were tested on media with different ranges of pH (3 – 7) supplemented with 0.5 and 2 mg/L NAA. Resultantly, numerous rhizoids (~38) were produced from each explant at pH4 in dark conditions. Further incubation of each rhizoid under light conditions led to the formation of a novel structure - rhizoid tubers (RTBs) on MS media supplemented with 5 mg/L TDZ/BAP at pH4. We observed that only lower pH-induced rhizoids and RTBs regenerated into multiple individual shoots on media at normal pH (5.8). The RTBs led to a complete plantlets regeneration in 45 days compared to the conventional invitro morphogenesis (60 days).

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