scholarly journals Direct Somatic Embryogenesis: A Highly Efficient Protocol for In Vitro Regeneration of Habanero Pepper (Capsicum chinense Jacq.)

HortScience ◽  
2006 ◽  
Vol 41 (7) ◽  
pp. 1645-1650 ◽  
Author(s):  
Guadalupe López-Puc ◽  
Adriana Canto-Flick ◽  
Felipe Barredo-Pool ◽  
Patricia Zapata-Castillo ◽  
María del C. Montalvo-Peniche ◽  
...  

To induce somatic embryogenesis in habanero pepper (Capsicum chinense Jacq.), the cultivar BVll-03, belonging to the red type, was used. Different explants were evaluated, as were different culture media, the composition of which varied in the content of plant growth regulators. Results showed the formation of somatic embryos from cotyledons, zygotic embryos, germinated zygotic embryos, hypocotyls, and cotyledonary leaves. Explants were cultured on Murashige and Skoog medium supplemented with 2,4-D (9.05 μm). The somatic embryos always formed directly from the explant, without callus formation, and the greatest efficiency was obtained when segments of hypocotyls were cultured, obtaining 175 ± 20 somatic embryos per explant. Only the somatic embryos obtained on Murashige and Skoog medium containing 2,4-D (9.05 μm) and treated with abscisic acid (ABA) (1.89 μm) before their transfer to the germination media (Murashige and Skoog + 1.1 μm GA3) emitted their radicule and expanded their cotyledonary leaves (60%), whereas the remaining embryos did not achieve germination because of different causes (abnormalities, delayed development). Not only is this protocol of somatic embryogenesis the first to be reported for this species (C. chinense Jacq.), but it is also the most efficient reported so far, within the Capsicum genus.

HortScience ◽  
2007 ◽  
Vol 42 (2) ◽  
pp. 329-333 ◽  
Author(s):  
Patricia Yolanda Zapata-Castillo ◽  
Adriana-Canto Flick ◽  
Guadalupe López-Puc ◽  
Anabel Solís-Ruiz ◽  
Felipe Barahona-Pérez ◽  
...  

To induce the somatic embryogenesis of Habanero pepper, different culture media and different types of explants (node, internode, hypocotyl, half seeds, and fruit segments) were evaluated. For the induction of embryogenic callus, 9.05 μm of 2,4-dichlorofenoxiacetic acid, 3% sucrose, and 0.8% gelrite were added to the basic MS medium over a period of 30 days at 25 ± 2 °C under continuous light (40–50 μmol·m2·s−1). Once the callus formed, they were transferred to liquid medium using the same induction formulation. Somatic embryogenesis only occurred from explants of hypocotyl and in the presence of 3.4 μm thidiazuron. This constitutes the first proposal of a protocol for the “induction of somatic embryogenesis in Habanero pepper (Capsicum chinense Jacq.) from cell suspensions.”


HortScience ◽  
1994 ◽  
Vol 29 (3) ◽  
pp. 172-174 ◽  
Author(s):  
Mari Tahara ◽  
Takeshi Yasuda ◽  
Naotsugu Uchida ◽  
Tadashi Yamaguchi

Somatic embryos were regenerated from protoplasts isolated from embryogenic callus on young leaf explants from mature coffee trees. Embryos were regenerated on modified Murashige and Skoog medium supplemented with 5 μm BA. Somatic embryos developed into intact plants. Mannitol at 0.5 m was adequate as an osmoticum for isolating protoplasts, but subsequent culture required 0.3 m mannitol. A culture system in which osmolality was decreased gradually accelerated formation of colonies and somatic embryogenesis. Chemical name used: N-(phenylmethyl)-1H-purine-6-amine (BA).


HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Nancy Santana-Buzzy ◽  
Guadalupe López-Puc ◽  
Adriana Canto-Flick ◽  
Felipe Barredo-Pool ◽  
Eduardo Balam-Uc ◽  
...  

The ontogenesis of direct high-frequency somatic embryogenesis of C. chinense induced from hypocotyl was characterized through a histological analysis of the different phases in the histodifferentiation process during the development of the somatic embryo. The anatomical analysis was carried out since the hypocotyl segments were placed in the culture medium until 45 days of culture. The somatic embryos were induced and maintained in Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (9.5 μm). Samples of tissues and organs were taken every 24 h, fixed in formalin acetic alcohol, and embedded in plastic resin. They were cut into serial sections (5 μm) and stained with toluidine blue. The analysis revealed that the proembryogenic cells originated just from provascular hypocotyl cells. Provascular cells acquired the embryogenic competence 48 h after induction and an intense mitotic division was observed and embryogenic structures were generated first along the vascular strands, which subsequently evolved into somatic embryos. After 2 weeks, there were observed embryos at different stages of development (preglobular, globular, heart-shaped, torpedo-shaped, and cotyledonary). This is the first report dealing with the ontogenesis of the direct somatic embryogenesis of C. chinense, and it is the most complete histological characterization carried out on somatic embryogenesis in the Capsicum genus to date.


2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
J. Lema-Rumińska ◽  
K. Goncerzewicz ◽  
M. Gabriel

Having produced the embryos of cactusCopiapoa tenuissimaRitt. formamonstruosaat the globular stage and callus, we investigated the effect of abscisic acid (ABA) in the following concentrations: 0, 0.1, 1, 10, and 100 μM on successive stages of direct (DSE) and indirect somatic embryogenesis (ISE). In the indirect somatic embryogenesis process we also investigated a combined effect of ABA (0, 0.1, 1 μM) and sucrose (1, 3, 5%). The results showed that a low concentration of ABA (0-1 μM) stimulates the elongation of embryos at the globular stage and the number of correct embryos in direct somatic embryogenesis, while a high ABA concentration (10–100 μM) results in growth inhibition and turgor pressure loss of somatic embryos. The indirect somatic embryogenesis study in this cactus suggests that lower ABA concentrations enhance the increase in calli fresh weight, while a high concentration of 10 μM ABA or more changes calli color and decreases its proliferation rate. However, in the case of indirect somatic embryogenesis, ABA had no effect on the number of somatic embryos and their maturation. Nevertheless, we found a positive effect of sucrose concentration for both the number of somatic embryos and the increase in calli fresh weight.


2014 ◽  
Vol 23 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Dinesh Giri ◽  
Sushma Tamta

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013  (December)


Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 168
Author(s):  
Muhammad Ajmal Bashir ◽  
Cristian Silvestri ◽  
Amelia Salimonti ◽  
Eddo Rugini ◽  
Valerio Cristofori ◽  
...  

An efficient in vitro morphogenesis, specifically through somatic embryogenesis, is considered to be a crucial step for the application of modern biotechnological tools for genetic improvement in olive (Olea europaea L.). The effects of different ethylene inhibitors, i.e., cobalt chloride (CoCl2), salicylic acid (SA), and silver nitrate (AgNO3), were reported in the cyclic somatic embryogenesis of olive. Embryogenic callus derived from the olive immature zygotic embryos of the cultivar Leccino, was transferred to the expression ECO medium, supplemented with the ethylene inhibitors at 20 and 40 µM concentrations. Among these, the maximum number of somatic embryos (18.6) was obtained in media containing silver nitrate (40 µM), followed by cobalt chloride (12.2 somatic embryos @ 40 µM) and salicylic acid (40 µM), which produced 8.5 somatic embryos. These compounds interfered on callus traits: white friable embryogenic calli were formed in a medium supplemented with 40 µM cobalt chloride and salicylic acid; in addition, a yellow-compact embryogenic callus appeared at 20 µM of all the tested ethylene inhibitors. The resulting stimulatory action of silver nitrate among all the tested ethylene inhibitors on somatic embryogenesis, clearly demonstrates that our approach can efficiently contribute to the improvement of the current SE protocols for olive.


Author(s):  
Rina Arimarsetiowati

One of the propagation technique for coffee plant production is tissue culture. Tissue culture technique for Coffea arabica L. faces some problems, mainly in the planlet formation regenerated from explants. The objective of this experiment was to examine the effect 2,4-D and 2-ip combination on the formation of direct somatic embryogenesis of Coffea arabica L. in leaves explant. Auxin (2,4-D) and cytokinin (2-ip) concentrations of, respectively, 1; 5 µM and 5; 10; 15; 20 were used as treatments. This research was conducted using completely randomized design with 10 replications. Observation to induce somatic embryos was done by quantitatively on number of callus from explant and number of embryogenic callus. Beside that, observation by qualitative descriptive was also done on deve lopment of embryogenesis. The results showed that Arabica coffee leaves explant of AS 2K clones could be induced in all medium combination except 5µM 2,4-D and 20µM 2-ip combination. Arabica coffee leaves explant of S 795, Sigararutang and AS 1 varieties could be induced in all medium combination. The highest frequency of callus formation was found in AS 2K, Sigararutang and AS 1 varieties on medium containing 1µM 2,4-D in combination with 10µM 2-ip, whereas for the S 795 variety on medium containing 5µM 2,4-D in combination with 10µM 2-ip. The highest frequency of embriogenic callus in all Arabica coffee variety could be reached on medium containing 5µM 2,4-D in combination with 15µM 2-ip. Key words : Coffea arabica L., somatic embryogenesis, 2,4-D, 2-ip, tissue culture, leaves, callus embryogenic.


2020 ◽  
Vol 29 (1) ◽  
pp. eSC05
Author(s):  
Ander Castander-Olarrieta ◽  
Paloma Moncaleán ◽  
Itziar A. Montalbán

Aim of the study: To develop an efficient method to regenerate plants through somatic embryogenesis of an ecologically relevant tree species such as Pinus canariensis.Area of study: The study was conducted in the research laboratories of Neiker-Tecnalia (Arkaute, Spain).Material and methods: Green cones of Pinus canariensis from two collection dates were processed and the resulting immature zygotic embryos were cultured on three basal media. The initiated embryogenic tissues were proliferated testing two subculture frequencies, and the obtained embryogenic cell lines were subjected to maturation. Germination of the produced somatic embryos was conducted and acclimatization was carried out in a greenhouse under controlled conditions.Main results: Actively proliferating embryogenic cell lines were obtained and well-formed somatic embryos that successfully germinated were acclimatized in the greenhouse showing a proper growth.Research highlights: This is the first report on Pinus canariensis somatic embryogenesis, opening the way for a powerful biotechnological tool for both research purposes and massive vegetative propagation of this species.Keywords: acclimatization; Canary Island pine; micropropagation; embryogenic tissue; somatic embryo.Abbreviations used: embryogenic tissue (ET); established cell line (ECL);  somatic embryogenesis (SE); somatic embryos (Se’s).


2006 ◽  
Vol 1 (3) ◽  
pp. 1934578X0600100
Author(s):  
Bishnu P. Chapagain ◽  
Vinod Saharan ◽  
Dan Pelah ◽  
Ram C. Yadav ◽  
Zeev Wiesman

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.


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