scholarly journals Coconut Micropropagation in Mexico using Plumule and Floral Explants

CORD ◽  
2016 ◽  
Vol 32 (2) ◽  
pp. 6
Author(s):  
Carlos Oropeza

This paper focuses on the research efforts carried out by CICY in Mexico on micropropagation of coconut. They started during the nineties in collaboration with Wye College (UK) and ORSTOM-CIRAD (France), with the development of a protocol that was reproducible and more efficient than previous ones, based on plumule explants grown in different media based on Y3 medium added with activated charcoal, gelling agent and of particular importance growth regulators 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). Within the next decade basic research was carried out to study the process of somatic embryogenesis from plumule explants, with an approach including morpho-histological, physiological, biochemical and molecular points of view, in order to gain knowledge that could be useful to further improvement of the process. Also different practical approaches were tested including changes in the media formulation, embryogenic callus multiplication and secondary somatic embryogenesis. As a result a highly efficient protocol was developed that could potentially yield over a hundred thousand somatic embryos from a single plumule explant. Embryos were able to germinate and convert to plantlets, that after planting, successfully grew to sexual maturity and fruit production. This protocol is currently being scaled up to a semi-commercial level. Also within the past five years, a protocol using rachilla explants has been developed for the production of embryogenic callus and its multiplication, and embryos produced were able to germinate and convert to plantlets, setting the basis to develop a process for massive  propagation of coconut, such as the one already developed using plumule explants.

1984 ◽  
Vol 62 (6) ◽  
pp. 1245-1249 ◽  
Author(s):  
L. S. Kott ◽  
K. J. Kasha

Somatic embryogenesis was induced in callus previously initiated from immature embryos of barley. These cultures ranged in age from 6 weeks to 30 months. Embryoids were readily initiated from homogenized suspension-grown aggregates when plated on modified B5 media with 2,4-dichlorophenoxyacetic acid. Low concentrations (0.1 and 0.05 mg∙L−1) of abscisic acid promoted further maturation of embryoids, while gibberellic acid (1 mg∙L−1) and kinetin (0.1 mg∙L−1) were used in the media to encourage embryoid germination. The development of somatic embryoids from initiation through maturation and germination is described.


2016 ◽  
Vol 19 (2) ◽  
pp. 168
Author(s):  
Toni Herawan ◽  
Mohammad Na'iem ◽  
Sapto Indrioko ◽  
Ari Indrianto

Sandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood.


2015 ◽  
Vol 16 (1) ◽  
pp. 17
Author(s):  
Wardatus Sholeha ◽  
Bambang Sugiharto ◽  
Dwi Setyati ◽  
Parawita Dewanti

Induction of somatic embryogenesis in sugarcane requires the composition Plant Growth Hormone (PGH) appropriate. Utilizing of PGH (2,4-D and kinetin) is expected to induce sugarcane somatic embryogenesis. The purpose of this study was to obtain the concentration of 2,4-D and kinetin that effective for the multiplication of sugarcane var. NXI 1-3 through somatic embryogenesis. This study consists of four stages: callus induction, callus proliferation, regeneration of shoots and encapsulation. The plant material used is a spindle leaf sugarcane var. NXI 1-3. Callus induction used 2,4-D with different concentration (2 ppm, 3 ppm and 4 ppm). Callus proliferation used 2,4-D with concentration 1 ppm and 2 ppm. Regeneration of shoots used kinetin 0,5 ppm. The results are showed that the optimal induction of embryogenic callus on medium MS + sucrose 30 g / L + CH 300 ppm + 300 ppm PVP + 2,4-D 4 ppm as indicated by the high percentage of explants forming embryogenic callus that is equal to 40% and the respective concentration 2 ppm and 3 ppm is 33,3% and 37,5%. In proliferation stage, the development callus optimal on medium MS + sucrose 30 g / L + CH 300pm + PVP 300 ppm + 2,4-D 2 ppm and formulations for regeneration shoot on medium MS + sucrose 30 g / L + kinetin 0.5 ppm. The result of encapsulation can be shaped 100 sythetic seed. Keywords: Somatic embryogenesis, spindle leaf, kinetin, 2,4-D


1986 ◽  
Vol 16 (3) ◽  
pp. 664-668 ◽  
Author(s):  
P. Krogstrup

Embryos from imbibed ripe seeds and cotyledon expiants of 7-day-old Norway spruce (Piceaabies (L.) Karst.) seedlings produced the early stages of somatic embryogenesis. Using a modified Murashige and Skoog medium, a whitish, glossy callus was induced consisting of translucent cells embedded in a mucilaginous cloudy matrix. This embryogenic callus formed on the surface of explants treated first with N-6-benzyladenine followed by 2,4-dichlorophenoxyacetic acid + 6-furfurylaminopurine (kinetin) + N-6-benzyladenine. Transfer of this callus to media lacking growth regulators resulted in the formation of numerous bipolar embryoids with suspensorlike structures. These embryoids strongly resembled repressed embryos in polyembryonic seeds.


1986 ◽  
Vol 16 (2) ◽  
pp. 420-422 ◽  
Author(s):  
S. A. Merkle ◽  
H. E. Sommer

Tissue cultures of yellow poplar (Liriodendrontulipifera L.) were initiated from immature and mature zygotic embryos. Nodular embryogenic callus developed from a low percentage of the cultures initiated from immature embryos on solid media supplemented with 2,4-dichlorophenoxyacetic acid, 6-benzyladenine, and casein hydrolysate. Embryoids differentiated from these culture lines within 1 month following transfer of embryogenic callus to hormone-free solid media. Although most embryoids appeared abnormal, embryoids with well-formed cotyledons and radicles were capable of developing into normal plantlets.


1993 ◽  
Vol 23 (5) ◽  
pp. 835-838 ◽  
Author(s):  
Paula M. Pijut

Immature cotyledonary explants excised from developing fruits of Juglanscinerea L. were cultured in vitro to induce regeneration of somatic embryos. Somatic embryos were initiated directly on cotyledons collected 9 weeks postanthesis and cultured on a Driver and Kuniyuki medium supplemented with 250 mg/L L-glutamine, 0.01 mg/L indole-3-butyric acid, 1 mg/L 6-benzylaminopurine, and 2 mg/L kinetin for 3 weeks, prior to transfer to hormone-free Driverand Kuniyuki medium. Embryogenic callus was initiated on explants collected 8–11 weeks postanthesis and cultured on two different media formulations containing 0.25 mg/L 6-benzylaminopurine and 2 mg/L 2,4-dichlorophenoxyacetic acid for 3 weeks, prior to transfer to hormone-free media. Globular to mature somatic embryos were differentiated, and conversion of somatic embryos into whole plants was incomplete. Competence of embryogenic callus was maintained for 1 year with regular subculturing on hormone-free media.


Author(s):  
Ketut Anom Wijaya ◽  
Adi Prawoto ◽  
Syrril Ihromi

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance


1990 ◽  
Vol 68 (2) ◽  
pp. 236-242 ◽  
Author(s):  
F. M. Tremblay

White spruce (Picea glauca) embryogenic callus was obtained using 3- to 11-year-old seeds as a source of zygotic embryos. They were cultured on half-strength Litvay's medium supplemented with 10 μM 2,4-dichlorophenoxyacetic acid, 5 μM benzylaminopurine, 1 g/L casein hydrolysate, 500 mg/L glutamine, and 1% sucrose. The frequency of induction of embryogenic callus was significantly improved by incubation at 25 °C and by a 4-h imbibition of the seeds. The yield of embryogenic callus was significantly affected by the geographic provenance of the seeds and by their number of years in storage. A significant correlation was also found between the yield of embryogénie callus and the percentage of germination of the seedlot used. Even after 11 years of storage, 40% of the zygotic embryos could produce an embryogenic callus when dissected from seeds with a high germination rate. Somatic embryos were matured after transfer onto an embryo development medium composed of the same medium but including 6% sucrose, 1 μM 2,4-dichlorophenoxyacetic acid, and 5 μM kinetin. The somatic embryos developed further under in vitro conditions and were then transplanted into soil. The somatic embryoderived plantlets established in the greenhouse were similar to control plantlets obtained from germinated seeds. Mature embryos from stored seeds were shown to constitute a valuable source for white spruce somatic embryogenesis.


Author(s):  
Fitria Yuliasmara ◽  
Aris Wibawa ◽  
Adi Prawoto

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance


Aquilaria malaccensis Lam. and Aquilaria subintegra Ding Hou belong to the family of Thymelaeaceae which is commonly known as gaharu or agarwood. It is a commercially important tree and identified as a potential aromatic plant. The overwhelming responses in the lodging sector reduce gaharu species in the forest. Mass propagation through plant tissue culture technology will substitute this problem. The present study was conducted to investigate the embryogenic callus induction between these two species. The most optimum sterilization method for both species was sodium hypochlorite 5.0% which gave the highest percentage of aseptic culture (95%) with the absence of tissue browning. The leaves of both species were cultured on Murashige and Skoog, (1962) (MS) media supplemented with combination of various concentrations of 6-benzylaminopurine (BAP) (0.5, 1.0, 2.0 and 2.5 mg/L) and 2,4-dichlorophenoxyacetic acid (2, 4-D) (0.5, 1.0, 1.5 and 2.0 mg/L) and kept under dark condition. The explants produced embryogenic, white and compact callus at the end cut of the explants after two weeks of culture in all treatments. The highest frequency of embryogenic callus formation was observed in explants cultured on 2.0 mg/L BAP and 0.5 mg/L 2,4-D for both species. From the present study, the optimum sterilization technique and embryogenic callus induction for A. malaccensis Lam. and A. subintegra were established.


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