Improvement of soybean somatic embryo development and maturation by abscisic acid treatment

2000 ◽  
Vol 80 (2) ◽  
pp. 271-276 ◽  
Author(s):  
Lining Tian ◽  
Daniel C. W. Brown
Keyword(s):  
Somatic Embryogenesis ◽  
Abscisic Acid ◽  
Embryo Development ◽  
Development Stage ◽  
Normal Embryo ◽  
Maturation Stage ◽  
Globular Stage ◽  
Solid Media ◽  
Stage Embryo

Recovery of tissue culture-derived plants through somatic embryogenesis is a useful system for genetic engineering of soybean. The effect of abscisic acid (ABA) on soybean somatic embryogenesis, development, and maturation was investigated. ABA at 1, 10, 50, 100, and 500 µM were applied at different stages of embryo development; namely, at the globular stage in suspension culture, at the development stage and at the maturation stage on solid media. ABA promoted embryo growth and development when applied at the globular stage. Embryo size, after 15 d and after 1 mo on development medium, was significantly greater than that without exposure to ABA. ABA promoted normal embryo morphogenesis and 62% more normal embryos developed when embryos were treated with ABA at the globular stage. ABA treated-embryos showed an increased tolerance to partial desiccation (from 24% to 78%) and exhibited an increased germination capability relative to non-ABA-treated controls (54% versus 8%). Somatic embryos appeared to undergo a decreasing sensitivity to ABA during maturation. ABA did not show an effect when applied during embryo development and maturation stages. A protocol for more normal embryo formation and improved embryo germination is reported. Key words: Glycine max, somatic embryogenesis, in vitro culture

scholarly journals Influence of Abscisic Acid and Sucrose on Somatic Embryogenesis in CactusCopiapoa tenuissimaRitt. formamostruosa

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
J. Lema-Rumińska ◽  
K. Goncerzewicz ◽  
M. Gabriel
Keyword(s):  
Somatic Embryogenesis ◽  
Abscisic Acid ◽  
Somatic Embryos ◽  
Sucrose Concentration ◽  
Turgor Pressure ◽  
Direct Somatic Embryogenesis ◽  
Fresh Weight ◽  
High Concentration ◽  
Globular Stage ◽  
Indirect Somatic Embryogenesis

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.

scholarly journals GENE EXPRESSION DURING INDIRECT SOMATIC EMBRYOGENESIS OF PLANTS

2004 ◽  
Vol 42 (2) ◽  
Author(s):  
Tammy Estabrooks ◽  
Zhongmin Dong
Keyword(s):  
Gene Expression ◽  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Current Literature ◽  
Molecular Mechanisms ◽  
Indirect Somatic Embryogenesis ◽  
Experimental Tool ◽  
Plant Embryo ◽  
Plant Embryo Development

Somatic embryogenesis is the process by which somatic cells are induced into an embryogenic state, followed by differentiation into embryos. Somatic embryogenesis, in addition to being a method of propagation, can serve as an experimental tool for research into plant embryo development. This is a review of the current literature on in vitro plant somatic embryogenesis and the molecular advances made to identify genes expressed during the various stages of this process. Some factors hindering the elucidation of the molecular mechanisms underlying somatic embryogenesis are discussed.L’embryogenèse somatique est le processus par lequel les cellules somatiques passent à l’état embryogène et se différencient en embryons. En plus de constituer une méthode de propagation, elle peut servir d’outil expérimental de recherche pour développer des embryons de plantes. Le présent document est une revue de la documentation sur l’embryogenèse somatique végétale in vitro et sur les progrès réalisés à l’échelle moléculaire pour identifier les gènes exprimés au cours des divers stades du processus. On examine aussi certains facteurs qui rendent difficile l’élucidation des mécanismes moléculaires de l’embryogenèse somatique.

14 EQUINE CLONING AND EMBRYO AGGREGATION: EFFECT OF BOVINE, PORCINE, FELINE AND EQUINE OOPLAST

2012 ◽  
Vol 24 (1) ◽  
pp. 118
Author(s):  
A. Gambini ◽  
J. Jarazo ◽  
A. De Stefano ◽  
F. Karlanian ◽  
D. Salamone
Keyword(s):  
Embryo Development ◽  
Metaphase Plate ◽  
Donor Cell ◽  
Development Stage ◽  
Blastocyst Stage ◽  
Chi Square ◽  
Cell Stage ◽  
Aggregation Effect ◽  
Group I

The low number of horse slaughterhouses is one of the reasons for the limited availability of horse oocytes for research in cloning. The aim of our study was to assess the capability of equine, bovine, porcine, or feline ooplast to produce cloned embryos when equine cells are used as donor nuclei and to evaluate if embryo aggregation improves their development. Oocytes from mentioned species were collected from ovaries derived from slaughterhouses, except for cat ovaries that were obtained from ovariectomized queens. Oocytes were matured in TCM199 supplemented following standard protocols for each species. After maturation, cumulus and zona pellucida were removed. Enucleation was performed by aspiration of the metaphase plate under ultraviolet light. Donor cell and ooplast were attached by phytohemagglutinin treatment and then electrofused. Activation protocols were ionomycin for 4 min, except for porcine, which were electrically activated, followed by culture in 1.9 mM 6-DMAP for bovine, feline and porcine, except for equine: 1 mM 6-DMAP with 5 mg mL–1 of cycloheximide. Reconstructed embryos (RE) were cultured in SOF in the well of well system in 2 different groups: only one RE per well (1X) and three RE per well (3X, aggregated embryos, AE). Blastocysts derived from homospecific clones were transferred to synchronized mares. Cleavage and maximum development stage achieved of all experimental groups were assessed. In vitro development was compared using the chi-square test. In group 1X, a total of 64, 49, 38 and 145 RE were performed for porcine, bovine, feline and equine, respectively and in group 3X, 88, 48, 48 and 195 RE. Cleavage of cloned embryos ranged from 67 to 87%. Aggregated of homospecific equine clones showed the highest blastocyst rates (1X: 5.5%, 3X: 34%) and after embryo transfer (4 recipients for each group), an ongoing pregnancy (day 300, at the time of submission) was only achieved with aggregated embryo confirming the positive effect of embryo aggregation in these clones. The stages with higher developmental arrest of heterospecific nonaggregated embryos were 2 to 4 cells for porcine ooplast (23/64, 36%) and 4 to 8 cells for bovine and feline ooplast (37/49, 75% and 18/38, 47%, respectively). Blastocyst stage was only reached using feline ooplast (group I: 2/38, 5.26% and group II: 2/16, 12.5%). Heterospecific aggregated clones were able to achieve 16-cell stage, showing statistic differences compared with group 1X. As we reported previously, embryo aggregation shows benefits for homospecific equine clones, although more studies are needed to clarify if aggregation of heterospecific clones has the same effect. All heterospecific ooplasm was able to support embryo development. The stage of major developmental arrests was similar to embryonic genomic activation stage. Our results suggest that cat oocyte seems to be the best receptor to support equine cloned embryo development.

257. Activation of a calcium-activated chloride channel by paf is required for normal preimplantation mouse embryo development in vitro

2008 ◽  
Vol 20 (9) ◽  
pp. 57
Author(s):  
Y. Li ◽  
M. L. Day ◽  
C. O.'Neill
Keyword(s):  
Embryo Development ◽  
Mouse Embryo ◽  
Preimplantation Embryo ◽  
Platelet Activating Factor ◽  
Outward Current ◽  
Normal Embryo ◽  
Cell Stage ◽  
Preimplantation Embryo Development ◽  
Cl Channel

Platelet activating factor (paf) is an autocrine survival factor for preimplantation embryo. Binding of paf to its receptor activates PI3kinase, causing an IP3-dependent release of Ca2+ from intracellular stores as well as activation of Ca2+ influx via a dihydropyridine-sensitive Ca2+ channel. These actions result in the generation of a defined intracellular calcium ([Ca2+]i) transient in the 2-cell embryo[1]. By using combined whole-cell patch-clamp and real-time [Ca2+]i analyses, we have shown that paf also induces a concomitant hyperpolarisation of the membrane potential in 2-cell embryos, accompanied by an increased net outward ion current. Both the membrane hyperpolarisation and outward current were dependent upon the occurrence of the paf-induced [Ca2+]i transient[2]. The aim of this study was to investigate the characteristics of the paf-induced outward current in 2-cell embryos and to assess whether it has a role in normal mouse preimplantation development. We show that: (1) removal of extracellular anions or treatment with niflumic acid (NFA, 100 μM, a Ca2+-activated Cl- channel blocker) prevented activation of the outward current by paf but had no effect on the paf-induced [Ca2+]i transient; and (2) The culture of embryos with NFA (100 μM) from the 1-cell to late 2-cell stage significantly reduced their development to the blastocyst stage (P < 0.001), but treatment with NFA from the late 2-cell stage had no effect on development. The results show that paf induces an increase in [Ca2+]i which in turn activates a Ca2+-activated Cl- channel. The activity of this NFA-sensitive channel during the zygote to 2-cell stage is required for normal embryo development. (1) C. O’Neill (2008) The potential roles of embryotrophic ligands in preimplantation embryo development. Hum Reprod Update 14:275–288. (2) Y. Li, M.L. Day & C. O’Neill (2007) Autocrine activation of ions currents in the two-cell mouse embryo. Exp Cell Res. 313:2785–2794.

Abscisic Acid and Sucrose Control of Velvetleaf (Abutilon theophrasti) Ovule Development in Vitro

Weed Science ◽  
1984 ◽  
Vol 32 (6) ◽  
pp. 798-801 ◽  
Author(s):  
Laura K. Thompson ◽  
Gerald R. Leather ◽  
Maynard G. Hale
Keyword(s):  
Abscisic Acid ◽  
Embryo Development ◽  
Parental Control ◽  
Abutilon Theophrasti ◽  
Ovule Development ◽  
Precocious Germination ◽  
Development In Vitro ◽  
Days After Anthesis

The culture of ovules excised from velvetleaf (Abutilon theophrasti Medic., ♯4 ABUTH) capsules 5 days after anthesis was used to measure the effects of abscisic acid (ABA) and sucrose on embryo development and prevention of precocious germination. ABA at 1 × 10-7 M combined with 6% sucrose in the medium for the first 14 days of culture increased embryo development but prevented precocious germination. Higher concentrations of ABA inhibited embryo development. Without ABA, precocious germination increased directly with the concentration of sucrose in the medium, and embryos died. In vivo, ABA reached its highest concentration in ovules 5 days after anthesis but was undetectable after 16 days. Parental control of embryo development may involve ABA and an increasing concentration of osmoticum as seeds dehydrate during maturation.

scholarly journals The Effect of Different Factors upon the in vitro Propagation in Quercus robur and Q. frainetto by Somatic Embryogenesis

2011 ◽  
Vol 39 (1) ◽  
pp. 288 ◽  
Author(s):  
Adrian Ioan TIMOFTE ◽  
Doru PAMFIL ◽  
Magdalena PALADA-NICOLAU ◽  
Claudia Simona TIMOFTE
Keyword(s):  
Somatic Embryogenesis ◽  
Significant Influence ◽  
Quercus Robur ◽  
Culture Medium ◽  
Clonal Propagation ◽  
Germination Percentage ◽  
Development Stage ◽  
Ex Situ ◽  
Conservation Of Genetic Resources

The somatic embryogenesis is an advanced method for clonal propagation and a useful tool for ex situ conservation of genetic resources. In this paper, the results of an experiment to investigate the influence of development stage of explants and culture medium on the germination percentage in two oak species (three provenances of Quercus robur and two provenances of Q. frainetto), are presented. A high significant influence of the development stage of explants and a significant influence of the interaction provenance x stage on the germination percentage were recorded for Q. robur explants, whilst no significant differences between the germination percentages against the nutritive media used were fould for both oak species.

Transient expression of a reporter gene changes significantly during somatic embryogenesis in alfalfa

2000 ◽  
Vol 80 (4) ◽  
pp. 765-771 ◽  
Author(s):  
Lining Tian ◽  
Daniel C. W. Brown ◽  
John Webb
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Somatic Embryogenesis ◽  
Medicago Sativa ◽  
Transient Expression ◽  
Embryo Development ◽  
Reporter Gene ◽  
Reporter Gene Expression ◽  
Maturation Stage ◽  
Embryogenic Cell

Transient expression of the β -glucuronidase (GUS) gene driven by a 35S constitutive promoter was investigated during somatic embryo development in alfalfa (Medicago sativa L.) following micro-projectile bombardment. The level of gene expression varied with the different developmental stages and changed dramatically within the stages. During embryogenic callus induction (stage I), expression was low in explants freshly transferred to culture, but increased when cells started to divide and show competence for somatic embryogenesis. Gene expression, however, decreased when extensive callus formation began and reached the lowest observed level when callus was fully developed. Reporter gene expression was consistently low during embryogenic cell proliferation (stage II). In contrast, high levels of reporter gene expression were detected in non-embryogenic suspension cultures during cell proliferation. Extremely low reporter gene expression was detected in embryonal tissues and young embryos at the embryo development and maturation stage (stage III). Expression increased as the somatic embryos developed and reached the highest level when the embryos reached maturity. Transient expression levels were similar in excised petiole tissue across 10 alfalfa genotypes, which included both diploid and tetraploid genotypes. In general, reporter gene expression was found to be higher in more organised mature tissues and organs than in less-organised or young tissues. The developmental status of the cells and tissues appears to be an important factor in the degree of transient expression of the introduced gene. Changes of gene expression during embryogenesis and factors affecting gene expression are discussed. Key words: GUS expression, Medicago sativa, bombardment, forage

scholarly journals Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica

2020 ◽  
Vol 21 (12) ◽  
Author(s):  
FITRIA ARDIYANI ◽  
Edy Setiti Wida Utami ◽  
HERY PURNOBASUKI ◽  
SENJA APRILIA PARAMITA
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Somatic Embryos ◽  
Economic Value ◽  
Genetic Material ◽  
Clonal Plants ◽  
Ms Medium ◽  
Cotyledonary Stage ◽  
Coffea Liberica

Abstract. Ardiyani F, Utami ESW, Purnobasuki H, Paramita SA. 2020. Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica. Biodiversitas 21: 5829-5834. Coffea liberica is an important and potentially commercial plant with a high economic value from the Coffea genus. Therefore, the availability of planting material is needed to increase productivity and ensure the sustainability of its farming. Somatic embryogenesis is a powerful propagation method used to produce clonal plants from limited genetic material. In the present research, we have shown that C. liberica could be successfully regenerated in vitro via somatic embryogenesis from leaves derived embryogenic callus. These calli were cultured on Murashige Skoog (MS) medium added with 1 mgL-1 BAP or in combination with 2.4 D (0.5, 1.0, 1.5 and 2 mgL-1) for embryo development induction. Furthermore, the medium containing only BAP was best for embryo development induction after culturing for 12 weeks, with the highest number of cotyledonary stage embryos (17.8%) and producing a total of embryo (20.2). Following cotyledonary stage embryo were cultured on new MS medium containing 0.5 mgL-1 BAP, 0.5 mgL-1 IAA, 0.5 mgL-1 NAA only, and 0.5 mgL-1 BAP in combination with 0.5 mgL-1 IAA or 0.5 mgL-1 NAA. Interestingly, the results showed that cotyledonary stage embryos were converted into complete plants at all treatment, but the MS medium containing 0.5 mgL-1 BAP was found to be the most effective in promoting regeneration with 2.6 leaves per-plantlet and height of 5.2 mm. Based morphological analysis confirm that the development of somatic embryo from leaves-derived calli of Coffea liberica started with the formation of embryo globular, heart, torpedo, cotyledonary stages, and finally conversion of cotyledonary embryo into complete plant.

scholarly journals Somatic embryogenesis of Sandalwood (Santalum album L.)

2016 ◽  
Vol 19 (2) ◽  
pp. 168
Author(s):  
Toni Herawan ◽  
Mohammad Na'iem ◽  
Sapto Indrioko ◽  
Ari Indrianto
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Native Species ◽  
Economic Value ◽  
Dichlorophenoxyacetic Acid ◽  
Santalum Album ◽  
The Media ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Santalum Album L

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.

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