Callus Embryogenesis Research Articles

Embryogenesis Callus Induction of Carica pubescens Using Divine Smoke Particulates Containing Amino Acids

Carica pubescens is one of plants that can live in the highlands which has a potential as medicinal plant to treat many diseases. The plant propagation is still done conventionally. Embryogenic callus culture is the alternative. The success of callus culture depends on the role of auxin growth regulator (2,4-D) and Amino Acids. The aim of this study is to determine the effet of amino acids through Divine smoke particulation to the induction of embryogenesis callus carica pubescens. This research is an experimental study with Complete Randomized Design (CRD) method. It has 7 treatments consist of control and the Amino Acids (Threonine, Tryptophan, Serine, Histidine, Asparagine, and Methionine). The result of this study showed a significant effect of amino acids through Divine smoke particulation to the induction of embryogenesis callus Carica pubescens which is grown on ½ MS media + 3 mg/L of 2,4-D. Tryptophan is the fastest treatment to induce callus which is grown 24 days after planting (DAP). Serine influences to the percentage of callus area in explants is 98,8% and the fresh weight of callus 0,6733 gr. The observation of morphology and anatomy embryonic callus color is yellowish white, friable texture and has a large cell nucleus

Identification of genes encoding Nuclear factor-YB in cassava (Manihot esculenta Crantz) using bioinformatics

Nuclear factor-Y (NF-Y) transcription factor can be found in most of plant species and is known to play important role in plant growth, development and stress response. In this study, 17 genes encoding NF-YB subunit were identified in cassava (Manihot esculenta Crantz). These genes were located in chromosomes with an uneven ratio. Analysis of gene structure indicated that MeNF-YB genes were varied in length and exon/intron organization. Genes in the same clade shared similar exon/intron structure. Based on available microarray database, most of NF-YB genes were up-regulated in at least 1 major tissue. Interestingly, MeNF-YB12 was highly expressed in shoot apical meristem, while MeNF-YB2 was specifically accumulated in friable embryogenesis callus. Additionally, some genes, such as MeNF-YB4/8/13/5 were found to be strongly expressed in friable embryogenesis callus. Our results strongly indicated that NF-YB genes in cassava might be associated with various growth and developmental processes in meristem tissues. 

Morphological Variation of Somatic Embryos of Coffea arabica L. During Some Sub-Culture Periods

One of factors that affects the success of a plant cultivation using somatic embryogenesis method is the formation of somatic embryos from embryogenesis callus. This research aimed to study the effect of sub-culture period on quality and quantity of the somatic embryos of Coffea arabica. This research used explants of somatic embryos of Arabica coffee obtained from the leaves of 2K Andungsari clone. The embryos were taken during embryogenes is callus phase using Murashige-Skoog culture media added with B5 vitamin and auxin hormone (2,4dichlorophenoxy acetic acid) 0.5 mg/L and sitokinin (benzyl amino purin) 1 mg/L. Observation on somatic embryos obtained from the sub-culture period of 3, 6, 9, 12 and 15 weeks. The parameters observed in this study included quantity and quality of the somatic embryos during each sub-culture period. Observations on quantity of the somatic embryos were conducted based on number of embryos per cluster, while quality was measured from the percentage of normal embryos, histological analysis, and morphological measurement on weight and size of the normal embryos. The result showed that the best quantity of somatic embryos was obtained from the sub-culture period of nine weeks with 18.4 somatic embryos per cluster. The best quality embryos were also obtained in the sub-culture period of nine weeks with the percentage of normal embryos 71.4%. Histological analysis carried out on the somatic embryos obtained from sub-culture of three weeks period showed that the cells of the embryos were formed by living and solid cells which nucleus were clearly seen in the center of the cell, indicating that the embryos were formed by young tissues. Data of morphological parameters showed that normal embryos during the sub-culture period 3 to 15 weeks weighed around 0.23–0.78 mg and length of around 0.18–0.25 cm. The data can be used to predict number of explants and required time to produce certain number of normal embryos

Enhancement of Regeneration Efficiency through Callus Induction Media Using 2,4-Dicholorophenoxyacetic acid in Indica Rice (Oryza sativa L. var. Ciherang)

Indica rice variety Ciherang is the most planted variety in Indonesia, but the micropropagation technique is restricted because it is known has low regeneration frequency and included as recalcitrant cultivar for tissue culture and tranformation activities. One of solution to resolve that problem is developing a technique of somatic embryogenesis in callus of ciherang rice cultivar. The aims of study were to determine medium composition for inducting embryogenic callus in Ciherang rice and to know the effectivity of rice regeneration using callus as explant. The methods were included induction of embryogenic callus in callus induction media (CIM) containing MS basal, Proline 600 mg/l, Casein Hidrolisat 300 mg/l, phytagel 2,5 g/l, BAP 0.25 mg/l, sukrosa 30 %, and 2,4-D in different concentration, from 2,4-D 0 mg/l as control (CIM 1), 2,4-D 2 mg/l (CIM 2), 2,4-D 3 mg/l (CIM 3), and 2,4-D 4 mg/l (CIM 4). It then be continued to regenerate the calli in RM 1 medium containing MS basal, NAA 0,2 mg/l, Kinetin 2 mg/l, Agarose 10 g/l, and sukrosa 30 %, pH 5,8 for six days in dark and RM2 medium containing MS basal , NAA 0,2 mg/l, Kinetin 2 mg/l, Agarose 8 g/l, sukrosa 30 %, pH 5,8 in light room. Results showed CIM 4 medium using 2,4-D 4 mg/l gave optimum result in calli induction with procentage 57,63% and CIM 3 using 2,4-D 3 mg/l gave optimum result in embryonic calli induction with procentage 53,63%. Result of embryonic calli gave regeneration frequency procentage is 9,6%. The total planlet obtained after five weeks old in regeneration medum were 11 planlets ready for aclimatization.
 Keywords: Somatic embryo, Indica rice (Oryza sativa L. var. Ciherang), Callus

Biological Network Analysis of Genes Involve in Embryogenesis of Oil Palm using ClueGO

<p class="Els-Abstract-text">The application of DNA sequencing technologies has a major impact on molecular biology, especially in understanding genes interaction in a certain condition. Due to a large number of genes produced by this high-throughput technology, a proper analysis tool is needed for data interpretation. ClueGO is a bioinformatics tool, an easy to use Cytoscape plug-in that strongly improves biological function interpretation of genes. It analyzes a cluster or comparing two clusters and comprehensively visualizes their group functions. This tool is applied to identify biological networks of genes involved in embryogenesis of oil palm, the most critical phase in oil palm tissue culture process. Two ESTs sequencing data from the GenBank database under accession number EY396120-EY413718 and DW247764-DW248770 were used in this study. Fifty-two and one hundred eight groups of genes were identified using biological process in Gene Ontology setting from the database of EY396120-EY413718 and DW247764-DW248770, respectively. Thirty-one groups of genes were consistently occurred in both ESTs. According to the literature, these genes play an important role in cell formations and developments, stresses and stimulus responses, photosynthesis and metabolic processes that indicate the involvement of these groups of genes in oil palm embryogenesis processes. ClueGO is the appropriate tool to analyze a large data set of genes in a specific condition, such as embryogenesis of oil palm.</p><div><p class="Els-keywords"><em> </em></p></div><strong>Keywords:</strong> callus embryogenesis; Cytoscape plug-in; DNA sequencing; expressed sequence tag; KEGG pathways

Short Communication: Organogenesis and somatic embryogenesis in callus derived from HBsAg-transgenic tomato mutant

Guan, Z.-j., Guo, B., Huo, Y.-l., Guan, Z.-p., Dai, J.-k. and Wei, Y.-h. 2012. Short Communication: Organogenesis and somatic embryogenesis in callus derived from HBsAg-transgenic tomato mutant. Can. J. Plant Sci. 92: 747–756. In the process of hepatitis B surface antigen (HBsAg) gene transformation, a transgenic cherry tomato (Lycopersivon esculentum Mill.) mutant N244 was occasionally obtained. Based on its obvious variation in ploidy and phenotypic characteristics, morphogenesis and development in the mutant in vitro were studied by cytological analysis and ultrastructural examination. The adventitious shoot primordia first emerged in the mutant N244 leaf explants after 14 d of culture, and the somatic embryo pathway developed after 35 d. Transmission electron microscopy and scanning electron microscopy further indicated that during the explant's development, there were significant changes in morphology and quantity of some organelles in N244 callus cells. On the 7th day of culture, there was little cy...

Influences of the coconut water and sucrose on the embryogenic callus induction and the clonal embryo formation of Phalaenopsis amabilis (L.) Blume

Phalaenopsis amabilis (L.) Blume (Orchidaceae) is one of many commercial valuable orchids as cut flowers and potted plants throughout the world. Considerable difficulties have been encountered in the clonal propagation of this orchid due to the characteristics ununiform and the limited plantlets number. In this paper, an established method of the Phalaenopsis amabilis propagation through embryogenesis callus was described. The in vitro leaves emerging from the flower stalk nodes were used for protocorm-like bodies (PLBs) induction. These PLBs were used for the embryogenesis callus (callus) induction. About 1455 mg calli without any morphological change were harvested from 70 mg calli on 30 ml MS medium supplemented with 2 mg/l BA, 0.5 mg/l NAA, 20% (v/v) CW, 1 g/l activated charcoal (AC), 30 g/l sucrose and 9 g/l agar after 5 culture weeks of. On the same medium without sucrose, 70 mg calli produced 1841 mg PLB (approximately 200 PLBs) after 5 weeks of culture; data also showed that PLB fresh weight and quantity in the nylon film culture system (NF) were almost equal to that in conventional system (C). The hyponex medium supplemented with 0.5 mg/l NAA, 2 mg/l BA, 30 g/l sucrose, 1 g/l AC and 15% (v/v) CW was used for the plantlet regeneration from PLB. The results showed that plantlet quantity obtained in NF system (185 plantlets) was higher than that in C system (109 plantlets) after 2 culture months. Phalaenopsis amabilis plantlets in NF system had much high survival rate (95%) than that in C system (70%) after 4 weeks transferred in the greenhouse.

Indirect Somatic Embryogenesis from Petal Explant of Endangered Wild Population of Fritillaria imperialis

Fritillaria imperialis is an endangered bulbous plant and therefore in vitro micropropagation of this plant will have a great importance for germplasm conservation and commercial production. Petal explants, for the first time, were cultured on media containing various concentrations of plant growth regulators. In addition, the effects of cold pretreatment and light on induction and regeneration of somatic embryogenesis trough callus were studied in detail. Cold pretreatment had inhibitory effects on somatic embryogenesis pathway. Among the different combinations of 6-Bnzylaminopurine (BAP), alpha-naphthaleneacetic acid (NAA) and indole-3-aceticacid (IAA) tested, B5 medium supplemented with 0.1 mg L(-1) BAP + 0.6 mg L(-1) NAA + 0.4 mg L(-1) IAA was the best treatment for bulblet production (6 bulblets per somatic embryogenesis callus). This research presents petal as a reliable material for micropropagation and germplasm conservation of Fritillaria imperialis.

Somatic Embryogenesis of Rosa Hybrida L.

ABSTRACTWe present two regeneration protocols via somatic embryogenesis in three cut roses—Rosa hybrida cvs. “Anny”, “Trimontsium” and “Saltze Gold”. The first regeneration protocols are based on regeneration via somatic embryogenesis in callus from leaf and petiole segments of Rosa hybrida cvs. “Trimontsium” and “Saltze Gold”. The other protocol is via somatic embryogenesis in callus culture from petals of Rosa hybrida cv. “Anny”. Both regeneration protocols were developed on solid medium under subculturing on every 35–40 days. MS-medium including MS (1962—Murashige & Skoog) or QL (Quorin & Lepoivre) vitamins were used. For the embryo induction of callus derived from leaf and petiole explants were used BAP, NAA and kinetin, while for the combination of dicamba, kinetin and 2-ip are the most efficient for the embryo induction of callus derived from petals. A part of these embryos was converted to intact plants and an other part had been multiplicated and maintained in vitro for two years in order to check their regeneration capacity. All the regenerants that had been obtained from the three cut roses were adapted under in vitro conditions and cultivated in a greenhouse. Regenerated plantlets were morphologically true to the type of the initial donor-genotype.

Somatic embryogenesis and plant regeneration from callus cultures of Aconitum heterophyllum Wall

Plants were obtained via somatic embryogenesis in callus derived from in vitro raised leaf and petiole explants of Aconitum heterophyllum Wall. Callus was induced on a Murashige-Skoog medium supplemented with either 2,4-dichlorophenoxy acetic acid (2,4-d 1 mg l-1) and kinetin (KN 0.5 mg l-1) with coconut water (CW 10% v/v) or naphthalene acetic acid (NAA 5 mg l-1) and benzylaminopurine (BAP 1 mg l-1). Somatic embryos appeared after 2–3 months or 2 subculture passages when 2,4-d or NAA induced source of the callus was transferred to a MS medium containing BAP (1 mg l-1) and NAA (0.1 mg l-1). For successful plantlet formation, the somatic embryos were transferred to a medium containing 1/4 strength MS nutrient with indole-3-butyric acid (IBA 1 mg l-1). Alternatively, the somatic embryos were dipped in a concentrated solution of IBA for 5 min and placed on a hormone free medium. Complete plantlets were formed after 4 weeks and were transferred successfully to soil.

Origin and Ontogenesis of Somatic Embryos in Hevea brasiliensis (Euphorbiaceae)

In Hevea, currently described culture conditions allow the induction of two modes of embryogenesis in callus formed on excised portions of the internal integument of immature seeds. One mode is of unicellular origin and is transitory, only resulting in the production of globular proembryos. The other is of multicellular origin and produces embryos. Specific culture conditions appear to favor one or the other mode of development in a given genotype. The ontogenesis of embryos of multicellular origin bypasses the classical stages of zygotic embryogenesis. The structural abnormalities observed in most somatic embryos are probably responsible for the low germination rates obtained.

ORIGIN AND ONTOGENESIS OF SOMATIC EMBRYOS IN HEYEA BRASILIENSIS (EUPHORBIACEAE)

In Hevea, currently described culture conditions allow the induction of two modes of embryogenesis in callus formed on excised portions of the internal integument of immature seeds. One mode is of unicellular origin and is transitory, only resulting in the production of globular proembryos. The other is of multicellular origin and produces embryos. Specific culture conditions appear to favor one or the other mode of development in a given genotype. The ontogenesis of embryos of multicellular origin bypasses the classical stages of zygotic embryogenesis. The structural abnormalities observed in most somatic embryos are probably responsible for the low germination rates obtained.

Induction of somatic embryogenesis in leaf-derived callus of Vicia narbonensis L.

A method for the induction of somatic embryogenesis in callus cultures, using explants from mature leaves of Vicia narbonensis L., is described. Callus developed on a solid medium (Murashige and Skoog 1962), which was supplemented with low concentrations of picloram and benzylaminopurine. Subsequent culture was carried out in different liquid media (culture length four months). The gradual reduction of auxin and cytokinin concentrations, and the addition of glutamine and pyridoxal·HCl were favourable. Somatic embryos appeared on solid media without phytohormones.

Embryogenesis in callus derived from rice microspores

Differentiating calli derived from rice (Oryza sativa L.) microspores were examined histologically. Shoot and root meristems were observed to be arising by both organogenesis as well as embryogenesis. Embryoid attachment to callus (as well as other embryoids) was at the scutellum adjacent to the mesocotyl and radicle. These observations could be interpreted as an indication of the totipotent plasticity of that tissue.

Effect of Carbohydrates on Somatic Embryogenesis in Subcultured Nucellar Callus of Citrus Cultivars

Summary The effect of several carbohydrates — sucrose, galactose, lactose, glucose and fructose — at 8 to 256 mM, on growth and embryogenesis of nucellar calli from five Citrus cultivars, was studied. While galactose and lactose over a broad concentration range enhanced embryogenesis in all calli studied, sucrose supported embryogenesis only at the lower levels tested (8–32 mM with various calli). Glucose and fructose were least effective in stimulating embryogenesis. Galactose and even more so lactose were also very effective in enhancing development of large embryos. All the sugars except fructose supported good callus growth over the concentration range tested. Addition of IAA, kinetin or GA3 to galactose, especially above 32 mM sugar, resulted in varying degrees of inhibition of embryogenesis. After 10 weeks in culture, compared with 5 weeks, the inhibition caused by IAA diminished significantly.

Asexual embryogenesis in callus derived from palm inflorescences: John F. Reynolds, Experimental Agricultural Sciences, The Upjohn Company, Kalamazoo, MI 49001

Asexual embryogenesis in callus derived from palm inflorescences: John F. Reynolds, Experimental Agricultural Sciences, The Upjohn Company, Kalamazoo, MI 49001