Somatic embryogenesis in callus culture of Mussaenda erythrophylla cvs. Queen Sirikit and Rosea

Somatic embryogenesis was achieved from mid-rib and internodal calluses of Mussaenda erythrophylla L. cvs. Queen Sirikit and Rosea cultured on Murashige and Skoog basal medium containing 8.9 M BA+0.57 M IAA+10 mg l^-1 ascorbic acid. Clumps of somatic embryos were separated and grown into complete plantlets when transferred to 1/2 MS medium+37 M adenine sulphate with 2% (w/v) sucrose.     

Changes of 6-benzylaminopurine and 2,4-dichlorophenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal

Somatic embryos and embryogenic callus were initiated from immature zygotic embryos of ginseng (Panax ginseng C.A. Meyer). These somatic embryos were multiplied by adventitious (secondary and tertiary) embryogenesis and their growth and development were dependent on growth hormones in the medium. Auxins, 2,4-d, NAA, and IAA at 1.0 mg l^-1 were effective in inducing secondary and tertiary somatic embryos, which proliferated directly from the apical or cotyledonary portions of the primary somatic embryos. Single somatic embryos or clusters or embryos developed from the explanted primary embryos. Cytokinin (Kn, BA) inhibited adventitious embryogenesis. Secondary somatic embryos developed to maturation and later regenerated into plantlets in two stage process; firstly elongation of the shoot axes on MS +1.0 mg l^-1 Kn, secondly formation of root on 1.0 mg l^-1 Kn+1.0 mg^-1 GA₃ medium.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - NAA naphthaleneacetic acid - IAA in-doleacetic acid - Kn kinetin - BA benzylaminopurine - PSE primary somatic embryo - SSE secondary somatic embryo - TSE tertiary somatic embryo     

Arrest of somatic embryo development in grapevine: histological characterization and the effect of ABA,BAP and zeatin in stimulating plantlet development

In an effort to understand the causes of arrest of somatic embryo development, generally observed in grapevine (Vitis sp.), histological studies were undertaken, using two cultivars (CH76 and 41B) which differ in their ability to develop into plants. Embryos with a high conversion rate (70%; CH76) formed a well-structured and functional shoot apex between two thread-like cotyledons. In contrast, embryos with a low conversion rate (10%; 41B) formed a normal root apex but lacked a well-structured shoot apex and developed a wide range of aberrant forms in the intercotyledonary area: uncontrolled cellular proliferation, formation of adventitious buds, over-growth of cotyledonary or leaf meristems. ABA increased the conversion rate of 41B embryos from 10% to 20%, but failed to improve embryo morphology. Zeatin and BAP promoted growth of 41B somatic embryos, but generated a high level of abnormalities and failed to improve conversion rate. Applied in combination with ABA, these PGRs increased the frequency of cotyledonary embryos, but decreased the conversion rate.     

Influence of some exogenous amino acids on the production of maize embryogenic callus and on endogenous amino acid content

The effects of four exogenous amino acids (proline, glycine, asparagine and serine) on the production of maize embryogenic callus and on its endogenous amino acid content have been investigated. For this purpose, an established embryogenic line of Type 1 callus from the inbred W64Ao2 has been used. From the results it may be concluded that a concentration of proline exceeding 6 mM is negative for the production of embryogenic callus. When proline is eliminated from the medium, other amino acids tested in certain concentrations yield a percentage of embryogenic callus production that exceeds or equals that of proline. The endogenous free proline content in embryogenic callus is significantly higher than that in non-embryogenic callus regardless of proline presence in the medium. The only exception are the glycine-containing media, in which endogenous free alanine of embryogenic callus increases at the expense of endogenous free proline. This study suggest a positive role of endogenous free proline or alanine accumulation in the embryogenic callus production which might be related to an adaptation to the metabolic changes produced by in vitro culture and embryogenesis induction. Furthermore, these results indicate that treatments with amino acids that are different from proline can be used to improve the efficiency of embryogenic callus production from well established maize callus cultures.Abbreviations Ala alanine - Asn asparagine - 2,4-d 2,4-dichlorophenoxyacetic acid - EC embryogenic callus - nEC non-embryogenic callus - Gaba gamma-aminobutyric acid - Glu glutamic acid - Gly glycine - Pro proline - Ser serine     

On Paraptychostomum almae N. G., N. Sp., a Commensal Ciliate from the Digestive Tract of Oligochaetes of the Cameroons, in a New Subclass Hysterocinetia

ABSTRACT. Morphological and ultrastructural studies on a new ciliate, Paraptychostomum almae , from the digestive tract of an oligochaete ( Alma emini ) from the Cameroons are carried out. The flattened cell has a large size; its left lateral face bears an anterior thigmotactic zone that includes seven-nine short kinetal segments. The somatic cortex is composed of flattened alveoli, a thin epiplasm and a microfibrillar ecto-endoplasmic boundary. Kineties are made of monokinetids, each particularly characterized by a long anteriorly directed kinetodesmal fiber, and a hyperdivergent postciliary ribbon. The postero-ventral buccal apparatus consists of a short peristome and a deep longitudinal infundibulum. The paroral organelle is a long stichodyad. The three adoral organelles are of different types: ADI and AD3 are of the membranoid type, respectively with two and one rows of ciliated kinetosomes; AD2 is of the peniculus type with six-seven rows of ciliated kinetosomes. A microfibrillar network with nodes arises from all the buccal kinetosomes and extends under the naked wall. Mitochondria are small and numerous and dispersed throughout the whole cell. The existence of an AD2 with more than two rows of kinetosomes warrants the creation of the new genus Paraptychostomum and a new family, Ptychostomatidae. The presence of a distinct ecto-endoplasmic boundary and of somatic kinetids exclusive without transversal dense tractus, hyperdivergent postciliary ribbons, and dispersed numerous mitochondria, added to particularities of the stomatogenesis, allow us to clearly separate hysterocinetians from the scuticociliates and to set up for them the new subclass Hysterocinetia, within the class Oligohymenophorea, with a single new order Hysterocinetida.     

Amplified somatic embryogenesis from male flowers of triploid banana and plantain cultivars (Musa spp.)

Summary Somatic embryogenesis and plant regeneration of banana and plantain cultivars (Musa spp.) were obtained by culturing young male flowers. Multiplication and maintenance of embryogenic cultures were achieved by culturing somatic embryos in a temporary immersion system (SIT). A multiplication rate of 40 allowed us to obtain more than 6000 somatic embryos after 6 mo. of subculture. Plant recovery frequencies were 60 to 70%. This method was expanded to different banana and plantain genomic groups.     

Nutrient utilization during bioreactor culture,and maturation of somatic embryo cultures ofPicea mariana andPicea glauca-engelmannii

Summary Embryogenic cell cultures ofPicea mariana (black spruce) and the species complexPicea glauca-engelmannii (interior spruce) were maintained either as suspensions in liquid medium in 250 or 500-ml-capacity shake-flasks, 7-liter-capacity airlift or mechanically stirred bioreactors, or on agar-solidified medium. Cultures from each of the maintenance conditions were subsequently transferred to agar-solidified LP medium containing 40μM (±) -abscisic acid for maturation into cotyledonary stage embryos. For both species, the highest maturation frequency resulted from cultures grown in the airlift bioreactor. With black spruce cells grown in the airlift bioreactor containing LP medium with 60 mM sucrose, a maximum of 7.1 g·liter⁻¹ dry weight and 2892 embryos·ml⁻¹ were obtained after 15 days. For interior spruce cells, a maximum dry weight of 5.9 g·liter⁻¹ and 2698 embryos·ml⁻¹ were obtained after 21 to 30 days. During culture over 2 wk, ammonia was almost completely utilized by both species, wherease nitrate was depleted to 40% of the initial concentration. Sucrose was rapidly hydrolyzed to glucose and fructose by both species. Black spruce cultures preferentially metabolized glucose, whereas interior spruce preferentially metabolized fructose. Improved growth of interior spruce cells in mechanically stirred bioreactors occurred when cultured in LP medium with 60 mM fructose as the sole carbon source. NRCC no. 36479     

Improvement of somatic embryogenesis inFeijoa sellowiana berg (Myrtaceae) by manipulation of culture media composition

Summary Somatic embryos could be induced from the cotyledons of zygotic embryos from immature fruits ofFeijoa sellowiana Berg (Feijoa) in the presence of a wide range of concentrations of fructose, glucose, maltose, and sucrose. Mannitol or sorbitol alone were ineffective. The highest frequencies of induction (99%) and the greatest number of somatic embryos per explant (134) were obtained with 0.4M fructose and 0.3M sucrose, respectively. This sucrose concentration also showed greater induction capacity than equimolar combinations of its monosaccharide constituents combined. Somatic embryo development was arrested at the globular stage at concentrations higher than 0.5M of all the sugars tested. When transferred to solid germination medium containing 2.0 mg/liter (5.77μM) gibberellic acid, 0.5 mg/liter (2.32μM) kinetin, and 0.029M sucrose, somatic embryos formed under 0.3 or 0.4M sucrose had better germination capacity than those induced under lower (0.1 and 0.2M) concentrations, as assessed by the frequency of explants presenting germinated embryos and by the number of plants obtained from those explants. On liquid media of similar composition somatic embryos did not germinate. Our data suggest that high (0.3 to 0.4M) carbohydrate levels improve somatic embryogenesis by acting both as carbon source and as osmotic regulator.     

Modulation of somatic embryogenesis in hypocotyl-derived cultures of geranium (Pelargonium X hortorum bailey) CV ringo rose by a bacterium

Summary The Ringo Rose cultivar of zonal geranium (Pelargonium x hortorum Bailey) has been shown to be morphogenetically unresponsive. Attempts to improve somatic embryogenesis using various seed stress treatments before germination proved ineffective. However, bacterial contamination of one of the seed-stress treatments led to infected explants that had a significant increase in frequency of high-quality somatic embryos. The co-cultivation of explants with the isolated bacterium (tentatively identified asBacillus sp.) was found to be repeatable, and potentially represents a novel way to improve morphogenesis in geranium and possibly other species.