Calli Ultrastructure of Globularia trichosantha ssp. trichosantha

Background and Purpose: This study aimed to produce calli with explants of aseptic seedlings after germination of G. trichosantha ssp. trichosantha seeds by plant tissue culture method and to examine the ultrastructure of the produced calli with electron microscope preparation.Materials and Methods: Seeds of G. trichosantha ssp. trichosantha were germinated in hormone-free Murashige and Skoog in in vitro conditions. Hypocotyl, epicotyl, cotyledon, young primer leaf, apical meristem and root explants taken from 30-day aseptic seedlings were transferred to Murashige and Skoog media for callus production which contained varying concentrations of 6-benzilamynopurine, indole acetic acid and 2,4 dichlorophenoxyacetic acid.Results: Two types of calli were determined: Yellow calli (Type 1) and Black calli (Type 2) with darkened colour and appearance that have not lost their development properties. Following lead staining, thin sections were examined by transmission electron microscope. The best callus production occurred at the Murashige and Skoog medium containing indole acetic acid and 6-benzilamynopurine and in root explants. The cells of Type 1 calli were spherical and large. The cells contained usually one nucleus and nucleolus. Also the cells contained a very large vacuole, endoplasmic reticulum, golgi complex, mitochondria, ribosomes, plastids. Deformed cells and spherical cells were determined in Type 2 calli. The cells were observed to have smaller vacoules and higher numbers of mitochondria different from Type 1 calli. Type 1 and Type 2 calli showed bulging mitochondrial cristae. Electron-dense droplets were observed in vacuoles of both Type 1 and Type 2 calli.</p

Progress in Biotechnological Applications of Diverse Species in Boraginaceae Juss

WOS: 000431815600014During the past four decades, plant cell biotechnology has evolved as a promising new area within the field of biotechnology, focusing on production of secondary metabolites and in vitro propagation of plants. Boraginaceae is one of the family that biotechnological tools were applied extensively because of their economically, ornamentally and medicinally valuable seconder metabolites as well as their endangered species. The Boraginaceae family is known as Borage or Forget-me-not, contains more than 156 genera and about 2000 species including annual, perrenial herbs, shrubs and trees. Members of the family were distributed mostly in sandy, -drier regions of the world. The most well-known members of the family are Forget me not (Myosotis sp.), Borage (Borago sp.), Comfreys (Symphytum sp.), and Heliotrope (Heliotropium sp.). A good number of the family members are used as a source of dye in cosmetics, food, textile and also in medical field. Selected species of the family utilized for obtaining secondary metabolites including naphtaquinone derivatives, rosmarinic acid and pyrrolizidine alkaloids. Due to their medicinal, economic and ecological importance, biotechnological tools such as plant tissue culture, metabolic engineering and in vitro micropropagation have been applied to produce biologically active compounds, pigments and to increase the population of the endangered species. The purpose of this chapter is to review studies performed utilizing biotechnological methods in diverse members of this family. Botanical aspects, traditional usage, chemical constituents and production of secondary metabolites in cultures and via metabolic engineering in some of the family members are also reviewed in brief

Ultrastructural observations in somatic embryogenesis of natural tetraploid Trifolium pratense L.

WOS: 000379975600006Previous reports of plant regeneration of natural tetraploid T. pratense L. 'Elci' could be realized only through the apical meristem calli. In order to proceed to the production stage, other regeneration methods need to be tried. Aseptic seedlings were used for the production of somatic embryos through various 2,4-D and kinetin trials. Nonuniform external callus cells with translucent cytoplasm were observed in various developmental stages of somatic embryos. Beneath these cells, there were uniformly aligned, dark-stained embryo cells with dense cytoplasm. Despite the similar developmental stages and cell characteristics of zygotic and somatic embryos, the walls of somatic embryo cells revealed a highly wavy pattern. The nucleus generally contained only one nucleolus, which was spherical, dark stained, and electron-dense. Electron-dense droplets were seen in vacuoles. The cytoplasm consisted of starch-containing amyloplasts, mitochondria, plastids, ribosomes, endoplasmic reticulum, dictyosomes, lipid, and protein bodies. In some of the somatic embryos at the globular and heart stages, vacuole or electron-translucent zones were observed in the nucleolus. Additionally, a few embryo degenerations were recorded during developmental stages of the zygotic embryo. For the first time, the somatic embryos of natural tetraploid T. pratense were produced from hypocotyl (85%), cotyledon (75%), and apical meristem (60%) explants in 0.3 mg/L 2,4-D and 2 mg/L kinetin-containing MS medium. Our study developed an effective and efficient in vitro production method for using natural tetraploid T. pratense in biotechnological studies.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108T830]This study was supported by the Scientific and Technological Research Council of Turkey (TUBITAK; project number: 108T830; TBAG). The authors would like to extend their gratitude to Prof Dr Sahabettin Elci for kindly providing the T. pratense seeds for this study

Influence of different sterilization methods on callus initiation and production of pigmented callus in Arnebia densiflora Ledeb.

WOS: 000294698400014We analyzed the effects of sodium hypochlorite and Plant Preservative Mixture (PPM) on surface sterilization. We also examined the effects that the addition of an antibiotic-antimycotic solution to the culture medium had on callus induction. Explains were initially sterilized with different concentrations of sodium hypochlorite and cultured on MS media containing kinetin (0.29 mu M) and naphthalene acetic acid (NAA, 10 mu M). No calluses were produced, either because of contamination of the explains, or loss of explants as a result of the high levels of sodium hypochlorite. The application of PPM and antibiotics at different concentrations reduced contamination and led to callus induction from shoot apexes and young root explants. The best callus responses were obtained using PPM at 1%-2%, whereas callus induction on shoot apexes diminished at higher concentrations (4% PPM). This is the first report of successful sterilization and reduced contamination of explains from naturally field grown A. densiflora by PPM. Moreover, established callus cultures produced pigmented calluses, which were analyzed spectrophotometrically.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [105S343]This study was supported by the Scientific and Technological Research Council of Turkey (TUBITAK), Project number 105S343 (Kariyer)

Pollen characteristics and in vitro pollen germination of Cedrus libani A. Rich.

WOS: 000273718700011This study aims to determine the germination characteristics, pollen tube developments, effects of germination media and temperature and incubation durations of the pollens obtained from the four clones (11342, 11344, 11345 and 11351) of Cedrus libani A. Rich. (Lebanon Cedrus) obtained from clonal seed orchard (with national registration no: Eskisehir 117) between 2004 and 2006 as well as of those taken from clone no: 11351 in 2004. They were stored at 3 degrees C for 13 months till they were investigated experimentally. MS medium was preferred for pollen germination for its relative superiority. Three-day incubation period at 33 degrees C temperature, in dark was applied along with the MS medium. The highest germination rate in MS medium was achieved in clone no. 11342 with 84.77% among the pollen samples of 2005. On the other hand the germination rate of the pollens taken from clone no: 11351 was determined as 49.95%

Comprehensive evaluation of phytoestrogen accumulation in plants and in vitro cultures of Medicago sativa L. 'Elci' and natural tetraploid Trifolium pratense L.

WOS: 000341523900009The main goal of this study was to establish callus cell suspension cultures of Medicago sativa L. 'Elci' and natural tetraploid Trifolium pratense L. and to compare the isoflavone production of the cultures to the original plants. The callus culture was transferred to liquid Murashige and Skoog media (MS3 and MS5) in order to establish the cell suspension cultures. The extracts were then analyzed by liquid chromatography-mass spectroscopy (LC-MS) for their isoflavones (phytoestrogen), mainly formononetin, biochanin A, daidzein, and genistein. The production of daidzein and formononetin was higher in cell suspension culture than in callus and herba of M. sativa L. 'Elci,' while biochanin A and genistein content could not be detected. On the other hand, the production of phytoestrogens was more successful in the herba of T. pratense L. than in both of the cultures. It might be suggested that T. pratense L. can be grown in larger fields, whereas M. sativa L. can be utilized to establish in vitro cultures in order to produce isoflavone compounds for pharmaceutical purposes.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108T830]This study was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) with project number 108T830 (TBAG). The authors would like to extend their gratitude to Prof Dr Sahabettin Elci for kindly providing the seeds for this study

Use of Red Clover (Trifolium pratense L.) Seeds in Human Therapeutics

WOS: 00031122240011

ANALYSIS OF PHENOLIC COMPOUNDS FOR DETERMINATION OF CAMBIUM DIFFERENTIATION AND TRACHEAL ELEMENTS IN OLIVE GRAFT COMBINATIONS

WOS: 000393033400004The aim of this investigation was to evaluate the effect of phenolic compounds during differentiation of cambium and tracheal elements in olive cultivars 'Ayvalik', 'Domat', 'Gemlik', 'Memecik', 'Nizip Yaglik' and 'Sari Ulak. These cultivars were grafted onto one year-old cv. 'Gemlik' rootstock. This rootstock has been propagated by cuttings. According to the results of histological studies for two years; new cambium cells were initiated in first three months while the formation of vascular bundles and sclerenchyma cells were initiated in six months. Large numbers of undifferentiated parancymatic cells were also determined in both side grafts of the cultivars 'Ayvalik', 'Domat' and 'Nizip Yaglik' after 3, 6 and 12 months of grafting. Further, High levels of 4 hydroxyphenylacetic acid, vanillic and ferulic acids content were determined in the scion of the cultivars Ayvalik, Nizip Yaglik and Domat. As a result, during recovery of graft zone, development of new cambium tissues, vascular connections and sclerenchyma tissues occurred imperfectly in 'Ayvalik', 'Domat' and 'Nizip Yaglik' cultivars and graft zones of these combinations were found weak. Moreover level of 4Hydroxyphenylacetic acid and Ferulic acidphenolic compounds had high concentration in scions of 'Ayvalik' and 'Domat', respectively. These results illustrated existence of problem in differentiation of cambium and vascular systems in graft interface of 'Ayvalik' and 'Domat' cultivars.Ministry of Food, Agriculture and LivestocksThis work was supported by Ministry of Food, Agriculture and Livestocks for financial. We are very grateful to Edremit Olive Nursery Plant Station and Bornova Olive Research Institute for their kind helps during grafting and maintain

Ultrastructural observations in somatic embryogenesis of natural tetraploid Trifolium pratense L.

Previous reports of plant regeneration of natural tetraploid T. pratense L. `Elci' could be realized only through the apical meristem calli. In order to proceed to the production stage, other regeneration methods need to be tried. Aseptic seedlings were used for the production of somatic embryos through various 2,4-D and kinetin trials. Nonuniform external callus cells with translucent cytoplasm were observed in various developmental stages of somatic embryos. Beneath these cells, there were uniformly aligned, dark-stained embryo cells with dense cytoplasm. Despite the similar developmental stages and cell characteristics of zygotic and somatic embryos, the walls of somatic embryo cells revealed a highly wavy pattern. The nucleus generally contained only one nucleolus, which was spherical, dark stained, and electron-dense. Electron-dense droplets were seen in vacuoles. The cytoplasm consisted of starch-containing amyloplasts, mitochondria, plastids, ribosomes, endoplasmic reticulum, dictyosomes, lipid, and protein bodies. In some of the somatic embryos at the globular and heart stages, vacuole or electron-translucent zones were observed in the nucleolus. Additionally, a few embryo degenerations were recorded during developmental stages of the zygotic embryo. For the first time, the somatic embryos of natural tetraploid T. pratense were produced from hypocotyl (85\%), cotyledon (75\%), and apical meristem (60\%) explants in 0.3 mg/L 2,4-D and 2 mg/L kinetin-containing MS medium. Our study developed an effective and efficient in vitro production method for using natural tetraploid T. pratense in biotechnological studies

Mericarp micromorphology and anatomy of Salvia hedgeana Donmez, S. huberi Hedge and S. rosifolia Sm. (section Salvia Hedge, Lamiaceae)

Mericarp (nutlet) micromorphology and pericarp structure of three morphologically similar endemic Salvia species; Salvia hedgeana, S. huberi and S. rosifolia were investigated using LM, SEM and TEM. Salvia hedgeana has larger mericarps and abscission scars than S. huberi and S. rosifolia. Mericarp length to width ratio ranges from 1.11 in S. hedgeana to 1.60 in S. huberi. Mericarp shape is mainly ovoid, rarely broadly ovoid in S. hedgeana, and oblong in S. huberi. The mericarp surface sculpturing pattern in all species is colliculate. However, exocarp cells are pentangular-hexangular in S. hedgeana, irregular in S. huberi and rounded and smaller in S. rosifolia. In Salvia huberi anticlinal walls are undulate whereas in S. hedgeana and S. rosifolia anticlinal walls are straight. Salvia hedgeana was distinguished from the others by the thickest pericarp (146–185 μm). The sclerenchymatous region significantly varied between the species. It was 84–99 μm in S. hedgeana, 56–82 μm in S. huberi and 27–61 μm in S. rosifolia. The mesocarp was also thicker in S. hedgeana. The wetted mericarps produced mucilage, but S. huberi differed from the others in having translucent-milky white opaque mucilage with fibres or radiating cordons