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Sample records for fruit callus cultures

  1. High Triterpenic Acids Production in Callus Cultures from Fruit Pulp of Two Apple Varieties.

    Science.gov (United States)

    Verardo, Giancarlo; Gorassini, Andrea; Ricci, Donata; Fraternale, Daniele

    2017-01-01

    Very rarely fruit pulp has been used in in vitro culture to produce secondary metabolites useful in promoting health. The aims of this work were the study of the best conditions to obtain the callus cultures from the pulp of two varieties of apples, Golden Delicious (GD) and "Mela Rosa Marchigiana" (MRM), and the quali-quantitative analysis of secondary metabolites produced by the two in vitro callus cultures. Callus was induced on both Murashige and Skoog and Gamborg B5 media containing various combinations of supplements. To achieve the maximum recovery of secondary metabolites produced, preliminary extraction tests were carried out on GD apple culture using two different organic solvents (MeOH and EtOAc). The quali-quantitative analysis of the methanolic extract of both cultures was carried out by ESI-MS n and GC-MS techniques. The GC-MS analysis revealed the presence of triterpenic acids, in particular, oleanolic, ursolic, maslinic, pomolic, tormentic, corosolic and annurcoic acid along with a phytosterol, β-sitosterol. In addition, GD callus culture produced phloridzin, absent in the MRM culture. In this last culture, however, the total amount of secondary metabolites was markedly higher. The in vivo production of these bioactive compounds were also quantified in the GD and MRM apple pulps. Apple pulps produced higher amounts of triterpenic acids in vitro than in vivo. The present work can be considered a method to amplify the production of important secondary metabolites which exert beneficial effects on human health. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  2. Callus induction and biomass accumulation in vitro in explants from chokeberry (Aronia melanocarpa (Michx. Elliot fruit

    Directory of Open Access Journals (Sweden)

    Tatiana I. Calalb

    2014-09-01

    Full Text Available In this study, the following features were determined: biological (the optimal histogen as explant and the optimal age of donor fruit, biotechnological (type, dosage and combination of growth regulators supplements in culture medium Murashige and Skoog as well as sucrose dosage, and physical (light regime, to induce callusing and biomass accumulation in vitro from the succulent chokeberry (Aronia melanocarpa (Michx. Elliot fruit. It turned out that it was much easier to induce callus from explants composed of the epicarp and hypoderm cut from fruits at 50–60 days after flowering. The role of light regime and varied supplementation of the basic MS medium with different doses of growth regulators was established; they resulted in four pigmented carpomass: violet, cream-pink, cream-white and green. The best combinations for the proliferation of fruit callus were culture media with 0.2–2.5 mg × dm-3 2,4-D+0.5 mg × dm-3 KIN +60 g × dm-3sucrose, while for fruit biomass accumulation enriched with phenolic substances – 2.5–3.5 mg × dm-3 NAA+0.5 mg × dm-3 KIN+60 g × dm-3sucrose. The chemical study of phenolic compounds by HPLC coupled with the mass spectrometry method identified chlorogenic acid, hiperozide, quercetrin, isoquercitrin and rutozide quantitatively and qualitatively in all pigmented carpomass and fruits; an exception is p-coumaric present only qualitatively in green carpomass and absent in fruit and quercetol absent in green carpomass.

  3. The durative use of suspension cells and callus for volatile oil by comparative with seeds and fruits in Capparis spinosa L.

    Directory of Open Access Journals (Sweden)

    Yongtai Yin

    Full Text Available Capparis spinosa is one of the most important eremophytes among the medicinal plants, and continued destruction of these plants poses a major threat to species survival. The development of methods to extract compounds, especially those of medicinal value, without harvesting the whole plant is an issue of considerable socioeconomic importance. On the basis of an established system for culture of suspension cells and callus in vitro, Gas Chromatograph-Mass Spectrometer (GC-MS was used for the volatile oil composition analyzing in seed, fruit, suspension cells and callus. Fatty acids were the major component, and the highest content of alkanes was detected in seed, with <1.0% in suspension cells and callus. Esters, olefins and heterocyclic compounds were significantly higher in fruit than in the other materials. The content of acid esters in the suspension cells and callus was significantly higher than in seed and fruit. This indicated that the suspension cells and callus could be helpful for increasing the value of volatile oil and replacing seeds and fruit partially as a source of some compounds of the volatile oil and may also produce some new medical compounds. The above results give valuable information for sustainable use of C. spinosa and provide a foundation for use of the C. spinosa suspension cells and callus as an ongoing medical resource.

  4. Accumulation of the sesquiterpenes nootkatone and valencene by callus cultures of Citrus paradisi, Citrus limonia and Citrus aurantium.

    Science.gov (United States)

    Del Río, J A; Ortuño, A; Puig, D G; Iborra, J L; Sabater, F

    1991-10-01

    The production of the sesquiterpenes nootkatone and valencene by callus cultures of Citrus species is described. The levels of these compounds were examined by gas chromatography-mass spectrometry and their yields were compared with the amounts found in mature fruits. A simultaneous increase and decrease in the levels of nootkatone and valencene, respectively, were observed with the aging of callus cultures of Citrus paradisi. These results suggest that valencene might be a possible precursor of nootkatone in this species. The high level of nootkatone detected in 9-month-old callus cultures of Citrus paradisi might be associated with the corresponding cell morphological changes observed.

  5. EFFECT OF PHYSIOLOGICAL AGE AND GROWTH REGULATORS ON CALLUS BROWNING OF COCONUT ENDOSPERM CULTURE IN VITRO

    Directory of Open Access Journals (Sweden)

    LAZARUS AGUS SUKAMTO

    2011-01-01

    Full Text Available The possibility of physiological age and growth regulators affecting callus browning ofcoconut endosperm was investigated. Solid endosperm explants of four coconut fruits fromsame brunches of two coconut cultivars “Samoan Dwarf ” were grown on modified Murashigeand Skoog (MS formula with addition of 10 mg l putresine, 2.50 g l activated charcoal (AC,1.70 g l phytagel, 0, 10 , 10 , 10 , 10 M 2,4-dichlorophenoxyacetic acid (2,4-D or 4-amino-3,5,6-trichloropicolinic acid (Picloram combined with 10 M 6-benzylaminopurine (BA.Callogenesis occurred on 98.83% of explants. Callus browning between different physiologicalages (antipodal and micropylar tissues of coconut endosperm at 9, 26 and 31 weeks of culture(WOC was significantly different, but not at 16 and 21 WOC. Auxins of 2,4-D and Picloramdid not affect significantly callus browning of endosperm cultures. Auxin doses at 10 , 10 , and10 M decreased significantly callus browning at 9 and 16 WOC, respectively, but at 10 Mbrowning was less significant compared to other doses at 21 WOC. Auxin dose at 10 M causedless significant browning compared to other doses at 31 WOC. The addition of BA decreasedsignificantly callus browning at 9 WOC, but did not affect callus browning thereafter.

  6. Growth and accumulation of flavan-3-ol in Camellia sinensis through callus culture and suspension culture method

    Directory of Open Access Journals (Sweden)

    Sutini Sutini

    2017-02-01

    Full Text Available This study was aimed to assess flavan-3-ol biomass in C. sinensis through callus cultures and suspension cultures derived from leaf explants. Callus initiation of both cultures were using Murashige and Skoog medium were enriched with plant growth regulators Naphtha-lene Acetic Acid 3.0 mg/L and kinetin 2.0 mg/L. The procedures in this study were: (1 callus initiation by cutting the leaves of C. sinen-sis shoots then planted on Murashige and Skoog medium that were enriched with plant growth regulators, (2 sub callus culture on fresh medium that enriched with the same growth regulators, (3 suspension culture initiation of liquid callus, (4 growth examination of callus and suspension cultures in week 12, (5 examination of qualitative-quantitative content of flavan-3-olin suspension cultures at week 4. The results show that suspension cultures contain biomass flavan-3-ol that increase in the same manner of the increase of callus age and weight

  7. Antioxidant and antimicrobial activities of callus culture and leaf ...

    African Journals Online (AJOL)

    The callus culture extract (CCE) gave the lowest MIC value of 0.78 mg/ mL for most of the bacteria and fungi and the lowest MBC values of 0.78 mg/ mL and 1.56 mg/ mL against bacteria and fungi, respectively. ... Keywords: Crotalaria retusa; In vitro propagation; Callus culture; Antimicrobial activity; Antioxidant activity ...

  8. Factors controlling phenol content on Theobroma cacao callus culture

    International Nuclear Information System (INIS)

    Quiñones-Galvez, Janet; HernándezTorre, Martha de la; Quirós Molina, Yemeys; Capdesuñer Ruiz, Yanelis; Trujillo Sánchez, Reinaldo

    2016-01-01

    Theobroma cacao L. is known in folk medicine as an antiseptic, diuretic and antiparasitic. Foods derived from this plant are rich in natural products of high added value, including phenolic compounds. As in vitro cultivation handle is an alternative source for the production of these metabolites. The present study was conducted to obtain phenolic compounds from callus culture with embryogenic structures. Culture conditions (agitation, light and glucose) were established to increase the concentration of phenols in calluses and elicitors to achieve the increase in callus and excretion into the culture area. The accumulation of phenolic compounds was favored with the additional supplement of glucose, growth in agitation and darkness. The addition of random hydroxylated cyclodextrins allowed the increase in the specific yield of phenols and biomass. (author)

  9. [Flavonoid glycosides from callus cultures of Dysosma versipellis].

    Science.gov (United States)

    Chen, Ri-Dao; Duan, Rui-Gang; Zou, Jian-Hua; Li, Jun-Wei; Liu, Xiao-Yue; Wang, Hai-Yan; Li, Qiu-Hong; Dai, Jun-Gui

    2016-01-01

    Various chromatographic techniques, including silica gel column chromatography, Sephadex LH-20, preparative thin-layer chromatography, and preparative HPLC, were employed to isolate the chemical constituents from callus cultures of Dysosma versipellis. Structures of the compounds were elucidated based on UV, IR, MS and NMR spectroscopic data analysis. Totally, seven flavonoid glycosides were isolated from the 95% ethanol extract of the callus cultures and identified as kaempferol-3-O-[6″-(3″'-methoxy)-malonyl]-β-D-glucopyranoside(1), kaempferol-3-O-(6″-O-acetyl)-β-D-glucopyranoside(2), kaempferide-3-O-β-D-glucopyranoside(3), kaempferol-3-O-β-D-glucopyranoside(4), isoquercitrin(5), quercetin-4'-O-β-D-glucopyranoside(6) and kaempferol-3-(6″-malonyl)-β-D-glucopyranoside(7), respectively.All these compounds were isolated from callus cultures of D. versipellis for the first time.Compounds 1, 2, 3, 6 and 7 were firstly obtained from plant materials of D. versipellis, and compound 1 was a new compound. Copyright© by the Chinese Pharmaceutical Association.

  10. Establishment of the callus and cell suspension culture of ...

    African Journals Online (AJOL)

    The objective of this work was the optimization of the conditions of callus and cell suspension culture of Elaeagnus angustifolia for the production of condensed tannins. The effects of different conditions on the callus growth and the production of condensed tannins were researched. The leaf tissue part of E. angustifolia was ...

  11. Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae).

    Science.gov (United States)

    Chen, J -T.; Chang, W -C.

    2000-12-07

    An efficient method was established for high frequency somatic embryogenesis and plant regeneration from callus cultures of a hybrid of sympodial orchid (Oncidium 'Gower Ramsey'). Compact and yellow-white embryogenic calli formed from root tips and cut ends of stem and leaf segments on 1/2 MS [11] basal medium supplemented with 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ, 0.1-3 mg/l), 2,4-dichlorophenoxyacetic acid (2,4-D, 3-10 mg/l) and peptone (1 g/l) for 4-7 weeks. Embryogenic callus was maintained by subculture on the same medium for callus induction and proliferated 2-4 times (fresh weight) in 1 month. Initiation of somatic embryogenesis and development up to the protocorm-like-bodies (PLBs) from callus cultures was achieved on hormone-free basal medium. Regenerants were recovered from somatic embryos (SEs) after transfer to the same medium and showed normal development. The optimized protocol required about 12-14 weeks from the initiation of callus to the plantlet formation. Generally, the frequency of embryo formation of root-derived callus was higher than stem- and leaf-derived calli. Combinations of naphthaleneacetic acid (NAA) and TDZ significantly promoted embryo formation from callus cultures. The high-frequency (93.8%) somatic embryogenesis and an average of 29.1 SEs per callus (3x3 mm(2)) was found in root-derived callus on a basal medium supplemented with 0.1 mg/l NAA and 3 mg/l TDZ. Almost all the SEs converted and the plantlets grew well with an almost 100% survival rate when potted in sphagnum moss and acclimatized in the greenhouse.

  12. Radio-sensitivity of callus and cell cultures, and RAPD characterization of variants in banana [Musa spp.

    International Nuclear Information System (INIS)

    Kulkarni, V.M.; Karmarkar, V.M.; Ganapathi, T.R.; Bapat, V.A.

    2000-01-01

    Although bananas and plantains are one of the most important fruit crops, gearing up the breeding programmes for these has always remained the most difficult task due to several inherent problems such as parthenocarpy, barriers in obtaining viable seeds and long life cycle etc. In this regard, incorporation of in vitro techniques such as shoot-tip / cell cultures along with conventional as well as non-conventional methods of genetic improvement is of utmost importance, especially in those vegetatively propagated species with long crop cycle and low in vivo proliferation rate. In order to understand the radio-sensitivity, the callus and cell cultures of banana were exposed to differential doses of gamma-rays. Growth of the callus cultures reduced with increasing dose of gamma-rays. Similar trend was noticed in irradiation of cell suspensions also where a dose of 40 Gy and more was completely lethal. The experience gained from previous and present experiments has yielded optimization of the procedures for gamma-irradiation and subsequent handling of banana in vitro cultures. The RAPD analysis of the selected variants was unable to detect adequate polymorphism, and further experimentation in these regards is being done. (author)

  13. Regeneration of Stevia Plant Through Callus Culture

    Science.gov (United States)

    Patel, R. M.; Shah, R. R.

    2009-01-01

    Stevia rebaudiana Bertoni that conventionally propagated by seed or by cuttings or clump division which has a limitation of quality and quantity seed material. In present study, callus culture technique was tried to achieve rapid plant multiplication for quality seed material. Callus induction and multiplication medium was standardized from nodal as well as leaf sagments. It is possible to maintain callus on Murashige and Skoog medium supplemented with 6-benzyl amino purine and naphthalene acetic acid. Maximum callus induction was obtained on Murashige and Skoog medium incorporated with 6-benzyl amino purine (2.0-3.0 mg/l) and naphthalene acetic acid (2.0 mg/l) treatments. However, Murashige and Skoog medium containing 2.0 mg/l 6-benzyl amino purine+2.0 mg/l naphthalene acetic acid was found to be the best for callus induction. Higher regeneration frequency was noticed with Murashige and Skoog medium supplemented with 2.0 mg/l 6-benzyl amino purine+0.2 mg/l naphthalene acetic acid. Regenerated plants were rooted better on ¼ Murashige and Skoog strength supplemented with 0.1 mg/l indole-3-butyric acid. The rooted plantlets were hardened successfully in tera care medium with 63 per cent survival rate. The developed protocol can be utilized for mass production of true to type planting material on large scale independent of season, i.e. external environmental conditions. PMID:20177455

  14. The effects of callus age, UV irradiation and incubation time on trans-resveratol production in grapevine callus culture

    International Nuclear Information System (INIS)

    Keskin, N.; Kunter, B.

    2009-01-01

    In this study, the effects of callus age, ultraviolet (UV) irradiation and incubation time were investigated for the induction of trans-resveratrol production in callus cultures of Kalecik karası (Clone 12) Vitis vinifera L. grape cultivar. Part of leaves (~1 cm 2 ) were taken from one year old grapevines grown in greenhouse and then were cultured in Gamborg B-5 media including 2% saccarose, 0.8% agar, 1.0 μM BAP (6-benzylaminopurine) and 0.1 μM 2, 4-D (2, 4-dichlorophenoxy-acetic acid). After the second subculture, 12 and 15 days old callus tissues were exposed to 254 nm UV light at 10 cm distance from the source for 10 and 15 min. Trans-resveratrol was measured at 0, 24, 48 and 72 hours of incubation by using High Pressure Liquid Chromatography (HPLC). Trans-resveratrol concentration of control callus ranged from 0.56 to 0.96 μg g fw -1 . The highest trans-resveratrol production was obtained from 12 and 15 days-old callus irradiated for 10 min. at the 48th hours of incubation (2.42 μg g fw -1 ). Considering the highest value of trans-resveratrol concentration in controls and experiments, it was determined that UV irraditation in Kalecik karası elicitated the trans-resveratrol production by 2.5 times. (author) [tr

  15. Morphohistobiochemical characteristics of embryogenic and nonembryogenic callus cultures of sweet potato (Ipomoea batatas L.).

    Science.gov (United States)

    Mukherjee, A; Debata, B K; Mukherjee, P S; Malik, S K

    2001-01-01

    Ipomoea batatas callus culture raised in a medium supplemented with 2,4-D (2,4-dichlorophenoxy acetic acid) alone or 2,4-D in combination with benzyl adenine, were found to be embryogenic. Supplementation of exogenous chemicals, such as 5 g/l NaCI or 0.7 g/l proline together with a mild dose of 0.2 mg/l 2,4-D, enhanced somatic embryogenesis significantly in all the genotypes tested. Morphological, growth, physiological, histological, and biochemical characteristics of the embryogenic callus were different from the nonembryogenic callus. The former was compact, slow growing, and nodular compared with the fast growing, fragile, nonembryogenic callus. The embryogenic callus tissue had more dry matter, protein and reducing sugar contents compared with the less embryogenic callus. The somatic embryogenic response remained steady in the cultures for up to 96 weeks.

  16. Embryogenic callus formation, growth and regeneration in callus and suspension cultures of Miscanthus x ogiformis Honda 'Giganteus' as affected by proline

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Krogstrup, Peter; Hansen, Jürgen

    1997-01-01

    .6 M 2,4-dichlorophenoxyacetic acid. Shoot apices and leaves from in vitro-propagated shoots, and immature inflorescences from greenhouse-grown plants were used as explants for callus induction and formation. Suspension cultures initiated from embryogenic callus of immature inflorescences were used...

  17. Regeneration from embryogenic callus and suspension cultures of ...

    African Journals Online (AJOL)

    ehab

    2012-04-25

    Apr 25, 2012 ... Micropropagation, through seed culture has been studied in C. martinii ... Effect of 2,4-D on total and embryogenic callus production ... ethanol for 1 min, followed by immersion in 20% Clorox (NaOCl ... Three flasks from each treatment were cultured. ... shaker at 110 rpm. ...... Plants cultivated in bioreactor of.

  18. Appraisal of beta-Phellandrene in Callus Cultures of Momordica charantia L. Cultivars, Jaunpuri and Jhalri

    International Nuclear Information System (INIS)

    Ali, S.; Tariq, A.; Ajaib, M.

    2015-01-01

    Secondary metabolite beta-phellandrene was analyze in callus cultures of two varieties of Momordica charantia L. i.e. Jaunpuri and Jhalri. Conditions for seed germination and callus induction were optimized. Seedlings grown under aseptic conditions served as explant sources. 6-Benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D) supplemented in Murashige and Skoog's (MS) medium were scrutinized as the most suitable combination of plant growth regulators with different concentrations for callus induction in both the varieties. Cotyledon explant of (cultivars) cv. Jaunpuri revealed maximum callus induction with 1.0 mgl/sup -1/ BAP and 1.5 mgl/sup -1/ 2,4-D in eight days as compared to internode, apical bud and leaf. Cotyledon and leaf explants of cv. Jhalri responded to 1.5 mgl/sup -1/ BAP and 1.0 mgl/sup -1/ 2,4-D in nine days for callus and internode and apical bud with 1.0 mgl-l BAP and 1.5 mgl-l 2,4-D. Best grown calli from different explants were analyzed through GC-MS for production of secondary metabolites. Along with other secondary metabolites beta-phellandrene was the most prominent secondary metabolites found in in vitro grown callus cultures of both the varieties. The callus cultures of cv. Jaunpuri produced substantial amount of beta-phellandrene i.e. up to 30 percent of the total secondary metabolites as compared to calli from cv. Jhalri explants. The callus cultures of M. charantia can prove the best alternative, rapid and uninterrupted source for natural beta-phellandrene production. (author)

  19. Callus and cell suspension cultures of carnation

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1972-01-01

    Callus cultures of carnation, Dianthus caryophyllus L. ev. G. J. Sim, were grown on a synthetic medium of half strength Murashige and Skoog salts, 3 % sucrose, 100 mg/l of myo-inositol, 0.5 mg/l each of thiamin, HCl, pyridoxin, HCl and nicotinic acid and 10 g/l agar. Optimal concentrations...

  20. callus culture

    African Journals Online (AJOL)

    steve

    flavonoids, anthocyanins, and other phenolic com-pounds. (Blando et al. .... TP content of the callus were estimated according to the method of. Slinkard and ..... quantification of phenolic compounds in grapes by HPLC-PDA-ESI-. MS on a ...

  1. Establishment of the callus and cell suspension culture of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-05

    Oct 5, 2009 ... Full Length Research Paper. Establishment of the callus ... study provided an efficient way for E. angustifolia cell suspension culture to produce secondary metabolite. .... was also observed that in these treatments the stem.

  2. Shoot Differentiation in Callus Cultures of Datura Innoxia

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1973-01-01

    promoted shoot differentiation. Gibberellic acid inhibited shoot formation weakly, but inhibited proper leaf blade formation. Root differentiation was rare. The callus cultures of Datura innoxia grew rapidly (100-fold in 4 weeks) on a slightly modified Murashige and Skoog medium (0.5 mg/l thiamin · HCl, p...

  3. Differential radiosensitivity of seeds, seedlings and callus cultures of Petunia inflata

    International Nuclear Information System (INIS)

    Bapat, V.A.; Rao, P.S.

    1976-01-01

    A comparative study of the effects of γ-irradiation on seeds, seedlings and callus cultures of Petunia inflata showed striking differences in radiosensitivity as reflected in differences in mean fresh and dry weights, seedling height and morphology. Seeds subjected to low doses (4-6 kR) of irradiation showed stimulation of seedling height. Direct exposure of seedlings to high doses (10 kR) of irradiation caused inhibition in their development. Callus cultures, however, were more radioresistant compared to seeds and seedlings. Tissues grown on either an irradiated nutrient medium or on a medium in which sucrose alone had been irradiated, showed a marked inhibition in their growth potential

  4. Additions of precursors and elicitors improve geranylgeraniol production in Croton stellatopilosus callus cultures

    Directory of Open Access Journals (Sweden)

    Juraithip Wungsintaweekul

    2015-02-01

    Full Text Available Strategies for enhancing GGOH production in Croton stellatopilosus callus culture included additions of precursors (sodium acetate-NA, sodium pyruvate-NP, mevalonic acid lactone-MVA and elicitors (methyl jasmonate-MJ, acetylsalicylic acid-ASA, yeast extract-YE. Treated cells were evaluated for their GGOH contents by GC-FID and compared with the nontreated cells as controls. Additions of NA (25 mg/L, NP (50 mg/L and MVA (100 mg/L resulted in an enhancement of GGOH productivity to 0.61 mg/g DW, 0.52 mg/g DW and 0.70 mg/g DW, respectively, compared to the control culture (0.29 mg/g DW. Callus cultures elicited with MJ at 30 mg/L for 24 h stimulated GGOH production to 0.35 mg/g DW compared to the control culture (0.07 mg/g DW. Cells also responded to ASA (20 mg/L, 2 days and YE (0.25 g/L, 4 days and produced GGOH contents of 0.46 mg/g DW and 1.37 mg/g DW, respectively. This study has shown that isoprenoid precursors and conventional elicitors enhanced GGOH production in the C. stellatopilosus callus culture.

  5. Evaluation of different carbon sources for high frequency callus culture with reduced phenolic secretion in cotton (Gossypium hirsutum L. cv. SVPR-2

    Directory of Open Access Journals (Sweden)

    G. Prem Kumar

    2015-09-01

    Full Text Available An efficient protocol was developed to control excessive phenolic compound secretion during callus culture of cotton. As cotton is naturally rich in phenolic compounds factors influencing the phenolic compound secretion, callus induction and proliferation were optimized for getting high frequency callus culture. Different carbon sources such as fructose, glucose, sucrose and maltose were tested at various concentrations to control phenolic secretion in callus culture. Among them, 3% maltose was found to be the best carbon source for effectively controlling phenolic secretion in callus induction medium. High frequency of callus induction was obtained on MSB5 medium supplemented with 3% Maltose, 2,4-D (0.90 μM and Kinetin (4.60 μM from both cotyledon and hypocotyl explants. The best result of callus induction was obtained with hypocotyl explant (94.90% followed by cotyledon explant (85.20%. MSB5 medium supplemented with 2,4-D (0.45 μM along with 2iP (2.95 μM gave tremendous proliferation of callus with high percentage of response. Varying degrees of colors and textures of callus were observed under different hormone treatments. The present study offers a solution for controlling phenolic secretion in cotton callus culture by adjusting carbon sources without adding any additives and evaluates the manipulation of plant growth regulators for efficient callus culture of SVPR-2 cotton cultivar.

  6. Shoot regeneration of callus culture from irradiated sheed of piper nigrum L by gamma rays

    International Nuclear Information System (INIS)

    Ishak; Hutabarat, D.

    1988-01-01

    Shoot regeneration was obtained from callus that induced by irradiated seed with 25 and 50 Gy of gamma-rays and then on M.S. medium containing NAA 1 ppm and 2-ip 0.5 ppm. Irradiated seed with a dose of 25 Gy produced normal root and failed to produce shoot, but rice callus. Irradiated seed with a dose of 50 Gy pruduce callus only. Shoot differentiation occured after the callus were cultured on M.S., medium containing 2-ip 1 ppm and Kinetin 2.5 ppm. (authors). 9 refs, 3 figs

  7. Factors affecting callus and protoplast production and regeneration of plants from garlic tissue cultures

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Nabulsi, I.

    2001-08-01

    Five cultivars of garlic, two explants, six callusing media, six regeneration media, two kinds of light and several doses of gamma irradiation were used to determine the best conditions for callus induction and plant regeneration from garlic tissue cultures. Also, some experiments were conducted to study the possibility to isolate protoplast and regenerate plants. The experiment showed that medium MS9 was good for regenerating plant directly from basal plate without going through callus phase. ANOVA exhibited significant differences among used cultivars in their ability to form callus. No significant difference was observed between 16 hr light and complete darkness in callus growth. However, appearance of callus was generally better on darkness. Cultivar varied in their ability to regenerate and interaction between cultivars and media was observed. Cultivar kisswany was the best in regeneration (38%) and medium MS47 was the best among used media (35%). Light type played a significant role in regeneration of plants where red light was much better than white light in inducing regeneration (68% vs 36%). ANOVA revealed significant effect of low doses of gamma irradiation on stimulation regeneration of plant whereas high doses prevented regeneration. Many experiments were conducted to isolate protoplast and regenerate plants. The best method for culturing was the droplet and the best conditions for incubation were complete darkness at 25 Degreed centigrade. This lead to formation of cell wall but no cell division was observed (author)

  8. EFFECT OF AUXIN AND CYTOKININ ON VINCRISTINE PRODUCTION BY CALLUS CULTURES OF CATHARANTHUS ROSEUS L. (APOCYNACEAE

    Directory of Open Access Journals (Sweden)

    Chinnamadasamy Kalidass

    2009-11-01

    Full Text Available Callus cultures of Catharanthus roseus L. were established to verify whether they produce vincristine as the intact plant. Different growth regulator combinations were applied to Murashige and Skoog (MS medium to influence the level of production of vincristine. The effects of various combinations (0.5 µM to 3.0 µM of auxin and cytokinin on the growth and accumulation of vincristine were investigated. MS medium supplemented with 2,4-Dichlorophenoxy acetic acid (2,4-D 1.0 µM and 6-furfurylaminopurine (Kinetin 1.0 µM was used to support the growth of callus cultures and the maximum amount of dry biomass (598.04 mg was produced after seven weeks of culture. High performance liquid chromatographic (HPLC analysis of methanol extracts from callus cultures of C. roseus revealed that the cultures produced vincristine. The concentrations of the growth regulators alpha-naphthalene acetic acid (NAA and kinetin played a critical role in the production of vincristine.

  9. Fatty acids, phenols content, and antioxidant activity in Ibervillea sonorae callus cultures

    OpenAIRE

    Estrada-Zúñiga, M.E.; Arano-Varela, H.; Buendía-González, L.; Orozco-Villafuerte, J.

    2012-01-01

    Ibervillea sonorae callus cultures were established in order to produce fatty acids (lauric, myristic, pentadecanoic, palmitic and stearic acids) and phenolic compounds. Highest callus induction (100%) was obtained in treatments containing 2.32 or 4.65 μM Kinetin (KIN) with 2.26 or 6.80 μM 2,4-Dichlorophenoxyacetic acid (2,4-D). Highest fatty acids (FA) production (48.57 mg g-1), highest total phenol content (TPC; 57.1 mg gallic acid equivalents [GAE] g-1) and highest antioxidant activity (EC...

  10. (Tongkat Ali) for callus

    African Journals Online (AJOL)

    hope&shola

    2010-12-06

    Dec 6, 2010 ... The induction of callus cultures using leaf, petiole, rachis, stem, tap root, fibrous root, cotyledon and ... other countries of the South-East Asia region and widely distributed ..... Indian ginseng plantlets from stem callus. Plant Cell ...

  11. Effect of gamma-radiation on callus initiation and oraganogenesis in the tissue culture of Nicotiana tabaccum L

    International Nuclear Information System (INIS)

    Shin, S. H.; Kim, J. G.; Song, H. S.

    2004-01-01

    It is generally agreed that ionizing radiations stimulate cell division, growth and development in various organisms including animals and plants. Differentiating tissues are the most sensitive to radiation. The present experiment was carried out to investigate the effects of ionizing radiation on callus initiation and organogenesis from the stem in the culture of Nicotiana tabaccum L. cv. When the stem segments were cultured on a Murashige and Skoog (MS) medium with 2 mg/L kinetin, with 1 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D), with 2 mg/L kinetin and 1 mg/L 2,4-D, the shoots and callus were differentiated 14 days after cultivation. Callus was especially formed on the MS medium with 2,4-D and/or kinetin and the formation was promoted by 1 Gy and 5 Gy of gamma radiation. The formation of the shoot clusters on the MS medium with 2 mg/L kinetin were prominent in the 5 Gy-irradiated groups. It is concluded that that gamma radiation enhanced the callus initiation and organogenesis in the tissue culture of Nicotiana tabaccum L

  12. The Extraction Process of Trimethyl Xanthina in Vitro Culture of Callus Camellia Sinensis with ethyl Acetate Solvent

    Directory of Open Access Journals (Sweden)

    Sutini

    2016-01-01

    Full Text Available Trimethyl xanthina is one of the compounds contained bioactive culture in vitro Cammelia sinensis callus which is widely used in the field of food, beverage, agriculture and health industries. The presence of trimethyl xanthina on food, beverages and health is needed in a certain amount depending on the use which is achieved by the user. To get a certain amount of trimethyl xanthina from callus culture of Cammelia sinensis, the extraction process is performed on the water solvent, as well as non-solvent water / organic solvent such as ethyl acetate. The purpose of this study was to obtain profile of trimethyl xanthina in the extraction of Cammelia sinensis callus. The experimental methods used consisted of dissolution, filtration, extraction with water solvent and ethyl acetate, then followed by identification of trimethyl xanthina using HPLC. The results shows the profile form of trimethyl xanthina of Cammelia sinensis callus have similarities with the standard form of trimethyl xanthina.

  13. Study of Androgenesis Ability and Callus Induction in Four Varieties of Tomato (Lycopersicon esculentum Mill by Anther Culture

    Directory of Open Access Journals (Sweden)

    R. Najib

    2016-02-01

    Full Text Available Introduction Tomato (Lycopersicon esculentum Mill is one of the most important vegetables which in addition of its importance as a food, is utilized as a model plant for cytological and cytogenetic studies. Tomato breeding programs are often based on the production and selection of hybrid plants. Producing hybrid plants and application of features that is needed to breed pure lines with high specific combining abilities, is highly required.New technologies such as doubled haploid can be an effective strategy to provide pure lines in tomato. Generation of homozygous doubled haploid lines through induction of androgenesis is a promising alternative method to the classical breeding programs. However, this technology is poorly developed in tomato so that some improvements in methodology are required. Genotype and stages of microspore development are critical factors for induction of androgenesis in tomato. Among them, the genotype is more important than other factors. The purpose of this study was to investigate the possibility of callus induction from anthers in some tomato genotypes. Materials and Methods: In order to investigate the androgenic response and callus induction through anther culture in tomato, four varieties including Mobil-Netherlands, Baker, U. S. Agriseed and Khoram were chosen. To determine the appropriate stage of microspore development for anther culture, cytologycal studies were accomplished at different size length of flower buds (2-7.9 mm. Flower buds were incubated at 4oC for 15 minutes and stained in acetocarmin %4 solution. Based on cytological studies in four tested cultivars, flower buds with size length 4-4.9 mm were chosen, as they had the highest frequency of meiotic microspores to microspores mid uninucleate. Pretreatments were colchicine solution (250 mgr/L at 4 °C for 48 h. The anthers were cultured on MS medium containing 2 mgr/L IAA and 1 mgr/L 2ip. All changes in frequency of callus induction and diameter of

  14. Biosynthesis of Gold and Silver Nanoparticles Using Extracts of Callus Cultures of Pumpkin (Cucurbita maxima).

    Science.gov (United States)

    Iyer, R Indira; Panda, Tapobrata

    2018-08-01

    The potential of callus cultures and field-grown organs of pumpkin (Cucurbita maxima) for the biosynthesis of nanoparticles of the noble metals gold and silver has been investigated. Biosynthesis of AuNPs (gold nanoparticles) and AgNPs (silver nanoparticles) was obtained with flowers of C. maxima but not with pulp and seeds. With callus cultures established in MS-based medium the biogenesis of both AuNPs and AgNPs could be obtained. At 65 °C the biogenesis of AuNPs and AgNPs by callus extracts was enhanced. The AuNPs and AgNPs have been characterized by UV-visible spectroscopy, TEM, DLS and XRD. Well-dispersed nanoparticles, which exhibited a remarkable diversity in size and shape, could be visualized by TEM. Gold nanoparticles were found to be of various shapes, viz., rods, triangles, star-shaped particles, spheres, hexagons, bipyramids, discoid particles, nanotrapezoids, prisms, cuboids. Silver nanoparticles were also of diverse shapes, viz., discoid, spherical, elliptical, triangle-like, belt-like, rod-shaped forms and cuboids. EDX analysis indicated that the AuNPs and AgNPs had a high degree of purity. The surface charges of the generated AuNPs and AgNPs were highly negative as indicated by zeta potential measurements. The AuNPs and AgNPs exhibited remarkable stability in solution for more than four months. FTIR studies indicated that biomolecules in the callus extracts were associated with the biosynthesis and stabilisation of the nanoparticles. The synthesized AgNPs could catalyse degradation of methylene blue and exhibited anti-bacterial activity against E. coli DH5α. There is no earlier report of the biosynthesis of nanoparticles by this plant species. Callus cultures of Cucurbita maxima are effective alternative resources of biomass for synthesis of nanoparticles.

  15. Production of parthenolide in organ and callus cultures of ...

    African Journals Online (AJOL)

    In addition, callusing capacity and weight of callus of different explants under different treatments were also determined. Parthenolide content was determined in the explants as well as in callus using RP-HPLC on Luna C18 column and SPD-10A UV detector. The mobile phase used was acetonitrile: water (55 : 45) and the ...

  16. Production of isoflavones, daidzein and genistein in callus cultures of Pueraria candollei Wall. ex Benth. var. mirifica

    Directory of Open Access Journals (Sweden)

    Sanha Panichajakul

    2006-03-01

    Full Text Available Callus cultures of Pueraria candollei var. mirifica were first established from various parts of explants with the objective of isoflavones, daidzein and genistein production. The cultures were studied on their growth and isoflavone production by various combinations of growth regulators, auxins and cytokinins at 25±2ºC. Daidzein and genistein accumulated in the cells were determined. The results revealed that callus of P. candollei var. mirifica was capable of producing high level of both isoflavones consistently. The culture temperature played an important role in the growth and isoflavone production. Over twofold of growth and threefold of isoflavone production were demonstrated at 32±2ºC. The callus established from the stems in MS medium supplemented with 4.5 µM 2,4-D and 0.46 µM kinetin produced the highest yield of daidzein (5.12 mg/g, DW and genistein (2.77 mg/g, DW, which was remarkably higher than the intact plants.

  17. Effects of plant growth regulators on seed germination and callus induction of hylocereus costaricensis

    International Nuclear Information System (INIS)

    Sheng, W.K.

    2016-01-01

    Dragon fruit (Hylocereus costaricensis) belongs to the family Cactaceae and are climbing vines which have received worldwide attention in recent years. However, there are still lack of information on the protocols for the establishment of In vitro culture system. In the present study, seed germination percentage were determined by culturing seeds on semi-solid Murashige and Skoog medium (MS) supplemented with 1 ppm 6-Benzylaminopurine (BAP) together with either 0, 0.5 or 0.8 ppm of Indole-3-butyric acid (IBA). Germination percentage was the highest by using plant growth regulators (PGRs) combination of 1 ppm BAP and 0 ppm IBA (93.33%). Subsequently, the cotyledons from seedlings of the germinated seeds were used for subsequent callus induction. Small pieces of cotyledons were excised and cultured on MS medium fortified with 0.45, 0.9, 1.8, 2.7, 3.6, and 4.5 ppm of 2,4-Dichlorophenoxyacetic acid (2,4-D) together with either 0, 0.9 or 1.8 ppm of BAP. Callus induction percentage was highest using the plant growth regulators (PGRs) combination of 3.6 ppm 2,4-D and 1.8 ppm BAP (75%). Hence, 3.6 ppm of 2,4-D and 1.8 ppm BAP was the best combination for callus induction of Hylocereus costaricensis. (author)

  18. Plants regeneration of a papaya hybrid (IBP 42-99 from callus obtained from apexes of in vitro plants.

    Directory of Open Access Journals (Sweden)

    Jorge Gallardo Colina

    2004-07-01

    Full Text Available In Cuba there realizes innumerable efforts to increase the food production and especially the fruit trees Inside which the papaya has great importance. In this sense studies are realized to obtain plants resistant to virus that they need of tools that they support and increase the indexes of obtaining transgenic line in the events of transformation specifically in a papaya hybrid. As main objective was to develop a protocol for the regeneration of plants of papaya hybrid from callus obtained of in vitro plant apexes.plan to develop a tool that supports and increases the indexes of obtaining line transgenic in the events of transformation in a hybrid of papaya.In vitro plants of the hybrid IBP 42-99 were used as plant material. The culture medium Nitsh and Nitsh was used and the growth regulators that permitted the obtaining of the best callus with embryogenic structures were studied, and also, the concentrations in which they were more efficient were adjusted. The capacity of callus formation from different parts of the stem of the in vitro plants was studied. Different culture medium for the regeneration of papaya plants from the obtained callus was studied. It was possible to obtain callus by combining 6-BAP with ANA and AIA. The best results are obtained when segments from in vitro plants shoot, 1 cm length from the meristem, were used. Also, by eliminating the meristem in the apexes, an increase in the callus formation capacity of the explant was achieved. Plants were obtained from callus using the culture medium MS supplement with 0.09mg.l-1 of AIA, 0.01mg.l-1 of AG3 and 2mg.l-1 of Zeatin and the best percentage was achieved with the callus coming from the treatments with less concentration of 6-BAP and AIA. Key words: culture medium, micropropagation, organogenesis, segments

  19. An attempt to conserve whatnot somniferum (L.) dual - a highly essential medicinal plant, through in vitro callus culture

    International Nuclear Information System (INIS)

    Rout, J.R.; Sahoo, S.

    2011-01-01

    A simple effective protocol was developed for conservation and plant propagation through callus cultures of Withania somnifera (Ashwagandha). Seed germination percentage reached a maximum value of 64.3% on half MS + GA3 0.25 mg/l at third week of culture. Three different basal media compared for seed germination, MS was most effective. Out of 25 combinations of growth regulators evaluated, MS + 1.0 mg/l BA + 1.0 mg/l 2, 4-D found to be best for callus induction and proliferation regardless to explants. Among the four different explants tested, In vivo leaf explant was found most suitable for callus induction, proliferation and fresh weight gain. The highest callus induction frequency percentage 86.4% was recorded with In vivo leaf explant whereas, 43.4% in In vitro leaf explant at day 30 on MS augmented with 1.0 mg/l BA + 1.0 mg/l 2,4-D. Among different growth regulator combinations tested in augmentation with MS for shoot initiation and elongation, 2.0 mg/l BA + 1.0 mg/l NAA was the best eliciting a maximum of 82.3% shoot induction with highest shoot number 4.8 shoots/callus. The original callus was sub-cultured 2 times on fresh shoot multiplication medium after each harvest of the shoots. Of three different auxins tested for In vitro rooting, IBA was most effective compared to IPA and NAA. Half-strength MS medium containing IBA at an optimum concentration of 2.0 mg/l induced rooting in 83.1% of the In vitro derived shoots. The rooted plantlets were acclimated and eventually established in soil. (author)

  20. Effective salt criteria in callus-cultured tomato genotypes.

    Science.gov (United States)

    Dogan, Mahmut; Tipirdamaz, Rukiye; Demir, Yavuz

    2010-01-01

    Na+, Cl-, K+, Ca2+, and proline contents, the rate of lipid peroxidation level in terms of malondialdehyde (MDA) and chlorophyll content, and the changes in the activity of antioxidant enzymes, such as superoxide dismutase (SOD: EC 1.15.1.1), catalase (CAT: EC 1.11.1.6), ascorbate peroxidase (APX: EC 1.11.1.11), and glutathione reductase (GR: EC 1.6.4.2), in tissues of five tomato cultivars in salt tolerance were investigated in a callus culture. The selection of effective parameters used in these tomato genotypes and to find out the use of in vitro tests in place of in vivo salt tolerance tests were investigated. As a material, five different tomato genotypes during a 10-day time period were used, and 150 mM NaCl was applied at callus plant tissue. The exposure to NaCl induced a significant increase in MDA content in both salt-resistant and salt-sensitive cultivars. But the MDA content was higher in salt-sensitive cultivars. The chlorophyll content was more decreased in salt-sensitive than in salt-resistant ones. The proline amount was more increased in salt-sensitive than in salt-resistant ones. It has been reported that salt-tolerant plants, besides being able to regulate the ion and water movements, also exhibit a strong antioxidative enzyme system for effective removal of ROS. The degree of damage depends on the balance between the formation of ROS and its removal by the antioxidative scavenging system that protects against them. Exclusion or inclusion of Na+, Cl-, K+, and Ca2+, antioxidant enzymes and MDA concentration play a key protective role against stress, and this feature at the callus plant tissue used as an identifier for tolerance to salt proved to be an effective criterion.

  1. Synthesis of deuterium-labelled substrates for the study of oleuropein biosynthesis in Olea europaea callus cultures.

    Science.gov (United States)

    Serrilli, Anna Maria; Maggi, Agnese; Casagrande, Valentina; Bianco, Armandodoriano

    2016-01-01

    We propose the cell culture approach to investigate oleuropein (1) biogenesis in Olea europaea L. We suggest employing olive callus cultures to identify the iridoidic precursor of oleuropein. In fact, we confirmed that callus cells from olive shoot explants are able to produce key secoiridoid as 1. To enable this approach, we synthesised and characterised deuterium-labelled iridoidic precursors belonging both to the loganin and the 8-epiloganin series. These iridoids are [7,8-(2)H2]-7-deoxy-8-epi-loganin (2(D)), [8,10-(2)H2]-8-epi-loganin (4(D)) and [7,8-(2)H2]-7-deoxy-loganin (3(D)).

  2. Primary characterization and evaluation of anti ulcerogenic activity of an aqueous extract from callus culture of Cereus peruvianus Mill. (Cactaceae).

    Science.gov (United States)

    Jayme, Milena O; Ames, Franciele Q; Bersani-Amado, Ciomar A; Machado, Maria de Fatima P S; Mangolin, Claudete A; Goncalves, Regina A C; de Oliveira, Arildo J B

    2015-01-01

    In the current study we reported cultivation, extraction procedure, analysis and preliminary characterization of the aqueous extract from Cereus peruvianus callus culture and evaluated its anti ulcerogenic activity in vivo models of experimental ulcers in Wistar rats. The obtained aqueous extract from callus (AC) was dialyzed and subjected to freeze-thaw process, providing a possible polysaccharide. The carbohydrate and protein contents of the aqueous extract were estimated at 53.4% and 0.66%, respectively, composed primarily of galactose, arabinose and galacturonic acid, with minor amounts of glucose. This appeared heterogeneous when analyzed by high-performance size-exclusion chromatography and a multiangle laser light scattering detector (HPSEC-MALLS). The AC was found to be significantly effective against ethanol-induced lesions but was ineffective against indomethacin-induced lesions. The callus culture of C. peruvianus is an alternative source for the synthesis of substances originally produced by plants. The calluses grown indefinitely in vitro under controlled conditions are stable tissues, and the aqueous extract from calluses may be used instead of fully developed plants using the protocols described in this study.

  3. Responses of some selected Malaysian rice genotypes to callus ...

    African Journals Online (AJOL)

    user

    2011-01-17

    Jan 17, 2011 ... embryogenic callus culture (Yin et al., 1993). In this study, overall, the callus from the MS medium had a good texture. Furthermore, some genotypes also produced better callus in other concentrations of 2,4-D treatment, however, the quality of callus was not as good as those in. 10 µM 2,4-D. Notably, the ...

  4. The Reserpine Production and Callus Growth of Indian Snake Root (Rauvolfia serpentina (L. Benth. Ex Kurz Culture by Addition of Cu2+

    Directory of Open Access Journals (Sweden)

    NUNUNG NURCAHYANI

    2008-07-01

    Full Text Available The objectives of this research were to study the effects of Cu2+ addition on the reserpine production and callus growth from in vitro culture indian snake root (Rauvolfia serpentina (L. Benth. Ex Kurz. This research frame work was based on the potency of snake root which was many exploited as anti-hypertension. The addition of elicitor Cu2+ in the form of CuCl2 would influence the ion transport of cell and changed of cytoplasm pH, and also has effects on synthesis and activity of enzymes which role in reserpine production and callus growth. The research was conducted in two steps, using Completely Randomized Design. The first step was the callus initiation to promote callus growth. Second step was the treatment to induce reserpine production. The callus was divided into five groups: 0; 5; 10; 20; 40; and 80 µM. Morphology, wet weight, dry weight, growth rate, and reserpine content of callus were determined after 15 treatment day. Data were analyzed using ANOVA and continued by DMRT 5%. The result showed that reserpine production increased in addition of 5 µM and 10 µM Cu2+ in callus culture of R. serpentina and reduced in addition of Cu2+ more than 10 µM. The callus growth significantly decreased by increasing concentration of Cu2+.

  5. Stimulation effects of γ-irradiation combined with colchicine on callus formation and green plant regeneration in rice anther culture

    International Nuclear Information System (INIS)

    Jin Wei; Chen Qiufang; Wang Cailian; Lu Yimei

    1999-09-01

    The ability of callus formation and green plant regeneration was very different for various rice types and varieties in rice anther culture. It was quite effective that rice anthers were irradiated with 10-40 Gy of γ-rays after 30 d incubation on induction medium and calli were treated on differentiation medium contained 10-75 mg/L of colchicine for increase of callus formation and green plant regeneration. Among these treatments, 10 Gy of γ-rats was the best for callus formation, and 20 Gy of γ-rays or 30 mg/L of colchicine was the most favourable for green plant regeneration. The simulation effect of 20 Gy of γ-irradiation combined with 30 mg/L of colchicine on green plant regeneration was much better than that of their separate use in rice anther culture

  6. Biosynthesis of phenolic compounds in hypocotyl callus cultures of fenugreek (Trigonella foenum graecum L. )

    Energy Technology Data Exchange (ETDEWEB)

    Dhandapani, M; Antony, A; Subba Rao, P V [Indian Inst. of Science, Bangalore. Dept. of Biochemistry

    1977-03-01

    Hypocotyl callus cultures of fenugreek were studied to determine their potential for synthesizing phenolics, particularly those which are intermediates in lignin and flavonoid biosynthesis. The cultures were found to be capable of synthesizing an array of phenolic compounds characteristic of higher plants. Both phenylalanine-U-/sup 14/C and cinnamic acid-U-/sup 14/C were found to be efficient precursors of these phenolics.

  7. Regenerative callus induction and biochemical analysis of Stevia rebaudiana Bertoni

    Directory of Open Access Journals (Sweden)

    Dhurva P. Gauchan

    2014-07-01

    Full Text Available Stevia Leaves are the principal source of stevioside, which is estimated to be 100-300 times sweeter than table sugar. Stevioside has clinical significance as they are reported to maintain glucose levels in human blood. Owing to the difficulties in propagation of stevia through seeds and vegetative methods, callus culture has been an efficient alternative for generation of stevioside. The aim of this study is to develop an efficient and standardized protocol for maximum induction and multiplication of callus from a leaf. Callus culture was established from leaves in MS basal media fortified with various combinations (BAP, NAA, 2,4-D, KN, IBA and concentrations of phytohormones. The best callusing (100% was recorded in MS media supplemented with (2,4-D 1.0mg/l + NAA 1.0mg/l. The callus was harvested after 4 weeks and screened for the presence of various bioactive compounds. The qualitative results showed that the extracts of callus contained bioactive compounds like flavonoids, glycosides, phenol, tannins, sterols and saponins thereby making callus one of the sources for extraction of various secondary metabolites.

  8. The influence of auxins on the biosynthesis of isoprene derivatives in callus cultures of Vaccinium corymbosum var. bluecrop.

    Science.gov (United States)

    Migas, Piotr; Luczkiewicz, Maria; Cisowski, Wojciech

    2006-01-01

    Callus cultures of Vaccinium corymbosum var. bluecrop were optimized for their isoprene derivatives production by supplementing Schenk-Hildebrandt (SH) medium with constant concentration of kinetin (2.32 microM) and two different amounts of selected auxins. Every auxin, except for IBA, used in 10-time higher concentration (2,4D, NAA, IAA, NOA) stimulated biosynthesis of beta-sitosterol and inhibited triterpene synthesis. Quantitative analysis of isoprene derivatives in callus biomass collected on the 25th day of the experiment proved that the analyzed callus of Vaccinium corymbosum var. bluecrop synthesized the highest amount of isoprene derivatives after subculturing on SH medium modified with 22.6 microM of 2,4D and 2.32 microM of kinetin.

  9. Callus regeneration from stem explants of Pseudarthira viscida (L ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-01

    Sep 1, 2009 ... regeneration frequency have come from tissue culture work done in ... Table 1. In vitro responses from stem callus of Psudarthria viscida Wight & Arn. Growth .... plantlets regeneration from cotyledonary callus of Tomato.

  10. Bioactivity of Neem (Azadirachta indica) callus extract

    International Nuclear Information System (INIS)

    Ahmed, I.M.

    2008-04-01

    This study was conducted in order to explore the possibility of utilizing plant tissue culture techniques for production of secondary metabolites from callus culture of Azadirachta indica (Neem) and to investigate the bioactivity of the established callus extract in comparison with the extract from the intact leaves. The presence of secondary metabolites in the extracts was detected by Thin Layer Chromatography (TLC). Both the callus and leaf extracts eluted five fraction of compounds and it were observed that callus extract had a good resolution. various extract concentration (5.10. and 20 mg/ml) were determined for the rate and extent of inhibition kinetics against staphylococcus aureus. Escherichia coli, and candida albicans. Results showed that callus extract of A. indica wiped out all viable cells of C. albicans within 18 hours and the subsequent concentration 5 and 10 mg/ m1 retard the growth after 24 h. A higher concentration of 20 mg/ ml had the same effect on S. aureus after 6 h and the E. coli cells were completely inhibited by the extracts after 24 h. Similar kinetics were showed by leaf extract but in slight rate as compared to the callus extract. In general both extract posses antimicrobial activity with notable efficient rates. For assaying of the inhibitory effect on some phyto pathogens the effect of different concentrations of the callus and leaf extracts on the radial growth of Drechslera rostrata. Fusarium oxysporum and Alterneria alternata were in vitro assessed. Obvious inhibitory effect was observed on the mycelia radial growth of the three treated fungi. The level of inhibition increased with the increase of te extract concentration. The maximum inhibitory effect (84%) was recorded with Drechslera rostrata when inoculated in media contain 20 mg/ ml of callus while the inhibition rate of mycelia growth of the same species reaches 61% when inoculated in a medium contain the same concentration of the neem leaf extract. The subsequent

  11. Biomass Yield and Steviol Glycoside Production in Callus and Suspension Culture of Stevia rebaudiana Treated with Proline and Polyethylene Glycol.

    Science.gov (United States)

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay

    2015-06-01

    Enhanced production of steviol glycosides (SGs) was observed in callus and suspension culture of Stevia rebaudiana treated with proline and polyethylene glycol (PEG). To study their effect, yellow-green and compact calli obtained from in vitro raised Stevia leaves were sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of proline (2.5-10 mM) and PEG (2.5-10 %) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension culture biomass (i.e. both fresh and dry weight content) was increased with 5 mM proline and 5 % PEG, while at further higher concentrations, they got reduced. Further, quantification of SGs content in callus (collected at 15th day) and suspension culture (collected at 10th and 15th day) treated with and without elicitors was analysed by HPLC. It was observed that chemical stress enhanced the production of SGs significantly. In callus, the content of SGs increased from 0.27 (control) to 1.09 and 1.83 % with 7.5 mM proline and 5 % PEG, respectively, which was about 4.0 and 7.0 times higher than control. However, in the case of suspension culture, the same concentrations of proline and polyethylene glycol enhanced the SG content from 1.36 (control) to 5.03 and 6.38 %, respectively, on 10th day which were 3.7 times and 4.7 times higher than control.

  12. Effect of salts (NaCl and Na2CO3) on callus and suspension culture of Stevia rebaudiana for Steviol glycoside production.

    Science.gov (United States)

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay

    2014-03-01

    Steviol glycosides are natural non-caloric sweeteners which are extracted from the leaves of Stevia rebaudiana plant. Present study deals the effect of salts (NaCl and Na2CO3) on callus and suspension culture of Stevia plant for steviol glycoside (SGs) production. Yellow-green and compact calli obtained from in vitro raised Stevia leaves sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of NaCl (0.05-0.20%) and Na2CO3 (0.0125-0.10%) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension biomass cultured on salts showed less growth as well as browning of medium when compared with control. Quantification of SGs content in callus culture (collected on 15th day) and suspension cultures (collected at 10th and 15th days) treated with and without salts were analyzed by HPLC. It was found that abiotic stress induced by the salts increased the concentration of SGs significantly. In callus, the quantity of SGs got increased from 0.27 (control) to 1.43 and 1.57% with 0.10% NaCl, and 0.025% Na2CO3, respectively. However, in case of suspension culture, the same concentrations of NaCl and Na2CO3 enhanced the SGs content from 1.36 (control) to 2.61 and 5.14%, respectively, on the 10th day.

  13. Establishment of callus from Opuntia robusta Wendl., a wild and ...

    African Journals Online (AJOL)

    coco santos

    2013-04-22

    Apr 22, 2013 ... plants produce tender cladodes, consumed as vegetable and fruits (prickly pear) .... The highest callus amount was obtained in media supplemented ... CO2 during carboxylation and decarboxylation reactions, improves the ...

  14. In Vitro Culture and Phytochemical Analysis of Passiflora tenuifila Killip and Passiflora setacea DC (Passifloraceae).

    Science.gov (United States)

    Sozo, Jenny Sumara; Cruz, Daniel Cuzziol; Pavei, Ana Flavia; Pereira, Isadora Medeiros da Costa; Wolfart, Marcia; Ramlov, Fernanda; Fiuza Montagner, Daiane; Maraschin, Marcelo; Viana, Ana Maria

    2016-01-01

    We have developed reproducible micropropagation, callus culture, phytochemical, and antioxidant analysis protocols for the wild passion fruit species P. tenuifila, and P. setacea, native to the Brazilian endangered biomes Atlantic Forest, Cerrado, and Caatinga, by using seeds and explants from seedlings and adult plants. Genotype and explant origin-linked differences are visible amongst the Passiflora species concerning callus production, total phenolics, and antioxidant activity. The protocols developed for screening phytochemicals and antioxidants in P. tenuifila and P. setacea callus extracts have shown their potential for phenolic production and antioxidant activity. The high level of phenolic compounds seems to account for the antioxidant activity of methanolic extracts of P. tenuifila derived from 45-day-old immature seed callus. The methanolic extracts of callus derived from P. setacea seedling leaf node and cotyledonary node explants have shown the highest antioxidant activity despite their lower content of phenolics, as compared to cotyledon callus extracts. The optimized micropropagation and callus culture protocols have great potential to use cell culture techniques for further vegetative propagation, in vitro germplasm conservation, and secondary metabolite production using biotic and abiotic elicitors.

  15. Influence of thiamine on the post-irradiation effect caused by fast neutrons or gamma radiation in callus cultures of Haplopappus gracilis (Nutt) A. Gray

    International Nuclear Information System (INIS)

    Wajda, L.; Korzonek, M.

    1979-01-01

    Influence of thiamine within the investigated range of concentrations 1.5 to 12.0 mg/l on the postirradiation effect caused by fast neutrons or gamma irradiation was found in cultures of Haplopappus gracilis callus. The lowest sensitivity to fast neutrons was noticed in callus grown on nutrient combination 1.5 to 3.0 and 12.0 to 3.0 mg/l of thiamine. On the contrary the highest sensitivity to gamma radiation was shown by cultures grown on nutrient media: 1.5 to 3.0 mg/l; all the other investigated thiamine concentrations caused a marked decrease in the sensitivity of the callus. (author)

  16. An efficient method for in vitro callus induction in Myrciaria dubia (Kunth Mc Vaugh "Camu Camu"

    Directory of Open Access Journals (Sweden)

    Ana M. Córdova

    2014-03-01

    Full Text Available Due to the high variability in vitamin C production in Myrciaria dubia "camu camu", biotechnological procedures are necessary for mass clonal propagation of promising genotypes of this species. The aim was to establish an efficient method for in vitro callus induction from explants of M. dubia. Leaf and knot sex plants were obtained from branches grown in the laboratory and from fruit pulp collected in the field. These were desinfected and sown on Murashige-Skoog (1962 medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, benzylaminopurine (BAP and kinetin(Kin. The cultures were maintained at 25±2°C in darkness for 2 weeks and subsequently with a photoperiod of 16 hours in light and 8 hours in dark for 6 weeks. Treatment with 2 mg/L 2,4-D and 0.1 mg/L BAP allowed major callus formation in the three types of explants. Calluswere generated from the first week (knots, fourth week (leaves and sixth week (pulp and these were friable (leaves and nodes and non-friable (pulp. In conclusion, the described method is efficient for in vitro callus induction in leaves, knots and pulp of M. dubia, been leaves and knots explants more suitable for callus obtention

  17. Using callus culture to study the drought tolerance of wheat genotypes

    Directory of Open Access Journals (Sweden)

    Kondić Ankica

    2000-01-01

    Full Text Available In this paper, we studied the drought tolerance under in vitro conditions of two winter wheat (T. aestivum L. cultivars, Košuta and Renesansa. The tolerance was tested on a modified MS (Murashige and Skoog, 1962 nutrient medium to which polyethylene glycol (PEG was added at three different concentrations: 10%, 20% and 30%. Calluses from the control group were grown on a medium without PEG. After four months of growing on these mediums, fresh weight and dry matter content in the callus tissue were determined. We found significant differences in genotype response to different PEG concentrations The highest concentration (30% was lethal to the isolated embryos, so at this concentration no calluses formed in any of the genotypes. At 10 and 20% PEG, there was a significant decrease in the fresh callus weight in both genotypes. Thus, at the lowest concentration ( 10% PEG, the fresh weight decreased by 90% in Košuta and by 93% in Renesansa relative to the control. Due to callus dehydration in the presence of PEG, the dry matter content in the calluses of both genotypes increased with increasing PEG concentrations in the medium. In Košuta, the dry matter content increased from 8.7% (control to 24.9% (20% PEG, while in Renesansa it increased from 8.6% (control to 39.7% (20% PEG.

  18. Enlargement of induced variations by combined method of chronic irradiations with callus culture in sugarcane

    International Nuclear Information System (INIS)

    Nagatomi, Shigeki

    1993-01-01

    The present study was conducted to elucidate the effects of gamma ray irradiation and callus culture upon induced variation of the regeneratives. The populations regenerated from young leaf tissue of chronic irradiated plnats grown under a gamma field receiving a total dose of 300 and 100 Gy, showed rather wider variation on quantitative characters than plants from populations of the non-irradiated. This variation extended in both negative and positive directions. Analysis of variance also revealed that variation and heritability in broad sense of most agronomic characters increased significantly among the subclones as the irradiation done rose. Principal component analysis also indicated that the subclones from the irradiated population were more variable than the non-irradiated. Such variation with higher heritability could be transmitted to the following generations by clonal propagation and utilized as genetic sources in mutation breeding. The combined method with chronic irradiation followed by tissue culture is evaluated as an effective method of widening mutation spectrum and increasing mutation frequency in regenerated plants. In addition, this method is valid to improve any crop species which can regenerate plants through callus culture. (author)

  19. Interaction of callus selection media and stress duration for in vitro ...

    African Journals Online (AJOL)

    Callus culture is a novel approach addressing cultured cells as selection units independent of whole plant. Natural variations for drought tolerance existing among cell lines can be exploited in vitro in the presence of suitable concentration of osmoticum and stress duration. The study was aimed to standardized callus ...

  20. Light-induced biochemical variations in secondary metabolite production and antioxidant activity in callus cultures of Stevia rebaudiana (Bert).

    Science.gov (United States)

    Ahmad, Naveed; Rab, Abdur; Ahmad, Nisar

    2016-01-01

    Stevia rebaudiana (S. rebaudiana) is a very important species with worldwide medicinal and commercial uses. Light is one of the major elicitors that fluctuate morphogenic potential and biochemical responses. In the present study, we investigated the effect of various spectral lights on biomass accumulation and secondary metabolite production in callus cultures of S. rebaudiana. Leaf explants were placed on Murashige and Skoog (MS) medium and exposed to various spectral lights. 6-Benzyle adenine (BA) and 2, 4-dichlorophenoxy acetic acid (2, 4-D; 2.0 mgl(-1)) were used for callus induction. The control light (16/8h) produced optimum callogenic response (92.73%) than other colored lights. Compared to other colored lights, control grown cultures displayed maximum biomass accumulation (5.78 gl(-1)) during a prolonged log phase at the 18th day of growth kinetics. Cultures grown under blue light enhanced total phenolic content (TPC; 102.32 μg/g DW), total flavonoid content (TFC; 22.07 μg/g DW) and total antioxidant capacity (TAC; 11.63 μg/g DW). On the contrary, green and red lights improved reducing power assay (RPA; 0.71Fe(II)g(-1) DW) and DPPH-radical scavenging activity (DRSA; 80%). Herein, we concluded that the utilization of colored lights is a promising strategy for enhanced production of antioxidant secondary metabolites in callus cultures of S. rebaudiana. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. The effect of gamma radiation and N-ethyl-N-nitrosourea on cultured maize callus growth and plant regeneration

    International Nuclear Information System (INIS)

    Moustafa, R.A.K.; Duncan, D.R.; Widholm, J.M.

    1989-01-01

    Regenerable maize calli of two inbred lines were exposed to 0 to 100 Gy of gamma rays or treated with 0 to 30 mM of N-ethyl-N-nitrosourea (ENU) to determine their effect on growth and plant regeneration capability. Both growth and plant regeneration capacity decreased with increasing levels of either gamma radiation or ENU; however, plant regeneration capacity was more sensitive to either agent than growth. The 50% inhibition dose (I 50 ) for callus growth (fresh-weight gain) was approximately 100 Gy of gamma radiation and 30 mM ENU. The I 50 for plant regeneration capacity of treated callus was approximately 25 Gy of gamma radiation and 2.5 mM ENU. The decrease in plant regeneration capacity correlated with a change in tissue composition of the treated callus from a hard, yellow and opaque tissue to a soft, grayish-yellow and translucent tissue. This change was quantified by measuring the reduction of MnO 4 - to MnO 2 (PR assay) by the callus. These results suggest that the effect of gamma radiation or ENU on plant regeneration capacity must be taken into consideration if these potentially mutagenic agents are to be used on maize callus cultures, for the purpose of producing useful mutations at a whole plant level. The data also suggest that the PR assay may be useful for predicting the actual plant regeneration capacity of maize callus. (author)

  2. In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells.

    Directory of Open Access Journals (Sweden)

    Arifullah Mohammed

    Full Text Available Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l-1 IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13 and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW and Echioidinin (ED. Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time- and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

  3. In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells

    Science.gov (United States)

    Mohammed, Arifullah; Chiruvella, Kishore K.; Rao, Yerra Koteswara; Geethangili, Madamanchi; Raghavan, Sathees C.; Ghanta, Rama Gopal

    2015-01-01

    Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l–1 IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time- and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents. PMID:26488879

  4. Agrobacterium-mediated genetic transformation of Coffea arabica (L.) is greatly enhanced by using established embryogenic callus cultures

    Science.gov (United States)

    2011-01-01

    Background Following genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures, co-cultivation with LBA1119 OD600 = 0.6 for five days at 20 °C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation) and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%). At the histological level, successful transformation was related to the number of proembryogenic masses present. All the selected plants were proved to be transformed by PCR and Southern blot hybridization. Conclusion Most progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. This is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. Our results make Agrobacterium

  5. Agrobacterium-mediated genetic transformation of Coffea arabica (L. is greatly enhanced by using established embryogenic callus cultures

    Directory of Open Access Journals (Sweden)

    Lashermes Philippe

    2011-05-01

    Full Text Available Abstract Background Following genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures, co-cultivation with LBA1119 OD600 = 0.6 for five days at 20 °C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%. At the histological level, successful transformation was related to the number of proembryogenic masses present. All the selected plants were proved to be transformed by PCR and Southern blot hybridization. Conclusion Most progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. This is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. Our

  6. Optimization of induction, subculture conditions, and growth kinetics of Angelica sinensis (Oliv.) Diels callus.

    Science.gov (United States)

    Huang, Bing; Han, Lijuan; Li, Shaomei; Yan, Chunyan

    2015-01-01

    Angelica sinensis (Oliv.) Diels is an important traditional Chinese medicine, and the medicinal position is its root. This perennial herb grows vigorously only in specific areas and the environment. Tissue culture induction of callus and plant regeneration is an important and effective way to obtain large scale cultures of A. sinensis. The objective was to optimize the inductive, subculture conditions, and growth kinetics of A. sinensis. Tissue culture conditions for A. sinensis were optimized using leaves and petioles (types I and II) as explants source. Murashige and Skoog (MS) and H media supplemented with 30 g/L sucrose, 7.5 g/L agar, and varying concentrations of plant growth regulators were used for callus induction. In addition, four different basal media supplemented with 1.0 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), 0.2 mg/L 6-benzyladenine (BA) and 30 g/L sucrose were optimized for callus subculture. Finally, growth kinetics of A. sinensis cultured on different subculture media was investigated based on callus properties, including fresh weight, dry weight, medium pH, callus relative fresh weight growth, callus relative growth rate (CRGR), and sucrose content. MS medium supplemented with 5 mg/L α-naphthaleneacetic acid, 0.5 mg/L BA, 0.7 mg/L 2,4-D, 30 g/L sucrose and 7.5 g/L agar resulted in optimal callus induction in A. sinensis while petiole I was found as the best plant organ for callus induction. The B5 medium supplemented with 1.0 mg/L 2,4-D, 0.2 mg/L BA and 30 g/L sucrose displayed the best results in A. sinensis callus subculture assays. The optimized conditions could be one of the most potent methods for large-scale tissue culture of A. sinensis.

  7. In vitro anti-microbial activity of extracts from the callus cultures of some Nigella species

    Czech Academy of Sciences Publication Activity Database

    Landa, P.; Maršík, Petr; Vaněk, Tomáš; Rada, V.; Kokoška, L.

    2006-01-01

    Roč. 61, č. 3 (2006), s. 285-288 ISSN 0006-3088 R&D Projects: GA ČR(CZ) GA525/02/0257; GA MŠk(CZ) 1P04OC926.001 Institutional research plan: CEZ:AV0Z40550506 Keywords : Nigella * callus culture * antimicrobial activity Subject RIV: CC - Organic Chemistry Impact factor: 0.213, year: 2006

  8. UV-radiation and the flavonoid content in callus culture of Ononis arvensis L

    International Nuclear Information System (INIS)

    Tumowa, L.; Psotowa, R.

    1998-01-01

    The paper discussed a possible influence on the production of secondary metabolites - the flavonoids, by the method of elicitation in the callus cultures of Ononis arvensis L., the elicitor employed being the UV 254 and 366 nm and the sun-lamp. In some cases there was an increase in the production of flavonoids particularly 60, 120, 240 and 300 s after sun-lamp irradiation and in case of 15 and 30 min irradiation with UV-254 nm

  9. Concentration on Callus Induction in Sugarcane

    African Journals Online (AJOL)

    protocols for in vitro micro propagation of sugarcane through callus culture from foliage leaves (Bugun et al., ... protocol for specific genotypes. Sucrose is the most important type of sugar produced from sugarcane. ... laboratories via plant tissue culture technique. (Lorenzo and Ganzalez, 1998). This allows for an extended ...

  10. Influence of hydroxyurea on nucleic acids content and 3H-uridine incorporation in callus and tumorous tobacco tissues cultured in vitro

    Directory of Open Access Journals (Sweden)

    A. Bielecka

    2015-01-01

    Full Text Available In callus and tumor tissues of Nicotiana tabacum cultured for 39 days in media supplemented with various concentrations of hydroxyurea (1.3 x 10-4 M - 1.3 x 10-3 M a decrease of DNA content (ca. 24 per cent in callus tissue and ca. 23 per cent in tumour tissue and a decrease of RNA content (over 10 per cent and ca. 9 per cent in callus and tumour tissue, respectively was observed. The autoradiographic method showed that a long-lasting action of this com-pound inhibits RNA synthesis. A stronger inhibitory influence of hydroxyurea upon incorporation of 3H-uridine from the incubation medium was revealed.

  11. Secondary Metabolite Content in Roots and Callus of Paeonia Anomala L.

    Directory of Open Access Journals (Sweden)

    A.A. ZARIPOVA

    2014-06-01

    Full Text Available Taking into account the fact that in the process of introduction in vitro culture the change of secondary metabolite content may take place we compared the chemical composition of plant material of wild-growing plants of Paeonia anomala introduced and produced using the methods of clonal micropropagation, callus tissue.The content of phenolic compounds, that is catechins and gallatos was compared. Paeoniflorin content was estimated by direct spectrophotometry of methanol extracts (λ = 231,7 nm, ε 1% 1 sm = 265.4. Integral characteristic of the obtained extracts was received by comparison of absorption spectra using spectrophotometer SP - 121 within wave diapason 300 - 460 nm.Light absorption curves of methanol extracts had two distinct peaks at λ = 232 nm and λ = 275 nm typical of paeoniflorin. Paeoniflorin content was 80 % higher in young peony roots than in control plant. This glycoside content in callus culture was 44 % higher than in wild-growing plant roots and 26 % lower than in plantlet roots.The use of ethanol as extragent showed a higher content of extracted substances in callus tissue. The comparison of the obtained spectra in the region corresponding to phenolic compound absorption shows the highest phenolic compound content in callus tissue and young plant roots. Judging by light absorption maximum it may be phenolic acids. The lowest phenolic compound content was determined in adult wild-growing plant roots, where a high phlobaphene content was visually observed.The conducted research confirms the fact that callus culture of Paeonia anomala L. is a perspective producer of monoterpene glycosides and phenolic compounds. Extracts from plantlets and callus culture exceed in biological active substance content rootstock extracts of open air plants.

  12. Metabolic changes associated with shoot formation in tobacco callus cultures

    Energy Technology Data Exchange (ETDEWEB)

    Grady, K.L.

    1982-08-01

    Callus tissue derived from Nicotiana tabacum L. stem pith parenchyma cells was grown either on medium which maintains the callus in an undifferentiated state, or on medium which induces the formation of shoots. Two complementary types of studies were performed with the goal of establishing metabolic markers for the initiation of shoot formation: one designed to characterize the flow of radioactive sucrose into various metabolic pools, and one which allowed measurement of intermediary metabolite concentrations. In the former, callus tissue was incubated in (U-/sup 14/C)sucrose for periods up to one hour, and patterns of metabolite labelling in tissue grown on shoot-forming and non-shoot-forming media were compared. In the latter studies, tissue was grown for an entire subculture period on non-shoot-forming medium labelled with (U-/sup 14/C)sucrose, then subcultured to labelled non-shoot-forming or shoot-forming media, and sampled at intervals during the first week of growth. 189 references.

  13. Metabolic changes associated with shoot formation in tobacco callus cultures

    International Nuclear Information System (INIS)

    Grady, K.L.

    1982-08-01

    Callus tissue derived from Nicotiana tabacum L. stem pith parenchyma cells was grown either on medium which maintains the callus in an undifferentiated state, or on medium which induces the formation of shoots. Two complementary types of studies were performed with the goal of establishing metabolic markers for the initiation of shoot formation: one designed to characterize the flow of radioactive sucrose into various metabolic pools, and one which allowed measurement of intermediary metabolite concentrations. In the former, callus tissue was incubated in [U- 14 C]sucrose for periods up to one hour, and patterns of metabolite labelling in tissue grown on shoot-forming and non-shoot-forming media were compared. In the latter studies, tissue was grown for an entire subculture period on non-shoot-forming medium labelled with [U- 14 C]sucrose, then subcultured to labelled non-shoot-forming or shoot-forming media, and sampled at intervals during the first week of growth. 189 references

  14. Effects of Low Dose Gamma Radiation on the Formation of Shikonon Derivatives in Callus Cultures of Lithospermum erythrorhizon

    International Nuclear Information System (INIS)

    Hwang, H.Y.; Lee, Y.B.; Kim, J.S.

    2003-01-01

    The effects of low dosage r-radiation on the cell growth and the formation of shikonin derivatives were investigated in callus cultures of Lithospermum erythrorhizon under different medium and light conditions. Gamma radiation significantly affected the cell growth and formation of shikonin derivatives, depending on the culture conditions. In the cell cultures grown on M09 medium, 2 Gy and 16 /Gy of r-radiation increased the calli growth and the formation of shikonin derivatives, respectively under 16 hr day light condition

  15. Callus induction and flavonoid production on the immature seed of Stelechocarpus burahol

    Science.gov (United States)

    Habibah, N. A.; Moeljopawiro, S.; Dewi, K.; Indrianto, A.

    2018-03-01

    Stelechocarpus burahol [(Bl.) Hook. f. & Th.] is one of the medicinal plants. In vitro callus induction studies on S. burahol were carried out to determine phytohormone requirement for optimum callus induction. Immature seed explants were cultured on MS medium by adding different kinds and different concentrations of plant growth regulators (picloram and 2,4-D) under light and dark conditions. The results showed that callus formation was initiated on the 18,50th to the 55th days. The best condition for optimum callus induction was found on MS medium, which was supplemented with 7.5 mg/L picloram and was maintained in the dark condition. The callus induction varied from 60% to 100%. The callus that produced the highest flavonoid was grown on the medium with the addition of 10 mg/L of 2,4-D. In conclusion, the results represented a suitable medium for S.burahol callus induction.

  16. Callus induction and formation of Anthurium andraeanum variety ‘Lambada’ starting from foliar sections of in vitro plants

    Directory of Open Access Journals (Sweden)

    Nydia del Rivero

    2006-01-01

    Full Text Available Foliar sections of in vitro plants of Anthurium andraeanum variety ‘Lambada’ were used as explants for the induction and formation of callus. The formation of callus was evaluated in two culture mediums (MS and Nitsch, modified to which the growth regulators 2,4-Dichlorophenoxyacetic acid (2,4-D were added in four concentrations 2.26, 4.52, 6.78 and 9.04 μM and the addition of 6-Benzilaminopurine (6-BAP to a concentration of 2.22 μM keeping it constant for both culture mediums. The treatments were placed in darkness at 28±2ºC during 45 days. Characteristics of the callus, such as colour and form, were also described . The results showed, in the statistical analysis, a significant interaction among the factors in study (concentrations of the growth regulator and type of culture medium. The calluses obtained presented translucent cream yellow colour with nodular structures. Callus were formed in the two culture medium used, however, the biggest explants percentage that allowed the formation of these structures was observed in the culture medium MS modified when a concentration of 6.78 μM 2,4-D was used . It was demonstrated that it is possible to use foliar sections of in vitro plants to form callus in Anthurium andraeanum variety ‘Lambada’ in the culture medium MS with the nitrates reduced in a half and 2,4-D. Key words: callus formation, culture medium, in vitro culture, micropropagation, growth regulators

  17. Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage.

    Science.gov (United States)

    Palama, Tony L; Menard, Patrice; Fock, Isabelle; Choi, Young H; Bourdon, Emmanuel; Govinden-Soulange, Joyce; Bahut, Muriel; Payet, Bertrand; Verpoorte, Robert; Kodja, Hippolyte

    2010-05-05

    Vanilla planifolia is an important Orchid commercially cultivated for the production of natural vanilla flavour. Vanilla plants are conventionally propagated by stem cuttings and thus causing injury to the mother plants. Regeneration and in vitro mass multiplication are proposed as an alternative to minimize damage to mother plants. Because mass production of V. planifolia through indirect shoot differentiation from callus culture is rare and may be a successful use of in vitro techniques for producing somaclonal variants, we have established a novel protocol for the regeneration of vanilla plants and investigated the initial biochemical and molecular mechanisms that trigger shoot organogenesis from embryogenic/organogenic callus. For embryogenic callus induction, seeds obtained from 7-month-old green pods of V. planifolia were inoculated on MS basal medium (BM) containing TDZ (0.5 mg l(-1)). Germination of unorganized mass callus such as protocorm -like structure (PLS) arising from each seed has been observed. The primary embryogenic calli have been formed after transferring on BM containing IAA (0.5 mg l(-1)) and TDZ (0.5 mg l(-1)). These calli were maintained by subculturing on BM containing IAA (0.5 mg l(-1)) and TDZ (0.3 mg l(-1)) during 6 months and formed embryogenic/organogenic calli. Histological analysis showed that shoot organogenesis was induced between 15 and 20 days after embryogenic/organogenic calli were transferred onto MS basal medium with NAA (0.5 mg l(-1)). By associating proteomics and metabolomics analyses, the biochemical and molecular markers responsible for shoot induction have been studied in 15-day-old calli at the stage where no differentiating part was visible on organogenic calli. Two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization time-of-flight-tandem mass spectrometry (MALDI-TOF-TOF-MS) analysis revealed that 15 protein spots are significantly expressed (P tissue culture, phenolic compounds such

  18. In vitro salt stress induced production of gymnemic acid in callus ...

    African Journals Online (AJOL)

    ... is presented in this investigation. The highest efficiency of callus formation was observed in the medium containing different concentrations of 2,4-D. The gymnemic acid content increased with increasing concentration of 2,4-D along with NaCl. Key words: In vitro, Gymnema sylvestre, callus culture, gymnemic acid, salt ...

  19. Metabolomic homeostasis shifts after callus formation and shoot regeneration in tomato

    Science.gov (United States)

    Kumari, Alka; Ray, Kamalika; Sadhna, Sadhna; Pandey, Arun Kumar; Sreelakshmi, Yellamaraju; Sharma, Rameshwar

    2017-01-01

    Plants can regenerate from a variety of tissues on culturing in appropriate media. However, the metabolic shifts involved in callus formation and shoot regeneration are largely unknown. The metabolic profiles of callus generated from tomato (Solanum lycopersicum) cotyledons and that of shoot regenerated from callus were compared with the pct1-2 mutant that exhibits enhanced polar auxin transport and the shr mutant that exhibits elevated nitric oxide levels. The transformation from cotyledon to callus involved a major shift in metabolite profiles with denser metabolic networks in the callus. In contrast, the transformation from callus to shoot involved minor changes in the networks. The metabolic networks in pct1-2 and shr mutants were distinct from wild type and were rewired with shifts in endogenous hormones and metabolite interactions. The callus formation was accompanied by a reduction in the levels of metabolites involved in cell wall lignification and cellular immunity. On the contrary, the levels of monoamines were upregulated in the callus and regenerated shoot. The callus formation and shoot regeneration were accompanied by an increase in salicylic acid in wild type and mutants. The transformation to the callus and also to the shoot downregulated LST8 and upregulated TOR transcript levels indicating a putative linkage between metabolic shift and TOR signalling pathway. The network analysis indicates that shift in metabolite profiles during callus formation and shoot regeneration is governed by a complex interaction between metabolites and endogenous hormones. PMID:28481937

  20. Callus remodelling model

    Science.gov (United States)

    Miodowska, Justyna; Bielski, Jan; Kromka-Szydek, Magdalena

    2018-01-01

    The objective of this paper is to investigate the healing process of the callus using bone remodelling approach. A new mathematical model of bone remodelling is proposed including both underload and overload resorption, as well as equilibrium and bone growth states. The created model is used to predict the stress-stimulated change in the callus density. The permanent and intermittent loading programs are considered. The analyses indicate that obtaining a sufficiently high values of the callus density (and hence the elasticity) modulus is only possible using time-varying load parameters. The model predictions also show that intermittent loading program causes delayed callus healing. Understanding how mechanical conditions influence callus remodelling process may be relevant in the bone fracture treatment and initial bone loading during rehabilitation.

  1. Ultraviolet-B (UV-B) radiation as an elicitor of flavonoid production in callus cultures of jatropha (Jatropha curcas L.)

    International Nuclear Information System (INIS)

    Alvero-Bascos, E.M.; Ungson, L.B.

    2012-01-01

    Callus cultures of jatropha (Jatropha curcas L.) grown in Murashige and Skoog's (MS) medium supplemented with naphthalene-acetic acid (NAA, 20 microM) and 6-furfurylaminopurine (kinetin, 20 microM) were exposed to ultraviolet-B (UV-B) radiation to investigate its potential as an abiotic elicitor of flavonoid production. Prior to irradiation, the levels of the flavonoids, apigenin, vitexin and isovitexin in the leaf and callus extracts were determined through high performance liquid chromatography (HPLC). Results showed that vitexin and isovitexin were the dominant flavonoids in the leaves while only apigenin was detected in the calli, suggesting a correlation between the degree of differentiation and biosynthesis of flavonoids in plant tissues. Irradiation of callus cultures for 7 d using two UV-B doses (12.6 and 25.3 kJ/sq m) induced synthesis of all three flavonoids (up to 780 micro g/g dw increase) to levels similar to or higher than those found in whole leaves. The combined levels of the three flavonoids in the cultures treated with the higher UV-B dose were 20-fold higher than the control and were comparable to concentrations found in leaves while a 10-fold increase in combined flavonoid levels was observed in calli irradiated with the lower UV-B dose. Furthermore, random amplified polymorphic DNA (RAPD) analyses of DNA extracts from the leaves and calli revealed that UV-B irradiation enhanced flavonoid synthesis without altering DNA sequence. These results further support the supposed involvement of UV-B in the transcriptional regulation of the expression of flavonoid biosysnthetic genes. Overall, the findings showed that elicitation through UV-B irradiation is an effective strategy to induce flavonoid production in dedifferentiated J. curcas cultures that have lost their capacity to produce the flavonoids normally synthesized in intact organs. (author)

  2. Callus induction and plant regeneration from different explant types of Miscanthus x ogiformis Honda 'Giganteus'

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Petersen, Karen Koefoed

    1996-01-01

    . The explants were cultured on urashige and Skoog medium supplemented with 4.5, 13.6, 22.6 or 31.7 μM 2,4-dichlorophenoxyacetic acid. Three types of callus were formed but only one was embryogenic and regenerated plants. Callus induction and formation of embryogenic callus depended on the type and developmental......-propagated shoots and older leaves of greenhouse-grown plants. Immature inflorescences smaller than 2.5 cm produced a higher percentage of embryogenic callus than larger more mature inflorescences. Embryogenic callus derived from immature inflorescences had the highest regeneration capacity. Differences in 2......,4-dichlorophenoxyacetic acid concentrations had no significant effect on callus induction, embryogenic callus formation and plant regeneration....

  3. Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae: proteomic and metabolic responses at early stage

    Directory of Open Access Journals (Sweden)

    Payet Bertrand

    2010-05-01

    Full Text Available Abstract Background Vanilla planifolia is an important Orchid commercially cultivated for the production of natural vanilla flavour. Vanilla plants are conventionally propagated by stem cuttings and thus causing injury to the mother plants. Regeneration and in vitro mass multiplication are proposed as an alternative to minimize damage to mother plants. Because mass production of V. planifolia through indirect shoot differentiation from callus culture is rare and may be a successful use of in vitro techniques for producing somaclonal variants, we have established a novel protocol for the regeneration of vanilla plants and investigated the initial biochemical and molecular mechanisms that trigger shoot organogenesis from embryogenic/organogenic callus. Results For embryogenic callus induction, seeds obtained from 7-month-old green pods of V. planifolia were inoculated on MS basal medium (BM containing TDZ (0.5 mg l-1. Germination of unorganized mass callus such as protocorm -like structure (PLS arising from each seed has been observed. The primary embryogenic calli have been formed after transferring on BM containing IAA (0.5 mg l-1 and TDZ (0.5 mg l-1. These calli were maintained by subculturing on BM containing IAA (0.5 mg l-1 and TDZ (0.3 mg l-1 during 6 months and formed embryogenic/organogenic calli. Histological analysis showed that shoot organogenesis was induced between 15 and 20 days after embryogenic/organogenic calli were transferred onto MS basal medium with NAA (0.5 mg l-1. By associating proteomics and metabolomics analyses, the biochemical and molecular markers responsible for shoot induction have been studied in 15-day-old calli at the stage where no differentiating part was visible on organogenic calli. Two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization time-of-flight-tandem mass spectrometry (MALDI-TOF-TOF-MS analysis revealed that 15 protein spots are significantly expressed (P Conclusion The

  4. Callus formation of american Persea Mill. cultivar Catalina starting from segments of leaves of in vitro plants

    Directory of Open Access Journals (Sweden)

    Marisol Freire Seijo

    2004-04-01

    Full Text Available The importance of establishing protocols for the regeneration of fruit-trees through out the organogenesis and the somatic embryogenesis is based on the possibility to use them in the propagation of the species or in programs of genetic improvement. Specifically the Catalina cultivar is very coveted in Cuba, however it has not been introduced in vitro. The present work had the objective of forming callus starting from leaves of in vitro plants from avocado tree of the Catalina cultivar. The phase of multiplication of the in vitro plants was carried out in temporary immersion systems in order to favour the expansion of the leaves. Erlenmeyers 1.0l were used In the TIS and immersions every four and eight hours with a minute of duration were carried out. The effect of the Picloram, the 6 BAP and the 2,4-D on leaves of in vitro plants for the formation of callus was studied. The plants with more quality were obtained when immersions every 8 hours were carried out. It was also possible to form callus using sections of leaves of in vitro plants as explants in a culture medium with 0.1 Picloram mgl-1. Key words: in vitro plant leaves, temporary immersion system

  5. Effects of Low Dose DD Radiation on Callus Growth of Lithospermum erythrorhizon S

    International Nuclear Information System (INIS)

    Hwang, H.Y.; Kim, J.S.

    2001-01-01

    The effect of low dose DD-radiation on the callus growth of Lithospermum erythrorhizon S. cultured on different medium and lighting condition was investigated. The 8 Gy irradiation stimulated callus growth on LS medium supplemented with BA 2 mg/L and NAA 2mg/L, however, the growth of callus was more effective on LS medium supplemented with BA 1 mg/L and NAA 1 mg/L under 16 hrs day light

  6. Plant regeneration system from cotyledons-derived calluses cultures ...

    African Journals Online (AJOL)

    Administrator

    2011-09-26

    Sep 26, 2011 ... The objective of this study was to successfully establish plant regeneration system with cotyledons of. Stylosanthes guianensis Sw. cv. 'Reyan 2' as explants. In this study, the following results were obtained; (1) the highest rates of callus induction on medium MS with 3.0 mg L-1 2, 4-D with cotyledons.

  7. Effects of auxins and cytokinins on tomato callus from anthers

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    Janina H. Rogozińska

    2015-01-01

    Full Text Available An investigation was carried out on growth substance requirements of tomato callus derived from anthers for culture in vitro. Linsmaier and Skoog (1965 medium was used with various levels of auxins (IAA and NAA and cytokinins (K and BAP. The results show that cytokinin is an absolute requirement for callus growth irrespective of the auxin level. The optimum concentration of auxin in combination with cytokinin was found to be 5 μM of NAA or 25 μM of IAA, with 5 μM of K or BAP. Callus growth on media with NAA and cytokinin was superior to that on IAA, amounting to 6.05 g per piece on medium with 5 μM of NAA and BAP. Tissues grown on this medium have the highest water content. At the onset of culture the tissue is characterized by weak growth and attains its maximal increase in fresh weight after 6 weeks.

  8. Enhancement of callus induction and regeneration efficiency from ...

    African Journals Online (AJOL)

    Administrator

    2011-09-05

    Sep 5, 2011 ... efficiency from embryo cultures of Datura stramonium ... cycle (callus induction and plant regeneration) for Datura stramonium by adjusting carbon sources and ... induction and development in various species, it is not.

  9. Isoprene derivatives from the leaves and callus cultures of Vaccinium corymbosum var. bluecrop.

    Science.gov (United States)

    Migas, Piotr; Cisowski, Wojciech; Dembińska-Migas, Wanda

    2005-01-01

    The phytochemical analysis of Vaccinium corymbosum var bluecrop leaves and callus biomass revealed ursolic acid, oleanolic acid, alpha-amyrin and beta-amyrin in both plant materials. Beta-sitosterol was determined only in callus biomass. The structure of isolated compounds was elucidated by TLC co-chromatography with standards and with spectroscopic methods (1H NMR, 13C NMR, EI-MS).

  10. Observation of antioxidant activity of leaves, callus and suspension ...

    African Journals Online (AJOL)

    GREGORY

    2011-12-16

    Dec 16, 2011 ... Antioxidant activity and phenolic compound was found in Justicia gendarussa via total phenolic content (TPC) and α,α-diphenyl-β-pycrilhydrazil hydrate (DPPH) radical scavenging assays. The assays were applied on aqueous and methanolic extracts of leaves, callus culture and cell suspension culture.

  11. Regeneration of three sweet potato (Ipomea batatas (L.)) accessions via meristem, Nodal and callus induction

    International Nuclear Information System (INIS)

    Addae-Frimpomaah, F.

    2012-11-01

    In vitro regeneration of three sweet potato accessions UE007, UK-BNARI and SA-BNARI using meristem, nodal cuttings or callus induction was studied. Meristematic explants cultured on Murashige and Skoog (1962) basal medium supplemented with low concentration of benzylaminopurine (BAP) or kinetin resulted in callus with or without shoot development which delayed shoot emergence. The degree of callus development increased as the concentration of the cytokinin in the culture medium increased. Although, callus development was comparatively lower on kinetin amended medium than BAP amended medium, Murashige and Skoog medium supplemented with 0.25mg/1BAP had the highest shoot induction (80%). For further differentiation of callus or shoots into distinct stem and leaves, the culture were transferred into fresh MS medium supplemented with 0.25mg/1 BAP, 0.1 mg/1 NAA and 0.1 mg/1 Gibberellic acid (GA 3 . To overcome the delay in shoot initiation using meristem culture, nodal cuttings of sweet potato were used as explants and cultured on MS medium amended with 0.3 - 0.9mg/1 BAP. All explants cultured on 0.3 or 0.6mg/1 BAP developed shoots. Furthermore, liquid MS medium amended with 0.25mg/1 BAP, 0.1mg/I NAA, and 0.1mg/1 GA 3 also enhanced early shoot development from nodal cutting explants compared to solid culture. Post flask acclimatisation of meristem or nodal cutting-derived plantlets showed that meristem derived plantlets were better acclimatised than nodal cutting plants due to vigorous root development leading to higher percentage survival in pots and subsequent tuber production. Callusogenesis was achieved when leaf lobe explants were cultured on CLC/ Ipomoea medium supplemented with 1.0 - 4.0mg/1 2,4-D with 4.0mg/1 2,4-D being the optimal concentration. However, the calli were non-embryogenic and therefore could not produce embryos when transferred to 0.1mg/1 BAP amended medium but rather produced either single or multiple shoots. The highest percentage shoot (83

  12. Comparison of fracture site callus with iliac crest bone marrow as the source of plastic-adherent cells

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    Achmad Zaki

    2013-05-01

    Full Text Available Background: Red marrow has been described as the main source of mesenchymal stem cells although its aspiration and isolation from bone marrow was reported to have significant donor site morbidity. Since secondary bone healing occurs through formation of callus as the result of proliferation and differentiation of mesenchymal stem cells, callus may become alternative source for mesenchymal stem cells. In this study, we compared the number of plastic-adherent cells from fracture site callus and bone marrow of iliac crest after two and four weeks of culture.Methods: Sixteen New Zealand rabbits were fracturized at the femoral shaft. Then, these rabbits were taken care. After two weeks of fracturization, 3 mL iliac crest bone marrow aspiration and callus extraction of eight rabbits were cultured (group I. The other eight rabbits were treated equally after four weeks of fracturization (group II. Simultaneously, the cultures were observed after one and two weeks. Four weeks later, they were harvested. Cells were counted using Neubauer hemocytometer. The average number of cells between the sources and groups were statistically analyzed using the unpaired t-test. Results: In group I, there were 2.6 ± 0.1 x 104 cells in the culture of iliac crest bone marrow aspirate and 2.5 ± 0.1 x 104 cells in culture of callus extract from fracture site (p = 0.34. In group II, there were 2.7 ± 0.1 x 104 cells and 2.1 ± 0.1 x 104 cells, respectively (p < 0.001.Conclusion: Fracture site callus at the second week post-fracturization may be potential as source of plastic-adherent cells compared with iliac crest bone marrow. (Med J Indones. 2013;22:70-5Keywords: Bone marrow, fracture site callus, iliac crest, long bone, mesenchymal stem cell, plastic-adherent cells

  13. Sugar alcohols-induced oxidative metabolism in cotton callus culture

    African Journals Online (AJOL)

    Sugar alcohols (mannitol and sorbitol) may cause oxidative damage in plants if used in higher concentration. Our present experiment was undertaken to study physiological and metabolic responses in cotton (Gossypium hirsutum L.) callus against mannitol and sorbitol higher doses. Both markedly declined mean values of ...

  14. Induction and multiplication of callus from endosperm of Cycas ...

    African Journals Online (AJOL)

    The usage of medicinal plants in traditional medication has gained the attraction from global and local markets, mainly to cure diseases or simply for health maintenance. Callus cultures were initiated from the endosperm of the medicinal plant Cycas revoluta, cultured on half-strength Murashige and Skoog (MS) medium ...

  15. Label-Free Quantitative Proteomics of Embryogenic and Non-Embryogenic Callus during Sugarcane Somatic Embryogenesis.

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    Angelo Schuabb Heringer

    Full Text Available The development of somatic cells in to embryogenic cells occurs in several stages and ends in somatic embryo formation, though most of these biochemical and molecular changes have yet to be elucidated. Somatic embryogenesis coupled with genetic transformation could be a biotechnological tool to improve potential crop yields potential in sugarcane cultivars. The objective of this study was to observe somatic embryo development and to identify differentially expressed proteins in embryogenic (E and non-embryogenic (NE callus during maturation treatment. E and NE callus were cultured on maturation culture medium supplemented with different concentrations (0.0, 0.75, 1.5 and 2.0 g L(-1 of activated charcoal (AC. Somatic embryo formation and differential protein expression were evaluated at days 0 and 21 using shotgun proteomic analyses. Treatment with 1.5 g L(-1 AC resulted in higher somatic embryo maturation rates (158 somatic embryos in 14 days in E callus but has no effect in NE callus. A total of 752 co-expressed proteins were identified through the SUCEST (The Sugarcane EST Project, including many housekeeping proteins. E callus showed 65 exclusive proteins on day 0, including dehydrogenase, desiccation-related protein, callose synthase 1 and nitric oxide synthase. After 21 days on maturation treatment, 14 exclusive proteins were identified in E callus, including catalase and secreted protein. NE callus showed 23 exclusive proteins on day 0 and 10 exclusive proteins after 21 days on maturation treatment, including many proteins related to protein degradation. The induction of maturation leads to somatic embryo development, which likely depends on the expression of specific proteins throughout the process, as seen in E callus under maturation treatment. On the other hand, some exclusive proteins can also specifically prevent of somatic embryos development, as seen in the NE callus.

  16. Physiological and biochemical effects of morphactin IT 3233 on callus and tumour tissues of Nicotiana tabacum L. cultured in vitro III. Transamination processes catalysed by aminotransferase L-alanine: 2-oxoglutarate

    Directory of Open Access Journals (Sweden)

    Z. Chirek

    2015-01-01

    Full Text Available An active alanine transaminase was found both in callus and tumour tissues of tobacco. The enzyme is more active in the latter tissue, and the reaction balance is strongly shifted towards alanine production, while in callus tissue towards glutamic acid formation. Morphactin applied to the tissue cultures stimulates markedly the enzyme activity only in callus. A negative correlation was observed between the intensity of transamination processes and enhanced synthesis of proteins in the tissues studied. Morphactin disturbs nitrogen metabolism in the callus tissue. Tumour tissue is more resistant to the action of this substance. The different hormonal activities in these tissues may be the cause of the different effects of morphactin.

  17. Divergent regeneration-competent cells adopt a common mechanism for callus initiation in angiosperms.

    Science.gov (United States)

    Hu, Bo; Zhang, Guifang; Liu, Wu; Shi, Jianmin; Wang, Hua; Qi, Meifang; Li, Jiqin; Qin, Peng; Ruan, Ying; Huang, Hai; Zhang, Yijing; Xu, Lin

    2017-06-01

    In tissue culture, the formation of callus from detached explants is a key step in plant regeneration; however, the regenerative abilities in different species are variable. While nearly all parts of organs of the dicot Arabidopsis thaliana are ready for callus formation, mature regions of organs in monocot rice ( Oryza sativa ) and other cereals are extremely unresponsive to tissue culture. Whether there is a common molecular mechanism beyond these different regenerative phenomena is unclear. Here we show that the Arabidopsis and rice use different regeneration-competent cells to initiate callus, whereas the cells all adopt WUSCHEL-RELATED HOMEOBOX 11 ( WOX11 ) and WOX5 during cell fate transition. Different from Arabidopsis which maintains regeneration-competent cells in mature organs, rice exhausts those cells during organ maturation, resulting in regenerative inability in mature organs. Our study not only explains this old perplexity in agricultural biotechnology, but also provides common molecular markers for tissue culture of different angiosperm species.

  18. Callus Induction of Gendarussa (Justicia gendarussa by Various Concentration of 2,4-D, IBA, and BAP

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    Dwi Kusuma Wahyuni

    2017-12-01

    Full Text Available Justicia gendarussa Burm.f., a medicinal plant, is Acanthaceae that has many functions. Furthermore, the compounds in gendarussa must be produced in high quantity and quality by applying callus culture method. Accordingly, it is important to study the effects of plant growth regulators (2,4-D, IBA, and BAP on callus induction of gendarussa leaves. This research design utilized a factorial design with two factors (2,4-D and IBA: 0.5, 1, 1.5 mg/L and BAP: 0.5, 1, 1.5, 2 mg/L. The experiment consisted of 24 treatments, each of which was repeated 3 times. Observation was carried out in 6 weeks. Data on the time of callus formation, percentage of explants formed callus, and callus morphology were analyzed descriptively,while data on fresh and dry weight were analyzed by Two-Way ANOVA (α = 0.5. Interestingly, the results showed that various concentration of plant growth regulators (2,4-D, IBA, and BAP affected callus induction from leaf explants of gendarussa. We concluded that the most optimal treatment combination of concentration of plant growth regulators in inducing callus from leaf explants of gendarussa is 1.5 mg/L 2,4-D and 2 mg/L BAP with a relatively long period of callus formation at the earliest, i.e. on day 5, 2.247 g of fresh weight, 0.108 gof dry weight, white callus translucent, and friable. Moreover, the optimum treatment will be used to produce secondary metabolite and seed synthetic by cell suspension culture.

  19. Effects of gamma irradiation on nucellar callus production of the Valencia sweet orange (Citrus Sinensis Osb.) in vitro

    International Nuclear Information System (INIS)

    Pasqual, M.; Ando, A.

    1990-01-01

    Nucelli extracted from developing fruits with twelve weeks after pollination were cultured in vitro, on MS medium supplemented (in mg/l) by: thiamine H Cl 0.2; piroxidine H Cl 1.0; nicotinic acid 1.0; meso inositol 100; malt extract; sucrose 50.000; and agar 8.000 with ph = 5.7. The irradiation at 0.0; 0.5; 1.0; 2.0; 4.0; 8.0; and 12.0 kR doses was applied on, only medium, only nucellus, and medium and nucellus together. Irradiation of only nucellus and both nucellus and medium, at the same time, showed similar effects. Low doses of irradiation (until 2 kR) decrease the number of differential embryoids. The irradiation of culture medium, mainly at high doses, increases callus proliferation. (author)

  20. Correlation of different spectral lights with biomass accumulation and production of antioxidant secondary metabolites in callus cultures of medicinally important Prunella vulgaris L.

    Science.gov (United States)

    Fazal, Hina; Abbasi, Bilal Haider; Ahmad, Nisar; Ali, Syed Shujait; Akbar, Fazal; Kanwal, Farina

    2016-06-01

    Light is one of the key elicitors that directly fluctuates plant developmental processes and biosynthesis of secondary metabolites. In this study, the effects of various spectral lights on biomass accumulation and production of antioxidant secondary metabolites in callus cultures of Prunella vulgaris were investigated. Among different spectral lights, green light induced the maximum callogenic response (95%). Enhanced fresh biomass accumulation was observed in log phases on day-35, when callus cultures were exposed to yellow and violet lights. Yellow light induced maximum biomass accumulation (3.67g/100ml) from leaf explants as compared to control (1.27g/100ml). In contrast, violet lights enhanced biomass accumulation (3.49g/100ml) from petiole explant. Maximum total phenolics content (TPC; 23.9mg/g-DW) and total flavonoids content (TFC; 1.65mg/g-DW) were observed when cultures were grown under blue lights. In contrast, green and yellow lights enhanced total phenolics production (TPP; 112.52g/100ml) and total flavonoids production (TFP; 9.64g/100ml) as compared to control. The calli grown under green, red and blue lights enhanced DPPH-free radical scavenging activity (DFRSA; 91.3%, 93.1% and 93%) than control (56.44%) respectively. The DFRSA was correlated either with TPC and TFC or TPP and TFP. Furthermore, yellow lights enhanced superoxide dismutase (SOD), peroxidase (POD) and protease activities, however, the content of total protein (CTP) was higher in control cultures (186μg BSAE/mg FW) as compared to spectral lights. These results suggest that the exposure of callus cultures to various spectral lights have shown a key role in biomass accumulation and production of antioxidant secondary metabolites. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Growth, proline and ion accumulation in sugarcane callus cultures ...

    African Journals Online (AJOL)

    Calli obtained from two sugarcane cultivars (R570 and CP59-73) were exposed to different osmotic stress intensities followed by a period of stress relief. Relative rate growth, callus water content and changes in organic and inorganic solutes were determined at the end of stress and relief periods. After the stress period, ...

  2. Effect of sucrose, erythrose-4-phosphate and phenylalanine on biomassa and flavonoid content of callus culture from leaves of Gynura procumbens Merr.

    Science.gov (United States)

    Nurisa, Aryana; Kristanti, Alfinda Novi; Manuhara, Yosephine Sri Wulan

    2017-08-01

    The aims of this study were to know the effect of concentration of sucrose, erythrose-4-phosphate and phenylalanine on biomass and flavonoid content of callus cultures from leaves of sambung nyawa (Gynura procumbens Merr.). This study was experimental research with complete randomized design. Callus induction was treated in MS medium supplemented with NAA 2 mg/L, BAP 1 mg/L and sucrose concentration (10 g/L, 30 g/L and 50 g/L) respectively were combined with erythrose-4-phosphate (0 µM, 2,5 µM and 5 µM) and phenylalanine (0 mg/L, 2 mg/L and 3 mg/L), each treatment were repeated four times. After six weeks of culture, fresh and dry weight of calli were measured and extracted with ethanol absolut. Crude extract ethanolic of callus was analyzed used by a modified colorimetric with spectrophotometer method. The best yield of calli biomass (0,672 ± 0,112 gram of fresh weight and 0,033 ± 0,009 gram of dry weight) was obtained in treatment of 30 g/L sucrose of and 5 µM erythrose-4-phosphate. The highest total flavonoid content was obtained of calli treated with 30 g/L of sucrose and 3 mg/L of phenylalanine (3633,4 ppm quercetin/gram dry weight and 15777,8 ppm kaempferol/gram dry weight).

  3. Microcolony formation from embryogenic callus-derived protoplasts of oil palm

    Directory of Open Access Journals (Sweden)

    Sompong Te-chato

    2005-07-01

    Full Text Available Embryogenic callus of oil palm induced from young leaves of seedlings DxP was used as initial material for protoplast isolation. Various combinations of cellulase Onozuka RS and macerozyme R-10 were tested. Isolated protoplasts were cultured by various methods in MS medium supplemented with different phytohormones. The results revealed that 2% cellulase RS in combination with 2% macerozyme R-10 (adjusted osmoticum to 0.4 M by manitol yielded the highest number of viable protoplasts (1x107 per gram fresh weight. Dicamba at concentration 2 mg/l with 1 mg/l 6-benzyladenin (BA containing in phytagel semisolidified MS medium promoted the highest division of 2.3-4.0%. First division of the protoplasts was observed at 4 days after culture. Microcolony formation (8-10 cells was seen after three weeks of culture. Unfortunately, neither callus formation nor plantlet regeneration were obtained.

  4. Micropropagation and non-steroidal anti-inflammatory and anti-arthritic agent boswellic acid production in callus cultures of Boswellia serrata Roxb.

    Science.gov (United States)

    Nikam, Tukaram D; Ghorpade, Ravi P; Nitnaware, Kirti M; Ahire, Mahendra L; Lokhande, Vinayak H; Chopra, Arvind

    2013-01-01

    Micropropagation through cotyledonary and leaf node and boswellic acid production in stem callus of a woody medicinal endangered tree species Boswellia serrata Roxb. is reported. The response for shoots, roots and callus formation were varied in cotyledonary and leafy nodal explants from in vitro germinated seeds, if inoculated on Murshige and Skoog's (MS) medium fortified with cytokinins and auxins alone or together. A maximum of 8.0 ± 0.1 shoots/cotyledonary node explant and 6.9 ± 0.1 shoots/leafy node explants were produced in 91 and 88 % cultures respectively on medium with 2.5 μM 6-benzyladenine (BA) and 200 mg l(-1) polyvinylpyrrolidone (PVP). Shoots treated with 2.5 μM IBA showed the highest average root number (4.5) and the highest percentage of rooting (89 %). Well rooted plantlets were acclimatized and 76.5 % of the plantlets showed survival upon transfer to field conditions. Randomly amplified polymorphic DNA (RAPD) analysis of the micropropagated plants compared with mother plant revealed true-to-type nature. The four major boswellic acid components in calluses raised from root, stem, cotyledon and leaf explants were analyzed using HPLC. The total content of four boswellic acid components was higher in stem callus obtained on MS with 15.0 μM IAA, 5.0 μM BA and 200 mg l(-1) PVP. The protocol reported can be used for conservation and exploitation of in vitro production of medicinally important non-steroidal anti-inflammatory metabolites of B. serrata.

  5. Genotype dependent callus induction and shoot regeneration in ...

    African Journals Online (AJOL)

    This study aims to observe the effect of genotype, hormone and culture conditions on sunflower (Helianthus annuus L.) callus induction and indirect plant regeneration. Calli were obtained from hypocotyl and cotyledon explants of five different sunflower genotypes; Trakya 80, Trakya 129, Trakya 259, Trakya 2098 and ...

  6. Selection of valine-resistance in callus culture of Arabidopsis thaliana (L. Heynh. derived from leaf explants

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    Małgorzata D. Gaj

    2014-01-01

    Full Text Available The selection of valine-resistant mutants was carried out in leaf explant cultures of three Arabidopsis thaliana (L. Heynh. ecotypes: C-24, RLD and Columbia. The valine concentration used for in vitro selection, lethal for seed-growing plants, has not affected callus formation and growth. However, strong inhibition of shoot regeneration ability of calli growing under selection pressure was noticed. In total, 1043 explants were cultured on valine medium and 18 shoots were regenerated with an average frequency of 1.7 shoots per 100 calli. Most R1 shoots were sterile and seeds were collected from 3 plants. The transmission of valine-resistance to the sexual progeny of these plants was scored and the increased level of valine-resistance was found in progeny of one line - 61 C. This line originated from the culture of Columbia leaf explant and displayed tetraploid chromosome number.

  7. Embryogenesis in gamma-irradiated habituated ovular callus of the 'Shamouti' orange as affected by auxin and by tissue age

    International Nuclear Information System (INIS)

    Kochba, J.; Spiegel-Roy, P.

    1977-01-01

    The response of habituated embryogenic ovular callus of the 'Shamouti' orange, Citrus sinensis, to gamma irradiation was studied. Stimulation of embryogenesis was observed when only callus, but not when the medium was irradiated. Age of callus prior to subculture modified the response to irradiation as regards optimal dose for embryogenesis and radiosensitivity. Radiation intensities also modified callus response to radiation dose. Addition of unirradiated IAA to irradiated callus cultures increased embryo formation and furthered development of embryos into plantlets, with some combinations of radiation dose and IAA concentration. Addition of IAA also decreased radiosensitivity at high doses (32 kR). (author)

  8. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

    Directory of Open Access Journals (Sweden)

    Tugba Ercetin

    2012-09-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  9. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

    Directory of Open Access Journals (Sweden)

    Tugba Ercetin

    2012-10-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  10. Development of highly regenerable callus lines and biolistic transformation of turf-type common bermudagrass [Cynodon dactylon (L.) Pers.].

    Science.gov (United States)

    Li, L; Qu, R

    2004-01-01

    Common bermudagrass, Cynodon dactylon, is a widely used warm-season turf and forage species in the temperate and tropical regions of the world. Improvement of bermudagrass via biotechnology depends on improved tissue culture responses, especially in plant regeneration, and a successful scheme to introduce useful transgenes. When the concentration of 6-benzylaminopurine was adjusted in the culture medium, yellowish, compact calluses were observed from young inflorescence tissue culture of var. J1224. Nine long-term, highly regenerable callus lines (including a suspension-cultured line) were subsequently established, of which six were used for biolistic transformation. Five independent transgenic events, with four producing green plants, were obtained following hygromycin B selection from one callus line. Three transgenic events displayed resistance to the herbicide glufosinate, and one of these showed beta-glucuronidase activity since the co-transformation vector used in the experiments contained both the gusA and bar genes.

  11. Callus culture development of two varieties of Tagetes erecta and carotenoid production

    Directory of Open Access Journals (Sweden)

    Israel Benítez-García

    2014-05-01

    Conclusions: WF callus appeared to be a suitable candidate as a source of different carotenoids, and tested varieties could represent an alternative for further studies about in vitro pigment production.

  12. Transcriptome analysis of Carica papaya embryogenic callus upon De-etiolated 1 (DET1 gene suppression

    Directory of Open Access Journals (Sweden)

    Diyana Jamaluddin

    2017-06-01

    Full Text Available Papaya is considered to be one of the most nutritional fruits. It is rich in vitamins, carotenoids, flavonoids and other phytonutrient which function as antioxidant in our body [1]. Previous studies revealed that the suppression of a negative regulator gene in photomorphogenesis, De-etiolated 1 (DET1 can improve the phytonutrient in tomato and canola without affecting the fruit quality [2,3]. This report contains the experimental data on high-throughput 3′ mRNA sequencing of transformed papaya callus upon DET1 gene suppression.

  13. Diallel analysis of anther culture response in wheat ( Triticum ...

    African Journals Online (AJOL)

    The four wheat (Triticum aestivum L.) genotypes differing in their ability to produce embryogenic callus from anther culture were reciprocally crossed and inheritance of anther culture response [callus induction frequency (CIF, %), embryogenic callus induction frequency (ECIF, %), regeneration capacity of callus (RCC, %) ...

  14. Optimization of callus induction of Zataria multiflora under the effect of different plant growth regulators and explant source

    Directory of Open Access Journals (Sweden)

    N. Mosavat*

    2017-11-01

    Full Text Available Background and objectives: The Lamiaceae family is rich in favorable secondary metabolites which have different medicinal properties and also use in food, cosmetic and sanitary industry. Zataria multiflora Boiss. is an aromatic and bushy plant containing specific pharmaceutical components which is only distributed in certain regions of Iran. Tissue culture technologies could be suitable for in vitro production of Zataria. Methods: In this study, callus production and callus related traits of Zataria was evaluated at in vitro condition. Callus induction was performed on Murashige and Skoog (MS medium containing different levels of plant growth regulators including different cytokinins (Kinetin, benzyl amino purine and auxins (2,4 dichlorophenoxyacetic acid and naphtalen acetic acid and two different explant (hypocotyl and leaf. Results: The friable calli with yellow-green color only appeared from leaf explants on three different treatments including: 1: 2.5 (mg/L 2,4-D; 2: 2 (mg/L  2,4-D; 3: [2 (mg/L 2,4-D+ 1 (mg/L Kin].  The best callus induction (75% was obtained at 2,4-D (2 mg/L + Kin (1 mg/L after 2 month of incubation under the photoperiod of 16/8 (light/dark. The highest callus growth rate (CGR (0.072 mm/day and callus fresh weight (0.135 g were denoted to the treatment of 2 mg/L (2,4-D.  Conclusion: The benefits of the protocol described here include the possibility of its use throughout the callus culture for commercial production of suitable secondary metabolites of Zataria in rapid time and huge scale.

  15. Search for alkaloids on callus culture of Passiflora alata

    Directory of Open Access Journals (Sweden)

    Michelli Wesz Machado

    2010-08-01

    Full Text Available Preliminary work on Passiflora alata leaves failed to detect harmane alkaloids using LC. The aim of this work was to investigate the production of harmane alkaloids through the cell culture of P. alata, inducing its precursor (L-tryptophan. The leaf explants presented satisfactory results after disinfection, and the callus formation was initiated in MS media with adequate quantities of phytohormones. Sixty days after inoculation, calli were inoculated in the optimized semi-solid MS media, with and without the addition of L-tryptophan (50, 100, 200 mg/L and kept in standard conditions for 90 days. Calli were collected on days 6, 16, 26, 36, and 90, followed by acid-base extraction, and analysed by LC. The results showed an absence of harmane, harmin, harmol, harmalol, and harmaline. With L-tryptophan feeding, two peaks were detected, collected and analysed through positive mode electrospray [ESI(+-MS] and sequential analysis in tandem ESI(+-MS/MS. The spectra obtained were very similar, with a repetition of the more intense ions, and consecutive loss of 68 Da units, attributed to the heterocycle pyrazole. It appeared that this transformation was not related to any enzymatic pathway previously described for the plant from L-tryptophan, and the biosynthesis of β-carboline alkaloids in callus culture of P. alata were not observed in this work.As folhas de varias espécies de Passiflora são utilizadas como ansioliticas e sedativas. Passiflora alata Curtis, Passifloraceae consta em três edições da farmacopéia brasileira, porem não há muitos estudos sobre sua composição química. No passado, enfatizava-se a ação conjunta de alcalóides e flavonóides. Em trabalho anterior, não foi detectada a presença de alcalóides harmanicos através de CLAE. Assim, decidiu-se investigar a produção dos mesmos através de cultivo celular, introduzindo seu precursor metabólico L-triptofano. Os explantes foliares apresentaram resultados satisfatorios

  16. Effect of plant growth regulators on callus induction and plant ...

    African Journals Online (AJOL)

    The present study was conducted to investigate the effects of different concentrations and combinations of growth regulators on callus induction and plant regeneration of potato (Solanum tuberosum L.) cultivar Diamant. The tuber segments were used as explants and cultured on Murashige and Skoog (MS) medium ...

  17. Effect of irradiation and colchicine on callus and somatic embryo formation in cassava (Manihot esculenta Crantz)

    International Nuclear Information System (INIS)

    Dzimega, D. A.

    2012-06-01

    A study was conducted to assess the mutagenic effect of gamma radiation on sprouting and height in four local cassava accessions. The four cassava accessions were assessed for their callus induction and somatic embryo formation ability from leaf lobes from gamma irradiated stakes as well as colchicine treated leaf lobes on different concentrations of plant growth regulators, incorporated into Murashige and skoog, (1962) (MS) basal medium. The cassava accessions were irradiated at 0, 32, 35, 45 and 50 Gy and planted in pots filled with loamy soil. The height of the shoots was measured with rule after sprouting. The leaf lobes were collected from the shoots and cultured on MS medium supplemented with 8 mg/l 2, 4-D and 16 mg/l Picloram. Another set of leaf lobes were treated with 0.0, 0.05, 0.1, 0.2, 0.25 g/l colchicine for one hour and thereafter culture on MS medium supplemented with 8 mg/l 2,4-D and 16 mg/l Picloram as described above. Callus induction from leaf lobes in 45 and 50 Gy were significantly (p≤0.05) affected by the irradiation. On the other hand, Callus induction from leaf lobes in 0.1-0.25 g/l colchicine were significantly (p≤0.05) affected by the mutagenic treatment whereas callus induced from leaf lobes in 0.05 g/l colchicine was not significantly (p≤0.05) affected. Callus induced on 8 mg/l 2, 4-D and 16 mg/l picloram gave the best response in Ankrah and all control tested while Tomfa recorded the least. Colchicine at a concentration of 0.05 g/l and radiation dose of 32 Gy treatments gave the best response of callusing. Callus induction decreased with increasing colchicine concentration and gamma irradiation. Callus derived from irradiated and colchicine leaf lobes appeared soft but friable and tiny, compact, respectively, predominately with creamy to brown colouration. Calli obtained were sub-cultures on embryo regeneration medium consisting of MS supplemented with 0.01mg/1 NAA and o.1 mg/1 BAP. There was no plantlet regeneration. Instead

  18. Aleuria aurantia - indole metabolites of fruit bodies, mycelial culture and culture medium

    Directory of Open Access Journals (Sweden)

    Janina Węgiel

    2014-08-01

    Full Text Available The aim of present study was to investigate and compare indole metabolites of fruit bodies, mycelium cultivated in vitro and culture medium of the fungus Aleuria aurantia (Fr. Fuck. By use of a number of chromatographic and spectroscopic methods several indole metabolites have been detected and identified among other the 3-indolebutyric acid was produced and extracted to the culture medium. Furthermore 3-indoleatonitrile and tryptophane degradative products have been found both in fruit bodies and mycelium.

  19. In vitro studies on callus induction in gamma-irradiated velvet bean seeds (Mucuna pruriens L.)

    International Nuclear Information System (INIS)

    Gupta, Ankit; Misra, Pragati; Shukla, Pradeep K.

    2014-01-01

    Gamma rays are often used for developing plants varieties that are agriculturally and economically important and have high productivity potential with the minimum input. Ionizing radiations are currently a very important way to create genetic variability that is not exists in nature or that is not available to the breeder. Irradiation treatments performed at in vitro culture has been also employed to increase genetic variability and mutants as a potential source of new commercial cultivars. Large number of research reports suggests also that mutagenesis in combination with tissue culture has high potential in plant breeding programs. Mucuna pruriens L., also known as velvet bean, contains L-DOPA, a precursor to the neurotransmitter dopamine and formulation of the seed power has been studied for management of Parkinson's disease. Seeds were exposed to different doses of gamma radiation (10 kGy, 20 kGy and 30 kGy) using 60 Co as source, at National Botanical Research Institute (NBRI) Lucknow. Gamma treated and untreated seeds (control) were inoculated in MS media supplemented with different phytohormone concentrations and combinations. The best callus induction was observed in control seeds on MS media supplemented with 0.5 mg/l kinetin, 2 mg/l NAA and 10 mg/l adenine sulphate, whereas gamma treated seeds showed poor callus induction in the same phytohormone concentrations. The callus induction was poor in control seeds on MS media supplemented with 1.0 mg/l kinetin, 2.0 mg/l NAA and 10 mg/l adenine sulphate, whereas gamma treated seeds showed even poor callus induction under the same phytohormone concentrations. The callus induction frequency was in declined gradually with the increasing dose of gamma radiation. Gamma treated seeds developed greenish and fragile callus and also showed decreased weight as compare to control which was white greenish, compact and heavier. (author)

  20. In vitro callus culture of Heliotropium indicum Linn. for assessment of total phenolic and flavonoid content and antioxidant activity.

    Science.gov (United States)

    Kumar, Muthusamy Senthil; Chaudhury, Shibani; Balachandran, Srinivasan

    2014-12-01

    The total phenolic and flavonoid content and percentage of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of callus and in vivo plant parts of Heliotropium indicum Linn. were estimated. Murashige and Skoog (MS) basal medium supplemented with α-naphthaleneacetic acid (NAA) 2.0 mg/l with benzyladenine (BA) 0.5 mg/l showed the highest amount of callus biomass (1.87 g/tube). The morphology of callus was significantly different according to the plant growth regulators and their concentrations used in the medium. The highest amount of total phenolic (21.70 mg gallic acid equivalent per gram (GAE/g)) and flavonoid (4.90 mg quercetin equivalent per gram (QE/g)) content and the maximum percentage (77.78 %) of radical scavenging activity were estimated in the extract of inflorescence. The synergistic effect of NAA (2.0 mg/l) and BA (0.5 mg/l) enhances the synthesis of total phenolic (9.20 mg GAE/g) and flavonoid (1.25 mg QE/g) content in the callus tissue. The callus produced by the same concentration shows 45.24 % of free radical scavenging activity. While comparing the various concentrations of NAA with 2,4-dichlorophenoxyacetic acid (2,4-D) for the production of callus biomass, total phenolic and flavonoid content and free radical scavenging activity, all the concentrations of NAA were found to be superior than those of 2,4-D.

  1. [Influence of genotype, explant type and component of culture medium on in vitro callus induction and shoot organogenesis of tomato (Solanum lycopersicum L.)].

    Science.gov (United States)

    Khaliluev, M R; Bogoutdinova, L R; Baranova, G B; Baranova, E N; Kharchenko, P N; Dolgov, S V

    2014-01-01

    The influence of explant type as well as of the type of growth regulators and concentration on callus induction processes and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties possess a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/l of zeatin in combination with 0.1 mg/l of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

  2. Optimization of Callus Induction and Regeneration in Two Fenugreek Landraces as a Medicinal Plant during in vitro Condition

    Directory of Open Access Journals (Sweden)

    Hasan Hasani Jifroudi

    2017-10-01

    Full Text Available Introduction: Fenugreek (Trigonella foenum- graecum is a medicinal plant extensively distributed in most regions of the world. Fenugreek is an annual plant from the family of papilionaceae, leguminosae. Fenugreek leaves and seeds have been used extensively to prepare extracts and powders for medicinal uses. Its root, leaf and seed contain a number of important medicinal compounds such as polysaccharide, galactomannan, different saponins such as diosgenin, yamogenin, mucilage, volatile oil and alkaloids such as choline and trigonelline. Plant tissue culture is fundamental to most aspects of biotechnology of plants. Establishment of an efficient callus induction and direct regeneration protocol is an essential prerequisite in harnessing the advantage of cell and tissue culture for genetic improvement. For the successful application of the tissue culture technique in plant breeding, callus induction and plant regeneration potential of each plant must be determined. The present study was performed in order to determine the optimum concentration of plant growth regulators (IBA + TDZ for producing of in vitro plantlet using cotyledon and hypocotyl as an explant for two different Iranian genotypes (Ardestani and Neyshabouri. Materials and Methods: In this investigation, Ardestani and Neyshabouri genotypes were used for callus induction and direct shoot regeneration. The medium used in this investigation was MS (Murashige and Skoog basal medium. Then seeds were germinated on MS medium. For callus induction and direct shoot regeneration, cotyledon and hypocotyl explants were excised from 8-day-old sterile seedlings and cultured on MS medium containing various concentrations of IBA and TDZ. In this experiment, two combinations (TDZ + IBA were used. In the first composition, IBA had four levels (0, 0.1, 0.3, 0.5 mg l-1 and TDZ had five levels (0, 0.2, 0.4, 0.6, 0.8 mg l-1 and in the second composition, IBA had four levels (0, 0.05, 0.1, 0.15 mg l-1 and TDZ

  3. Project on production of mutants by irradiation of in vitro cultured tissues of coconut and banana and their mass propagation by the tissue culture technique

    International Nuclear Information System (INIS)

    Guzman, E.V. de

    1975-01-01

    Fruit pulp tissue, ovary segments with or without ovules and sections from shoot tips of banana were used for studies on growth stimulating or morphogenetic effects of irradiation. Irradiation at 0.1-1.0 kR tended to induce faster callus growth in the otherwise slow-growing cultures. The physical condition and composition of the culture media especially with respect to growth regulators were studied, as were techniques to overcome discoloration of explants, the best choice of plant tissue for explant, and radiation effects on growth and morphogenesis. Due to the difficulty of callus induction with coconut, only the effects of irradiation on embryos cultured in vitro were studied. They were irradiated at various stages of development, i.e. during the early and final stage of liquid culture, and several days after transfer to a solid medium. Adverse effects of irradiation became evident only during the subsequent growth in solid, during the latter stage of which morphological changes were observed. Whereas irradiation of the liquid as well as solid media up to 50 kR had no adverse effect; survival and development became adversely affected at a dose of 1 kR

  4. Induction of embryogenic callus and plantlet regeneration from young leaves of high yielding mature oil palm

    Directory of Open Access Journals (Sweden)

    Yeedum, I.

    2004-09-01

    Full Text Available Callus induction and plantlet regeneration from young leaves of high-yielding mature oil palm were carried out using 10-year and 20-year-old trees from Thepa Research Station, Faculty of Natural Resources,Prince of Songkla University, Hat Yai, and Trang Agricultural College, respectively. Culture media used in this experiment were Murashige and Skoog (1962 and Oil Palm supplemented with various concentrations of α-naphthaleneacetic acid (NAA or 2,4- dichlorophenoxy acetic acid (2,4-D or dicamba (Di and antioxidants.Young leaves from 6th to 11st frond were excised, sterilized, cut into 5x5 mm pieces and cultured in the dark at 26±4ºC or 28±0.5ºC for 3 months. The results revealed that MS medium with 200 mg/l ascorbic acid (As and 1 mg/l Di (MS-AsDi gave the highest callus induction percentage (7.93 after culture for 3 months at 28±0.5ºC. Leaf segments from 6th - 8th frond yielded callus forming percentage at 10% (averaged from 1, 2.5 and 5 mg/l Di containing MS medium. Ascorbic acid as an antioxidant at concentration of 200 mg/l supplemented in MS medium in the presence of 2.5 mg/l Di produced the highest callus induction percentage (11.2 and number of nodules (7.06. A high percentage of embryogenic callus formation (66.67 was obtained when the calli were transferred to the same medium component supplemented with 0.5 mg/l Di and 1,000 mg/l casein hydrolysate (CH (MS-AsDiCH. Haustorial-staged embryos were observed to be isolated as an individual embryo and germinated on MS medium without plant growth regulator (MS-free. Development of root could be classified into two distinct types, fibrous and tap root.

  5. Citrus tissue culture employing vegetative explants.

    Science.gov (United States)

    Chaturvedi, H C; Singh, S K; Sharma, A K; Agnihotri, S

    2001-11-01

    Citrus being a number one fruit of the world due to its high nutritional value, huge production of fruits and fruit products, the citrus industry may be considered a major fruit industry. Though citrus orchard area in India is comparable to USA, the produce is far less, while its export is nil. Biotechnology has played an outstanding role in boosting the citrus industry, e.g., in Spain, which is now the biggest exporter of citrus fruit with the application of micrografting. Amongst the fruit trees, perhaps the maximum tissue culture research has been done in citrus during the past four decades, however, the results of practical value are meagre. The shortfalls in citrus tissue culture research and some advancements made in this direction along with bright prospects are highlighted, restricting the review to vegetative explants only. Whilst utilization of nucellar embryogenesis is limited to rootstocks, the other aspects, like, regeneration and proliferation of shoot meristems measuring 200 microm in length--a global breakthrough--of two commercially important scion species, Citrus aurantifolia and C. sinensis and an important rootstock, C. limonia, improvement of micrografting technique, cloning of the same two scion species as well as some Indian rootstock species, employing nodal stem segments of mature trees, of immense practical value have been elaborated. A rare phenomenon of shift in the morphogenetic pattern of differentiation from shoot bud differentiation to embryoid formation occurred during the long-term culture of stem callus of C. grandis. Stem callus-regenerated plants of C. aurantifolia, C. sinensis and C. grandis showed variation in their ploidy levels and a somaclonal variant of C. sinensis, which produced seedless fruits was isolated. Tailoring of rooting in microshoots to a tap root-like system by changing the inorganic salt composition of the rooting medium, resulting in 100% transplant success, and germplasm preservation through normal growth

  6. Absence of arabinan in the side chains of the pectic polysaccharides strongly associated with cell walls of Nicotiana plumbaginifolia non-organogenic callus with loosely attached constituent cells.

    Science.gov (United States)

    Iwai, H; Ishii, T; Satoh, S

    2001-10-01

    When leaf disks from haploid plants of Nicotiana plumbaginifolia Viv. were transformed with T-DNA and cultured on shoot-inducing medium, nonorganogenic callus. designated nolac (for non-organogenic callus with loosely attached cells), appeared on approximately 7% of leaf disks. In contrast, normal callus was generated on T-DNA-transformed leaf disks from diploid plants and on non-transformed leaf disks from haploid and diploid plants. Transmission electron microscopy revealed that the middle lamellae and the cell walls of one line of mutant callus (nolac-H14) were barely stained by ruthenium red. even after demethylesterification with NaOH, whereas the entire cell wall and the middle lamella were strongly stained in normal callus. In cultures of nolac-H14 callus, the level of sugar components of pectic polysaccharides in the hemicellulose fraction was reduced and that in the culture medium was elevated, as compared with cultures of normal callus. These results indicate that pectic polysaccharides are not retained in the cell walls and middle lamellae of nolac-H14 callus. In nolac-H14, the ratio of arabinose to galactose was low in the pectic polysaccharides purified from all cell wall fractions and from the medium, in particular, in the hemicellulose fractions. The low levels of arabinofuranosyl (T-Araf, 5-Araf, 2,5-Araf, and 3,5-Araf) residues in the pectic polysaccharides of the hemicellulosic fraction of nolac-H,14 indicated that no neutral-sugar side chains, composed mainly of linear arabinan. were present in nolac-H14. Arabinose-rich pectins. which are strongly associated with cellulose-hemicellulose complexes, might play an important role in intercellular attachment in the architecture of the cell wall.

  7. Effect of plant growth regulators on callus induction and plant ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... toum state and later spread to other part of the country ..... Effect of different concentrations of IBA and MS salt strength on rooting percentage, ... study for tissue culture of potato can get enough callus and plant regeneration efficiency to perform transgenic operation. Moreover, as the potentiality of shoot ...

  8. Effects of plant growth regulators on callus, shoot and root formation ...

    African Journals Online (AJOL)

    Root and stem explants of fluted pumpkin were cultured in medium containing different types and concentrations of plant growth regulators (PGRs). The explants were observed for callus, root and shoot formation parameters after four months. Differences among explants, plant growth regulators and their interaction were ...

  9. Distribution of Trans-Anethole and Estragole in Fennel (Foeniculum vulgare Mill of Callus Induced from Different Seedling Parts and Fruits

    Directory of Open Access Journals (Sweden)

    Abd El-Moneim Mohamed Radwan AFIFY

    2011-03-01

    Full Text Available In the present study, seeds from local cultivar of fennel were germinated on Murashige and Skoog medium (MS without plant growth regulators. Different types of explants from the growing seedling such as cotyledonal leaves, hypocotyls, epicotyls and roots were cultured on MS medium, contained different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D either alone or with kinetin. Differential responses in the essential oil constituents were observed in the induction and development of callus. The major components of essential oils includes estragole, trans-anethole, limonene and fenchone were studied under different conditions to find out the best methods which could be used to reduce the amount of estragole (not favorite for human consumption and increase the amount of trans-anethole.

  10. The effect of plant growth regulators on callus induction somatic embryogenesis of hybird tomato

    International Nuclear Information System (INIS)

    Jan, S. A.; Shah, S. H.; Ali, S.; Ali, G. H.

    2015-01-01

    Efficient tissue culture system is important for transformation of important genes in hybrid tomato cultivars. The present study was undertaken to develop an efficient tissue culture system for hybrid tomato cultivar Peto-86. The young primary leaves and stems were inoculated into five different MS media having different concentrations of plant growth regulators in different combinations for callus induction, somatic embryogenesis and for both direct and indirect regeneration. Maximum callus induction frequency 90 percentage was achieved with MS media containing 2,4-D 4 mg L-1 and BAP 0.5 mg L-1. The direct somatic embryogenesis was found highest on MS media supplemented with 2,4-D 4 mg L-1 and BAP 0.5 mg L-1. Maximum indirect regeneration frequency 87 percentage was achieved from primary leaves explants with MS media containing IAA 0.5 mg L-1 and BAP 3 mg L-1 and highest direct regeneration frequency 77% was obtained from primary leaves explants with MS media containing NAA 1 mg L-1 and BAP 3 mg L-1. The high concentration of 2,4-D increased callus induction and somatic embryogenesis frequencies while the high concentration of BAP increased regeneration frequency. An improved tissue culture system of hybrid tomato cultivar Peto-86 was established and it may be recommended for further transformation experiments. (author)

  11. Experimental study upon the effect of irradiation on callus formation of fracture. Observation of vascular alteration and callus formation

    Energy Technology Data Exchange (ETDEWEB)

    Saigusa, F [Nippon Dental Coll., Tokyo

    1981-02-01

    Irradiation effects on callus formation after bone fracture were studied in rats with fractured right lower extremity. Follow-up study was continued for 112 days since 3000 rad was irradiated to the fractured site 3 days after bone fracture. Callus formation was noted in both of the outer and inner part (bone marrow) of the diaphysis before 14 days after bone fracture, but it was slow and sparse compared with that of non-irradiated group. Callus formation tended to disappear gradually from the outside of the diaphysis after 28 days after bone fracture. Strong disturbance was found in the surrounding vascular system at this time. Inside of the diaphysis, callus formation was restricted the end of the fracture, where lamellar calluses fused together. Changes in vascular system remained until 56 days after bone fracture. Vascular distribution was most dense 28 days after bone fracture. In many of the calluses which have established fusion, findings suggested excessive calcification in the trabeculae. Vascular distribution at this time was sparse, vascular formation was markedly suppressed in the bone marrow, and very little vascular formation was found in the fractured edges of the bone.

  12. Development of technology for plantlet propagation by tissue culture. (3). Strawberry callus growth and changes in pH in liquid medium; Soshiki baiyo ni yoru shubyo tairyo zoshoku gijutsu no kaihatsu. (3). Ekitai baiyo ni okeru ichigo callus no zoshoku to baichi no pH henka

    Energy Technology Data Exchange (ETDEWEB)

    Yoshihara, T; Hanyo, H [Central Research Inst. of Electric Power Industry, Tokyo (Japan)

    1991-02-01

    Development of strawberry cultivation plants is described as a part of night-time power utilization activities for power load levelling. Strawberry calluses (undifferent tissue mass) cultured in a liquid medium reach the fastest growth period in one week and the steady state in four weeks. The callus growth shows the maximum value at this time, which was 20 times as much of the seedling. The medium pH changed in a range from 4 to 7. If the initial pH is 4.0 or higher, no difference is created in the callus propagation in the steady state period, but at 3.0, no propagation whatsoever. The pH after the fastest growth period converged to a range from 6.0 to 7.0, with the exception of initial pH at 3.0. The medium pH decreased as a result of pre-culture heating sterilization, formation of iron phosphate due to light irradiation, and organic acid release during the initial growth phase. The pH increased because of difference in the speed of absorbing ammonium and nitric acid during the later growth phase. The growth efficiency of 20 times is about the same as other plants. Since the pH change is maintained within the range from 4 to 7, which causes no difference in in growth, there is no need of adjusting the pH within this range. 18 refs., 15 figs., 3 tabs.

  13. INDUKSI KALUS JERUK KASTURI (CITRUS MICROCARPA MENGGUNAKAN HORMON 2,4-D DAN BAP DENGAN METODE IN VITRO

    Directory of Open Access Journals (Sweden)

    Imam Mahadi

    2016-08-01

    Full Text Available Calamansi fruits (Citrus microcarpa or musk lime is one horticulture crops has been known as beverage and food flavorings. This research aimed to determine the effect of 2,4-D and BAP on callus induction of explants calamansi fruits (Citrus microcarpa. Parameters observed is percentage of explants, time appeared callus, callus width, and texture of the callus. Research results and a growing percentage of the width callus explants were analyzed by ANOVA and a further test DMRT at 5% level. The next to appear callus timing parameters and texture of callus analyzed in description. The result shows that a combination of hormones D4B1 and D4B2 produces a callus emerged fastest of 3.3 days after culture. The highest percentage of callus formation that is 100% contained in the treatment of D2B0, D2B0.5-D4B0. The mean width of callus was highest in treatment D4B2with a width of 0.97 cm. Embryogenic callus resulting by treatment of D2B1, D2B2, and D3B0. However, the best a combination of hormones is treatment D2B2 (2 mg/l, 2,4-D, dan 2 mg/l BAP producing embryogenic callus edvanced to suspension cultures.

  14. The combination effect of auxin and cytokinin on in vitro callus formation of Physalis angulata L. - A medicinal plant

    Science.gov (United States)

    Mastuti, Retno; Munawarti, Aminatun; Firdiana, Elok Rifqi

    2017-11-01

    Physalis angulata L. (Ciplukan) is one member of Solanaceae that has a potential as herbal medicine. This plant grows wild in the crop fields, forest edges, etc. However, ciplukan is increasingly difficult to find recently. In vitro callus is an alternative source to produce secondary metabolite production as well as to regenerate plants through indirect organogenesis. This study aims to identify the response of hypocotyl explants on in vitro callus formation induced by a combination of auxin and cytokinins. Two types of cytokinins, Kinetin and BAP (0.5 ppm) were combined with three types of auxin, i.e. 2.4-D, IBA and IAA, at three concentrations 0.5, 1.0 and 1.5 ppm. In all combinations of cytokinin and auxin, 50-100% of hypocotyl explants derived from in vitro seedling were able to produce callus either in a compact or watery friable texture. In MS medium supplemented with 2.4-D, callus FW (fresh weight) began to decline in the fourth week after culture. Callus FW that increased until 5 weeks of culture was obtained in medium IAA 0.5 + Kin 0.5, IBA 1.0 + Kin 0.5 and IBA 1 + BA 0.5. Almost all calli induced on a medium + Kinetin also produced roots. While medium + BAP was able to induce shoots regeneration.

  15. In vitro assay of the inhibitory effect of neem callus and leaf extracts on some phytopathogenes

    International Nuclear Information System (INIS)

    El Gaali, E.; Mukhtar, I.

    2006-01-01

    The effect of different concentrations of neem (Azadirachta indica) callus and leaf extracts on the radial growth of drechslera rostrata, fusarium oxysporum and alterneria alternata was assessed. Obvious inhibitory effect was observed on the mycelia radial growth of the three treated fungi. The level of inhibition increased with the increase of the extract concentration. The maximum inhibitory effect (84%) was recorded with drechslera rostrata when inoculated in media containing 20 mg/ml of neem callus extract, while the inhibition rate of the mycelial growth of the same species reached 61% when inoculated in a medium containing the same concentration of neem leaf extract. The subsequent concentrations of the callus and leaf extracts gave similar trends of inhibition on the fungi cultured on extract amended agar plates. (Author)

  16. The influence of different hormone concentration and combination on callus induction and regeneration of Rauwolfia serpentina L. Benth.

    Science.gov (United States)

    Salma, U; Rahman, M S M; Islam, S; Haque, N; Jubair, T A; Haque, A K M F; Mukti, I J

    2008-06-15

    The influence of media composition on callus induction and subsequent regeneration of Rauwolfia serpentina L. Benth has been studied. High frequency (96.43%) callus induction was obtained when nodal segments from in vitro raised shoots were cultured on MS medium supplemented with 0.5 mg L(-1) BA and 2.0 mg L(-1) NAA. The callus differentiated into adventitious shoots when it was subcultured on MS medium supplemented with 2.0 mg L(-1) BA with 0.2 mg L(-1) NAA. Regenerated shoots were best rooted on half-strength MS medium with 1.0 mg L(-1) each of IBA and IAA.

  17. A survey of the effect of explants type, plant growth regulators and activated charcoal on callus induction in Papaver bracteatum

    Directory of Open Access Journals (Sweden)

    Bahman Hosseini

    2015-09-01

    Full Text Available Callus culture is necessary for production of suspension cell culture in plant breeding programs. Regarding to the application of Papaver bracteatum as an important medicinal plant in production of benzophenantridine alkaloids, this study was performed to find the most suitable hormone combination and explant type for achieving to high percentage of callus induction fresh weight and somatic embryogenesis in this plant. For this purpose, hypocotyl explants were cultured in ½MS media containing active charcoal (2 and 4 mgL-1 in combination of different concentrations of NAA, 2,4-D (0, 1, 2, 3 and 5 mgL-1 and BA (0, 0.1 and 0.5 mgL-1. The seed explants were cultured in same treatments without active charcoal. Also, somatic embryogesis induction using seed explants in ½MS media containing different concentrations of NAA and 2,4-D (0, 0.5, 1 and 2 mgL-1 with BA 0.5 mgL-1 were investigated. The results showed that the highest percentage of callus induction (43.6%, 54% in hypocotyls explants were obtained in the ½MS media containing 2 mgL-1 active charcoal and 2 mgL-1 2,4-D and 5 mgL-1 NAA in companion with BA 0.5 mgL-1 respectively. The maximum callus induction (84% was obtained in ½MS medium with 1 mgL-1 2,4-D without active charcoal. The highest callus fresh weight (0.35% was obtained in MS media with 0.5 mgL-1 2,4-D andthe maximum rate of somatic embryogenesis induction (77% was observed in ½MS media containing 1 mgL-1 2,4-D with 0.5 mgL-1 BA.

  18. Propagation of jarrah (Eucalyptus marginata) by organ and tissue culture

    Energy Technology Data Exchange (ETDEWEB)

    Bennett, M.J.; McComb, J.A.

    1982-01-01

    Micropropagation methods are described for the production of clonal lines from Eucalyptus marginata (jarrah) seedlings. Nodal explants from mature trees can also yield shoot cultures, but a high frequency of contamination occurs among such explants. Uncontaminated callus cultures can be produced from mature trees by culturing stamen filaments and shoots can subsequently be regenerated from this callus. The rooting percentage of shoot cultures from either nodes or stamen callus of mature trees is low compared with that from seedling explants. Considerable variation was observed between trees in the ability of stamen callus to regenerate shoots and in the frequency of rooting. (Refs. 27)

  19. A computational technique to measure fracture callus in radiographs.

    Science.gov (United States)

    Lujan, Trevor J; Madey, Steven M; Fitzpatrick, Dan C; Byrd, Gregory D; Sanderson, Jason M; Bottlang, Michael

    2010-03-03

    Callus formation occurs in the presence of secondary bone healing and has relevance to the fracture's mechanical environment. An objective image processing algorithm was developed to standardize the quantitative measurement of periosteal callus area in plain radiographs of long bone fractures. Algorithm accuracy and sensitivity were evaluated using surrogate models. For algorithm validation, callus formation on clinical radiographs was measured manually by orthopaedic surgeons and compared to non-clinicians using the algorithm. The algorithm measured the projected area of surrogate calluses with less than 5% error. However, error will increase when analyzing very small areas of callus and when using radiographs with low image resolution (i.e. 100 pixels per inch). The callus size extracted by the algorithm correlated well to the callus size outlined by the surgeons (R2=0.94, p<0.001). Furthermore, compared to clinician results, the algorithm yielded results with five times less inter-observer variance. This computational technique provides a reliable and efficient method to quantify secondary bone healing response. Copyright 2009 Elsevier Ltd. All rights reserved.

  20. Determination of abscisic acid and its glucosyl ester in embryogenic callus cultures of Vitis vinifera in relation to the maturation of somatic embryos using a new liquid chromatography-ELISA analysis method.

    Science.gov (United States)

    Prado, María Jesús; Largo, Asier; Domínguez, Cristina; González, María Victoria; Rey, Manuel; Centeno, María Luz

    2014-06-15

    The levels of abscisic acid (ABA), its conjugate ABA-GE, and IAA were determined in embryogenic calli of Vitis vinifera L. (cv. Mencía) cultured in DM1 differentiation medium, to relate them to the maturation process of somatic embryos. To achieve this goal, we developed an analytical method that included two steps of solid-phase extraction, chromatographic separation by HPLC, ABA-GE hydrolysis, and sensitive ELISA quantification. Because the ABA immunoassay was based on new polyclonal antibodies raised against a C4'-ABA conjugate, the assay was characterized (detection limit, midrange, measure range, and cross-reaction) and validated by a comparison of the ABA data obtained with this ELISA procedure and with a physicochemical method (LC-ESI-MS/MS). Radioactive-labeled internal standards were initially added to callus extracts to correct the losses of plant hormones, and thus assure the accuracy of the measurements. The endogenous concentration of ABA in the embryogenic callus cultured in DM1 medium was doubled at the fifth week of culture, concurring with the maturation process of somatic embryos, as indicated by the accumulation of carbohydrates observed through histological analysis. The ABA-GE content was higher than ABA, decreasing at 21 days of culture in DM1 medium but increasing thereafter. The data suggest the involvement of the synthesis and conjugation of ABA in the final stages of development in grapevine somatic embryos from embryogenic callus. IAA levels were low, suggesting that auxin plays no significant role during the maturation of somatic embryos. In addition, the lower ABA levels in calli cultured in DM differentiation medium with PGRs, a medium presenting high precocious germination and deficiencies in somatic embryo development indicate that an increase in ABA content during the development of somatic embryos in grapevine is necessary for their correct maturation. Copyright © 2014 Elsevier GmbH. All rights reserved.

  1. High frequency plant regeneration from leaf explants derived callus of evening primrose (oenothera biennis)

    International Nuclear Information System (INIS)

    Ghauri, E.G.; Shafi, N.; Ghani, S.; Fatima, A.

    2008-01-01

    The seeds of Evening primrose were aseptically grown and leaf explants were used for establishment of callus culture. The Excellent growth in callus biomass was achieved on MS medium supplemented with 2, 4, -D and TDZ. For optimal growth of bud and shoot regeneration, fortification of IAA along with TDZ, or BAP was found to be essential. Rooting (70%) could be inducted on hormone free MS-medium. This percentage improved to 98 when NAA was added to the medium. The plantlets thus obtained were transferred to the field successfully after passing through the process of hardening. (author)

  2. Cloning of the queen variety pineapple (Ananas comosus (L.) merr) through the callus

    International Nuclear Information System (INIS)

    Lapade, A.G.; Veluz, A.M.S.; Santos, I.S.

    1988-01-01

    Crown sections of pineapple were inoculated aseptically in Murashige and Skoog (MS) medium with varying levels of benzyl adenine (BA) in combination with napthalene acetic acid (NAA). Callus was induced from crown sections that was grown in MS with 0 ppm BA + 2 ppm NAA, 2 ppm BA + 6 ppm NAA. These calli when grown continuously in the same medium developed profusely into shoots. Callus and shoots developed profusely at medium containing 2 ppm BA + 2 ppm NAA. Calli that were further sub-cultured in MS with 2 ppm BA + 2 ppm NAA produced multiple shoots. Calli and shoots that were further sub-cultured in 0 to 4 ppm BA to 4 to 6 ppm NAA formed roots. Results showed that complete plantlets were produced from crown sections of pineapple variety Queen in MS medium supplemented with the different treatment combinations of BA and NAA. (Author). 9 refs.; 1 fig

  3. Establishment of Aquilaria malaccensis Callus, cell suspension and adventitious root systems

    International Nuclear Information System (INIS)

    Norazlina Noordin; Rusli Ibrahim

    2010-01-01

    Aquilaria malaccensis is a tropical forest tree from the family Thymelaeaceae, an endangered forest species and was listed in CITES since 1995. Locally known as Pokok Karas, this tree produces agar wood or gaharu, a highly valuable, resinous and fragrant forest product. Karas has been highly recognized for its vast medicinal values and gaharu has been widely use for perfumery, incense and religious purposes. The phyto chemical studies of agar wood showed that Sesqui terpenoid and Phenyl ethy chromone derivatives are the principal compounds that have anti allergic and anti microbe activities. Cell and organ culture systems provide large scale production of biomass and offers feasibilities for the production of secondary metabolites. This paper describes the work done for establishing reproducible systems for callus initiation and production of cell suspension cultures as well as production of adventitious roots that will later be amenable for the production of secondary metabolites of A. malaccensis. Hence, further manipulation with Methyl Jasmonate, a chemical elicitor could be done to induce secondary metabolites using callus, cell suspension and adventitious roots systems. (author)

  4. Heritability of regeneration in tissue cultures of sweet potato (Ipomoea batatas L.).

    Science.gov (United States)

    Templeton-Somers, K M; Collins, W W

    1986-03-01

    A population of open-pollinated progeny from 12 parents, and the 12 parents, was surveyed for in vitro growth and regeneration characteristics. Four different tissue culture procedures involving different media and the use of different explants to initiate the cultures were used. Petiole explants from young leaves were used as explants for initiation of callus cultures. These were evaluated for callus growth rate, friability, and callus color and texture, before transferring to each of three different regeneration media for evaluation of morphogenetic potential. Small shoot tips also were used to initiate callus cultures, which were evaluated for the same growth characteristics and transferred to growth-regulator free regeneration media. Regeneration occurred through root or shoot regeneration or through embryogenesis. Tissue culture treatment effects, as well as genotypic effects, were highly significant in determining: the types of callus produced, callus growth rates, color and texture on the two types of media used for the second and third subcultures. The family x treatment interaction was generally not statistically significant, affecting only callus color. Estimates of narrow sense heritability for callus growth rate in both the second and third subcultures were high enough (0.35 and 0.63, respectively) for the evaluation of parental lines for selection procedures. These characteristics were also the only early culture callus traits that were consistently correlated with later morphogenesis of the cultures. They were negatively correlated with root or shoot regeneration. The occurence of somatic embryogenesis was not correlated with early callus growth characteristics. Genetic and treatment effects were highly significant in the evaluation of morphogenetic potential, through root or shoot regeneration, or through embryogenesis. Regeneration of all types was of low frequency for all procedures, expressed in ≦ 11% of the cultures of the total population.

  5. Distribution of linker histone variants during plant cell differentiation in the developmental zones of the maize root, dedifferentiation in callus culture after auxin treatment

    Directory of Open Access Journals (Sweden)

    ANASTASIOS ALATZAS

    2008-01-01

    Full Text Available Although several linker histone variants have been studied in both animal and plant organisms, little is known about their distribution during processes that involve alterations in chromatin function, such as differentiation, dedifferentiation and hormone treatment. In this study, we identified linker histone variants by using specific anti-histone Hl antibodies. Each variant's ratio to total Hl in the three developmental zones of maize (Zea mays L. root and in callus cultures derived from them was estimated in order to define possible alterations either during plant cell differentiation or during their dedifferentiation. We also evaluated linker histone variants' ratios in the developmental zones of maize roots treated with auxin in order to examine the effects of exogenous applied auxin to linker histone variant distribution. Finally, immunohistochemical detection was used to identify the root tissues containing each variant and correlate them with the physiological status of the plant cells. According to the results presented in this study, linker histone variants' ratios are altered in the developmental zones of maize root, while they are similar to the meristematic zone in samples from callus cultures and to the differentiation zone in samples from roots treated with auxin. We propose that the alterations in linker histone variants' ratios are correlated with plant cell differentiation and dedifferentiation.

  6. The role of 2,4-D and auxin-binding proteins during the induction of embryogenic and non-embryogenic callus in Zea mays (L.)

    NARCIS (Netherlands)

    Bronsema, F.

    1998-01-01

    This thesis deals with the influence of the growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D) on the induction of callus in cultured immature embryos of Zea mays (L). In maize, two types of embryogenic callus can be induced in immature zygotic embryos.
    Type I

  7. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shirish C

    2016-06-02

    We present experimental evidence to show that acetylcholine (ACh) causes decrease in shoot formation in leaf explants of tomato (Lycopersicon esculentum Miller var Pusa Ruby) when cultured on shoot regeneration medium. The optimum response was obtained at 10(-4) M ACh-enriched medium. ACh also causes decrease in percentage of cultures forming callus and reduces the callus mass. Inhibitors of enzymatic hydrolysis of ACh, neostigmine and physostigmine, also suppresses callogenesis and caulogenesis. On the other hand, the breakdown products of Ach, choline and acetate, do not alter the morphogenic response induced on the shoot regeneration medium. Neostigmine showed optimal reduction in shoot formation at 10(-5) M. The explants cultured on neostigmine augmented medium showed decline in the activity of ACh hydrolyzing enzyme acetylcholinesterase. ACh and neostigmine added together showed marked reduction in callus mass. These results strongly support the role of ACh as a natural regulator of morphogenesis in tomato plants.

  8. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shirish C.

    2016-01-01

    ABSTRACT We present experimental evidence to show that acetylcholine (ACh) causes decrease in shoot formation in leaf explants of tomato (Lycopersicon esculentum Miller var Pusa Ruby) when cultured on shoot regeneration medium. The optimum response was obtained at 10−4 M ACh-enriched medium. ACh also causes decrease in percentage of cultures forming callus and reduces the callus mass. Inhibitors of enzymatic hydrolysis of ACh, neostigmine and physostigmine, also suppresses callogenesis and caulogenesis. On the other hand, the breakdown products of Ach, choline and acetate, do not alter the morphogenic response induced on the shoot regeneration medium. Neostigmine showed optimal reduction in shoot formation at 10−5 M. The explants cultured on neostigmine augmented medium showed decline in the activity of ACh hydrolyzing enzyme acetylcholinesterase. ACh and neostigmine added together showed marked reduction in callus mass. These results strongly support the role of ACh as a natural regulator of morphogenesis in tomato plants. PMID:27348536

  9. Development of Betalain Producing Callus Lines from Colored Quinoa Varieties (Chenopodium quinoa Willd).

    Science.gov (United States)

    Henarejos-Escudero, Paula; Guadarrama-Flores, Berenice; Guerrero-Rubio, M Alejandra; Gómez-Pando, Luz Rayda; García-Carmona, Francisco; Gandía-Herrero, Fernando

    2018-01-17

    Betalains are water-soluble plant pigments of hydrophilic nature with promising bioactive potential. Among the scarce edible sources of betalains is the grain crop quinoa (Chenopodium quinoa Willd), with violet, red, and yellow grains being colored by these pigments. In this work, callus cultures have been developed from differently colored plant varieties. Stable callus lines exhibited color and pigment production when maintained on Murashige and Skoog medium supplemented with the plant growth regulators 6-benzylaminopurine (8.88 μM) and 2,4-dichlorophenoxyacetic acid (6.79 μM) with a reduction of the nitrogen source to 5.91 mM. Pigment analysis by HPLC-DAD and ESI-MS/MS fully describes the content of individual pigments in the cell lines and allows the first report on the pigments present in quinoa seedlings. Phyllocactin and vulgaxanthin I are described as novel pigments in the species and show the potential of C. quinoa culture lines in the production of compounds of nutritional value.

  10. Signal transduction in artichoke [Cynara cardunculus L. subsp. scolymus (L.) Hayek] callus and cell suspension cultures under nutritional stress.

    Science.gov (United States)

    Lattanzio, Vincenzo; Caretto, Sofia; Linsalata, Vito; Colella, Giovanni; Mita, Giovanni

    2018-06-01

    Stimulated production of secondary phenolic metabolites and proline was studied by using cell cultures of artichoke [Cynara cardunculus L. subsp. scolymus (L.) Hayek] submitted to nutritional stress. Artichoke cell cultures accumulated phenolic secondary metabolites in a pattern similar to that seen in artichoke leaves and heads (capitula). This paper shows that both callus and cell suspension cultures under nutritional stress accumulated phenolic compounds and proline, at the same time their biomass production was negatively affected by nutrient deficiency. The results obtained strongly suggest that plant tissues respond to nutrient deprivation by a defensive costly mechanism, which determines the establishment of a mechanism of trade-off between growth and adaptive response. Furthermore, the results of this research suggest that perception of abiotic stress and increased phenolic metabolites are linked by a sequence of biochemical processes that also involves the intracellular free proline and the oxidative pentose phosphate pathway. The main conclusion of this paper is that, once calli and cell suspension cultures respond to nutrient deficiency, in acclimated cells the establishment of a negative correlation between primary metabolism (growth) and secondary metabolism (defence compounds) is observed. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  11. Callus Growth Kinetics of Physic Nut (Jatropha curcas L.) and Content of Fatty Acids from Crude Oil Obtained In Vitro.

    Science.gov (United States)

    da Luz Costa, Jefferson; da Silva, André Luís Lopes; Bier, Mário César Jucoski; Brondani, Gilvano Ebling; Gollo, André Luiz; Letti, Luiz Alberto Junior; Erasmo, Eduardo Andrea Lemus; Soccol, Carlos Ricardo

    2015-06-01

    The callus growth kinetics allows identifying the appropriate moment for callus pealing and monitoring the accumulation of primary and secondary metabolites. The physic nut (Jatropha curcas L.) is a plant species used for biofuel production due to its high oil content; however, this plant presents a great amount of bioactive compounds which can be useful for industry. The aim of this research was to establish a calli growth curve and to evaluate the fatty acid profile of crude oil extracted from callus. The callus growth kinetics presented a sigmoid standard curve with six distinct phases: lag, exponential, linear, deceleration, stationary, and decline. Total soluble sugars were higher at the inoculation day. Reducing sugars were higher at the inoculation day and at the 80th day. The highest percentage of ethereal extract (oil content) was obtained at the 120th day of culture, reaching 18 % of crude oil from the callus. The calli produced medium-chain and long-chain fatty acids (from 10 to 18 carbon atoms). The palmitic acid was the fatty acid with the highest proportion in oil (55.4 %). The lipid profile obtained in callus oil was different from the seed oil profile.

  12. Effects of gamma-rays and neutrons on the seedling and callus growth in rice seeds

    International Nuclear Information System (INIS)

    Baradjanegara, A.A.; Fujii, Taro; Amano, Etsuo

    1976-01-01

    Seeds of rice c.v. Norin-8 and two radiation induced dwarf mutants MGS-46 and -96 were used to investigate the effects of gamma- and 14 MeV neutron-radiations in different culture systems. Seedling growth of irradiated seeds in soil, and in two types of synthetic media, modified White's (M-W) or modified Eriksson's (M-E), as well as the callus growth on 2.4-D supplemented media were measured as an index of radiation damage. Comparing the seedling height about two types of media, the M-E promoted the plant growth than the M-W in irradiated as well as non-irradiated lots of the three strains studied. The callus growth on M-E surpassed by more 10 times that obrained on the M-W in all the lots. The M-E medium seem to be appropriate for both the seedling and callus growth of rice. (auth.)

  13. Effects of gamma rays and neutrons on the seedling and callus growth in rice seeds

    International Nuclear Information System (INIS)

    Baradjanegara, A.A.; Fujii, T.; Amano, E.

    1976-01-01

    Seeds of rice c.v. Norin-8 and two radiation induced dwarf mutants MGS-46 and -96 were used to investigate the effects of gamma and 14 MeV neutron-radiations in different culture systems. Seeding growth of irradiated seeds in soil, and in two types of synthetic media, modified White's (M-W) or modified Erickson's (M-E), as well as the callus growth on 2,4-D supplemented media were measured as an index radiation damage. Comparing the seeding height about two types of media, the M-E promoted the plant growth than the M-W in irradiated as well as non irradiated lots of the three strains studied. The callus growth on M-E surpassed by more 10 times that obtained on the M-W in all the lots. The M-E medium seems to be appropriate for both the seeding and callus growth of rice. (author)

  14. Premilinary studies for optimiziing a protocol for obtaining embryogenic calluses in two rubber (Hevea brasiliensis Mull. Arg clones from different geographical origins

    Directory of Open Access Journals (Sweden)

    Santiago Cadavid Ruiz

    2006-01-01

    Full Text Available The influence of growth regulators on obtaining friable rubber (Hevea brasiliensis Müll. Arg. calluses with no plant regeneration as investigated. Two clones having different geographical origin were used in all trails carried out in this study: FX 3864 (South-American and PB 254 (Asian. Young leaves and eight- to ten-week-old seed integument from both clones were used as explants in several experiments; they were initially cultured in MH medium (Carron, et ál., 1989, modified MH medium (Montoro, et ál., 1993, 2000 and modified MS medium (Carron, et at,. 1992, no positive response being obtained by days 25 or 50. However, other trials were carried out with the integument in modified MS medium (1962, 0.67 mg/L BAP and 0.66 mg/L 2-4 D being added as medium for initiating embryogenesis, the formation of white, friable calluses being observed by day 25 in the two selected clones. These calluses were sub-cultured in MS supplemented with 0.35 mg/L BAP and 0.2 mg/L 2-4 D as callogenesis expression medium, embryogenic expression being observed in both clones by day 50. Equally friable white calluses were obtained from young leaves in the two clones in MS medium supplemented with 1.0 mg/L BAP, 1.0 mg/L ANA but without IBA and kinetin by day 25. Calluses sub-cultured in the same medium supplemented with 0.5 mg/L BAP and 0.5 mg/L ANA began to show increased friability after 50 days. culture. This work is a partial report of a macro-project for optimising a protocol for rubber (Hevea brasiliensis  multiplication by somatic embryogenesis.

  15. Callus features of regenerate fracture cases in femoral lengthening in achondroplasia

    International Nuclear Information System (INIS)

    Devmurari, Kamlesh N.; Song, Hae Ryong; Modi, Hitesh N.; Venkatesh, K.P.; Ju, Kim Seung; Song, Sang Heon

    2010-01-01

    We studied the callus features seen in cases of regenerate fracture in femoral lengthening using a monolateral fixator in achondroplasia to determine whether callus types and shapes can predict the probability of callus fracture. The radiographs of 28 cases of femoral lengthening in 14 patients, 14 cases of callus fracture, and 14 cases without callus fracture were retrospectively analyzed by four observers and classified into different shapes and types in concordance with the Ru Li classification. The average lengthening of 9.4 cm (range 7.5-11.8 cm) was achieved, which was 41% (range 30-55%) of the original length and the average timing of callus fracture was 470 days (range 440-545 days) after surgery in the callus fracture group. While the average lengthening of 9.1 cm (range 8-9.7 cm) was achieved, this was 30% (range 28-32%) of the original length in the group of patients without callus fracture. The callus was atypically shaped, there was a 48% average (range 30-72%) reduction of the callus width compared with the natural width of the femur, and a lucent pathway was present in all cases of regenerate fracture. A lucent pathway was seen in all fracture cases with concave, lateral, and atypical shapes, and there was more than 30% lengthening and 30% reduction of the callus width compared with the natural width of the femur, which are the warning signs for regenerate fractures. These signs help the surgeon to predict the outcome and guide him in planning for any additional interventions. The Ru Li classification is an effective method for the evaluation of the chance of callus fracture. (orig.)

  16. Callus features of regenerate fracture cases in femoral lengthening in achondroplasia

    Energy Technology Data Exchange (ETDEWEB)

    Devmurari, Kamlesh N.; Song, Hae Ryong; Modi, Hitesh N.; Venkatesh, K.P.; Ju, Kim Seung; Song, Sang Heon [Korea University Medical College, Institute for Rare Diseases and Department of Orthopedic Surgery, Seoul (Korea)

    2010-09-15

    We studied the callus features seen in cases of regenerate fracture in femoral lengthening using a monolateral fixator in achondroplasia to determine whether callus types and shapes can predict the probability of callus fracture. The radiographs of 28 cases of femoral lengthening in 14 patients, 14 cases of callus fracture, and 14 cases without callus fracture were retrospectively analyzed by four observers and classified into different shapes and types in concordance with the Ru Li classification. The average lengthening of 9.4 cm (range 7.5-11.8 cm) was achieved, which was 41% (range 30-55%) of the original length and the average timing of callus fracture was 470 days (range 440-545 days) after surgery in the callus fracture group. While the average lengthening of 9.1 cm (range 8-9.7 cm) was achieved, this was 30% (range 28-32%) of the original length in the group of patients without callus fracture. The callus was atypically shaped, there was a 48% average (range 30-72%) reduction of the callus width compared with the natural width of the femur, and a lucent pathway was present in all cases of regenerate fracture. A lucent pathway was seen in all fracture cases with concave, lateral, and atypical shapes, and there was more than 30% lengthening and 30% reduction of the callus width compared with the natural width of the femur, which are the warning signs for regenerate fractures. These signs help the surgeon to predict the outcome and guide him in planning for any additional interventions. The Ru Li classification is an effective method for the evaluation of the chance of callus fracture. (orig.)

  17. The Reserpine Production and Callus Growth of Indian Snake Root (Rauvolfia serpentina (L.) Benth. Ex Kurz) Culture by Addition of Cu2+

    OpenAIRE

    NUNUNG NURCAHYANI; SOLICHATUN; ENDANG ANGGARWULAN

    2008-01-01

    The objectives of this research were to study the effects of Cu2+ addition on the reserpine production and callus growth from in vitro culture indian snake root (Rauvolfia serpentina (L.) Benth. Ex Kurz). This research frame work was based on the potency of snake root which was many exploited as anti-hypertension. The addition of elicitor Cu2+ in the form of CuCl2 would influence the ion transport of cell and changed of cytoplasm pH, and also has effects on synthesis and activity of enzymes w...

  18. Effect of applied pressure on callus formation and its relevance in ...

    African Journals Online (AJOL)

    ... effect of applied pressure on callus formation, cell differentiation and its importance in grafting. Callus was formed earlier in pressure-free explants. The importance of applied pressure in graft formation was indicated by the fusion of callus cells in capped explant experiments. Where callus grew from the edges, a cambium ...

  19. Alpha-glucosidase Inhibitory and Antioxidant Potential of Antidiabetic Herb Alternanthera sessilis: Comparative Analyses of Leaf and Callus Solvent Fractions.

    Science.gov (United States)

    Chai, Tsun-Thai; Khoo, Chee-Siong; Tee, Chong-Siang; Wong, Fai-Chu

    2016-01-01

    Alternanthera sessilis is a medicinal herb which is consumed as vegetable and used as traditional remedies of various ailments in Asia and Africa. This study aimed to investigate the antiglucosidase and antioxidant activity of solvent fractions of A. sessilis leaf and callus. Leaf and callus methanol extracts were fractionated to produce hexane, chloroform, ethyl acetate, butanol, and water fractions. Antiglucosidase and 1,1-diphenyl-2-picrylhydrazyl scavenging activities as well as total phenolic (TP), total flavonoid (TF), and total coumarin (TC) contents were evaluated. Lineweaver-Burk plot analysis was performed on leaf and callus fractions with the strongest antiglucosidase activity. Leaf ethyl acetate fraction (LEF) had the strongest antiglucosidase (EC 50 0.55 mg/mL) and radical scavenging (EC 50 10.81 μg/mL) activity among leaf fractions. Callus ethyl acetate fraction (CEF) and chloroform fraction had the highest antiglucosidase (EC 50 0.25 mg/mL) and radical scavenging (EC 50 34.12 μg/mL) activity, respectively, among callus fractions. LEF and CEF were identified as noncompetitive and competitive α-glucosidase inhibitors, respectively. LEF and CEF had greater antiglucosidase activity than acarbose. Leaf fractions had higher phytochemical contents than callus fractions. LEF had the highest TP, TF, and TC contents. Antiglucosidase and antioxidant activities of leaf fractions correlated with phytochemical contents. LEF had potent antiglucosidase activity and concurrent antioxidant activity. CEF had the highest antiglucosidase activity among all fractions. Callus culture is a promising tool for enhancing production of potent α-glucosidase inhibitors. Leaf ethyl acetate fraction (LEF) had the strongest antiglucosidase (EC 50 0.55 mg/mL) and radical scavenging (EC 50 10.81 μg/mL) activity among leaf fractionsCallus ethyl acetate fraction (CEF) and chloroform fraction had the highest antiglucosidase (EC 50 0.25 mg/mL) and radical scavenging (EC 50 34.12

  20. Callus induction and plant regeneration of Ulex europaeus

    OpenAIRE

    Ramírez,Ingrid; Dorta,Fernando; Cuadros-Inostroza,Álvaro; Peña-Cortés,Hugo

    2012-01-01

    A callus induction and plant regeneration protocol was developed from leaf and thorn explants for the plant Ulex europaeus. Explants were incubated on 2% sucrose half-strength Murashige and Skoog Medium (MS) with various combinations of plant growth regulators and antioxidants. The best frequency of callus and shoot formation was obtained with 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg/l x kinetin (Kin) 0.2 mg/l (DK Medium; callus induction) and zeatin (Z) 1 mg/l (DK medium; shoot induction)...

  1. Factors influencing callus induction and plant regeneration of ...

    African Journals Online (AJOL)

    ajl yemi

    2012-01-12

    ). Effect of basal medium on callus induction and plant regeneration. Three different kinds of basal mediums (MS, N6 and SH) were used to investigate their effects on callus induction and regeneration. Significant differences ...

  2. Nitrogen-15 uptake by whole plants and root callus cultures of inbred maize lines and their F1 hybrids

    International Nuclear Information System (INIS)

    Mladenova, Y.; Karadimova, M.

    1981-01-01

    The uptake of nitrogen-15 by 3 maize genotypes was investigated. Comparative analysis of N15 assimilation and distribution in the organs of intact plants of two self-pollinated lines and their F1 hybrid and also in a callus tissue of roots of the same genotypes was made. From the results the conclusion is drawn that the N-use efficiency of the female line is higher than that of the male line both in intact plants and callus tissues from roots. This fact indicates that the N-use efficiency is determined not only by the functions of the cells in the shoots, suggesting the participation of the photosynthetic carboxylases but also by the functions of cells without a photosynthesizing apparatus. The N-use efficiency in the F1 hybrid manifests ''heterosis'', in spite of the intact plants or root callus tissues are being studied. (author)

  3. Improvement of Caper (Capparis spinosa L) propagation using in vitro culture and gamma irradiation

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Elias, R.

    2010-01-01

    We studied some of the factors influencing seed germination and shoot growth of caper (Capparis spinosa L) and callus formation and regeneration from different parts of the plant (leaf, shoots, fruits) in order to establish a protocol for propagating this plant on a commercial scale. For the dormant seeds, we studied scratching the seed coat using iron particle filings prior to culture on nutrient medium or peatmos. Scratched and non-scratched seeds were also treated with concentrated sulfuric acid for various periods (20, 30, 45, 60 min). Seeds were additionally treated with ultrasonic waves for different periods (15, 30, 45, 60 min). The seeds were also irradiated with 50, 100, and 150 gray of gamma rays to study the effects of irradiation on the germination of caper seeds. Irradiation of the seed with 100 gray dose led to 50% germination one month after culturing in vitro, whereas, no seeds germinated in the control. Irradiation with 150 gray dose led to 18% germination of the seed. As for irradiated seed cultured in the peatmos, 70% of the seed germinated at the 100 gray dose whereas, only 5% of the seed germinated in the control. Treatment of the seed with H sub 2 SO sub 4 for 20 mints with scratching was very effective in stimulating germination, where the percentage of germinated seed reached 46% compared with 0% in the control. Treatment with just H sub 2 SO sub 4 for 20 mints resulted in 32% seed germination and treatment with just scratching resulted in only 20% germination. On the other hand, treating the seed with ultrasonic waves did not result in any seed germination. The effect of irradiation was also studied on the growth of caper shoots in vitro. The shoots were irradiated with 10, 15, and 20 gray and subsequently cultured on the MS medium without any growth regulators. The 10 gray dose stimulated shoot growth where average leaf area increased from 1.1 to 2.4 cm sup 2. The same irradiation dose, also, led to an increase in shoot length from 1.5 to

  4. Improvement of Caper (Capparis spinosa L) propagation using in vitro culture and gamma irradiation

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Elias, R.

    2009-05-01

    We studied some of the factors influencing seed germination and shoot growth of caper (Capparis spinosa L) and callus formation and regeneration from different parts of the plant (leaf, shoots, fruits) in order to establish a protocol for propagating this plant on a commercial scale. For the dormant seeds, we studied scratching the seed coat using iron particle filings prior to culture on nutrient medium or peatmos. Scratched and non-scratched seeds were also treated with concentrated sulfuric acid for various periods (20, 30, 45, 60 min). Seeds were additionally treated with ultrasonic waves for different periods (15, 30, 45, 60 min). The seeds were also irradiated with 50, 100, and 150 gray of gamma rays to study the effects of irradiation on the germination of caper seeds. Irradiation of the seed with 100 gray dose led to 50% germination one month after culturing in vitro, whereas, no seeds germinated in the control. Irradiation with 150 gray dose led to 18% germination of the seed. As for irradiated seed cultured in the peatmos, 70% of the seed germinated at the 100 gray dose whereas, only 5% of the seed germinated in the control. Treatment of the seed with H 2 SO 4 for 20 mints with scratching was very effective in stimulating germination, where the percentage of germinated seed reached 46% compared with 0% in the control. Treatment with just H 2 SO 4 for 20 mints resulted in 32% seed germination and treatment with just scratching resulted in only 20% germination. On the other hand, treating the seed with ultrasonic waves did not result in any seed germination. The effect of irradiation was also studied on the growth of caper shoots in vitro. The shoots were irradiated with 10, 15, and 20 gray and subsequently cultured on the MS medium without any growth regulators. The 10 gray dose stimulated shoot growth where average leaf area increased from 1.1 to 2.4 cm 2 . The same irradiation dose, also, led to an increase in shoot length from 1.5 to 2.2 cm. This was

  5. 21 CFR 358.510 - Corn and callus remover active ingredients.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 5 2010-04-01 2010-04-01 false Corn and callus remover active ingredients. 358.510 Section 358.510 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... USE Corn and Callus Remover Drug Products § 358.510 Corn and callus remover active ingredients. The...

  6. Morphological Characterization and Identification of Coffea liberica Callus of Somatic Embryogenesis Propagation.

    Directory of Open Access Journals (Sweden)

    Fitria Ardiyani

    2015-08-01

    Full Text Available Compared with other types of coffee, Liberica coffee is more difficult to be propagates using clonal methods. Meanwhile, demand for planting materials and consumption of this type of coffee is increasing lately. The objective of this paper is to present results of the work on morpological characterization of Liberica coffee (Coffea liberica callus produced by somatic embryogenesis propagation. This research used C. liberica Arruminensis clone. This clone was one of Liberica coffee clones which had superior taste. Main activitis carried out in this experiment were explant sterilization, explant induction and histological analysis on the callus produced. The result of this research showed that non embryogenic callus was higher (72% than embryogenic callus (28%. The callus description can be used to identify type and characteristic of the callus. Therefore, it can be a parameter to choose type of callus for propagation material. This is important because choosing the right callus is determine of the succesfully process of Liberica somatic embryogenesis. Keywords: somatic embryogenesis, Liberica, embryogenic, non-embryogenic

  7. Studies in tissue culture of some indigenous rice (Oryza glaberrima Steud.) accessions in Ghana

    International Nuclear Information System (INIS)

    Diawuoh, R.G.

    2011-01-01

    A study was conducted with the aim of developing separate protocols for callus induction and plant regeneration from different parts of three O. glaberrima accessions indigenous to Ghana. The three O. glaberrima accessions, Guame, N/4 and SARI 1 were assessed for their callus induction and plant regeneration ability from leaf segments, mature dehusked seeds and anthers on different concentrations of plant growth regulators, incorporated into Murashige and Skoog, (1962) (MS) basal medium. For leaf segments, callus was induced on MS supplemented with (0-10) mg/l 2,4-D. Callus induction frequency was significantly (p≤0.05) different among accessions, as well as among the 2,4-dichlorophenoxyacetic acid (2,4-D) levels tested. Highest callus induction frequency was exhibited at a concentration of 6 mg/l 2,4-D for all accessions tested. Callus obtained was sub-cultured on regeneration medium consisting of MS supplemented with (1:0-5) mg.l NAA:BAP. Plant regeneration was nil. Instead, prolific root formation was observed. For mature dehusked seeds, callus induction medium consisted of MS supplemented with (0-6) mg/l 2,4-D. All tested accessions exhibited highest callus frequency at 4 mg/l 2,4-D. Similarly callus induction frequency was significantly (p≤0.05) different among accessions, as well as among concentrations of 2,4-D tested. Calli obtained were sub-cultured on MS medium supplemented with (0-2.5) mg/l 6-benzylaminopurine (BAP) and exhibited the highest regeneration frequency on medium containing 2.0 mg/l BAP. However, callus induced on a concentration of 3 mg/l 2,4-D and sub-cultured on a concentration of 2 mg/l BAP gave the best response n terms of shoot proliferation, growth and root development and therefore were considered to be the optimum concentrations for callus induction and plant regeneration respectively. Plantlet regeneration was achieved only in accession N/4 while Guame and SARI 1 exhibited poor regeneration response. Among the three rice

  8. Phytochemical and biological study of callus cultures of Tulbaghia violacea Harv. Cultivated in Egypt.

    Science.gov (United States)

    Eid, Hanaa H; Metwally, Ghada F

    2017-08-01

    As in vitro plant cultures are used extensively to produce bioactive metabolites, our goal was to establish calli from Tulbaghia violacea Harv. flowers and assess the tissue phytochemically and biologically. Murashige & Skoog medium(MS) + 22.6 μM 2,4-dichlorophenoxyacetic acid +2.2 μM benzylaminopurine induced callus from flowers. Gas chromatography/mass spectrometry(GC/MS) analyses of n-hexane extracts of calli(HC) and flowers(HF) revealed 33 and 32 components(92.6 and 98.5%, respectively). Hydrocarbons were predominant in HC (55.0%), whereas a higher percentage of oxygenated compounds was found in HF(74.6%). Trans(E)-anethole(39.1%) and 16-hentriacontanone (30.3%) dominated in HF and HC, respectively. However, sulphur compounds were only detected in HF. Quantitative estimation of thiosulphinates, phenolics, flavonoids and saponins in ethanolic extracts of calli(EC) and flowers(EF) showed much higher contents in EF. Antioxidant, antimicrobial and cytotoxic screening of extracts demonstrated that EF was the most potent, followed by HF and EC; conversely, HC was inactive. Although HC and EC were less biologically active, these calli could be an alternative source of bioactive metabolites.

  9. STUDIES REGARDING CULTURE MEDIUM INFLUENCE ON IN VITRO REGENERATION FROM WHEAT IMATURE EMBRYOS

    Directory of Open Access Journals (Sweden)

    M. DANCI

    2008-05-01

    Full Text Available Surnamed „embryos’ saving method”, embryos culture is an in vitro technique used for over half of the century for saving the distant hybridization products, that would have degenerate in other conditions. Immature embryos culture is used for initiation of in vitro cultures imposed by the impossibility of using other explants for some of the plant species. Wheat is one of the crops that immature embryos culture technique is suitable for. This methods principle is based on aseptic embryos excision and their inoculation to an adequate culture medium. In vitro culture results depend in a greater manner of the basic culture medium and the hormonal balance used. Immature embryos isolated from two Romanian wheat cultivars – Dropia and Lovrin 41 – were inoculated for callus production on two types of basic media added with 2,4 D. The selected calluses were transferred on MS basic medium and several parameters were registered. Both cultivars emphasized a good callusing capacity, in a different percentage depending on the culture media used, such as 71,09 – 94,45%.. big differences between the cultivars regarding embriogenic callus frequency, shooting callus frequency and regenerated plants percentage were registered.

  10. Production of friable embryogenic callus and regeneration of ...

    African Journals Online (AJOL)

    Generation of embryogenic callus is a key step in genetic engineering of many crop species, including cassava. Protocols for generation of friable embryogenic callus (FEC) have been lacking for Ugandan cassava genotypes, thereby delaying their genetic engineering for agronomic and other desirable traits. The objective ...

  11. Locked plating of distal femur fractures leads to inconsistent and asymmetric callus formation.

    Science.gov (United States)

    Lujan, Trevor J; Henderson, Chris E; Madey, Steven M; Fitzpatrick, Dan C; Marsh, J Lawrence; Bottlang, Michael

    2010-03-01

    Locked plating constructs may be too stiff to reliably promote secondary bone healing. This study used a novel imaging technique to quantify periosteal callus formation of distal femur fractures stabilized with locking plates. It investigated the effects of cortex-to-plate distance, bridging span, and implant material on periosteal callus formation. Retrospective cohort study. One Level I and one Level II trauma center. Sixty-four consecutive patients with distal femur fractures (AO types 32A, 33A-C) stabilized with periarticular locking plates. Osteosynthesis using indirect reduction and bridge plating with periarticular locking plates. Periosteal callus size on lateral and anteroposterior radiographs. Callus size varied from 0 to 650 mm2. Deficient callus (20 mm2 or less) formed in 52%, 47%, and 37% of fractures at 6, 12, and 24 weeks postsurgery, respectively. Callus formation was asymmetric, whereby the medial cortex had on average 64% more callus (P=0.001) than the anterior or posterior cortices. A longer bridge span correlated minimally with an increased callus size at Week 6 (P=0.02), but no correlation was found at Weeks 12 and 24 postsurgery. Compared with stainless steel plates, titanium plates had 76%, 71%, and 56% more callus at Week 6 (P=0.04), Week 12 (P=0.03), and Week 24 (P=0.09), respectively. Stabilization of distal femur fractures with periarticular locking plates can cause inconsistent and asymmetric formation of periosteal callus. A larger bridge span only minimally improves callus formation. The more flexible titanium plates enhanced callus formation compared with stainless steel plates.

  12. Determination of suitable microspore stage and callus induction from anthers of kenaf (Hibiscus cannabinus L.).

    Science.gov (United States)

    Ibrahim, Ahmed Mahmood; Kayat, Fatimah Binti; Hussin, Zeti Ermiena Surya Mat; Susanto, Dwi; Ariffulah, Mohammed

    2014-01-01

    Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6-8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.

  13. Determination of Suitable Microspore Stage and Callus Induction from Anthers of Kenaf (Hibiscus cannabinus L.

    Directory of Open Access Journals (Sweden)

    Ahmed Mahmood Ibrahim

    2014-01-01

    Full Text Available Kenaf (Hibiscus cannabinus L. is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6–8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.

  14. Determination of Suitable Microspore Stage and Callus Induction from Anthers of Kenaf (Hibiscus cannabinus L.)

    Science.gov (United States)

    Binti Kayat, Fatimah; Ermiena Surya Mat Hussin, Zeti; Susanto, Dwi; Ariffulah, Mohammed

    2014-01-01

    Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6–8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA. PMID:24757416

  15. An environment with strong gravitational and magnetic field alterations synergizes to promote variations in Arabidopsis thaliana callus global transcriptional state

    Data.gov (United States)

    National Aeronautics and Space Administration — Using diamagnetic levitation we have exposed A. thaliana in vitro callus cultures to five environments with different levels of effective gravity (from levitation...

  16. Assessment of silver nitrate on callus induction and in vitro shoot regeneration in tomato (solanum lycopersicum mill.)

    International Nuclear Information System (INIS)

    Shah, S.H.; Ali, S.; Jan, S.A.

    2014-01-01

    In vitro morphogenesis is greatly influenced by a gaseous plant growth regulator (ethylene). The effect of silver nitrate (AgNO/sub 3/) and different plant growth regulators were assessed on callus induction, In vitro shoot regeneration and multiple primordial shoots per explant in tomato. The maximum callus induction frequency was recorded culturing hypocotyls, while In vitro shoot regeneration frequency and the number of primordial shoots per explant were significantly higher when leaf discs were used as explants. The callus induction frequency was improved by the supplementation of 10-15 mg/l AgNO/sub 3/ in MS basal media along with 2.0 mg/l IAA, 2.5 mg/l BAP and yielded the highest callus induction frequency (91.33%) in cv. Rio Grande, followed by Roma (88.33%) and Moneymaker (82.66%). Similarly, the highest In vitro shoot regeneration frequency (96.66, 92.66 and 90%) was recorded in Rio Grande, Roma and Moneymaker on MS media fortified with 0.1 mg/l IAA, 1.0 mg/l ZEA and 2.0 mg/l BAP along with 8-10 mg/l AgNO/sub 3/. AgNO/sub 3/ also had promotive effect on induction of multiple shoots per explant. These findings indicate that ethylene is linked with the suppression of In vitro morphogenesis in tomato and AgNO3 interacts with ethylene and enhances callus induction and In vitro shoot regeneration in tomato. (author)

  17. Experimental Paper. In vitro synthesis of mucilage in Plantago ovata Forsk affected by genotypes and culture media

    Directory of Open Access Journals (Sweden)

    Golkar Pooran

    2017-03-01

    Full Text Available Introduction: Psyllium (Plantago ovata Forsk is medicinally used mainly for its mucilage content. Objective: In the present study, an attempt was made to improve mucilage yield under in vitro callus culture using different genotypes, explants and culture media. Methods: The effects of a range of concentrations of plant growth regulators including 2,4-dichlorophenoxyacetic acid (2,4-D and kinetin (Kin were evaluated on mucilage synthesis under in vitro culture using cotyledon, hypocotyl and seed explants. Fourteen genotypes originating from different geographical regions of Iran were used to evaluate their response to in vitro mucilage synthesis. Results: The highest rate of callus induction (76% and callus growth rate CGR (0.38 mm/day were induced on MS medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l Kin and the hypocotyl explant. The results of analysis of variance showed significant genotypic differences for callus induction, CGR and mucilage content of callus and seeds. The mucilage content ranged from 0.38 to 0.08 (g/g DW and 0.13 to 0.042 (g/g DW for callus and seed, respectively. The superior callus induction (73%, CGR (0.45 mm/day and mucilage content of callus (0.38 g/g DW was denoted to Po1 genotype. The callus produced nearly three times more mucilage than the seeds using superior genotype (Po1. Conclusion: The results of this study revealed that high efficiency of callus culture of P. ovata using hypocotyl explant accompanied by the exploration of genetic diversity are important to improve the yield of mucilage synthesis by in vitro callus culture.

  18. The biological effect of gamma radiation on in vitro culture in rice

    International Nuclear Information System (INIS)

    Wang Cailian; Xu Gang; Shen Mei; Chen Qiufang

    1994-08-01

    Radiobiological effects of gamma radiation on different types of rice before or during in vitro culture, combined treatments of 137 Cs γ-rays and NaN 3 on mature embryo culture, and irradiation on growth of calli derived from anther in rice were studied. The dose-effects relations of callus induction rate and callus growth rate could be fitted according to the multi-target and single-hit model. Effect of somatic cultures of different types in rice was different. Increase in plant regeneration capacity was found with 100, 150 Gy gamma rays. Decrease of callus induction rate, callus growth rate and callus differentiation rate (especially in the 1st culture) were observed in combined treatments of γ-rays and NaN 3 . However, mutagenic effects of treatments with γ-rays were much higher than those of combined treatment of γ-rays and NaN 3 in the 2nd and the 3rd culture. Combined treatments of 137 Cs γ-rays with 200 Gy and 2 mmol NaN 3 were suitable for explant in rice before culture. To irradiate the calli derived from anther in rice with 30 Gy gamma rays can rise plant regeneration capacity during continuing culture

  19. Study on the secondary metabolic regulation of callus of Panax quinquefolium L

    International Nuclear Information System (INIS)

    Zhang Meiping; Wang Yi; Sun Chunyu; Li Xianggao

    2003-01-01

    The effects of precursors with different kind, concentration, growth phase, time and different ATP, nicotinic acid concentration on the callus growth and saponin formation with the buds of Panax quinquefolium L. has been studied. The results showed that 0.1% mevalonic acid inhibited the callus growth but promoted the saponin formation; L-leucine of 100 mg/L had no obvious effects on callus growth, but promoted the saponin formation; Mg(Ac) 2 of 444 mg/L promoted both the callus growth and saponin content. The optimum time for precursor feeding is at the beginning of regeneration. The optimum time for mevalonic acid is 6h; ATP of 160 mg/L promoted both the callus growth and saponin formation. Nicotinic acid of 1.0 mg/L promoted callus growth and saponin formation. With 4000 Gy of 60 Co γ-rays, the saponin content has been increased 1 time compared with nonirradiation

  20. Vitamin C and total phenols quantification in calli of native passion fruit induced by combinations of Picloram and Kinetin

    Directory of Open Access Journals (Sweden)

    Fabiane Aparecida Artioli-Coelho

    2015-08-01

    Full Text Available Brazil is one of the center of origin of passion fruit and has an important natural variability of the genus Passiflora. Several wild species of this genus are resistant to some pests and diseases and many are considered as medicinal. The aim of this research was to induce callus from in vitro Passiflora gibertii leaf explants for quantification of vitamin C and total phenols. Once the appropriate auxin/cytokine balance promotes callus formation and may optimize the production of secondary compounds and vitamins, calli were induced using a half-strength MS medium with a combination of the auxin Picloran (PIC and the cytokine Kinetin (KIN. The vitamin C and total phenols were quantified by colorimetric methods from calli after different culture periods. The calli induction was strongly dependent of the combination PIC/KIN. It was observed high vitamin C content (94.8mg 100g-1 during the calli induction period in MS+4.14µM PIC+ 0.207µM KIN. Higher PIC/KIN concentrations promoted an increase in the vitamin C content after three subcultures. The higher PIC (8.28µM/KIN (0.828µM concentration was the higher was the total phenols production (66mg tannic acid 100g-1 of fresh callus during the calli induction period.

  1. De novo assembly and comparative analysis of the transcriptome of embryogenic callus formation in bread wheat (Triticum aestivum L.).

    Science.gov (United States)

    Chu, Zongli; Chen, Junying; Sun, Junyan; Dong, Zhongdong; Yang, Xia; Wang, Ying; Xu, Haixia; Zhang, Xiaoke; Chen, Feng; Cui, Dangqun

    2017-12-19

    During asexual reproduction the embryogenic callus can differentiate into a new plantlet, offering great potential for fostering in vitro culture efficiency in plants. The immature embryos (IMEs) of wheat (Triticum aestivum L.) are more easily able to generate embryogenic callus than mature embryos (MEs). To understand the molecular process of embryogenic callus formation in wheat, de novo transcriptome sequencing was used to generate transcriptome sequences from calli derived from IMEs and MEs after 3d, 6d, or 15d of culture (DC). In total, 155 million high quality paired-end reads were obtained from the 6 cDNA libraries. Our de novo assembly generated 142,221 unigenes, of which 59,976 (42.17%) were annotated with a significant Blastx against nr, Pfam, Swissprot, KOG, KEGG, GO and COG/KOG databases. Comparative transcriptome analysis indicated that a total of 5194 differentially expressed genes (DEGs) were identified in the comparisons of IME vs. ME at the three stages, including 3181, 2085 and 1468 DEGs at 3, 6 and 15 DC, respectively. Of them, 283 overlapped in all the three comparisons. Furthermore, 4731 DEGs were identified in the comparisons between stages in IMEs and MEs. Functional analysis revealed that 271transcription factor (TF) genes (10 overlapped in all 3 comparisons of IME vs. ME) and 346 somatic embryogenesis related genes (SSEGs; 35 overlapped in all 3 comparisons of IME vs. ME) were differentially expressed in at least one comparison of IME vs. ME. In addition, of the 283 overlapped DEGs in the 3 comparisons of IME vs. ME, excluding the SSEGs and TFs, 39 possessed a higher rate of involvement in biological processes relating to response to stimuli, in multi-organism processes, reproductive processes and reproduction. Furthermore, 7 were simultaneously differentially expressed in the 2 comparisons between the stages in IMEs, but not MEs, suggesting that they may be related to embryogenic callus formation. The expression levels of genes, which

  2. GABA shunt in the callus cells derived from soybean cotyledon

    Energy Technology Data Exchange (ETDEWEB)

    Tokunaga, M; Nakano, Y; Kitaoka, S [Osaka Prefectural Univ., Sakai (Japan). Coll. of Agriculture

    1975-01-01

    In the growing callus cells from soybean cotyledon, the activities of glutamate decarboxylase and GABA transaminase were increased in the early phase of the callus growth on the Miller agar medium. Succinate dehydrogenase activity was also changed in a similar manner. From these and the additional evidences that GABA transaminase was probably localized in the mitochondria, it has been made clear that the GABA shunt (GABA by-pass pathway) is operative and contributes to the respiratory metabolism in growing callus cells. Feeding young callus cells with GABA-U-/sup 14/C for 24 hr actually resulted in finding 53% of the taken up radioactivity in released carbon dioxide. Considerable parts of the taken up radioactivity were found in amino acids and proteins which should have been formed via the GABA shunt also.

  3. Study on Effect of Type of Explant and Hormone on Callus Induction and Regeneration in Saffron (Crocus sativus L.

    Directory of Open Access Journals (Sweden)

    Mohsen Sajjadi

    2015-10-01

    Full Text Available Saffron (Crocus sativus L. is one of the medicinal plants that contain active components and medicinal materials. Tissue culture of saffron can improve the quality and quantity of the saffron product, increase its export and the farmers’ income. In this study, 36 different types of hormone combinations in the dark and 9 different treatments of hormone combinations in cold (4°C, using different saffron explants (bulb, leaf, scales around leaf and distal parts of the leaf were studied in tissue culture. To investigate the growth of corms, the callus formation and the regeneration rate, three replications for each treatment were used and the length of shoot (cm, the callus formation percentage and the regeneration percentage were measured and statistical analysis was performed. Among the types of explants, only explants from bulbs produced the callus on MS medium containing 2 mg.l-1 BAP and 1 mg.l-1 IBA in both the dark and cold conditions. The highest percentage of regeneration was obtained in MS medium with hormonal composition of 0.3 mg.l-1 TDZ, 1 mg.l-1 BAP, 2 mg.l-1 IBA and 0.01 mg.l-1 GA3 in the cold conditions.

  4. C4 photosynthesis in Euphorbia degeneri and E. remyi: a comparison of photosynthetic carbon metabolism in leaves, callus cultures and regenerated plants

    International Nuclear Information System (INIS)

    Ruzin, S.E.

    1984-04-01

    Based on analysis of 14 CO 2 fixation kinetics and assays of enzymes related to C 4 metabolism (NAD-ME, NADP-ME, NAD-MDH, NADP-MDH, AST, ALT), leaves and regenerated plants of Euphorbia degeneri exhibit a modified NADP-ME-type photosynthesis. Apparently, both aspartate and malate are used for transport of CO 2 to bundle sheath cells. Callus grown on either non-shoot-forming or shoot-forming media fixes CO 2 into RPP-cycle intermediates and sucrose, as well as malate and aspartate. 14 CO 2 pulse/chase kinetics show no significant loss of label from C 4 acids throughout a one minute chase. Analysis of PEPCase revealed the presence of 2 isoenzymes in both leaf and regenerated plant tissues (K/sub m/ [PEP] = 0.080 and 0.550) but only one isoenzyme in callus (K/sub m/ = 0.100). It appears that C 4 photosynthesis does not occur in callus derived from this C 4 dicot but is regenerated concomitant with shoot regeneration, and β-carboxylation of PEP in callus, mediated by the low K/sub m/ isoenzyme of PEPCase, produces C 4 acids that are not involved in the CO 2 shuttle mechanism characteristic of C 4 photosynthesis. 161 references, 19 figures, 12 tables

  5. Availability of commonly consumed and culturally specific fruits and vegetables in African-american and Latino neighborhoods.

    Science.gov (United States)

    Grigsby-Toussaint, Diana S; Zenk, Shannon N; Odoms-Young, Angela; Ruggiero, Laurie; Moise, Imelda

    2010-05-01

    Although the importance of culture in shaping individual dietary behaviors is well-documented, cultural food preferences have received limited attention in research on the neighborhood food environment. The purpose of this study was to assess the availability of commonly consumed and culturally specific fruits and vegetables in retail food stores located in majority African-American and Latino neighborhoods in southwest Chicago, IL. A cross-sectional survey of 115 stores (15% grocery stores, 85% convenience/corner stores) in African-American neighborhoods and 110 stores (45% grocery stores, 55% convenience/corner stores) in Latino neighborhoods was conducted between May and August of 2006. chi(2) tests were used to assess differences in the availability (presence/absence) of commonly consumed (n=25) and culturally specific fruits and vegetables for African Americans (n=16 varieties) and Latinos (n=18 varieties). Stores located in neighborhoods in which the majority of residents were African American or Latino were more likely to carry fresh fruits and vegetables that were culturally relevant to the dominant group. For example, grocery stores located in Latino neighborhoods were more likely to carry chayote (82.0% vs 17.6%, P<0.05), whereas grocery stores located in African-American neighborhoods were more likely to carry black-eyed peas (52.9% vs 20%, P<0.05). Most stores, however, carried fewer than 50% of commonly consumed or culturally specific fruits and vegetables. Findings from this study highlight that limited availability of culturally specific as well as commonly consumed fruits and vegetables in the neighborhood may be a barrier to fruit and vegetable consumption among African Americans and Latinos. Copyright 2010 American Dietetic Association. Published by Elsevier Inc. All rights reserved.

  6. Studies on Callus Induction and Regeneration of Medicinal Plant Chicory (Cichorium intybus L. from Leaf and Petiole Explants

    Directory of Open Access Journals (Sweden)

    H. Hadizadeh

    2016-07-01

    Full Text Available Introduction: Chicory (Cichorium intybus L. belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant contain these volatile oils, with the majority of the toxic components concentrated in the plant's root. In folk medicine, the plant is used for the treatment of diarrhea, spleen enlargement, fever, and vomiting. Antihepatotoxic activity on damaged rat’s liver sections and anti-bacterial activity of this crop has been recently reported. In vitro regeneration from leaf explants with various hormonal combinations has been reported previously. Moreover, in vitro regeneration of Chicory from cotyledon explants using different combinations of plant growth regulators has been studied. Also, a protocol for the regeneration of plantlets from leaf and petiole explants of witloof chicory has been developed. The aim of the present investigation was optimization of callus induction and shoot regeneration from leaf and petiole tissues of Chicory (Esfahan genotype. Materials and Methods: In this investigation, Esfahan genotype was used for callus induction and direct shoot regeneration. Seeds were first washed with running tap water for 30 min then seeds were surface sterilized by dipping in 70% ethanol for 90 s and rinsed with sterile distilled water, followed by immersing in 5% sodium hypochlorite solution for 25 min and thereafter rinsed for 30 min with sterile distilled water. The basal medium used in this investigation was MS. For shoot regeneration, leaf and petiole explants (5 mm segments were excised from 4-week-old sterile seedlings and cultured on MS medium containing different combinations of NAA / BA and KIN / BA in two separate experiments. Experiments were performed factorial based on completely randomized design. Cultures were incubated at 25

  7. Hot callusing for propagation of American beech by grafting

    Science.gov (United States)

    David W. Carey; Mary E. Mason; Paul Bloese; Jennifer L. Koch

    2013-01-01

    To increase grafting success rate, a hot callus grafting system was designed and implemented as part of a multiagency collaborative project to manage beech bark disease (BBD) through the establishment of regional BBD-resistant grafted seed orchards. Five years of data from over 2000 hot callus graft attempts were analyzed using a logistic regression model to determine...

  8. Experimental study upon the effect of irradiation on callus formation of fracture

    International Nuclear Information System (INIS)

    Saigusa, Fujio

    1981-01-01

    Irradiation effects on callus formation after bone fracture were studied in rats with fractured right lower extremity. Follow-up study was continued for 112 days since 3000 rad was irradiated to the fractured site 3 days after bone fracture. Callus formation was noted in both of the outer and inner part (bone marrow) of the diaphysis before 14 days after bone fracture, but it was slow and sparse compared with that of non-irradiated group. Callus formation tended to disappear gradually from the outside of the diaphysis after 28 days after bone fracture. Strong disturbance was found in the surrounding vascular system at this time. Inside of the diaphysis, callus formation was restricted the end of the fracture, where lamellar calluses fused together. Changes in vascular system remained until 56 days after bone fracture. Vascular distribution was most dense 28 days after bone fracture. In many of the calluses which have established fusion, findings suggested excessive calcification in the trabeculae. Vascular distribution at this time was sparse, vascular formation was markedly suppressed in the bone marrow, and very little vascular formation was found in the fractured edges of the bone. (Ueda, J.)

  9. Factors Associated With Callus in Patients with Diabetes, Focused on Plantar Shear Stress During Gait.

    Science.gov (United States)

    Hamatani, Masako; Mori, Taketoshi; Oe, Makoto; Noguchi, Hiroshi; Takehara, Kimie; Amemiya, Ayumi; Ohashi, Yumiko; Ueki, Kohjiro; Kadowaki, Takashi; Sanada, Hiromi

    2016-11-01

    The aim of this study is to identify whether plantar shear stress in neuropathic patients with diabetes with callus is increased compared with those without callus. The differences in foot deformity, limited joint mobility, repetitive stress of walking, and ill-fitting shoes between patients with callus and those without callus were also determined. Subjects were recruited from the Diabetic Foot Outpatient Clinic. A newly developed in-shoe measurement system, which has flexible and thin insoles, enabled measurement of both plantar pressure and shear stress simultaneously when subjects walked as usual on a 10 m walkway. It was found that plantar shear stress adjusted for weight during the push-off phase was increased by 1.32 times in patients with callus compared with those without callus (mean ± SD: 0.0500 ± 0.0160 vs 0.0380 ± 0.0144, P = .031). Moreover, hallux valgus deformity, reduction in dorsiflexion of the ankle joint and increase in plantar flexion were showed in feet with callus. Increased plantar shear stress may be caused by gait change that patients having callus push off with the metatarsal head instead of the toe as a result of foot deformity and limited joint mobility. It was found that plantar shear stress adjusted for weight during the push-off phase was increased in patients with callus compared with those without callus by using the newly developed measurement system. These results suggest that reduction of plantar shear stress during the push-off phase can prevent callus formation in neuropathic patients with diabetes. © 2016 Diabetes Technology Society.

  10. In vitro antibacterial, antifungal, and antioxidant activities of the essential oil and methanol extracts of herbal parts and callus cultures of Satureja hortensis L.

    Science.gov (United States)

    Güllüce, M; Sökmen, M; Daferera, D; Ağar, G; Ozkan, H; Kartal, N; Polissiou, M; Sökmen, A; Sahin, F

    2003-07-02

    The present study was designated to evaluate the antimicrobial and antioxidant activities of the essential oil, obtained by using a Clevenger distillation apparatus, water soluble (polar) and water insoluble (nonpolar) subfractions of the methanol extracts from aerial parts of Satureja hortensis L. plants, and methanol extract from calli established from the seeds using Gamborg's B5 basal media supplemented with indole-3-butyric acid (1.0 ppm), 6-benzylaminopurine (N(6)-benzyladenine) (1.0 ppm), and sucrose (2.5%). The antimicrobial test results showed that the essential oil of S. hortensis had great potential antimicrobial activities against all 23 bacteria and 15 fungi and yeast species tested. In contrast, the methanol extract from callus cultures and water soluble subfraction of the methanol extract did not show antimicrobial activities, but the nonpolar subfraction had antibacterial activity against only five out of 23 bacterial species, which were Bacillus subtilis, Enterococcus fecalis, Pseudomonas aeruginosa, Salmonella enteritidis, and Streptococcus pyogenes. Antioxidant studies suggested that the polar subfractions of the methanol extract of intact plant and methanol extract of callus cultures were able to reduce the stable free radical 2,2-diphenyl-1-picrylhydrazyl to the yellow-colored diphenylpicrylhydrazine. In this assay, the strongest effect was observed for the tissue culture extract, with an IC(50) value of 23.76 +/- 0.80 microgram/mL, which could be compared with the synthetic antioxidant agent butylated hydroxytoluene. On the other hand, linoleic acid oxidation was 95% inhibited in the presence of the essential oil while the inhibition was 90% with the chloroform subfraction of the intact plant. The chemical composition of a hydrodistilled essential oil of S. hortensis was analyzed by gas chromatography (GC)/flame ionization detection (FID) and a GC-mass spectrometry system. A total 22 constituents representing 99.9% of the essential oil were

  11. Optimization of callus and cell suspension cultures of Barringtonia racemosa (Lecythidaceae family for lycopene production Otimização de culturas de suspensões de calos e células de Barringtonia racemosa (família Lecythidaceae para produção de licopeno

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    Mandana Behbahani

    2011-02-01

    Full Text Available Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS, Woody Plant Medium (WPM and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D. Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.O licopeno está presente numa série de frutas frescas e hortaliças principalmente na folhas de Barringtonia racemosa. A extra

  12. The GABA shunt in the callus cells derived from soybean cotyledon

    International Nuclear Information System (INIS)

    Tokunaga, Masao; Nakano, Yoshihisa; Kitaoka, Shozaburo

    1975-01-01

    In the growing callus cells from soybean cotyledon, the activities of glutamate decarboxylase and GABA transaminase were increased in the early phase of the callus growth on the Miller agar medium. Succinate dehydrogenase activity was also changed in a similar manner. From these and the additional evidences that GABA transaminase was probably localized in the mitochondria, it has been made clear that the GABA shunt (GABA by-pass pathway) is operative and contributes to the respiratory metabolism in growing callus cells. Feeding young callus cells with GABA-U- 14 C for 24 hr actually resulted in finding 53% of the taken up radioactivity in released carbon dioxide. Considerable parts of the taken up radioactivity were found in amino acids and proteins which should have been formed via the GABA shunt also. (auth.)

  13. Fully automated segmentation of callus by micro-CT compared to biomechanics.

    Science.gov (United States)

    Bissinger, Oliver; Götz, Carolin; Wolff, Klaus-Dietrich; Hapfelmeier, Alexander; Prodinger, Peter Michael; Tischer, Thomas

    2017-07-11

    A high percentage of closed femur fractures have slight comminution. Using micro-CT (μCT), multiple fragment segmentation is much more difficult than segmentation of unfractured or osteotomied bone. Manual or semi-automated segmentation has been performed to date. However, such segmentation is extremely laborious, time-consuming and error-prone. Our aim was to therefore apply a fully automated segmentation algorithm to determine μCT parameters and examine their association with biomechanics. The femura of 64 rats taken after randomised inhibitory or neutral medication, in terms of the effect on fracture healing, and controls were closed fractured after a Kirschner wire was inserted. After 21 days, μCT and biomechanical parameters were determined by a fully automated method and correlated (Pearson's correlation). The fully automated segmentation algorithm automatically detected bone and simultaneously separated cortical bone from callus without requiring ROI selection for each single bony structure. We found an association of structural callus parameters obtained by μCT to the biomechanical properties. However, results were only explicable by additionally considering the callus location. A large number of slightly comminuted fractures in combination with therapies that influence the callus qualitatively and/or quantitatively considerably affects the association between μCT and biomechanics. In the future, contrast-enhanced μCT imaging of the callus cartilage might provide more information to improve the non-destructive and non-invasive prediction of callus mechanical properties. As studies evaluating such important drugs increase, fully automated segmentation appears to be clinically important.

  14. The effects of gamma-ray irradiation to strawberry (Fragaria x ananassa Duch.) calli on shoot regeneration, achene formation and morphological variations of regenerants

    International Nuclear Information System (INIS)

    Kasumi, Masakazu

    2002-01-01

    Strawberry (Fragaria x ananassa 'Nyoho') calli, derived from anthers, were irradiated with gamma -ray at dosages from 100 to 1,600 Gy (10 Gy · hr - 1) and their effects on callus growth, plant regeneration, and somaclonal variation were analyzed. The growth of callus and percentage of plant regeneration from the callus were reduced slightly at dosages less than 200 Gy, moderately at 400 Gy, and markedly above 800 Gy. Fruit growth associated with seed fertility (>50 % ) was morphologically normal at less than 100 Gy, whereas achene formation was significantly inhibited above 200 Gy. The percentages of morphological variation in regenerants formed from the irradiated callus were:13.7 (no irradiation), 18.6 (100 Gy), 66.0 (200 Gy), 75.7 (400 Gy), and 97.0 (800 Gy). Furthermore, higher dosages yielded wider variations, e.g., thick and small leaf, light leaf color, white flesh, and long fruit. In some regenerants, more than one morphological aberrations developed, some of which were transmissible to daughter plants. These results revealed that the irradiating strawberry callus with gamma - ray resulted in a higher degree and wider spectrum of somaclonal variation than did simple callus culture. (author)

  15. Progressive erosion of genetic and epigenetic variation in callus-derived cocoa (Theobroma cacao) plants.

    Science.gov (United States)

    Rodríguez López, Carlos M; Wetten, Andrew C; Wilkinson, Michael J

    2010-06-01

    *Relatively little is known about the timing of genetic and epigenetic forms of somaclonal variation arising from callus growth. We surveyed for both types of change in cocoa (Theobroma cacao) plants regenerated from calli of various ages, and also between tissues from the source trees. *For genetic change, we used 15 single sequence repeat (SSR) markers from four source trees and from 233 regenerated plants. For epigenetic change, we used 386 methylation-sensitive amplified polymorphism (MSAP) markers on leaf and explant (staminode) DNA from two source trees and on leaf DNA from 114 regenerants. *Genetic variation within source trees was limited to one slippage mutation in one leaf. Regenerants were far more variable, with 35% exhibiting at least one mutation. Genetic variation initially accumulated with culture age but subsequently declined. MSAP (epigenetic) profiles diverged between leaf and staminode samples from source trees. Multivariate analysis revealed that leaves from regenerants occupied intermediate eigenspace between leaves and staminodes of source plants but became progressively more similar to source tree leaves with culture age. *Statistical analysis confirmed this rather counterintuitive finding that leaves of 'late regenerants' exhibited significantly less genetic and epigenetic divergence from source leaves than those exposed to short periods of callus growth.

  16. Biotransformation of isofraxetin-6-O-β-d-glucopyranoside by Angelica sinensis (Oliv.) Diels callus.

    Science.gov (United States)

    Zhou, Di; Zhang, Yuhua; Jiang, Zhe; Hou, Yue; Jiao, Kun; Yan, Chunyan; Li, Ning

    2017-01-15

    Isofraxetin-6-O-β-d-glucopyranoside, identified from traditional medicinal herbal Xanthoceras sorbifolia Bunge, has been demonstrated to be a natural neuroinflammatory inhibitor. In order to obtain more derivatives with potential anti-neuroinflammatory effects, biotransformation was carried out. According to the characteristics of coumarin skeleton, suspension cultures of Angelica sinensis (Oliv.) Diels callus (A. sinensis callus) were employed because of the presence of diverse phenylpropanoids biosynthetic enzymes. As a result, 15 products were yielded from the suspension cultures, including a new coumarin: 8'-dehydroxymethyl cleomiscosin A (1), together with 14 known compounds. Their structures were elucidated by extensive spectroscopic analysis. Furthermore, the biotransformed pathways were discussed. Among them, compound 13 was transformed from isofraxetin-6-O-β-d-glucopyranoside, while compounds 1-6, 10-12, 14-15 were derived from the culture medium stimulated by the substrate. The biotransformation processes include hydroxylation, oxidation and esterification. Furthermore, their inhibitory effects on lipopolysaccharide (LPS)-activated nitric oxide (NO) production were evaluated in BV2 microglial cells. It is worth noting that, 1, 1'-methanediylbis(4-methoxybenzene) (3), obtucarbamates A (5), 2-nonyl-4-hydroxyquinoline N-oxide (10) and 1H-indole-3-carbaldehyde (11) exhibited significant inhibitory effect against neuroinflammation with IC 50 values at 1.22, 10.57, 1.02 and 0.76μM respectively, much stronger than that of the positive control minocycline (IC 50 35.82μM). Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Shear Stress-Normal Stress (Pressure) Ratio Decides Forming Callus in Patients with Diabetic Neuropathy

    Science.gov (United States)

    Noguchi, Hiroshi; Takehara, Kimie; Ohashi, Yumiko; Suzuki, Ryo; Yamauchi, Toshimasa; Kadowaki, Takashi; Sanada, Hiromi

    2016-01-01

    Aim. Callus is a risk factor, leading to severe diabetic foot ulcer; thus, prevention of callus formation is important. However, normal stress (pressure) and shear stress associated with callus have not been clarified. Additionally, as new valuables, a shear stress-normal stress (pressure) ratio (SPR) was examined. The purpose was to clarify the external force associated with callus formation in patients with diabetic neuropathy. Methods. The external force of the 1st, 2nd, and 5th metatarsal head (MTH) as callus predilection regions was measured. The SPR was calculated by dividing shear stress by normal stress (pressure), concretely, peak values (SPR-p) and time integral values (SPR-i). The optimal cut-off point was determined. Results. Callus formation region of the 1st and 2nd MTH had high SPR-i rather than noncallus formation region. The cut-off value of the 1st MTH was 0.60 and the 2nd MTH was 0.50. For the 5th MTH, variables pertaining to the external forces could not be determined to be indicators of callus formation because of low accuracy. Conclusions. The callus formation cut-off values of the 1st and 2nd MTH were clarified. In the future, it will be necessary to confirm the effect of using appropriate footwear and gait training on lowering SPR-i. PMID:28050567

  18. Shear Stress-Normal Stress (Pressure Ratio Decides Forming Callus in Patients with Diabetic Neuropathy

    Directory of Open Access Journals (Sweden)

    Ayumi Amemiya

    2016-01-01

    Full Text Available Aim. Callus is a risk factor, leading to severe diabetic foot ulcer; thus, prevention of callus formation is important. However, normal stress (pressure and shear stress associated with callus have not been clarified. Additionally, as new valuables, a shear stress-normal stress (pressure ratio (SPR was examined. The purpose was to clarify the external force associated with callus formation in patients with diabetic neuropathy. Methods. The external force of the 1st, 2nd, and 5th metatarsal head (MTH as callus predilection regions was measured. The SPR was calculated by dividing shear stress by normal stress (pressure, concretely, peak values (SPR-p and time integral values (SPR-i. The optimal cut-off point was determined. Results. Callus formation region of the 1st and 2nd MTH had high SPR-i rather than noncallus formation region. The cut-off value of the 1st MTH was 0.60 and the 2nd MTH was 0.50. For the 5th MTH, variables pertaining to the external forces could not be determined to be indicators of callus formation because of low accuracy. Conclusions. The callus formation cut-off values of the 1st and 2nd MTH were clarified. In the future, it will be necessary to confirm the effect of using appropriate footwear and gait training on lowering SPR-i.

  19. CALLUS INDUCTION FROM 15 CARNATION (DIANTHUS CARYOPHYLLUS L. CULTIVARS

    Directory of Open Access Journals (Sweden)

    Jaime A. TEIXEIRA DA SILVA

    2014-12-01

    Full Text Available Plant growth regulators (PGRs were used to induce callus in 15 carnation (Dianthus caryophyllus L.; Caryophyllaceae cultivars: Orange Sherbert, Avalanche, Magenta, La France, Stripe Red, Marie, Concerto PVP, Snap, Lucky Pierot, Cinnamon Tea, White Love, Siberia, Magesta, Spark Bruno, and Honono no Estejo. Seeds were initially sown on autoclaved moistened filter paper and internodes of surface-sterilized seedlings were used as explants. Most callus was induced in the presence of 0.5 mg/L α-naphthaleneacetic acid used together with 1 mg/L 6-benzyladenine or 1 mg/L 2,4-dichlorophenoxyacetic acid on basal Murashige and Skoog medium. Callus is not a desirable method to clonally propagate important germplasm but can serve as one possible way of deriving periclinal mutants as a result of somaclonal variation.

  20. Proliferation Potential of 18-Month-Old Callus of Ananas comosus L. cv. Moris

    Directory of Open Access Journals (Sweden)

    A.E. De Silva

    2006-01-01

    Full Text Available Differential effect of plant growth regulators and additives in proliferation of 18-month-old calli of Ananas comosus L. cv. Moris were assessed in vitro. The proliferation of callus relied on the growth regulators and additives. Of the different auxins supplemented in the Murashige and Skoog (MS media, 32.22 μM α-naphthaleneacetic acid (NAA gave the highest mean fresh weight of callus (46.817 g. Medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D was inferior to NAA, while b-naphthoxy acetic acid (BNOA and p-chlorophenoxy acetic acid (4-CPA were not effective in proliferating 18-months old callus. Addition of casein hydrolysate and coconut water to NAA supplemented medium showed better proliferation and production of callus. However, in terms of callus production, NAA at 32.22 μM was economically better.

  1. Plant tissue culture techniques

    Directory of Open Access Journals (Sweden)

    Rolf Dieter Illg

    1991-01-01

    Full Text Available Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus or organized tissues or organs put in culture, under controlled sterile conditions.

  2. Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants.

    Science.gov (United States)

    Chen, M H; Wang, P J; Maeda, E

    1987-10-01

    The regeneration potential of shoot tip, stem, leaf, cotyledon and root explants of two papaya cultivars (Carica papaya cv. 'Solo' and cv. 'Sunrise') were studed. Callus induction of these two cultivars of papaya showed that the shoot tips and stems are most suitable for forming callus, while leaves, cotyledons and roots are comparatively difficult to induce callus. Callus induction also varied with the varities. Somatic embryogenesis was obtained from 3-month-old root cultures. A medium containing half strength of MS inorganic salts, 160 mg/l adenine sulfate, 1.0 mg/1 NAA, 0.5 mg/1 kinetin and 1.0 mg/1 GA3 was optimal for embryogenesis. The callus maintained high regenerative capacity after two years of culture on this medium. Plants derived from somatic embryos were obtained under green-house conditions.

  3. C/sub 4/ photosynthesis in Euphorbia degeneri and E. remyi: a comparison of photosynthetic carbon metabolism in leaves, callus cultures and regenerated plants

    Energy Technology Data Exchange (ETDEWEB)

    Ruzin, S.E.

    1984-04-01

    Based on analysis of /sup 14/CO/sub 2/ fixation kinetics and assays of enzymes related to C/sub 4/ metabolism (NAD-ME, NADP-ME, NAD-MDH, NADP-MDH, AST, ALT), leaves and regenerated plants of Euphorbia degeneri exhibit a modified NADP-ME-type photosynthesis. Apparently, both aspartate and malate are used for transport of CO/sub 2/ to bundle sheath cells. Callus grown on either non-shoot-forming or shoot-forming media fixes CO/sub 2/ into RPP-cycle intermediates and sucrose, as well as malate and aspartate. /sup 14/CO/sub 2/ pulse/chase kinetics show no significant loss of label from C/sub 4/ acids throughout a one minute chase. Analysis of PEPCase revealed the presence of 2 isoenzymes in both leaf and regenerated plant tissues (K/sub m/ (PEP) = 0.080 and 0.550) but only one isoenzyme in callus (K/sub m/ = 0.100). It appears that C/sub 4/ photosynthesis does not occur in callus derived from this C/sub 4/ dicot but is regenerated concomitant with shoot regeneration, and ..beta..-carboxylation of PEP in callus, mediated by the low K/sub m/ isoenzyme of PEPCase, produces C/sub 4/ acids that are not involved in the CO/sub 2/ shuttle mechanism characteristic of C/sub 4/ photosynthesis. 161 references, 19 figures, 12 tables.

  4. [Chromosome variability in the tissue culture of rare Gentiana species].

    Science.gov (United States)

    Tvardovs'ka, M O; Strashniuk, N M; Mel'nyk, V M; Adonin, V I; Kunakh, V A

    2008-01-01

    Cytogenetic analysis of plants and tissue culture of Gentiana lutea, G. punctata, G. acaulis has been carried out. Culturing in vitro was found to result in the changes of chromosome number in the calluses of the species involved. Species specificity for variation of the cultured cell genomes was shown. Contribution of the original plant genotypes to the cytogenetic structure of the tissue culture was established. Gentiana callus tissues (except for in vitro culture of G. punctata, derived from plant of Breskul'ska population) were found to exhibit modal class with the cells of diploid and nearly diploid chromosome sets.

  5. Callus formation impedes adventitious rhizogenesis in air layers of broadleaved tree species

    Directory of Open Access Journals (Sweden)

    Sanjay Singh

    2014-07-01

    Full Text Available Callusing and root induction in air layering was evaluated aiming at evolution of procedure for mass clonal propagation of mature ortets of five tropical broadleaf species differing in their potential for adventitious root formation in shoot cuttings as: Anogiessus latifolia < Boswellia serrata < Dalbergia latifolia < Gmelina arborea < Dalbergia sissoo. Two experiments were conducted in rainy season during consecutive years; without application of growth regulators in the first year and with growth regulators (T1 - water, T2- 100 ppm indole-3-acetic acid, T3-100 ppm thiamine-HCl and T4 -combination of T2 + T3 in the next year. Air layered branches were detached from the trees to record percentage of alive airlayers, callusing and rooting (% as well as root number and root length. Response to air layering was found to be highly variable in five tree species but appeared to be feasible procedure for clonal propagation of mature ortets of B. serrata and D. sissoo with 100% (in auxin + thiamine treatment and 83.3% (in auxin treatment success, respectively. Maximum callusing (% was found in D. latifolia while no callusing was observed in D. sissoo, which is most easy-to-root among all five species. Callus formation impedes adventitious rhizogenesis in air layers as significant negative correlation of callusing (% and adventitious root formation was recorded in air layers of five tropical broadleaved tree species. Application of exogenous auxin alone or in combination with thiamine circumvents callusing to ensure direct development of roots for successful air layering.

  6. Allelopathy of small everlasting (Antennaria microphylla) : Phytotoxicity to leafy spurge (Euphorbia esula) in tissue culture.

    Science.gov (United States)

    Hogan, M E; Manners, G D

    1990-03-01

    Media and media extracts from callus cultures of small everlasting (Antennaria microphylla) inhibited leafy spurge (Euphorbia esula L.) callus tissue and suspension culture growth (50 and 70% of control, respectively) and were phytotoxic in lettuce and leafy spurge root elongation bioassays (64 and 77% of control, respectively). Hydroquinone, a phytotoxic compound previously isolated from small everlasting, was also biosynthesized by callus and suspension cultures of this species. Exogenously supplied hydroquinone (0.5 mM) was toxic to leafy spurge suspension culture cells and was only partially biotransformed to its nontoxic water-soluble monoglucoside, arbutin, by these cells. This report confirms the chronic involvement of hydroquinone in the allelopathic interaction between small everlasting and leafy spurge.

  7. In Vivo Fast Induction of Homogeneous Autopolyploids via Callus in Sour Jujube (Ziziphus acidojujuba Cheng et Liu

    Directory of Open Access Journals (Sweden)

    Qinghua Shi

    2016-05-01

    Full Text Available Polyploidization has been demonstrated as a very effective approach in fruit tree improvement. Sour jujube (Ziziphus acidojujuba Cheng et Liu is a promising diploid wild, traditional fruit species (2n = 2x = 24 that is rich in vitamin C, which is the main rootstock of Chinese jujube (Z. jujuba Mill.. The novel method we developed for rapid in vivo induction of homogeneous autopolyploids (IVIHA via callus in Chinese jujube was first applied and further optimized in sour jujube. Under optimized conditions, an average of one pure autotetraploid shoot could be regenerated from one treated branch, thereby indicating a relatively high efficiency rate. A total of 9 pure autotetraploid genotypes were created, and one of these was released as a new cultivar named ‘Zhuguang’ in 2015. Moreover, unexpected octoploids and hexaploids were also simultaneously created and detected. The leaves of tetraploids were thicker, broader, and darker in color than those of the original diploids, whereas the leaf sizes of octoploids were much smaller compared to that of diploids. However, stoma size increased with the occurrence of ploidy, mainly from diploid to octoploid. The well grown ploidies of jujube included diploids, triploids, and tetraploids. Anatomical observation indicated that adventitious buds/shoots emerged from the callus that formed on the cut, which was then followed by the development of connective vascular tissues between the adventitious bud and the stock plant tissue. This study demonstrates the universality of the IVIHA method that was initially developed in Chinese jujube, as well as provides a foundation for high-efficiency pure polyploid induction in sour jujube.

  8. Uptake and fate of IAA in apple callus tissue using IAA-1-14C

    International Nuclear Information System (INIS)

    Epstein, E.; Lavee, S.

    1975-01-01

    Incubation of young growing and older non-growing apple callus tissues in a medium containing IAA-1- 14 C resulted in rapid disappearance of the IAA. In old calluses (3 months), the major portion of IAA was lost by decarboxylation (90% after 4 hr) and very little (1.4%) was maintained by the tissue. In young calluses, after 4 hr in light, decarboxylation reached 20% and absorption 35% of the labelled IAA. Some decomposition of IAA was caused by photolysis and autoclaving (19% and 3%, respectively) but the final distribution of radioactivity was not affected. Factors such as sucrose concentration in the incubation medium, distilled water as incubation medium, and cutting of the callus did not affect tissue behavior. Special precautions were taken to eliminate non-biological decomposition of IAA. Therefore, we believe that the rapid CO 2 evolution is of enzymatic nature. This theory is supported by the drop in decarboxylation after killing of the callus, and the increase of decarboxylation with age. No enzyme was secreted by the callus into the medium after 24 hr of incubation, and IAA decomposition in old tissues is done probably on the surface. Absorption of IAA increased with increasing callus size and decarboxylation decreased. (auth.)

  9. α(1) adrenergic receptor agonist, phenylephrine, actively contracts early rat rib fracture callus ex vivo.

    Science.gov (United States)

    McDonald, Stuart J; Dooley, Philip C; McDonald, Aaron C; Djouma, Elvan; Schuijers, Johannes A; Ward, Alex R; Grills, Brian L

    2011-05-01

    Early, soft fracture callus that links fracture ends together is smooth muscle-like in nature. We aimed to determine if early fracture callus could be induced to contract and relax ex vivo by similar pathways to smooth muscle, that is, contraction via α(1) adrenergic receptor (α(1) AR) activation with phenylephrine (PE) and relaxation via β(2) adrenergic receptor (β(2) AR) stimulation with terbutaline. A sensitive force transducer quantified 7 day rat rib fracture callus responses in modified Krebs-Henseliet (KH) solutions. Unfractured ribs along with 7, 14, and 21 day fracture calluses were analyzed for both α(1) AR and β(2) AR gene expression using qPCR, whilst 7 day fracture callus was examined via immunohistochemistry for both α(1) AR and β(2) AR- immunoreactivity. In 7 day callus, PE (10(-6)  M) significantly induced an increase in force that was greater than passive force generated in calcium-free KH (n = 8, mean 51% increase, 95% CI: 26-76%). PE-induced contractions in calluses were attenuated by the α(1) AR antagonist, prazosin (10(-6)  M; n = 7, mean 5% increase, 95% CI: 2-11%). Terbutaline did not relax callus. Gene expression of α(1) ARs was constant throughout fracture healing; however, β(2) AR expression was down-regulated at 7 days compared to unfractured rib (p contract. We propose that increased concentrations of α(1) AR agonists such as noradrenaline may tonically contract callus in vivo to promote osteogenesis. Copyright © 2010 Orthopaedic Research Society.

  10. Mevastatin-induced inhibition of cell growth in avocado suspension ...

    African Journals Online (AJOL)

    Cell suspension cultures were established using soft, friable callus derived from nucellar tissue of 'Hass' avocado (Persea americana Mill.) seed from fruit harvested 190 days after full bloom. Cell cultures were maintained in liquid medium supplemented with naphthalene acetic acid (NAA), isopentenyl adenine (iP) and ...

  11. CALLUS INDUCTION FROM 15 CARNATION (DIANTHUS CARYOPHYLLUS L.) CULTIVARS

    OpenAIRE

    Jaime A. TEIXEIRA DA SILVA

    2014-01-01

    Plant growth regulators (PGRs) were used to induce callus in 15 carnation (Dianthus caryophyllus L.; Caryophyllaceae) cultivars: Orange Sherbert, Avalanche, Magenta, La France, Stripe Red, Marie, Concerto PVP, Snap, Lucky Pierot, Cinnamon Tea, White Love, Siberia, Magesta, Spark Bruno, and Honono no Estejo. Seeds were initially sown on autoclaved moistened filter paper and internodes of surface-sterilized seedlings were used as explants. Most callus was induced in the presence of 0.5 mg/L α-n...

  12. Tissue Culture Technique and Gamma Irradiation Used in Evaluation of Five Genotypes of Bread Wheat to Salinity Tolerance

    International Nuclear Information System (INIS)

    AL Jibouri, A. A. M.; Sulaiman, A. A.; Dallul, R. A.

    2005-01-01

    Callus has been induced from immature embryos of five genotypes of Triticum aestivum L (i.e. Maxipak, IPA99, 113, Tamus 3 and Babylon) on MS culture media. The calli were irradiated by Gamma ray at doses (0,15, 30,45, And 60) Gray, and cultured on MS media supplemented with salt mixture consist of sodium, calcium and magnesium chlorides in portions 3; 3; 1 respectively.The results showed significant differences in genotype responses to irradiation and salinity . A significant reduction in fresh and dry weights with increasing of irradiation dose and salt concentration in culture media in all genotypes. The genotype IPA99 showed a highest fresh and dry weight than the other genotypes . The proline contents increased significantly in callus with irradiation doses and concentration increases. On the other hand callus carbohydrate content reduced significantly with increased salt concentration in culture media. The capability of callus regeneration was reduced by increasing irradiation doses and salt concentration in culture media. (author)

  13. Efficient callus formation and plant regeneration of goosegrass [Eleusine indica (L.) Gaertn.].

    Science.gov (United States)

    Yemets, A I; Klimkina, L A; Tarassenko, L V; Blume, Y B

    2003-02-01

    Efficient methods in totipotent callus formation, cell suspension culture establishment and whole-plant regeneration have been developed for the goosegrass [ Eleusine indica (L.) Gaertn.] and its dinitroaniline-resistant biotypes. The optimum medium for inducing morphogenic calli consisted of N6 basal salts and B5 vitamins supplemented with 1-2 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg l(-1) glycine, 100 mg l(-1) asparagine, 100 mg l(-1) casein hydrolysate, 30 g l(-1) sucrose and 0.6% agar, pH 5.7. The presence of organogenic and embryogenic structures in these calli was histologically documented. Cell suspension cultures derived from young calli were established in a liquid medium with the same composition. Morphogenic structures of direct shoots and somatic embryos were grown into rooted plantlets on medium containing MS basal salts, B5 vitamins, 1 mg l(-1) kinetin (Kn) and 0.1 mg l(-1) indole-3-acetic acid (IAA), 3% sucrose, 0.6% agar, pH 5.7. Calli derived from the R-biotype of E. indica possessed a high resistance to trifluralin (dinitroaniline herbicide) and cross-resistance to a structurally non-related herbicide, amiprophosmethyl (phosphorothioamidate herbicide), as did the original resistant plants. Embryogenic cell suspension culture was a better source of E. indica protoplasts than callus or mesophyll tissue. The enzyme solution containing 1.5% cellulase Onozuka R-10, 0.5% driselase, 1% pectolyase Y-23, 0.5% hemicellulase and N(6) mineral salts with an additional 0.2 M KCl and 0.1 M CaCl(2) (pH 5.4-5.5) was used for protoplast isolation. The purified protoplasts were cultivated in KM8p liquid medium supplemented with 2 mg l(-1) 2,4-D and 0.2 mg l(-1) Kn.

  14. Generation and multiplication of plantlets from callus derived from Haplopappus gracilus (Nutt.) Gray and their karyotype analysis

    Science.gov (United States)

    Kann, R. P.; O'Connor, S. A.; Levine, H. G.; Krikorian, A. D.

    1991-01-01

    Unopened flower heads of Haplopappus gracilis (2n = 4) provided primary explants for callus production and subsequent induction of organized growth. Callus was initiated from small (3-5 mm in length) floral buds with benzylaminopurine (BAP) (44.4 micromoles; 10 mg/l) and naphthalene acetic acid (NAA) (0.54 micromole; 0.1 mg/l). Lowering the BAP level to 4.44 micromoles (1 mg/l) but maintaining the NAA level, gave rise to organized but highly compressed shoot growing points from an otherwise undifferentiated callus mass. Shoots selected from such cultures were maintainable and could be proliferated by growing 1-1.5-cm stem tip cuttings on Murashige and Skoog basal medium (solidified with agar) containing 0.444 micromole (0.1 mg/l) BAP and 0.054 micromole (0.01 mg/l) NAA. The stem tip multiplication rates obtainable by these means permit reliable strategies for shoot multiplication or production of rooted plantlets. Prolonged subculture and maintenance of shoots on growth regulator-free medium leads to in vitro flowering and greatly reduces rooting capacity. Karyotype analysis of chromosomes from root tip cells at metaphase and chromosome measurements show that karyologically uniform plantlets (based on chromosome number and morphology) can be obtained.

  15. Primary structures of two ribonucleases from ginseng calluses - New members of the PR-10 family of intracellular pathogenesis-related plant proteins

    NARCIS (Netherlands)

    Moiseyev, GP; Fedoreyeva, LI; Zhuravlev, YN; Yasnetskaya, E; Jekel, PA; Beintema, JJ

    1997-01-01

    The amino acid sequences of two ribonucleases from a callus cell culture of Panax ginseng were determined, The two sequences differ at 26% of the amino acid positions, Homology was found with a large family of intracellular pathogenesis-related proteins, food allergens and tree pollen allergens from

  16. Studies on the effects of mutagens on cultured cells of higher plants, (2)

    International Nuclear Information System (INIS)

    Mori, Shigeyuki; Nakanishi, Hiroo; Shiojiri, Satoshi; Murakami, Michio

    1980-01-01

    A comparative study on the effects of 60 Co gamma radiation was carried out on callus tissue, seedlings and seeds of Phaseolus vulgaris L. cv. Kentucky Wonder. Callus was obtained from the hypocotyl pieces grown on an agar-solidified, modified Linsmaier and Skoog's medium supplemented with 2, 4-D (5 mg/l), kinetin (5 mg/l) and yeast extract (1,000 mg/l). Subculture was conducted on the same medium in the dark at 28 +- 1 0 C. Callus was exposed to various doses (2.5 - 15 KR) of gamma radiation at 20 KR/hr. Fresh and dry weight, potency of growth in subculture of irradiated callus, color and friability were measured. As compared with control, irradiated callus growth showed a significant decrease with increasing doses. Callus growth was classified into three types, i.e. callus began to grow from the early stage of culture at 5 KR and below, from 6.25 - 8.75 KR there was a delayed growth, at 10 KR and over there was little visual sign of growth. The potency of growth in subculture for 28 days of irradiated- and 24-day-cultured callus was recognized at 10 KR and below. With increasing doses, color of callus darkened, and the degree of friability increased. While 10-day-old seedlings were irradiated, striking inhibition of growth occurred at 2 KR, followed by degradation of the apical meristem and growth ceased completely at 5 KR. Severe inhibition of growth of irradiated seeds occurred at 20 KR and over. A remarkable difference in radiosensitivity was observed among callus tissue, seedlings and seeds, i.e. seedlings were most sensitive, followed by callus tissue, and seeds were most resistant. (author)

  17. Sugarcane in vitro culture technology: Opportunities for Kenya's ...

    African Journals Online (AJOL)

    free clonal materials. Successful protocols for shoot tip culture, callus culture, embryo culture, virus free plant production and somatic embryogenesis have already been established. Thus, in vitro technology can be used to enhance ...

  18. Statistically optimized biotransformation protocol for continuous production of L-DOPA using Mucuna monosperma callus culture.

    Science.gov (United States)

    Inamdar, Shrirang Appasaheb; Surwase, Shripad Nagnath; Jadhav, Shekhar Bhagwan; Bapat, Vishwas Anant; Jadhav, Jyoti Prafull

    2013-01-01

    L-DOPA (3,4-dihydroxyphenyl-L-alanine), a modified amino acid, is an expansively used drug for the Parkinson's disease treatment. In the present study, optimization of nutritional parameters influencing L-DOPA production was attempted using the response surface methodology (RSM) from Mucuna monosperma callus. Optimization of the four factors was carried out using the Box-Behnken design. The optimized levels of factors predicted by the model include tyrosine 0.894 g l(-1), pH 4.99, ascorbic acid 31.62 mg l(-1)and copper sulphate 23.92 mg l(-1), which resulted in highest L-DOPA yield of 0.309 g l(-1). The optimization of medium using RSM resulted in a 3.45-fold increase in the yield of L-DOPA. The ANOVA analysis showed a significant R (2) value (0.9912), model F-value (112.465) and probability (0.0001), with insignificant lack of fit. Optimized medium was used in the laboratory scale column reactor for continuous production of L-DOPA. Uninterrupted flow column exhibited maximum L-DOPA production rate of 200 mg L(-1) h(-1) which is one of the highest values ever reported using plant as a biotransformation source. L-DOPA production was confirmed by HPTLC and HPLC analysis. This study demonstrates the synthesis of L- DOPA using Mucuna monosperma callus using a laboratory scale column reactor.

  19. Methyl jasmonate differentially affects tocopherol content and tyrosine amino transferase activity in cultured cells of Amaranthus caudatus and Chenopodium quinoa.

    Science.gov (United States)

    Antognoni, F; Faudale, M; Poli, F; Biondi, S

    2009-03-01

    Tocopherols are lipid-soluble compounds synthesised exclusively by photosynthetic organisms. In this study, in vitro callus cultures were established from two plants that are naturally rich in tocopherols, Amaranthus caudatus and Chenopodium quinoa, in order to examine whether callus cultures were able to produce these compounds at levels comparable to those observed in planta. In both species, cotyledon explants produced the best callus induction and, once established, callus cultures were grown under two different hormonal treatments to check for effects of growth and to induce chloroplast differentiation in the cells. A rapid differentiation of chloroplasts occurred only in C. quinoa cell aggregates grown in the presence of benzyladenine, leading to the production of a homogeneous green callus. In both species, only alpha-tocopherol was produced by callus cultures, although levels were much lower than in planta, and the production was not influenced by the hormonal conditions. Interestingly, cell cultures of the two species responded in different ways to methyl jasmonate (MJ). In A. caudatus cultures, treatment with 100 mum MJ increased the production of alpha-tocopherol up to fivefold, and the inductive effect was influenced by the hormonal composition of the medium. This increase in alpha-tocopherol was associated with a proportional increase in tyrosine aminotransferase (TAT) activity, one of the key enzymes involved in tocopherol biosynthesis. By contrast, in C. quinoa cultures, elicitation with MJ did not have any effect, neither on tocopherol production, nor on TAT activity. These results are discussed in relation to chloroplast differentiation and the interplay between jasmonates and phytohormones.

  20. MRT letter: Contrast-enhanced computed tomographic imaging of soft callus formation in fracture healing.

    Science.gov (United States)

    Hayward, Lauren Nicole Miller; de Bakker, Chantal Marie-Jeanne; Lusic, Hrvoje; Gerstenfeld, Louis Charles; Grinstaff, Mark W; Morgan, Elise Feng-I

    2012-01-01

    Formation of a cartilaginous soft callus at the site of a bone fracture is a pivotal stage in the healing process. Noninvasive, or even nondestructive, imaging of soft callus formation can be an important tool in experimental and pre-clinical studies of fracture repair. However, the low X-ray attenuation of cartilage renders the soft callus nearly invisible in radiographs. This study utilized a recently developed, cationic, iodinated contrast agent in conjunction with micro-computed tomography to identify cartilage in fracture calluses in the femora of C57BL/6J and C3H/HeJ mice. Fracture calluses were scanned before and after incubation in the contrast agent. The set of pre-incubation images was registered against and then subtracted from the set of post-incubation images, resulting in a three-dimensional map of the locations of cartilage in the callus, as labeled by the contrast agent. This map was then compared to histology from a previous study. The results showed that the locations where the contrast agent collected in relatively high concentrations were similar to those of the cartilage. The contrast agent also identified a significant difference between the two strains of mice in the percentage of the callus occupied by cartilage, indicating that this method of contrast-enhanced computed tomography may be an effective technique for nondestructive, early evaluation of fracture healing. Copyright © 2011 Wiley Periodicals, Inc.

  1. In vitro elicitation, isolation, and characterization of conessine biomolecule from Holarrhena antidysenterica (L.) Wall. callus and its larvicidal activity against malaria vector, Anopheles stephensi Liston.

    Science.gov (United States)

    Kumar, Dinesh; Kumar, Gaurav; Das, Ram; Kumar, Ravindra; Agrawal, Veena

    2018-03-01

    In vitro elicitation of an important compound conessine has been done in the bark-derived callus culture of Holarrhena antidysenterica (L.) Wall. employing different elicitors. For induction of callus, green bark explants excised from field-grown plants were cultured on MS medium augmented with different concentrations (0, 1, 2.5, 5, and 10 μM) of various growth regulators such as BA, IBA, NAA, and 2,4-D either alone or in combinations. The maximum amount of conessine (458.18 ± 0.89 d μg/g dry wt.) was achieved in callus developed on MS medium supplemented with 5 μM BA and 5 μM 2,4-D through HPLC analysis. Elicitation in conessine content in the above callus was achieved employing a variety of organic (phenylalanine, tyrosine, chitosan, tryptophan, casein hydrolysate, proline, sucrose, and yeast extract) as well as inorganic elicitors (Pb(NO 3 ) 2 , As 2 O 3 , CuSO 4 , NaCl, and CdCl 2 ) in different concentrations. The optimum enhancement in conessine content (3518.58 ± 0.28 g  μg/g dry wt.) was seen at the highest concentration (200 mg/L) of phenylalanine. The enhancement was elicitor specific and dose dependent. The overall increment of the conessine content was seen in the order of phenylalanine > tryptophan > Pb(NO 3 ) 2 > sucrose > NaCl > As 2 O 3 > casein hydrolysate > CdCl 2 > chitosan > proline > yeast extract > CuSO 4 > tyrosine. The isolation and purification of conessine was done using methanol as a solvent system through column chromatography (CC) and TLC. The isolated compound was characterized by FT-IR, 1 H-NMR, and HRMS which confirmed with the structure of conessine. The bioassays conducted with the isolated compound revealed a strong larvicidal activity against Anopheles stephensi Liston with LC 50 and LC 90 values being 1.93 and 5.67 ppm, respectively, without harming the nontarget organism, Mesocyclops thermocyclopoides Harada, after 48 h of treatment. This is our first report for the isolation and elicitation of conessine

  2. Use of culture filtrates of Ceratocystis ulmi as a bioassay to screen for disease tolerant Ulmus americana

    Science.gov (United States)

    Paula M. Pijut; Subash C. Domir; R. Daniel Lineberger; Lawrence R. Schreiber

    1990-01-01

    Callus cultures of elm (Ulmus americana L.) derived from Dutch elm disease susceptible, intermediate-resistant, and resistant genotypes were exposed to the culture filtrates of three pathogenic isolates of Ceratocystis ulmi, the causal agent of Dutch elm disease. Callus fresh weights, cell viability, and reactions of stem cuttings...

  3. Callus induction, regeneration and transformation of sugarcane ...

    African Journals Online (AJOL)

    Yomi

    2012-03-22

    Mar 22, 2012 ... dichlorophenoxyacetic acid (2,4-D), naphthalene acetic acid (NAA), yeast extract and coconut water .... transformed (control) plantlets using CTAB method (Doyle and .... Embryogenic Sugarcane Callus by Image Analysis.

  4. In vitro androgenetic cultures of Hyoscyamus niger L., H. albus L. and alkaloid content assay

    Directory of Open Access Journals (Sweden)

    Maria Wesołwska

    2014-01-01

    Full Text Available In vitro cultures of Hyoscyamus niger L. and H. albus L. anthers were initiated which resulted in obtaining androgenectic plants and callus cultures. The leaves of these pants and the callus cultures were subjected to analysis (TLC, GC for the presence of alkaloids, derivatives of tropane. In the studied material, alkaloids of different qualitative and quantitative composition from that of ground-grown plants were found.

  5. Investigation of radiosensitivity and growth dynamics for callus tissues Crepis Capillaris, Haplopappus gracilis, Phasolium vulgaris exposed to gamma radiation

    International Nuclear Information System (INIS)

    Gatsek, Eh.; Glinkova, E.; Ismailova, Eh.N.

    1983-01-01

    Radiosensitivity of three kinds of callus tissues (Crepis capillaris, Haplopappus gracilis, Phasolium vulgaris) manifested in the change of fresh weight after γ-irradiation has been investigated. Irradiated callus arowth showed decrease with increasing doses. It is shown that the radiosensitivity of ''young'' callus tissues is determined by the kind of the plant. Callus of Phaseolis has been found to have the highest radioresistance, while that of Crepis has the lowest one. Radiosensitivity of ''old'' callus tissues is the same for all kinds. Potential mechanism of radiosensitivity of callus tissUes are discussed

  6. Determining the pharmacological activity of Physalis peruviana fruit juice on rabbit eyes and fibroblast primary cultures.

    Science.gov (United States)

    Pardo, Juan Manuel; Fontanilla, Marta Raquel; Ospina, Luis Fernando; Espinosa, Lady

    2008-07-01

    The pharmacologic activity of compounds isolated from Physalis peruviana has been demonstrated. The use of this fruit juice for treating pterygium has been reported in Colombian traditional medicine. However, studies demonstrating the fruit juice's pharmacologic activity when used in this disease have not been published to date. In the present study the anti-inflammatory and cytostatic activities of P. peruviana fruit juice in a rabbit eye inflammatory model were investigated. A novel rabbit eye inflammation model was developed for studying the juice's anti-inflammatory activity (based on an adaptation of the Draize test). Cytostatic activity was evaluated by measuring and comparing growth rates of cultured fibroblasts exposed and not exposed to various fruit juice concentrations. P. peruviana fruit juice exhibited a mild anti-inflammatory activity compared with methylprednisolone, a known anti-inflammatory drug. An interesting dose-dependent cytostatic effect on cultured fibroblasts was also established. The data found suggest that the P. peruviana fruit juice anti-pterygium effect described in traditional medicine may be related to its inhibiting fibroblast growth. The present study contributes to the pharmacologic knowledge regarding a remedy commonly used in Colombian traditional medicine.

  7. Anther development stage and gamma radiation effects on tomato anther-derived callus formation

    International Nuclear Information System (INIS)

    Brasileiro, Ana Christina R.; Willadino, Lilia; Guerra, Marcelo; Colaco, Waldeciro; Meunier, Isabelle; Camara, Terezinha R.

    1999-01-01

    Two experiments were carried (I) to determine tomato anther development stage influence on callus production; and (II) to investigate gamma radiation effects on anther culture. In the first experiment, anthers of a tomato hybrid (IPA 5 x Rotam 4-F 1 ) were grown on three media. Although calli were induced at all stages of anther development, varying from prophase I to mono nucleate microspore, callus frequency decreased as anther development progressed and calli induction were not significantly affected by all media tested. Anthers containing prophase I meiocytes produced the highest calli frequency. Anther and flower bud length both were significantly correlated with anther development stage. In the second experiment, seed and floral buds of tomato hybrids IPA 5 x Rotam 4 (F 2 ), IPA 6 x Rotam 4 (F 2 ) and IPA 8 x 217.1 (F 2 ) were submitted to gamma-ray and anthers were plated on two media described by Gresshoff and Doy (1972) supplemented with 2.0 mg L -1 NAA + 5.0 mg L -1 KIN and 2.0 mg L -1 NAA + 1.0 mg L -1 KIN. No significant differences for genotype and dosage testes were found for calli formation. (author)

  8. Salinity-Induced Callus Browning and Re-Differentiation, Root Formation by Plantlets and Anatomical Structures of Plantlet Leaves in Two Malus Species

    International Nuclear Information System (INIS)

    Gou, W.; Zheng, P.; Zheng, P.; Wang, K.; Zhang, L.; Akram, N. A.

    2016-01-01

    Apple (Malus domestica L.) is widely grown in northern China. However, soil salinization has become one of the most severe factors limiting apple productivity in some regions including the Loess Plateau. In our study, the regeneration system of both rootstock Rehd (Malus robusta Rehd) and scion Fuji (Malus domestica Borkh. cv. Fuji) was established In vitro. The two Malus species were cultured on the MS medium containing 0 or 150 mM NaCl to examine salt-induced effects on callus browning and re-differentiation, root formation of plantlets and anatomical structures of plantlet leaves at 15 days old callus and plantlet stages. Salt stress caused a marked increase in callus browning rate, while a decrease in re-differentiation rate, rooting rate, root number and length in both species. Additionally, anatomical structures of plantlet leave showed salt-induced damage such as reduced palisade tissue and intracellular chloroplast, incomplete development of xylem and severe damage of the phloem tissue. Salt stress also caused a few adaptive structural features in leaves including increased thickness of upper and lower epidermis, elevated proportion of spongy tissue and formation of lignified vessels. The responses of the two Malus species did not differ significantly at the differentiation stage. However, they were more sensitive to salinity at the callus stage than those at the plantlet stage in each species. Therefore, callus stage has been found to be more suitable for evaluating responses of the two apple species to salt stress. The Fuji and Rehd could be treated as a good scion/rootstock combination of apple to adapt to soil salinity based on their similar degree of salt stress-tolerance. (author)

  9. Hyperplastic callus formation in osteogenesis imperfecta: CT and MRI findings

    International Nuclear Information System (INIS)

    Rieker, O.; Kreitner, K.F.; Karbowski, A.

    1998-01-01

    Hyperplastic callus formation is a noteworthy condition in patients with osteogenesis imperfecta because it often mimicks osteosarcoma on radiography. The findings of CT and MRI in hyperplastic callus formation have not been reported. In the presented case, MRI demonstrated contrast enhancement and edema of the surrounding soft tisssue, consistent with benign as well as malignant disease. Computed tomography showed a calcified rim of the lesion which may be a useful feature to rule out osteosarcoma in this condition. (orig.)

  10. Genetic Dissection of Maize Embryonic Callus Regenerative Capacity Using Multi-Locus Genome-Wide Association Studies

    Directory of Open Access Journals (Sweden)

    Langlang Ma

    2018-04-01

    Full Text Available The regenerative capacity of the embryonic callus, a complex quantitative trait, is one of the main limiting factors for maize transformation. This trait was decomposed into five traits, namely, green callus rate (GCR, callus differentiating rate (CDR, callus plantlet number (CPN, callus rooting rate (CRR, and callus browning rate (CBR. To dissect the genetic foundation of maize transformation, in this study multi-locus genome-wide association studies (GWAS for the five traits were performed in a population of 144 inbred lines genotyped with 43,427 SNPs. Using the phenotypic values in three environments and best linear unbiased prediction (BLUP values, as a result, a total of 127, 56, 160, and 130 significant quantitative trait nucleotides (QTNs were identified by mrMLM, FASTmrEMMA, ISIS EM-BLASSO, and pLARmEB, respectively. Of these QTNs, 63 QTNs were commonly detected, including 15 across multiple environments and 58 across multiple methods. Allele distribution analysis showed that the proportion of superior alleles for 36 QTNs was <50% in 31 elite inbred lines. Meanwhile, these superior alleles had obviously additive effect on the regenerative capacity. This indicates that the regenerative capacity-related traits can be improved by proper integration of the superior alleles using marker-assisted selection. Moreover, a total of 40 candidate genes were found based on these common QTNs. Some annotated genes were previously reported to relate with auxin transport, cell fate, seed germination, or embryo development, especially, GRMZM2G108933 (WOX2 was found to promote maize transgenic embryonic callus regeneration. These identified candidate genes will contribute to a further understanding of the genetic foundation of maize embryonic callus regeneration.

  11. Mineral crystal alignment in mineralized fracture callus determined by 3D small-angle X-ray scattering

    Energy Technology Data Exchange (ETDEWEB)

    Liu Yifei; Manjubala, Inderchand; Fratzl, Peter [Department of Biomaterials, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam (Germany); Roschger, Paul [4th Medical Department, Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1140 Vienna (Austria); Schell, Hanna; Duda, Georg N, E-mail: fratzl@mpikg.mpg.d [Julius Wolff Institut and Center for Musculoskeletal Surgery, Charite- University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin (Germany)

    2010-10-01

    Callus tissue formed during bone fracture healing is a mixture of different tissue types as revealed by histological analysis. But the structural characteristics of mineral crystals within the healing callus are not well known. Since two-dimensional (2D) scanning small-angle X-ray scattering (sSAXS) patterns showed that the size and orientation of callus crystals vary both spatially and temporally [1] and 2D electron microscopic analysis implies an anisotropic property of the callus morphology, the mineral crystals within the callus are also expected to vary in size and orientation in 3D. Three-dimensional small-angle X-ray scattering (3D SAXS), which combines 2D SAXS patterns collected at different angles of sample tilting, has been previously applied to investigate bone minerals in horse radius [2] and oim/oim mouse femur/tibia [3]. We implement a similar 3D SAXS method but with a different way of data analysis to gather information on the mineral alignment in fracture callus. With the proposed accurate yet fast assessment of 3D SAXS information, it was shown that the plate shaped mineral particles in the healing callus were aligned in groups with their predominant orientations occurring as a fiber texture.

  12. Mineral crystal alignment in mineralized fracture callus determined by 3D small-angle X-ray scattering

    International Nuclear Information System (INIS)

    Liu Yifei; Manjubala, Inderchand; Fratzl, Peter; Roschger, Paul; Schell, Hanna; Duda, Georg N

    2010-01-01

    Callus tissue formed during bone fracture healing is a mixture of different tissue types as revealed by histological analysis. But the structural characteristics of mineral crystals within the healing callus are not well known. Since two-dimensional (2D) scanning small-angle X-ray scattering (sSAXS) patterns showed that the size and orientation of callus crystals vary both spatially and temporally [1] and 2D electron microscopic analysis implies an anisotropic property of the callus morphology, the mineral crystals within the callus are also expected to vary in size and orientation in 3D. Three-dimensional small-angle X-ray scattering (3D SAXS), which combines 2D SAXS patterns collected at different angles of sample tilting, has been previously applied to investigate bone minerals in horse radius [2] and oim/oim mouse femur/tibia [3]. We implement a similar 3D SAXS method but with a different way of data analysis to gather information on the mineral alignment in fracture callus. With the proposed accurate yet fast assessment of 3D SAXS information, it was shown that the plate shaped mineral particles in the healing callus were aligned in groups with their predominant orientations occurring as a fiber texture.

  13. Influence of gamma radiation and fast neutrons on the growth of Haplopappus gracilis (Nutt) A. Gray callus

    International Nuclear Information System (INIS)

    Cebulska-Wasilewska, A.; Wajda, L.; Korzonek, M.; Polska Akademia Nauk, Krakow. Inst. Fizjologii Roslin)

    1979-01-01

    The sensitivity of the callus of Haplopappus gracilis to gamma radiation and fast neutrons was studied. High doses of radiation cause inhibition of callus growth. At small doses the effect is less pronounced. Stimulation of callus growth was seen. Apart from morphological changes, ionizing radiations lowered the fresh weight ratio of the callus. The RBE value for 5.5 MeV neutrons depended on the dose rate of radiation and the combination of growth medium. (author)

  14. Use of stem sliced from in vitro plants of papaya (hybrid IBP 42-99 by obtained callus with embryogenic structure

    Directory of Open Access Journals (Sweden)

    Jorge Gallardo Colina

    2004-10-01

    Full Text Available Inside in vitro propagation via, the somatic embriogenesis offers possibilities of obtaining top volumes of production in a minor period of time and a lower cost, which are a method potentially more efficient than the regeneration via organogenesis. In papaya the somatic embryogenesis could have developed from zigotic embryos and axis hipocotilos, nevertheless in case of the hybrids these methods cannot be used and it becomes necessary to develop it from a somatic fabric, without link with the sexual reproduction. This work chased as main objective Evaluated the use of in vitro plants stem sections of the Carica papaya IBP 42-99 hybrid for the formation of callus with embryogenic structures. As plant material were use in vitro plants of the papaya hybrid IBP 42-99. For it there took sections of different parts of the stem from the meristem up to the base of the in vitro plants, was use the culture medium Nitsh and Nitsh supplemented with 1.5 mg.l-1 of 6-BAP and 1.5 mg.l-1 of AIA. It was achieved to obtain callus from the stem sections with the culture medium used, nevertheless, in the treatments where used cylinders inside 1.0 cm from the apex down, the best results were achieved. The use of in vitro plants stem sections as explant for the formation of callus in this vegetable species it opens new possibilities for his in vitro propagation, specially in case of resultant hybrids of genetic improvement programs. Key words: apexes, Carica papaya, callogenesis, somatic embryogenesis

  15. Cold Response of Dedifferentiated Barley Cells at the Gene Expression, Hormone Composition, and Freezing Tolerance Levels: Studies on Callus Cultures

    Czech Academy of Sciences Publication Activity Database

    Vashegyi, I.; Marozsan-Toth, Z.; Galiba, G.; Dobrev, Petre; Vaňková, Radomíra; Toth, B.

    2013-01-01

    Roč. 54, č. 2 (2013), s. 337-349 ISSN 1073-6085 R&D Projects: GA ČR GA522/09/2058 Institutional research plan: CEZ:AV0Z50380511 Keywords : ABA * Barley * Callus Subject RIV: ED - Physiology Impact factor: 2.275, year: 2013

  16. Morphogenesis and tissue culture of sweet orange (Citrus sinensis (L.) Osb.): effect of temperature and photosynthetic radiation

    International Nuclear Information System (INIS)

    Duran-Vila, N.; Gogorcena, Y.; Ortega, V.; Ortiz, J.; Navarro, L.

    1992-01-01

    Both incubation temperature and photosynthetic radiation affected morphogenesis, callus culture and plantlet culture of sweet orange (Citrus sinensis) cultured in vitro. Bud culture from nodal stem segments, regeneration of shoots and buds from internode stem segments and induction of primary callus were near optimal at incubation temperatures between 21–30°C. The optimal temperature for root formation was 27°C with temperatures above and below being clearly deleterious. Incubation in the dark or under low photosynthetic photon flux density (PPFD) was beneficial for callus induction and growth and also favored the production of rooted plantlets from bud cultures. Incubation in the dark improved considerably the regeneration of shoots and buds from internode segments and the recovery of whole plants. No off-types, as determined by protein and isoenzyme analysis, were observed among plantlets recovered from bud cultures or from regeneration of shoots from internode stem segments

  17. Callus induction of leaf explant Piper betle L. Var Nigra with combination of plant growth regulators indole-3-acetic acid (IAA), benzyl amino purin (BAP) and kinetin

    Science.gov (United States)

    Junairiah, Zuraidassanaaz, Nabilah Istighfari; Izdihar, Fairuz Nabil; Manuhara, Yosephine Sri Wulan

    2017-09-01

    The purpose of this research was to determine the combination of plant growth regulators IAA, BAP and kinetin towards callus induction and growth of leaf explants Piper betle L. VarNigra. Explants from leaf of Piper betle L. VarNigra was cultured on MS medium with 24 treatment combinations of plant growth regulators IAA and BAP and 24 treatment combinations of plant growth regulators IAA and kinetin with 0.0;0.5;1.0;1.5;2.0 mg/L concentration respectively, the observed variable were the length of time the formation of callus, callus morphology, fresh and dry weight of callus. The results of this research showed that the combination of growth regulators IAA with BAP and kinetin had effects on leaf growth of Piper betle L. VarNigra. During 8 weeks observation, it indicated that the combination of concentration IAA 0.5 mg/L and BAP 2.0 mg/L showed fastest callus formation at 8.5 days. Combination of concentration IAA 1.0 mg/L and BAP 1.5 mg/L showed the highest of fresh weight at 0.6596 grams, and the highest dry weight was obtained from the combination of concentration IAA 0.5 mg/L and BAP 0.5 mg/L at 0.0727 grams. Combination of concentration IAA 1.0 mg/L and kinetin 1.5 mg/L had the highest of fresh weight at 0.2972 grams and the highest dry weight at 0.1660 grams. Callus of Piper betle L. VarNigra had two textures, that were compact and friable, and also showed various kind of colors, like white, greenish white, yellowish white, tanned white, brown and black. Based on this research, that concentration IAA 1.0 mg/L and 1.5 mg/L kinetin was the best combination for induction of callus from leaf of Piper betle L. Var Nigra.

  18. Effect of gamma rays in embryogenic callus of agave tequilana weber var. blue and in vitro selection for resistance to pathogens

    International Nuclear Information System (INIS)

    Rubluo, A.; Brunner, I.; Rodriguez-Garay, B.; Rodriguez-Dominguez, J.M.; Santacruz-Ruvalcava, F.; De la Cruz, E.; Gonzalez, J.

    1999-01-01

    Agave tequilana from which tequila is done, suffers genetic erosion leaving the plant susceptible to pathogens. Recently, a pest invades the tequilana fields. To face this problem we designed experiments to induce mutations for disease resistance. Our aims were: to analyse the in vitro responses of embryogenic callus exposed to gamma rays, to determine the LD50 and challenge surviving embryos to crude bacterial extracts (CBE). Axillary buds were cultured in vitro. Leaves from the former plant lets were used as explants to induce embryogenic callus. Samples of 1g embryogenic callus were irradiated from 5 to 40 Gy of CO 60 gamma rays. Effects were recorded at 15, 30 and 45 days after irradiation. The CBE was obtained from isolated bacteria collected from sick field plants. Embryogenic calli were irradiated at 0, 15, 25 and 40 Gy and challenged with the CBE at 0, 10 and 20 % v/v. Embryo expression (globular, white and green cucumber shaped) were affected as a function of the dose. However at low doses (5-15 Gy) globular forms increased its production. On the contrary , green cucumber shaped embryos were severely affected. However, even at high doses (30, 35 and 40 Gy) healthy green forms were recorded. When the embryogenic callus was challenged to the CBE, high mortality was recorded, although, some green forms survived at high radiation doses and high CBE concentrations. Because of the unicellular origin of the somatic embryos, solid mutants can be expected

  19. In vitro cultures of Salvia officinalis L. as a source of antioxidant compounds

    Directory of Open Access Journals (Sweden)

    Izabela Grzegorczyk

    2011-01-01

    Full Text Available The concentrations of carnosic acid, carnosol and rosmarinic acid in different materials from differentiated (multiple shoot cultures and regenerated plants and undifferentiated (callus and cell suspension in vitro cultures of Salvia officinalis were determined by HPLC. The results suggested that diterpenoid (carnosic acid and carnosol production is closely related to shoot differentiation. The highest diterpenoid yield (11.4 mg g-1 for carnosic acid and 1.1 mg g-1 for carnosol was achieved in shoots of 10-week-old micropropagated plants. The levels were comparable to those found in shoots of naturally growing plants. Undifferentiated callus and cell suspension cultures produced only very low amounts of carnosol (ca. 0.05 mg g-1 of dry weight. In contrast, content of rosmarinic acid in callus and suspension cultures as well as shoots growing in vitro and in vivo was similar and ranged between 11.2 and 18.6 mg g-1 of dry weight.

  20. Embryogenic competence acquisition in sugarcane callus is associated with differential H+ pump abundance and activity.

    Science.gov (United States)

    Passamani, Lucas Z; Bertolazi, Amanda A; Ramos, Alessandro C; Santa-Catarina, Claudete; Thelen, Jay J; Silveira, Vanildo

    2018-06-22

    Somatic embryogenesis is an important biological process in several plant species, including sugarcane. Proteomics approaches have shown that H + pumps are differentially regulated during somatic embryogenesis; however, the relationship between H + flux and embryogenic competence is still unclear. This work aimed to elucidate the association between extracellular H + flux and somatic embryo maturation in sugarcane. We performed a microsomal proteomics analysis and analyzed changes in extracellular H + flux and H + pump (P-H + -ATPase, V-H + -ATPase and H + -PPase) activity in embryogenic and non-embryogenic callus. A total of 657 proteins were identified, 16 of which were H + pumps. We observed that P-H + -ATPase and H + -PPase were more abundant in embryogenic callus. Compared with non-embryogenic callus, embryogenic callus showed higher H + influx, especially on maturation day 14, as well as higher H+ pump activity (mainly P-H+-ATPase and H+-PPase activity). H+-PPase appears to be the major H + pump in embryogenic callus during somatic embryo formation, functioning in both vacuole acidification and PPi homeostasis. These results provide evidence for an association between higher H + pump protein abundance and, consequently, higher H + flux and embryogenic competence acquisition in the callus of sugarcane, allowing for optimization of the somatic embryo conversion process by modulating the activities of these H + pumps.

  1. Hyperplastic callus formation in osteogenesis imperfecta: CT and MRI findings

    Energy Technology Data Exchange (ETDEWEB)

    Rieker, O.; Kreitner, K.F. [Klinik fuer Radiologie, Johannes-Gutenberg-Univ. Mainz (Germany); Karbowski, A. [Orthopaedische Abtl., Krankenhaus der Augustinerinnen, Koeln (Germany)

    1998-09-01

    Hyperplastic callus formation is a noteworthy condition in patients with osteogenesis imperfecta because it often mimicks osteosarcoma on radiography. The findings of CT and MRI in hyperplastic callus formation have not been reported. In the presented case, MRI demonstrated contrast enhancement and edema of the surrounding soft tisssue, consistent with benign as well as malignant disease. Computed tomography showed a calcified rim of the lesion which may be a useful feature to rule out osteosarcoma in this condition. (orig.) With 2 figs., 18 refs.

  2. Effect of Cold Stress on Fruiting Body Production by Medicinal Basidiomycetes in Submerged and Solid-phase Culture

    Directory of Open Access Journals (Sweden)

    E.P. Vetchinkina

    2017-05-01

    Full Text Available The ability of the medicinal xylotrophic basidiomycetes Lentinus edodes, Pleurotus ostreatus, Ganoderma lucidum and Grifola frondosa to produce typical and atypical fruiting bodies with viable basidiospores in submerged and solid-phase culture under stationary conditions in a beer wort-containing medium under cold stress was shown. The examined mushrooms, when not exposed to temperature stress, did not form fruiting bodies. In solid-phase culture in an agarized medium after cold treatment, the basidiome formation period was shortened by 1.5–2 times. Furthermore, the use of a mycelium subjected to temperature stress for inoculation induced and accelerated the formation of fruiting bodies on an industrial wood substrate, which is of great biotechnological importance.

  3. Optimization of callus induction and regeneration system for ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... 2Plant Biotechnology, NIGAB, National Agriculture Research Center Park Road Islamabad, Pakistan. Accepted 30 June ... transformation. MATERIALS AND METHODS ... Surface sterilization and callus induction. Seeds were ...

  4. Ziziphus mauritiana (masau) fruits fermentation in Zimbabwe: from black-box to starter culture development

    OpenAIRE

    Nyanga, L.K.

    2012-01-01

    This thesis reports on studies of microbiological and biochemical properties of masau (Ziziphus mauritiana) fruit fermentation and the development of starter cultures for the production of masau beverages. A survey to document the traditional processing techniques was conducted using a questionnaire and focus group discussions in each of the three districts, i.e., Mudzi, Mt Darwin and Muzarabani in Zimbabwe. The survey results showed that the masau fruit is usually gathered by women and chi...

  5. Cultural relevance of a fruit and vegetable food frequency questionnaire.

    Science.gov (United States)

    Paisley, Judy; Greenberg, Marlene; Haines, Jess

    2005-01-01

    Canada's multicultural population poses challenges for culturally competent nutrition research and practice. In this qualitative study, the cultural relevance of a widely used semi-quantitative fruit and vegetable food frequency questionnaire (FFQ) was examined among convenience samples of adults from Toronto's Cantonese-, Mandarin-, Portuguese-, and Vietnamese-speaking communities. Eighty-nine participants were recruited through community-based organizations, programs, and advertisements to participate in semi-structured interviews moderated in their native language. Data from the interviews were translated into English and transcribed for analysis using the constant comparative approach. Four main themes emerged from the analysis: the cultural relevance of the foods listed on the FFQ, words with multiple meanings, the need for culturally appropriate portion-size prompts, and the telephone survey as a Western concept. This research highlights the importance of investing resources to develop culturally relevant dietary assessment tools that ensure dietary assessment accuracy and, more important, reduce ethnocentric biases in food and nutrition research and practice. The transferability of findings must be established through further research.

  6. High efficiency protoplast isolation from in vitro cultures and hairy ...

    African Journals Online (AJOL)

    In vitro cultures of the medicinal plant Maesa lanceolata were established to enable the cultivation of plant material for the production of protoplasts. Callus cultures were initiated using leaves collected from shoot cultures and the root tips from hairy root cultures obtained upon Agrobacterium rhizogenes transformation.

  7. ( Linum usitatissimum L. cv. Modran cell suspension culture

    Directory of Open Access Journals (Sweden)

    Aleksandra Seta-Koselska

    2018-01-01

    Full Text Available Flax ( Linum usitatissimum L. is an ancient crop that is widely cultivated as a source of oil, fiber, and bioactive compounds. Flax fiber is traditionally used in textile industry, linseed oil is processed for industrial oils, paints, varnishes and bio-petroleum. Flaxseeds are also rich in α-linolenic acid and phytochemicals such as lignans. In addition to the commercial aspects, this species has been used widely and readily in biotechnological, developmental, and plant-pathogen interaction studies. Differences in the levels of endogenous hormones in various cultivars of flax significantly affected the intensity of callogenesis and determined the type and concentration of growth regulators necessary for callus production. The aim of our investigation was to optimize the culture conditions for callus formation and cell proliferation in liquid medium of the Polish cultivar of fiber flax – Modran. In the first step, 4 combinations of phytohormones in the medium were tested to obtain established callus tissue suitable for initiation of suspension culture. Next, we investigated the effect of chosen plant growth regulators on cell divisions, fresh and dry weight, and dispersal of callus cells in liquid medium. Fast growing and friable callus was obtained in a modified MS medium supplemented with 0.5 mg/l BAP and 0.1 mg/l NAA. We determined that for the initiation of cell suspension supplementation with 0.5 mg/l BAP and 0.5 mg/l NAA is optimal. The results obtained indicated that high concentration of cytokinin (BAP in liquid medium limited cell proliferation and decreased biomass formation.

  8. Myostatin (GDF-8) Deficiency Increases Fracture Callus Size, Sox-5 Expression, and Callus Bone Volume

    OpenAIRE

    Kellum, Ethan; Starr, Harlan; Arounleut, Phonepasong; Immel, David; Fulzele, Sadanand; Wenger, Karl; Hamrick, Mark W.

    2008-01-01

    Myostatin (GDF-8) is a negative regulator of skeletal muscle growth and mice lacking myostatin show increased muscle mass. We have previously shown that myostatin deficiency increases bone strength and biomineralization throughout the skeleton, and others have demonstrated that myostatin is expressed during the earliest phase of fracture repair. In order to determine the role of myostatin in fracture callus morphogenesis, we studied fracture healing in mice lacking myostatin. Adult wild-type ...

  9. Spatial and temporal variations of the callus mechanical properties during bone transport

    Energy Technology Data Exchange (ETDEWEB)

    Mora-Macias, J.; Reina-Romo, E.; Pajares, A.; Miranda, P.; Dominguez, J.

    2016-07-01

    Nanoindentation allows obtaining the elastic modulus and the hardness of materials point by point. This technique has been used to assess the mechanical propeties of the callus during fracture healing. However, as fas as the authors know, the evaluation of mechanical properties by this technique of the distraction and the docking-site calluses generated during bone transport have not been reported yet. Therefore, the aim of this work is using nanoindentation to assess the spatial and temporal variation of the elastic modulus of the woven bone generated during bone transport. Nanoindentation measurements were carried out using 6 samples from sheep sacrificed at different stages of the bone transport experiments. The results obtained show an important heterogeneity of the elastic modulus of the woven bone without spatial trends. In the case of temporal variation, a clear increase of the mean elastic modulus with time after surgery was observed (from 7±2GPa 35 days after surgery to 14±2GPa 525 days after surgery in the distraction callus and a similar increase in the docking site callus). Comparison with the evolution of the elastic modulus in the woven bone generated during fracture healing shows that mechanical properties increase slower in the case of the woven bone generated during bone transport. (Author)

  10. Palm fruit in traditional African food culture.

    Science.gov (United States)

    Atinmo, Tola; Bakre, Aishat Taiwo

    2003-01-01

    The centre of origin of the oil palm is the tropical rain forest region of West Africa. It is considered to be the 200-300 kilometre wide coastal belt between Liberia and Mayumbe. The oil palm tree has remained the 'tree of life' of Yoruba land as well as of other parts of southern West Africa to which it is indigenous. The Yoruba are adept at spinning philosophical and poetical proverbs around such ordinary things as hills, rivers, birds, animals and domestic tools. Hundreds of the traditional proverbs are still with us, and through them one can see the picture of the environment that contributed to the moulding of the thoughts of the people. Yoruba riddles or puzzles were also couched in terms of the environment and the solutions to them were also environmental items. They have a popular saying: A je eran je eran a kan egungun, a je egungun je egungun a tun kan eran: 'A piece of meat has an outer layer of flesh, an intermediate layer of bone and an inner layer of flesh'. What is it? A palm fruit: it has an outer edible layer, the mesocarp; then a layer of shell, inedible, and the kernel inside, edible. The solution to this puzzle summarises the botanical and cultural characteristics of the palm fruit.

  11. Strontium Is Incorporated into the Fracture Callus but Does Not Influence the Mechanical Strength of Healing Rat Fractures

    DEFF Research Database (Denmark)

    Brüel, Annemarie; Olsen, Jakob; Birkedal, Henrik

    2011-01-01

    in callus bone mineral content (P\\0.05). However, after 8 weeks of healing, no difference was found in either callus volume or bone mineral content. SrR did not influence maximum load or stiffness of the fractures after either 3 or 8 weeks of healing. EDX showed that Sr was incorporated into the callus...

  12. Callus Distraction Versus Single-Stage Lengthening With Bone Graft for Treatment of Brachymetatarsia: A Systematic Review.

    Science.gov (United States)

    Jones, Marc D; Pinegar, David M; Rincker, Sarah A

    2015-01-01

    Brachymetatarsia deformity is a cosmetically displeasing anomaly that can become physically symptomatic. The surgical techniques most commonly used to repair the anomaly include single-stage lengthening with a bone graft, callus distraction, or a combination of bone grafting and callus distraction. A systematic review of the published data was performed to compare the outcomes of these 3 surgical procedures. A total of 61 studies reporting the use of callus distraction or single-stage lengthening, or both, for the treatment of brachymetatarsia were included in the present review. The incidence of major postoperative complications after callus distraction, single-stage lengthening, and the combination procedure was 49 (12.62%), 13 (3.72%), and 3 (33.33%), respectively. The number of minor complications with callus distraction, single-stage lengthening, and the combination procedure was 152 (39.18%), 55 (15.76%), and 1 (11.11%); the mean percentage of the original length achieved was 37.36%, 25.98% and 36.00%; and the mean length achieved was 17.5, 13.2, and 14.0 mm, respectively. The healing index (mo/cm) and healing time was 2.31 and 16.04 weeks, 1.90 and 9.35 weeks, and 3.93 and 14.62 weeks for callus distraction, single-stage lengthening, and the combination procedure, respectively. Our findings indicate that the callus distraction technique is associated with greater length gained but results in greater complication rates and requires almost twice the time to heal. Single-stage lengthening with a bone graft was associated with fewer complications and faster healing times than callus distraction but with lesser gains in length. From the information reported in the studies we reviewed, the prevalence of bilateral brachymetatarsia was 44.52%, and the female/male ratio was 13.7:1. Both of these findings seem to contradict the usual data given (72% for bilateral brachymetatarsia and a female/male ratio of 25:1). Copyright © 2015 American College of Foot and

  13. An efficient method for the establishment of cell suspension cultures in potato (Solanum tuberosum L.)

    International Nuclear Information System (INIS)

    Sajid, Z.A.

    2016-01-01

    Cell suspension cultures offers an In vitro system that can be used as a tool for various studies involving mutant selection, mass propagation, protoplast isolation, gene transfer and selection of cell-lines which are resistant to various biotic or abiotic stresses. Research work on the development of cell suspension cultures was carried out to establish the most efficient method in Potato (cv. Desiree). Healthy, well-proliferating tissues from different types of callus cultures (compact, friable, embryogenic or non-embryogenic) were inoculated on various media combinations, i.e., MS, MS2 or AA liquid medium containing 18.09 micro M 2, 4-D. A fixed quantity (0.5-1.0 g) of callus tissue from 60-day-old callus cultures was transferred to 10-25 ml of liquid medium in 100 ml Erlenmeyer flask. Cultures were placed on an orbital shaker and agitated at different speeds (75, 100 or 125 rpm) under 16-h photoperiod at 25 ± 2 degree C. Medium was changed after every 3 days and fractionated tissue was filtered after every 6 days through sterile mesh (100-800 micro m) to develop a cell-line by transferring resulting suspension to fresh medium under the same conditions. Results indicated that eight-week-old translucent, friable, off-white callus cultures were an excellent starting material for the initiation of homogeneous cell suspension cultures as compared to other tested sources. Of the three tested media (MS, MS2 or AA medium containing 18.09 micro M 2, 4-D), MS2 was found to be a better medium for the initiation of cell suspension cultures. Cell suspension cultures, placed in 16-h photoperiod at 25 ± 2 degree C and agitated at 120 rpm using a gyratory shaker showed excellent results. Several other factors influencing quick establishment of cell suspension cultures in this cultivar are also discussed in this communication. (author)

  14. Production of friable embryogenic callus and regeneration of ...

    African Journals Online (AJOL)

    sunny t

    2015-06-03

    Jun 3, 2015 ... Generation of embryogenic callus is a key step in genetic engineering of many crop species, including cassava. ... CBSD is caused by two Ipomovirus; Cassava .... through a 1 mm2 pore-sized steel wire mesh using a spatula.

  15. Picroside I and Picroside II from Tissue Cultures of Picrorhiza kurroa

    Science.gov (United States)

    Ganeshkumar, Yamjala; Ramarao, Ajmera; Veeresham, Ciddi

    2017-01-01

    Background: Picrorhiza kurroa (PK) belongs to Scrophulariaceae family and is a representative endemic, medicinal herb, widely distributed throughout the higher altitudes of alpine Himalayas from west to east, between 3000 and 4500 m above mean sea level. Objective: The objective of the present study is to assess the production of picroside I and picroside II from tissue cultures of PK. Materials and Methods: Auxiliary shoot tips of PK were incubated in Murashige and Skoog medium supplemented with indole-3-butyric acid and kinetin phytohormones. The callus produced was collected at different time intervals and was processed for extraction of picroside I and picroside II followed by thin layer chromatography and high-performance liquid chromatography HPLC analysis. Results: The maximum growth index was found to be 5.109 ± 0.159 at 16-week-old callus culture. The estimation of picroside-I and picroside-II was carried out by (HPLC) analysis; quantity of secondary metabolite found to be 16.37 ± 0.0007 mg/g for PK-I and 6.34 ± 0.0012 mg/g for PK-II. Conclusion: This is the first attempt to produce the Picroside-I and II in large amount by the tissue culture technique. It can be observed that the method of callus culture can be used in production of secondary metabolites Picroside-I and II from PK SUMMARY Picrorhiza kurroa is a high value medicinal herb due to rich source of hepatoprotective metabolites, Picroside-I and Picroside-II. The medicinal importance of P. kurroa is due to its pharmacological properties like hepatoprotective, antioxidant (particularly in liver), antiallergic and antiasthamatic, anticancer activity particularly in liver and immunomodulatory. Shoot apices which were produced a good response was inoculated on selected medium i.e., on MS medium containing 2, 4 D (mg/l) + KN (1mg/l) for induction of callus. The initiation of callus was observed after 4weeks and it was light green and fragile Maximum growth was observed with 3% w/v of sucrose

  16. Comparison of auxin activty in tumourous and normal callus cultures from sunflower and tobacco plants

    Directory of Open Access Journals (Sweden)

    Z. Chirek

    2015-01-01

    Full Text Available In normal and tumourous calluses of sunflower and tobacco the level of extractable auxins was determined by Avena coleoptile straight growth test. Auxin activity was detected practically in two zones: I - at position with Rf 0.2-0.4 and II - at position with Rf 0.6-0.9. The tumour tissues of sunflower and tobacco plants, representing different types of neoplastic growth exhibit a 3 times higher auxin activity as compared with that of the corresponding normal tissues. Tobacco tissues, on the other hand, had a higher auxin level than the corresponding sunflower tissues and they exhibited different proportions in the activity of zones I and II, which points to a dominance of genetic regulation of hormone metabolism in these plants.

  17. Responses of meristematic callus cells of two Cynodon dactylon genotypes to aluminium.

    Science.gov (United States)

    Ramgareeb, Sumita; Cooke, John A; Watt, M Paula

    2004-11-01

    Responses to Al3+ of embryogenic callus cells of an Al-sensitive (Al-S) and Al-resistant (Al-R) Cynodon dactylon genotype were evaluated with regard to Al3+ toxicity and resistance. A chemical equilibrium speciation model (MINTEQA2) was used to ensure the availability of the Al3+ ion in culture media, which was supplied as 0.08-2.3 mM Al3+ for 2-8 weeks. Increasing Al3+ concentration and exposure time had a greater negative impact on the Al-S than on the Al-R genotype, in terms of callus growth rate and frequency of non-embryogenic cells. Exposure to 0.8 mM Al3+ for 2 weeks resulted in an 88% reduction in the Al-S meristematic cell number, whereas that of the Al-R genotype remained unaffected. In addition, the Al-S cells accumulated three times more Al in the nucleus than did the Al-R cells, suggesting that Al interfered with mitosis. The Al-R cells appeared to exclude Al3+ from its cells through an increase in extracellular pH (4.34 in Al-R and 4.08 in Al-S) and by the immobilisation of Al in the cell wall (33% more in Al-R). The results showed that by studying the cellular responses to Al3+ it is possible to discriminate between the Al-S and Al-R C. dactylon genotypes.

  18. Hyperplastic callus formation in osteogenesis imperfecta. A case report

    International Nuclear Information System (INIS)

    Burchardt, A.J.; Wagner, A.A.; Basse, P.

    1994-01-01

    We report a case of bilateral hyperplastic callus formation as a complication of fracture in a patient with osteogenesis imperfecta. The clinical and radiographic findings and the differential diagnosis are discussed. (orig.)

  19. Hyperplastic callus formation in osteogenesis imperfecta. A case report

    Energy Technology Data Exchange (ETDEWEB)

    Burchardt, A.J. (Depts. of Radiology and Pediatric Orthopedic Surgery, Rigshospitalet, Copenhagen Univ. (Denmark)); Wagner, A.A. (Depts. of Radiology and Pediatric Orthopedic Surgery, Rigshospitalet, Copenhagen Univ. (Denmark)); Basse, P. (Depts. of Radiology and Pediatric Orthopedic Surgery, Rigshospitalet, Copenhagen Univ. (Denmark))

    1994-09-01

    We report a case of bilateral hyperplastic callus formation as a complication of fracture in a patient with osteogenesis imperfecta. The clinical and radiographic findings and the differential diagnosis are discussed. (orig.).

  20. Optimization of embryogenic-callus induction and embryogenesis of ...

    African Journals Online (AJOL)

    Optimization of embryogenic-callus induction and embryogenesis of Glycyrrhiza glabra. Fu Chunhua, Cheng Lei, Lu Gan, Maoteng Li, Ying Yang, Longjiang Yu. Abstract. Glabridin is a major biologically active flavonoid isolated specifically from the root of Glycyrrhiza glabra, which has many pharmacological activities.

  1. Production of secondary metabolites trimethyl xanthina by Camellia sinensis L suspension culture

    Science.gov (United States)

    Sutini, Sodiq, Mochamad; Muslihatin, Wirdhatul; Indra, Mochamad Rasjad

    2017-06-01

    Bioactive trimethyl xanthina can be obtained from the plant Camellia sinensis L. To obtain bioactive plant of which there are several hurdles for instance to wait up to five years to be harvested, also it needs land at a certain height from the sea level. Therefore, the production of secondary metabolites trimethyl xanthina need to be developed with suspense culture techniques. The purpose of this study obtained the production of bioactive trimethyl xanthina way culturally suspense in large scale with a relatively short time, potentially as anti-oxidants. Research methods include: (1) initiation of callus from pieces of leaves, shoots the youngest of the plant Camellia sinensis L in the media MS with the optimization of the addition of growth regulators, (2) the subculture of callus on media and plant growth regulator that is equal to the stage of initiation, (3) initiation of suspension culture using explants of callus Camellia sinensis L, (4) Analysis of secondary metabolites trimethyl xanthina growth in suspension culture, (5) the isolation and identification of trimethyl xanthina qualitatively and quantitatively using thin layer chromatography/high performance chromatography column. The results of the study suspension cultures containing bioactive trimethyl xanthina candidates that can be used as an antioxidant.

  2. Axenic Seed Culture and in vitro mass propagation of Malaysian Wild Orchid Cymbidium finlaysonianum LINDL

    International Nuclear Information System (INIS)

    Islam, T.; Bhattacharjee, B.; Islam, S. M. S.; Uddain, J.; Subramaniam, S.

    2015-01-01

    Under this study an efficient protocol on mass propagation of Cymbidium finlaysonianum an epiphytic Malaysian wild orchid has been established using axenic culture. To obtain an axenic seed culture, it is important to perform an adequate a disinfection procedure in tissue culture. Four nutrient media viz. MS, 0.5MS, KC and VW were evaluated on In vitro seed germination with callus initiation. The maximum seed germination with callus initiation (100 percentage) was recorded in MS basal medium with a short span of time (40 days after culture). After 45 days of callus initiation the effect of eight different treatments (T /sub 1/-T /sub 8/) on callus size and nature were also studied. The experiment revealed that in T /sub 3/ (MS + 2.0 mgl /sup -1/ BAP + 0.5 mg /sup -1/ NAA) was found to be the best for callus development (1.98 cm length and 1.01 cm breadth). The effect of different concentration of BAP was evaluated on protocorm formation and its proliferation. Maximum number (7.75) and percentage (81.40) of PLBs was recorded in MS medium supplemented with 1.5 mg-1 BAP. Very good PLBs development was recorded also in MS + BAP 1.0 mg-1 + NAA 0.5 mg /sup -1/. The highest elongation of shoot (3.80 cm) was observed in MS + 1.0 mg-1 BAP + 0.50 mg /sup -1/ NAA. For root induction 1.0 mg-1 NAA has proven to the best in 0.5 MS medium. The In developed seedlings were finally transferred to pots by successive phases of acclimatization. (author)

  3. Investigations on some metabolites of Tecoma stans Juss. callus tissue. Part III. Chromatographical search for iridoids, phenolic acids, terpenoids and sugars

    Directory of Open Access Journals (Sweden)

    Barbara Dohnal

    2015-01-01

    Full Text Available Tissus cultures of Tecoma stans Juss. cultivated on modified Murashige-Skoog medium (RT-k were phytochemically analysed by means of chromatographical methods (PC, TLC. The following products were found as metabolites: phenolic acids - chlorogenics, caffeic, ferulic, vanillic, o-coumaric and sinapic; steroids - β-sitosterol; triterpenes - ursolic and oleanolic acids, α-amyrine; sugars - glucose, fructose, sucrose, xylose. Meso-inositol was isolated in 0.8% yield. In intact plant leaves, some differences concerning the content and/or number of individual compounds were observed, namely: lack of sinapic acid and occurrence of p-coumaric acid, lower content of β-sitosterol, lack of oleanolic acid, occurrence of β-amyrine and of one unidentified triterpenoid, lack of xylose, occurrence of maltose, raffinose, and stachiose. The level of mesoinositol inn leaves was distincly lower than in the callus tissues. Neither in callus tissues nor in leaves iridoid glycosides were found.

  4. Nano-copper-bearing stainless steel promotes fracture healing by accelerating the callus evolution process

    Directory of Open Access Journals (Sweden)

    Wang L

    2017-11-01

    Full Text Available Lei Wang,1,* Guoyuan Li,1,* Ling Ren,2,* Xiangdong Kong,1 Yugang Wang,1 Xiuguo Han,1 Wenbo Jiang,3 Kerong Dai,1 Ke Yang,2 Yongqiang Hao11Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, 2Special Materials and Device Research Department, Institute of Metal Research, Chinese Academy of Sciences, Shenyang, 3Medical 3D Printing Innovation Research Center, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: Treatment for fractures requires internal fixation devices, which are mainly produced from stainless steel or titanium alloy without biological functions. Therefore, we developed a novel nano-copper-bearing stainless steel with nano-sized copper-precipitation (317L-Cu SS. Based on previous studies, this work explores the effect of 317L-Cu SS on fracture healing; that is, proliferation, osteogenic differentiation, osteogenesis-related gene expression, and lysyl oxidase activity of human bone mesenchymal stem cells were detected in vitro. Sprague–Dawley rats were used to build an animal fracture model, and fracture healing and callus evolution were investigated by radiology (X-ray and micro-CT, histology (H&E, Masson, and safranin O/fast green staining, and histomorphometry. Further, the Cu2+ content and Runx2 level in the callus were determined, and local mechanical test of the fracture was performed to assess the healing quality. Our results revealed that 317L-Cu SS did not affect the proliferation of human bone mesenchymal stem cells, but promoted osteogenic differentiation and the expression of osteogenesis-related genes. In addition, 317L-Cu SS upregulated the lysyl oxidase activity. The X-ray and micro-CT results showed that the callus evolution efficiency and fracture healing speed were

  5. Callus formation in bone fractures combined with brain injury in rat

    Directory of Open Access Journals (Sweden)

    Yu-Ping Chen

    2017-01-01

    Full Text Available Objective: The objective of this study was to determine the speed of bony union and the serum levels of biomarkers in the setting of bone fractures combined with brain injury. Materials and Methods: In this study, Sprague–Dawley rats were randomized into four groups: sham, brain injury, bone fracture, and bone fracture plus brain injury groups. The serum levels of biochemical markers, namely, nerve growth factor (NGF, Wnt-3a, Dickkopf-related protein-1, receptor-activator of NF-κB ligand, and adrenocorticotropic hormone (ACTH, were measured on the days 1, 3, 7, and 14 following injury. Bony union was evaluated using radiographs every week for 6 weeks. Results: Compared with the brain injury group and bone fracture group, the radiographs of the bone fracture plus brain injury group revealed enhanced callus formations in week 2. From week 3, the callus formation did not differ significantly among the groups. The serum levels of the biomarkers varied at different time points. The serum levels of NGF on days 1 and 3, Wnt-3a on days 3 and 14, and ACTH on days 1, 3, and 7 were significantly higher in the bone fracture plus brain injury group than in the bone fracture group. Conclusions: Brain injury increases callus formation in simultaneous bone fracture. Considering the time point, early NGF, Wnt-3a, and ACTH elevation might be associated with early callus formation enhancement. The results indicate that these brain injury-induced biomarkers might play crucial role in accelerating bone healing.

  6. Polyamine patterns in haploid and diploid tobacco tissues and in vitro cultures

    Directory of Open Access Journals (Sweden)

    Sílvia Bicudo Carone

    2010-04-01

    Full Text Available The aim of this work was to determine PAs levels in pith tissues and callus cultures from haploid and diploid tobacco plants, explanted from the apical and basal regions of the stem. These explants were cultured in an RM-64 medium supplied with IAA and kinetin, under light or in the dark, during successive subcultures. PAs levels followed a basipetal decrease in diploid and an increase in haploid, pith tissues. A similar pattern of total PAs (free + conjugated was observed for the callus of diploid and haploid plants maintained in the light, and for the haploid callus in the dark, whereas the diploid callus in the dark showed a constant increase in total PAs levels until the end of culture. The PA increase in the diploid callus in the dark was related to free Put levels increase. The ploidy status of the plants could express different PA gradients together with the plant pith and in vitro callus cultures.O objetivo deste trabalho foi determinar os níveis de PAs em tecidos de medula e cultura de calos de plantas haplóides e diplóides de tabaco, obtidas da região apical e basal do caule. Estes explantes foram cultivados em meio RM-64 suplementado com AIA e cinetina, na luz e no escuro, durante vários subcultivos. Nos tecidos medulares, os níveis de PAs apresentam um decréscimo basípeto em diplóides e um aumento em haplóides.Um padrão similar nos níveis de PAs totais (livres+ conjugadas foi observado em calos haplóides e diplóides mantidos na luz, e haplóides no escuro, enquanto os diplóides cultivados no escuro mostraram um aumento constante até o final do cultivo. O aumento no conteúdo de PAs nos calos diplóides no escuro, foi devido ao aumento do conteúdo de Put livre. Foi observado que a ploidia da planta pode expressar diferentes gradientes de PA ao longo do tecido medular e nas culturas de calos in vitro.

  7. Gamma-irradiation activates biochemical systems: induction of nitrate reductase activity in plant callus.

    OpenAIRE

    Pandey, K N; Sabharwal, P S

    1982-01-01

    Gamma-irradiation induced high levels of nitrate reductase activity (NADH:nitrate oxidoreductase, EC 1.6.6.1) in callus of Haworthia mirabilis Haworth. Subcultures of gamma-irradiated tissues showed autonomous growth on minimal medium. We were able to mimic the effects of gamma-irradiation by inducing nitrate reductase activity in unirradiated callus with exogenous auxin and kinetin. These results revealed that induction of nitrate reductase activity by gamma-irradiation is mediated through i...

  8. Callus induction via different growth regulators from cotyledon ...

    African Journals Online (AJOL)

    Cicer arietinum L.) cultivars KK-1 and Hassan-2K on MS and B5 media containing different combinations and concentrations of growth regulators. Different MS and B5 callusing media containing varying level of 2, 4-D (2 and 4 mg/l), NAA (0.50 ...

  9. Ziziphus mauritiana (masau) fruits fermentation in Zimbabwe: from black-box to starter culture development

    NARCIS (Netherlands)

    Nyanga, L.K.

    2012-01-01

    This thesis reports on studies of microbiological and biochemical properties of masau (Ziziphus mauritiana) fruit fermentation and the development of starter cultures for the production of masau beverages.

    A survey to document the traditional processing

  10. Availability of commonly consumed and culturally specific fruits and vegetables in African-American and Latino neighborhoods

    OpenAIRE

    Grigsby-Toussaint, Diana S.; Zenk, Shannon N.; Odoms-Young, Angela; Ruggiero, Laurie; Moise, Imelda

    2010-01-01

    Although the importance of culture in shaping individual dietary behaviors is well documented, cultural food preferences have received limited attention in research on the neighborhood food environment. The purpose of this study was to assess the availability of commonly consumed and culturally specific fruits and vegetables in retail food stores located in majority African-American and Latino neighborhoods in southwest Chicago. A cross-sectional survey of 115 stores (15% grocery stores, 85% ...

  11. [Factors affecting bone regeneration in Ilizarov callus distraction].

    Science.gov (United States)

    Fink, B; Krieger, M; Schneider, T; Menkhaus, S; Fischer, J; Rüther, W

    1995-12-01

    We evaluated the X-rays of 36 patients who underwent 50 callus distractions. With the aid of a computerized digitalisation system for analogue films, the relative X-ray density of the distraction area was calculated for each X-ray. These relative X-ray densities were figured graphically for the duration of treatment for each patient. In the consolidation phase, the graph of each patient had a logarithmic relationship. The gradients of the logarithmic density curves were considered an indicator of the quantity of new bone formation. These gradients were correlated to the following clinical parameters: age of the patient, beginning of distraction after corticotomy, average speed of distraction, average weight bearing during the distraction and consolidation phase, location of corticotomy (distal femur versus proximal tibia) and diclofenac medication. Except for the location of the corticotomy and diclofenac, all parameters had an influence on osteoneogenesis by callus distraction. The parameters affecting new bone formation the most were the age of the patient and weight bearing. Patients aged under 18 years (p = 0.005), beginning of distraction later than 8 days (p = 0.109), an average distraction speed below 1 mm/day (p = 0.079), and average weight bearing of more than 30 kg (p = 0.068 for the distraction phase and p = 0.089 for the consolidation phase) showed a quantitatively higher rate of new bone formation by callus distraction than the patients in the other groups. Patients with a shorter leg due to poliomyelitis and one patient with an amniotic leg tie showed a slower increase in X-ray density graphs than the other patients.

  12. The Role of Ultrasound Imaging of Callus Formation in the Treatment of Long Bone Fractures in Children

    International Nuclear Information System (INIS)

    Wawrzyk, Magdalena; Sokal, Jan; Andrzejewska, Ewa; Przewratil, Przemysław

    2015-01-01

    In the process of diagnosis and treatment of fractures, an X-ray study is typically performed. In modern medicine very important is the development of new diagnostic methods without adverse effects on the body. One of such techniques is ultrasound imaging. It has a high value in imaging most areas of the body, including the musculoskeletal system. Reports on the use of ultrasound in the evaluation of the callus are rare and this could be a method equivalent to or even better than standard radiographs. The aim of the study was to analyze the correlation of ultrasound with radiographs in imaging of callus formation after fractures of long bones in children and to analyze the correlation of vascular resistance index (RI) and the degree of vascularization of the callus with a subjective radiological assessment of the bone union quality. The prospective study was planned to qualify 50 children treated for long bones fractures of the arm, forearm, thigh and lower leg. Ultrasound diagnosis was carried out using a Philips iU22 camera equipped with a linear probe with 17-5-MHz resolution and MSK Superficial program. During ultrasound examination measurements of the callus were performed. Using the Power Doppler callus vascularity was visualized and vascular resistance index (RI) was measured. The same measurements were made within the corresponding area of the healthy limb. The results obtained by ultrasound were compared with radiograph measurements and with the subjective assessment of the callus quality. Preliminary results were developed on a group of 24 patients, where 28 fractured bones and 28 corresponding healthy bones were examined. Fifteen boys and 9 girls participated in the study. The average age at injury was, respectively, 11 and 9 years. In both groups fractures without displacement were the most frequent. A similar frequency was observed in fractures requiring reposition and subperiosteal fractures. In contrast, fractures with a slight displacement of the

  13. The Role of Ultrasound Imaging of Callus Formation in the Treatment of Long Bone Fractures in Children.

    Science.gov (United States)

    Wawrzyk, Magdalena; Sokal, Jan; Andrzejewska, Ewa; Przewratil, Przemysław

    2015-01-01

    In the process of diagnosis and treatment of fractures, an X-ray study is typically performed. In modern medicine very important is the development of new diagnostic methods without adverse effects on the body. One of such techniques is ultrasound imaging. It has a high value in imaging most areas of the body, including the musculoskeletal system. Reports on the use of ultrasound in the evaluation of the callus are rare and this could be a method equivalent to or even better than standard radiographs. The aim of the study was to analyze the correlation of ultrasound with radiographs in imaging of callus formation after fractures of long bones in children and to analyze the correlation of vascular resistance index (RI) and the degree of vascularization of the callus with a subjective radiological assessment of the bone union quality. The prospective study was planned to qualify 50 children treated for long bones fractures of the arm, forearm, thigh and lower leg. Ultrasound diagnosis was carried out using a Philips iU22 camera equipped with a linear probe with 17-5-MHz resolution and MSK Superficial program. During ultrasound examination measurements of the callus were performed. Using the Power Doppler callus vascularity was visualized and vascular resistance index (RI) was measured. The same measurements were made within the corresponding area of the healthy limb. The results obtained by ultrasound were compared with radiograph measurements and with the subjective assessment of the callus quality. Preliminary results were developed on a group of 24 patients, where 28 fractured bones and 28 corresponding healthy bones were examined. Fifteen boys and 9 girls participated in the study. The average age at injury was, respectively, 11 and 9 years. In both groups fractures without displacement were the most frequent. A similar frequency was observed in fractures requiring reposition and subperiosteal fractures. In contrast, fractures with a slight displacement of the

  14. DNA MUTAGENESIS IN PANAX GINSENG CELL CULTURES

    Directory of Open Access Journals (Sweden)

    Kiselev K.V.

    2012-08-01

    Full Text Available At the present time, it is well documented that plant tissue culture induces a number of mutations and chromosome rearrangements termed “somaclonal variations”. However, little is known about the nature and the molecular mechanisms of the tissue culture-induced mutagenesis and the effects of long-term subculturing on the rate and specific features of the mutagenesis. The aim of the present study was to investigate and compare DNA mutagenesis in different genes of Panax ginseng callus cultures of different age. It has previously been shown that the nucleotide sequences of the Agrobacterium rhizogenes rolC locus and the selective marker nptII developed mutations during long-term cultivation of transgenic cell cultures of P. ginseng. In the present work, we analyzed nucleotide sequences of selected plant gene families in a 2-year-old and 20-year-old P. ginseng 1c cell culture and in leaves of cultivated P. ginseng plants. We analysed sequence variability between the Actin genes, which are a family of house-keeping genes; the phenylalanine ammonia-lyase (PAL and dammarenediol synthase (DDS genes, which actively participate in the biosynthesis of ginsenosides; and the somatic embryogenesis receptor kinase (SERK genes, which control plant development. The frequency of point mutations in the Actin, PAL, DDS, and SERK genes in the 2-year-old callus culture was markedly higher than that in cultivated plants but lower than that in the 20-year-old callus culture of P. ginseng. Most of the mutations in the 2- and 20-year-old P. ginseng calli were A↔G and T↔C transitions. The number of nonsynonymous mutations was higher in the 2- and 20-year-old callus cultures than the number of nonsynonymous mutations in the cultivated plants of P. ginseng. Interestingly, the total number of N→G or N→C substitutions in the analyzed genes was 1.6 times higher than the total number of N→A or N→T substitutions. Using methylation-sensitive DNA fragmentation

  15. Optimization of callus induction and plant regeneration from ...

    African Journals Online (AJOL)

    An efficient regeneration system was developed using germinating seeds of two cultivars of sweet sorghum, Sorghum bicolor 'Yuantian No.1' and 'M81E', as explants. We tested different media supplements effects on callus induction. The effects of combinations of 2,4-D, KT, sucrose, agar and proline at different ...

  16. Callus induction via different growth regulators from cotyledon ...

    African Journals Online (AJOL)

    Jane

    2011-06-17

    Jun 17, 2011 ... NAA, naphthylacetic acid α; BAP, benzyl aminopurine; µM, micro mole. Chickpeas are a good source of zinc, folate and are very high in dietary fiber and thus, .... auxin, the callus inductions rates increased and induction time decreased ..... clonal variation and giving birth to traits of agronomic importance.

  17. Determination of fruit characteristics, fatty acid profile and total antioxidant capacity of Mespilus germanica L. fruit

    Directory of Open Access Journals (Sweden)

    Hale Seçilmiş Canbay

    2015-11-01

    Full Text Available Objective: To determine fruit characteristics, fatty acid profile and total antioxidant capacitiy of first cultured Mespilus germanica L. Methods: A total of 15 fruits were taken randomly from four directions of adult trees. Then the physical and chemical properties of first cultured medlar fruit (Istanbul/Turkey were measured by using refractometer, colorimeter, spectrophotometer and gas chromatograph mass spectrometer, respectivly. Results: In the fruit studied, the results showed that palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid and behenic acid were the most abundant fatty acids (FAs, and the main FA was palmitic acid [(35.35 ± 1.20%]. The percentage of linoleic acid and stearic acid in this fruit oil were (29.10 ± 1.70% and (8.53 ± 0.25%, respectively. As a result of the analysis, the total antioxidant capacity of medlar fruit was (1.1 ± 0.2 mmol trolox equivalents/L. Conclusions: The present study has demonstrated the concentrations of FAs and the antioxidantive capacity of first cultured Istanbul medlar fruits by using many tested methods. It is proved that in our daily life, medlar fruit plays a significant role with its nutrition and health effect.

  18. Effect of gamma irradiation on Callus formation and regeneration of wheat immature embryos

    International Nuclear Information System (INIS)

    Saleh, O.M.

    2007-01-01

    Four Egyptian bread wheat cultivars; G164, G168, SK61 and Sids1, were tested for their response to six gamma irradiation treatments; 1, 2, 3 Gy (as low doses) and 10, 20, 30 Gy (as high doses) in addition to 0 Gy (as a control) in terms of callus formation and regeneration of immature embryos. Low doses of gamma irradiations ( 1, 2 and 3 Gy) showed favourable effects on both traits; number of regenerated calli and number of shoots per callus comparing with the control (0 Gy), while high doses; 10, 20 and 30 Gy, had the worst effect comparing with the control (0 Gy). G164 cultivar was shown to get the best response in terms of callus formation and regeneration when exposed to low doses of gamma irradiation. In conclusion, gamma irradiation can serve in increasing regeneration efficiency of Egyptian bread wheat cells when used in low doses

  19. Response of different genotypes of wheat, rice and black beans to anther, embryo and other tissue cultures

    International Nuclear Information System (INIS)

    Franco, E.; Amador, D.; Calderon, J.; Alvarez, G.; Alvarado, J.; Ramazzini, H.; Ramos, S.; Acuna, G.; Zuniga, B.

    1996-01-01

    The objective of the basic studies we have been conducting in our laboratory is to establish callus induction and in vitro plant regeneration protocols starting with several tissues of Guatemalan varieties of wheat (Triticum aesticum L.), rice (Oryza sativa L.) and especially black bean (Phaseolus vulgaris L.) in order to obtain disease resistance, earliness, and dwarf plants. Wheat anthers and immature embryos of varieties Patzun, Comalapa, Chocoyo, and Xequijel cultured in N 6 , Potato II, and MS basal media supplemented with auxin and cytokinin gave the best responses in callus induction and plant regeneration. Anthers and mature embryos of indica rice varieties Precozicta and Virginai, when cultured in MS, B 5 , N 6 , and Potato II basal media with different hormonal combinations gave a good response in callus induction. However, a satisfactory response in plant regeneration was not obtained. With black beans, when hypocotyls and mature embryos of black bean varieties Quinack Che and Parramos were cultured in MS basal medium supplemented with different concentrations of NAA and kinetin, more than 60% callus induction was produced. When Quinack Che calli were transferred to MS basal medium supplemented with 1 mg/l NAA plus 0.5 mg/l BAP, green points of regeneration were visible in these calli. (author). 34 refs, 28 tabs

  20. Do Bananas Have a Culture? United Fruit Company Colonies in Central America 1900-1960

    Directory of Open Access Journals (Sweden)

    Atalia Shragai

    2014-06-01

    Full Text Available This article is concerned with the processes underlying the development of the unique identifications and culture which evolved among the First Class Workers of the United Fruit Company - the vast majority of whom were citizens of the United States, working alongside Europeans and Central Americans - during the first half of the twentieth century. Examining the social and cultural practices widespread among the Company’s colonies, I trace the nature of the ‘Banana Culture’, a term coined by the members of this group.

  1. Improved embryogenic callus induction and plant regeneration in ...

    African Journals Online (AJOL)

    Pramod Pantha

    2016-09-28

    Sep 28, 2016 ... solidified with 4.0 g l-1. Gelzan (Phyto Technology Laboratory). Pantha et al. 2167. Callus induction was performed under dark at 28°C (Isotemp. Incubator, Fisher Scientific, Hanover Park, IL, USA). Primary shoots from seed were removed 10 days after inoculation and subculture was done 3 weeks after ...

  2. [Dynamics of genome changes in Rauwolfia serpentina callus tissue upon the switch to conditions of submerged cultivation].

    Science.gov (United States)

    Spiridonova, E V; Adnof, D M; Andreev, I O; Kunakh, V A

    2008-01-01

    Genome of Rauwolfia serpentina callus cells was found to fail undergo the noticeable changes for several early passages upon the switch from surface to submerged cultivation in the liquid medium of special composition. After subsequent 4-6 passages in submerged culture RAPD spectra polymorphism was revealed which may reflect the changes in DNA sequence as well as in the structure of cell population that forms the strain. Introduction of the intermediary passage on the agar-solidified medium of more simple composition prior to transfer into liquid medium appeared not to affect essentially the level and the pattern of genome changes.

  3. The use of tissue culture techniques to detect irradiated vegetables

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Sharabi, N.E.; Nabulsi, I

    2001-01-01

    the ability of two tissue culture methods, callus and vegetable growth induction, to detect irradiated vegetables was evaluated. Potato tubers, carrot roots, garlic cloves and onion bulbs were subjected to various gamma radiation doses (0, 25, 100, 150, 250, 500, 750, and 1000 Gy). Irradiated vegetables were cultured in vitro and in vivo (pots). Gamma irradiation significantly reduced callus-forming ability especially in carrot and potato where no callus was observed in doses higher than 50 Gy. Length of shoots and roots growing from irradiated garlic and onion explants was considerably reduced starting from the 25 Gy dose. No roots were formed on garlic explants at any irradiation dose. Garlic leaves growing from irradiated explants were spotted with purple to brown spots. The intensity of these spots increased as gamma ray dosage increased. In the pot experiment, potato plant appeared in the control only. On the contrary, a complete sprouting of garlic and onion was seen in all irradiation treatments. It was not possible to distinguish between the various irradiation treatments and the control 3 days after planting in pots. The two in vitro techniques, tested in our study, may effectively be used to detect irradiated vegetables and estimate the range of doses used. The callus formation method is more useful for potato and carrot, since regeneration of shoots in vitro from these two plants takes along time, making this method unpractical. The other technique is very useful in the case of onion and garlic since it is rapid. The two techniques can be used with most of the vegetables that can be cultured in vitro. (Author)

  4. Zinc tolerance and accumulation in stable cell suspension cultures and in vitro regenerated plants of the emerging model plant Arabidopsis halleri (Brassicaceae).

    Science.gov (United States)

    Vera-Estrella, Rosario; Miranda-Vergara, Maria Cristina; Barkla, Bronwyn J

    2009-03-01

    Arabidopsis halleri is increasingly employed as a model plant for studying heavy metal hyperaccumulation. With the aim of providing valuable tools for studies on cellular physiology and molecular biology of metal tolerance and transport, this study reports the development of successful and highly efficient methods for the in vitro regeneration of A. halleri plants and production of stable cell suspension lines. Plants were regenerated from leaf explants of A. halleri via a three-step procedure: callus induction, somatic embryogenesis and shoot development. Efficiency of callus proliferation and regeneration depended on the initial callus induction media and was optimal in the presence of 1 mg L(-1) 2,4-dichlorophenoxyacetic acid, and 0.05 mg L(-1) benzylaminopurine. Subsequent shoot and root regeneration from callus initiated under these conditions reached levels of 100% efficiency. High friability of the callus supported the development of cell suspension cultures with minimal cellular aggregates. Characterization of regenerated plants and cell cultures determined that they maintained not only the zinc tolerance and requirement of the whole plant but also the ability to accumulate zinc; with plants accumulating up to 50.0 micromoles zinc g(-1) FW, and cell suspension cultures 30.9 micromoles zinc g(-1) DW. Together this work will provide the experimental basis for furthering our knowledge of A. halleri as a model heavy metal hyperaccumulating plant.

  5. Meristem Culture of Aloe vera L

    International Nuclear Information System (INIS)

    Kay Thi Mya; Thida Myint; Myo Han; Khin Maung Sein

    2005-10-01

    Shoot tips of Aloe vera L were excised and inoculated in basal and modified MS (Murashige and Skoog, 1962) medium. Growth regulators and growth factors were supplemented to the basal medium. Development of explants, initiation of callus and multiplication of shoots in culture were recorded.

  6. Effect of x-ray irradiation on maize inbred line B73 tissue cultures and regenerated plants

    International Nuclear Information System (INIS)

    Wang, A.S.; Cheng, D.S.K.; Milcic, J.B.; Yang, T.C.

    1988-01-01

    In order to enhance variation induced by the tissue culture process and to obtain agronomically desirable mutants, friable embryogenic tissue cultures of maize (Zea mays L.) inbred line B73 were x-ray irradiated with 11 doses [0-8.4 kilorads (kR)]. Reductions in callus growth rate and embryogenic callus formation occurred with increasing x-ray doses 20 d and 3 months after irradiation. Callus irradiated with 0.8 kR showed a significant increase in growth rate and a 20% increase in embryogenic callus 9 months after irradiation. A total of 230 R 0 plants were regenerated for evaluation. Pollen fertility and seed set of R 0 plants decreased with increasing x-ray dosage. Days to anthesis and plant height of R 0 plants varied among x-ray treatments but were generally reduced with higher dosages. The number of chromosomal aberrations increased with x-ray dosage. The R 1 seeds taken from R 0 plants were also grown and tested for mutant segregation. Plants regenerated from irradiated calli had a two- to 10-fold increase in mutations over plants regenerated from unirradiated control callus. Germination frequency of seeds from R 0 plants decreased with increasing x-ray dosage. Although chlorophyll mutants were most frequently observed, a number of vigorous plants with earlier anthesis date were also recovered

  7. callus induction and proliferation from cotyledon explants in ...

    African Journals Online (AJOL)

    ACSS

    2013-07-19

    Jul 19, 2013 ... between the tested cytokinins and the 2,4-D in callus induction and growth index. Similar results have also been obtained in soybean (Glycine max L.) (Sairam et al., 2003). In contrast to this finding, calli were reportedly induced from cotyledon explants on MS basal medium containing 2,4-D in combination ...

  8. Morpho-anatomical characterization of embryogenic calluses from immature zygotic embryo of peach palm during somatic embryogenesis =

    Directory of Open Access Journals (Sweden)

    Simone de Alencar Maciel

    2010-04-01

    Full Text Available The objective of this study was to morpho-anatomically characterizenodular embryogenic calluses from zygotic embryos of peach palm during the induction of somatic embryogenesis. Immature zygotic embryos were pre-treated in MS medium added to Picloram and 2,4-D (25 μM and BAP (0, 5, 10 μM. After three months, primary calluses were transferred to MS induction medium added to Picloram and 2,4-D (450 μM. After six months, the embryogenic calluses were then histologically analyzed and cultivated in the maturation medium. The competent tissues of the zygotic embryos differentiated embryogenic calluses under action of both Picloram and 2,4-D auxins (450 μM, where the presence of multi-granular structures were observed. Histological observations showed that in the nodular embryogenic calluses, the outlying parenchymal cells exhibit cellular characteristics of high mitotic activity. Differentiation of tracheal elements exists in embryogenic calluses connecting the callus to the explant. The evaluated cytokinin/auxin interaction influences the development of embryogenic calluses and globular structures.O objetivo deste trabalho foi caracterizar morfoanatomicamente calos nodulares embriogênicos originados de embriões zigóticos de pupunheira durante a indução da embriogênese somática. Embriões zigóticos imaturos de pupunha foram inicialmente pré-tratados em meio de cultura MS, solidificado com 2,5 g L-1 de phytagel® e suplementado com Picloram e 2,4-D na concentração de 25 μM e BAP (0, 5, 10 μM. Após três meses, os calos primários foram transferidos para meio de indução, com Picloram e 2,4-D (450 μM. Após seis meses, os calosnodulares embriogênicos formados foram então analisados histologicamente e repicados para o meio de maturação para a progressão das estruturas multigranulares embriogênicas. Verificou-seque os tecidos competentes dos embriões zigóticos imaturos diferenciaram nódulos embriogênicos pela ação de ambas

  9. Growing, regeneration and radiosensibility of sugarcane callus (Saccharum spp. hybrid var. “SP 70-1284” treated with gamma radiation source 60Co.

    Directory of Open Access Journals (Sweden)

    Apolonio Valdez Balero

    2004-07-01

    Full Text Available Calluses in growth of sugar cane, variety “SP 70-1284”, were radiated with a source of 60Co. The studied doses were between 10 and 80 Gy. The affectation of the Gamma radiation source 60Co in the growth of the callus and the regeneration of plants was evaluated. The results indicated that the dose that diminished the growth of callus in a value under 50% was the one of 30 Gy, and also the dose that diminished the regenerative capacity of the callus below to the 50% of the population was the one of 30 Gy. The results showed that for this variety of sugar cane doses superior to 30Gy of Gamma radiations jeopardizes the growth and the later regeneration of the callus. Its application is recommended to induce genetic variability in the variety of sugar cane “SP 70-1284”, as well as in programs of genetic improvement by mutations, in susceptible varieties of the sugar cane red blight (Puccinia melanocephala Syd using calluses in growth. Key words: callus, hybrid, in vitro, improvement, rust, Saccharum sp, variability

  10. Hyperplastic callus formation in osteogenesis imperfecta type V mimicking osteosarcoma: 4-year follow-up with resolution

    Energy Technology Data Exchange (ETDEWEB)

    Vieira, R.L.V.; Amaral, D.T. [Federal University of Sao Paulo, Department of Radiology, Sao Paulo (Brazil); Jesus-Garcia, Filho R. [Federal University of Sao Paulo, Department of Orthopedic Surgery, Sao Paulo (Brazil); Saraiva, G. [Federal University of Sao Paulo, Department of Endocrinology, Sao Paulo (Brazil); Fernandes, A.R.C. [University of California San Diego, Department of MSK Radiology, San Diego, CA (United States); Resnick, D.

    2006-06-15

    We report a case of hyperplastic callus formation that occurred in both femurs in a patient with type V osteogenesis imperfecta (OI), with 4-year follow-up and resolution. The clinical, histological and imaging aspects of this condition are discussed. Recognition of the hyperplastic callus formation in this particular type of OI is important in order to avoid misdiagnosis. (orig.)

  11. Hyperplastic callus formation in osteogenesis imperfecta type V mimicking osteosarcoma: 4-year follow-up with resolution

    International Nuclear Information System (INIS)

    Vieira, R.L.V.; Amaral, D.T.; Jesus-Garcia, Filho R.; Saraiva, G.; Fernandes, A.R.C.; Resnick, D.

    2006-01-01

    We report a case of hyperplastic callus formation that occurred in both femurs in a patient with type V osteogenesis imperfecta (OI), with 4-year follow-up and resolution. The clinical, histological and imaging aspects of this condition are discussed. Recognition of the hyperplastic callus formation in this particular type of OI is important in order to avoid misdiagnosis. (orig.)

  12. Effect of 60Co γ-rays irradiation pretreatment on callus inductivity and differentiation of strawberry anther

    International Nuclear Information System (INIS)

    Zhang Huiqin; Xie Ming; Jiang Guihua; Sun Congbo; Huang Pule; Wu Yanjun; Wu Qing

    2007-01-01

    Strawberry buds, in which a majority of pollen had single nuclear keeping to the side, were pretreated by 60 Co γ-rays irradiation at different doses. The irradiation effect on the callus inductivity and plant differentiation rate of strawberry anther were studied. The results indicated that the pretreatment by 60 Co γ-rays could increase the anther callus inductivity and plant differentiation rate significantly, and 20 Gy was the best dose. The callus inductivity of Akihime and Toyonoka increased by 39.09% and 35.68%, respectively, compared with the no-pretreatment materials. On the same media, the plant differentiation rate of Akihime and Toyonoka were also increased by 6.67%-6.72% and 4.73%-6.45%, respectively, compared with the two varieties of no-pretreatment. (authors)

  13. Callus formation in vitro and internodal stem apices in savory = Calogênese in vitro de segmentos apicais caulinares e internodais em segurelha (Satureja hortensis L.

    Directory of Open Access Journals (Sweden)

    Marcio Carlos Navroski

    2012-12-01

    Full Text Available We sought to evaluate with this work different growth regulators on callus formation in shoot apical and internodal stem segments of Satureja hortensis. The explants were isolated from in vitro seedlings and cultured on MS nutrient medium supplemented with NAA (0 and 1 μM and BAP (0, 5, 10, 15 and 20 μM. The presence of auxin NAA gave higher marks to the calluses at 30 days of evaluation, these notes also increased with the addition of BAP. There were significant interactions between factors in evaluating NAA and BAP for 60 days, both in apical stem segments as in internodal stem segments. To stemapices percentage of friable callus tends to decrease with increasing concentration of BAP. As for compact calluses increased BAP leads to an increase in the percentage of this type of callus. The presence of NAA increased callus formation in both friable and compact calluses on. This trend was also observed in internodal stem segments. The callus formation was highly rhizogenic observed in the presence of NAA, hardly occurs in the absence of auxin. The use of BAP is recommended in case of regeneration of plants through micropropagation, if the goal is the production of metabolites, the use of BAP can be harmful by reducing the production of friable callus.= Buscou-se com este trabalho avaliar diferentes reguladores de crescimento sobre a calogênese em segmentos caulinares apicais e internodais de Satureja hortensis L.. Os explantes foram isolados de plântulas germinadas in vitro e cultivados em meio nutritivo MS acrescido de ANA (0 e 1 μM e de BAP (0; 5; 10; 15 e 20 μM. A presença da auxina ANA proporcionou notas mais altas para os calos aos 30 dias de avaliação, estas notas também aumentaram com o acréscimo da citocinina BAP. Houve interações significativas entre os fatores ANA e BAP na avaliação aos 60 dias, nos dois tipos de segmentos caulinares(apicais e internodais. Nos segmentos apicais caulinares a porcentagem de calos fri

  14. Comparative study of metals accumulation in cultured in vitro mycelium and naturally grown fruiting bodies of Boletus badius and Cantharellus cibarius.

    Science.gov (United States)

    Reczyński, Witold; Muszyńska, Bożena; Opoka, Włodzimierz; Smalec, Agata; Sułkowska-Ziaja, Katarzyna; Malec, Mirosław

    2013-06-01

    Cantharellus cibarius Fr. (chanterelle) and Boletus badius Pers. (bay bolete) harvested from natural sites in Poland were used to derive in vitro cultures. The optimal medium composition for cultures was developed. Concentrations of the chosen elements (Zn, Cu, Fe, Mg, Ni, and Cd) in mycelium samples were measured by means of atomic absorption spectrometry. Fe concentration in the analyzed mushroom materials was in the range 215.4-680.3 μg/g dry weight. Mean values of Mg were respectively (in micrograms per gram dry weight) 541.8 for mycelium of C. cibarius cultured in vitro and 1,004.1 for C. cibarius fruiting bodies and 928.9 for the mycelium of B. badius cultured in vitro and 906.4 for B. badius fruiting bodies. The mean concentrations of Zn were 442.7 μg/g dry weight in mycelium from in vitro cultures of B. badius and 172.1 in B. badius fruiting bodies and 131.9 in the case of C. cibarius in mycelium from in vitro cultures and 95.5 for the C. cibarius fruiting bodies. Cu exhibited a reversal tendency, i.e., the element concentrations in naturally grown mushrooms were significantly higher (43.57 μg/g dry weight for C. cibarius and 43.54 μg/g for B. badius) than in cultured in vitro mycelium (12.47 μg/g for C. cibarius and 4.17 μg/g for B. badius). Ni was found in lowest concentrations ranging from 0.33 to 1.88 μg/g dry weight. Toxic metal Cd was found in relatively high concentrations in naturally grown species (0.79 μg/g dry weight-1.02). The lowest was the concentration of Cd in C. cibarius mycelium from in vitro culture-0.06 μg/g dry weight-a bit higher than it was in the B. badius mycelium (0.21 μg/g).

  15. Explant age, auxin concentrations and media type affect callus ...

    African Journals Online (AJOL)

    The effects of explant age of oil palm (Elaeis huineensis) embryo axes, 15 and 18 weeks after anthesis (WAA), media type (Eeuwens and Murashige and Skoog) supplemented with various concentrations of 2,4-D on callus production employing standard in vitro techniques were investigated. The results of the study showed ...

  16. Short-term effects of carbon dioxide on carnation callus cell respiration

    International Nuclear Information System (INIS)

    Palet, A.; Ribas-Carbo, M.; Argiles, J.M.; Azcon-Bieto, J.

    1991-01-01

    The addition of potassium bicarbonate to the electrode cuvette immediately stimulated the rate of dark O 2 uptake of photomixotrophic and heterotrophic carnation (Dianthus caryophyllus L.) callus, of Elodea canadensis (Minchx) leaves, and of other plant tissues. This phenomenon occurred at pH values lower than 7.2 to 7.8, and the stimulation depended on the concentration of gaseous CO 2 in the solution. These stimulatory responses lasted several minutes and then decreased, but additional bicarbonate or gaseous CO 2 again stimulated respiration, suggesting a reversible effect. Carbonic anhydrase in the solution increased the stimulatory effect of potassium bicarbonate. The CO 2 /bicarbonate dependent stimulation of respiration did not occur in animal tissues such as rat diaphragm and isolated hepatocytes, and was inhibited by salicylhydroxamic acid in carnation callus cells and E. canadensis leaves. This suggested that the alternative oxidase was engaged during the stimulation in plant tissues. The cytochrome pathway was severely inhibited by CO 2 /bicarbonate either in the absence or in the presence of the uncoupler carbonylcyanide m-chlorophenyl hydrazone. The activity of cytochrome c oxidase of callus tissue homogenates was also inhibited by CO 2 /bicarbonate. The results suggested that high carbon dioxide levels (mainly free CO 2 ) partially inhibited the cytochrome pathway (apparently at the oxidase level), and this block in electron transport elicited a large transient engagement of the alternative oxidase when present uninhibited

  17. Unilateral Palmar Callus and Irritant Hand Eczema – Underreported Signs of Dependency on Crutches

    Directory of Open Access Journals (Sweden)

    Uwe Wollina

    2018-01-01

    Full Text Available Leg amputees who can’t use prostheses and patients with arthritis are often dependent on crutches. Their chronic use can exert significant friction forces. The palmar skin will respond by forming a hyperkeratotic callus. We report for the first time unilateral palmar callus formation caused by friction from using crutches. Another possible adverse effect is the triggering of irritant contact dermatitis by the handholes of crutches. We report two cases with hand dermatitis due to the chronic dependence on crutches and discuss treatment options.

  18. Enhanced production of phenolic acids in cell suspension culture of Salvia leriifolia Benth. using growth regulators and sucrose.

    Science.gov (United States)

    Modarres, Masoomeh; Esmaeilzadeh Bahabadi, Sedigheh; Taghavizadeh Yazdi, Mohammad Ehsan

    2018-04-01

    Salvia leriifolia Benth. (Lamiaceae) is an endangered medicinal plant with hypoglycemic, anti-inflammatory and analgesic properties. Many of the beneficial effects of Salvia spp. are attributed to the phenolic compounds. In the present study, an efficient procedure has been developed for establishment of cell suspension culture of S. leriifolia as a strategy to obtain an in vitro phenolic acids producing cell line for the first time. The effect of growth regulators and various concentrations of sucrose have been analyzed, to optimize biomass growth and phenolic acids production. The callus used for this purpose was obtained from leaves of 15-day-old in vitro seedlings, on Murashige and Skoog (MS) basal medium supplemented with different hormone balances including benzylaminopurine (BAP) and indole butyric acid (IBA); 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KIN); naphthaleneacetic acid (NAA) and BAP. Modified MS medium supplemented with 5 mg/L BAP and 5 mg/L NAA was the optimal condition for callus formation with the highest induction rate (100%), the best callus growth and the highest phenolic acids content. No callus induction was observed in combinations of IBA and BAP. Cell suspension cultures were established by transferring 0.5 g of callus to 30 mL liquid MS medium supplemented with 5 mg/L BAP and 5 mg/L NAA. Dynamics of phenolic acids production has been investigated during the growth cycle of the suspension cultures. The maximum content of caffeic acid and salvianolic acid B were observed on the 15th day of the cultivation cycle while the highest amount of rosmarinic acid was observed on the first day. In response to various sucrose concentrations, cell cultures with 40 g/L sucrose not only produced the highest dry biomass but also the highest induction of caffeic acid and salvianolic acid B. The highest amount of rosmarinic acid was observed in media containing 50 g/L sucrose. These prepared cell suspension cultures provided a useful

  19. Substituted Indoleacetic Acids Tested in Tissue Cultures

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1978-01-01

    Monochloro substituted IAA inhibited shoot induction in tobacco tissue cultures about as much as IAA. Dichloro substituted IAA inhibited shoot formation less. Other substituted IAA except 5-fluoro- and 5-bromoindole-3-acetic acid were less active than IAA. Callus growth was quite variable...

  20. Induction of bulb organogenesis in in vitro cultures of tarda tulip (Tulipa tarda Stapf.) from seed-derived explants.

    Science.gov (United States)

    Maślanka, Małgorzata; Bach, Anna

    2014-01-01

    A protocol for obtaining bulbs via in vitro organogenesis was developed for tarda tulip ( Tulipa tarda Stapf). Scale explants were obtained from bulbs formed at the base of seedlings or from adventitious bulbs that developed from callus tissue forming on stolons or on germinating seeds. Some explants were subjected to chilling at 5°C for 12 wk. The culture media contained 3 or 6% sucrose and was supplemented with either no growth regulators, either 0.5 μM 6-benzyl-aminopurine (BAP) or 18.9 or 94.6 μM abscisic acid (ABA). Cultures were maintained in the dark at 20°C. Callus tissue developed mainly on media without growth regulators or with BAP. Callus was formed from up to 96% of explants derived from non-chilled adventitious bulbs that were treated with 3% sucrose and 0.5 μM BAP. Less callus was formed from chilled explants compared with non-chilled explants. Newly formed adventitious bulbs appeared on the explants via direct and indirect organogenesis. The media with BAP promoted the formation of adventitious bulbs at a rate of 56-92% from non-chilled explants, whereas a maximum rate of 36% was observed from chilled explants. ABA inhibited the induction of adventitious bulbs and callus. The adventitious bulbs obtained in these experiments contained a meristem, which was evidence that they had developed properly.

  1. Micropropagation of Dalbergia sissoo Roxb. through tissue culture technique.

    Science.gov (United States)

    Sahu, Jyoti; Khan, Shagufta; Sahu, Ram Kumar; Roy, Amit

    2014-04-01

    Multiple shoots of Dalbergia sissoo Roxb. (Sissoo) were incited from seeds through indirect somatic embryogenesis method. Seeds were inoculated in Murashige and Skoog's medium without any growth hormone. Than cotyledonary leaves were struck and used for callus induction on MS medium amplified with 2, 4-dichlorophenoxyacetic acid (0.5 to 4 mg mL(-1)). After 3 to 4 weeks the embryogenic callus clumps was transferred to medium supplemented with cytokinin (BAP 1 to 5 mg L(-1), kinetin 1-5.0 mg L(-1)) for embryo maturation and germination. The high-frequency shoot proliferation (82%) and maximum number of shoots per explants were recorded in MS medium containing NAA (0.5)+BAP (0.5). The findings of recent investigations have shown that, it is possible to induce indirect somatic embryogenesis in Dalbergia sissoo and plant regeneration from callus cultures derived from cotyledonary leaves as explants.

  2. Effect of induced mutagenesis in rice tissue culture

    International Nuclear Information System (INIS)

    Maddumage, R.

    1994-01-01

    The influence of chemical mutagens and ionising radiation on growth, regenerative capacity of rice callus culture and the effect o9f mutagens on frequency and spectrum of mutant regenerants, derived from calli and determination of approximate semi-lethal dose of each mutagen on rice calli was studied. Intact mature de-husked grains and pieces of primordial particles of four varieties were used as explants in the experiment. Organogenesis was induced using MS media supplemented with agar. After thirty days calluses were subjected to varying concentrations/dosage of mutagens. The effect of mutagens on growth of callus was stimulative in low concentration/doses at short exposure, but in higher concentration/doses at longer exposure it was oppressive. In x-radiation treatment all the studied doses showed only stimulative effect on growth. The effect of mutagenic treatment on regenerative capacity was negative. No specificity was found even between two chemical mutagens of their action on studied characters

  3. In Vitro Callus Induction and Growth of Stevia (Stevia rebaudiana Bertoni M. with Difference Concentrations of PEG (Polyethylene Glycol and Light Conditions

    Directory of Open Access Journals (Sweden)

    Mohamad Ana Syabana

    2017-07-01

    Full Text Available Stevia (Stevia rebaudiana Bert. M. is known as a natural non-caloric sweetener. This plants contain glycoside such steviosida type, mainly on the leave contain sweetness level between 200-300 cane sugar but the calorie is very low. This research was aimed to determine difference effects of PEG (Polyethylene Glycol concentrations and the light conditions on Stevia (Stevia rebaudiana Bert. M. callus induction in vitro. This research was conducted from April to June 2016 at Biotechnology Laboratory Faculty of Agriculture, Sultan Ageng Tirtayasa University. This research used a completely randomized design (CRD, which consisted of two factors with three replications. Concentrations of PEG as first factor consisted of four levels (0 mg/L, 5 mg/L, 15 mg/L, and 25 mg/L. Light conditions as second factor consisted of two levels (Dark and Light. The results showed that the concentrations of PEG (Polyethylene Glycol did not significantly effect the time of callus appearance and diameter of callus on 4, 5 and 6 weeks after planting. Dark condition was the best conditions for callus induction of stevia. The texture of callus was compact on all treatments and the callus dominant color produced is golden brown.

  4. Effect of sorbitol in callus induction and plant regeneration in wheat

    African Journals Online (AJOL)

    STORAGESEVER

    2009-12-01

    Dec 1, 2009 ... Key words: Callus induction, plant regeneration, wheat, 2,4-D, sorbitol. INTRODUCTION ... regeneration is better on hormone-free medium or that .... AB (interaction). 15 ... element and creates osmotic stress as reported by.

  5. Biological effects of fast neutron irradiation on callus tissues of Tecoma stans Juss. and Ammi visnaga Lam

    International Nuclear Information System (INIS)

    Supniewska, J.H.; Dohnal, B.; Cebulska Wasilewska, A.; Huczkowski, J.

    1982-01-01

    Callus tissues of Tecoma stans Juss. and Ammi visnaga Lam. were subjected to fast neutron irradiation. Nine doses were applied within the range of 100 - 10.000 cGy. Small doses caused growth stimulation. Intermediate and high doses caused morphological changes, reduced growth and biosynthesis of biologically active substances (monoterpene alkaloids in T. stans, furanochromones in A. visnaga). In A. visnaga neutron irradiation considerably decreased the chlorophyll content in callus tissues. The radiosensitivity of A. visnaga at 50% growth reduction level was 1.5 times higher than that of the callus of T. stans. The recovery of the tissues takes place during a subculturing course. Three to 7 months after neutron exposure growth and biosynthesis reach the control level. (author)

  6. Passion fruit by-product and fructooligosaccharides stimulate the growth and folate production by starter and probiotic cultures in fermented soymilk.

    Science.gov (United States)

    Albuquerque, Marcela Albuquerque Cavalcanti; Bedani, Raquel; LeBlanc, Jean Guy; Saad, Susana Marta Isay

    2017-11-16

    Two starter cultures (Streptococcus (St.) thermophilus ST-M6 and TA-40) and five probiotic strains (St. thermophilus TH-4, Lactobacillus (Lb.) acidophilus LA-5, Lb. rhamnosus LGG, Lb. fermentum PCC, and Lb. reuteri RC-14) were used to ferment different soymilk formulations supplemented with passion fruit by-product and/or fructo-oligosaccharides (FOS) with the aim of increasing folate concentrations. Growth and folate production of individual strains were evaluated and the results used to select co-cultures. Both St. thermophilus ST-M6 and TH-4 were the best folate producers and were able to increase the folate content of all soymilk formulations when used alone or in co-culture with lactobacilli strains, especially in the presence of both passion fruit by-product and FOS. Thus, passion fruit by-product and FOS could be used as dietary ingredients to stimulate the folate production by selected bacterial strains during the fermentation of soymilk. It was also shown that vitamin production by microorganisms is strain-dependent and may also be influenced by nutritional and environmental conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Stimulation of reserpine biosynthesis in the callus of Rauvolfia ...

    African Journals Online (AJOL)

    So enhancing this alkaloid in the already available system is a beneficial approach. Tryptophan is the starting material in the biosynthesis of reserpine. Callus was induced from leaf explants of Rauvolfia tetraphylla L. on MS medium supplemented with the combination of 9 μM 2,4-D and 25, 50, 75 and 100 mg/l tryptophan.

  8. Regeneration from embryogenic callus and suspension cultures of ...

    African Journals Online (AJOL)

    ehab

    2012-04-25

    Apr 25, 2012 ... cultures of the wild medicinal plant Cymbopogon schoenanthus ... nardus, C. martini, C. citratus and C. flexuosus are the source of palmerosa, citronella, lemon-grass and oil of. Malabar ...... marketing of essential oils. J. Med.

  9. In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur.

    Science.gov (United States)

    Haffner-Luntzer, Melanie; Müller-Graf, Fabian; Matthys, Romano; Abaei, Alireza; Jonas, René; Gebhard, Florian; Rasche, Volker; Ignatius, Anita

    2017-11-14

    Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and µCT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

  10. Regeneration and acclimatization of salt-tolerant arachis hypogaea plants through tissue culture

    International Nuclear Information System (INIS)

    Ghauri, E.G.

    2006-01-01

    Excised embryos of Arachis hypogaea were cultured on Murashige and Skoog's medium (MS medium) supplemented with different combinations of growth hormones. The highest frequency of callus proliferation (80%) was recorded on MS medium mixed with 1.0 mg/1 of 2,4-D and 0.5 mg/1 of BAP. These cultures were treated with 0.65 mg/l of trans-4-hydroxy-L-proline (HyP) a:1d various concentrations (0.1-0.5%) of NaCl. In all cases the presence of salt reduced the fresh mass of callus. Shoot regeneration in the cultures took place when transferred to MS medium supplemented with 1.0 mg/1 of kinetin (Kin) and 0.5 mg/1 of 6-benzyl aminopurine (BAP). Percentage of shoot regeneration decreased with the increase of NaCl (0.1- 0.5%) in the shoot regeneration medium. Root formation in these cultures took place when the cultures were nurtured on MS medium free of growth hormones. Regeneration, hardening and acclimatization of the salt tolerant plants was conducted. (author)

  11. Endophytic Penicillium citrinum Thom. from Scoparia dulcis Linn.

    Science.gov (United States)

    Mathew, Annie J; Jayachandran, K; Mathew, Jyothis

    2010-10-01

    Scoparia dulcis of Scrophulariaceae is an annual herb distributed through out the tropics. Penicillium citrinum was obtained from apparently healthy roots, stem, leaves and fruits of this plant. Callus and multiple shoots produced during micropropagation from various explants were also symptomless but showed occurrence of Penicillium citrinum when cultured in Murashige & Skoog liquid medium for the production of secondary metabolites.

  12. Endophytic Penicillium citrinum Thom. from Scoparia dulcis Linn

    OpenAIRE

    Mathew, Annie J.; Jayachandran, K.; Mathew, Jyothis

    2010-01-01

    Scoparia dulcis of Scrophulariaceae is an annual herb distributed through out the tropics. Penicillium citrinum was obtained from apparently healthy roots, stem, leaves and fruits of this plant. Callus and multiple shoots produced during micropropagation from various explants were also symptomless but showed occurrence of Penicillium citrinum when cultured in Murashige & Skoog liquid medium for the production of secondary metabolites.

  13. In Vitro Callus Induction and Growth of Stevia (Stevia rebaudiana Bertoni M. with Difference Concentrations of PEG (Polyethylene Glycol and Light Conditions

    Directory of Open Access Journals (Sweden)

    Mohamad Ana Syabana

    2017-06-01

    Full Text Available Stevia (Stevia rebaudiana Bert. M. is known as a natural non-caloric sweetener. This plants contain glycoside such steviosida type, mainly on the leave contain sweetness level between 200-300 cane sugar but the calorie is very low. This research was aimed to determine difference effects of PEG (Polyethylene Glycol concentrations and the light conditions on  Stevia (Stevia rebaudiana Bert. M. callus induction  in vitro. This research was conducted from April to June 2016 at Biotechnology Laboratory Faculty of Agriculture, Sultan Ageng Tirtayasa University. This research used a completely randomized design (CRD, which consisted of two factors with three replications. Concentrations of PEG as first factor consisted of four levels (0 mg/L, 5 mg/L, 15 mg/L, and 25 mg/L. Light conditions as second factor consisted of two levels (Dark and Light. The results showed that the concentrations of PEG (Polyethylene Glycol did not significantly effect the time of callus appearance and diameter of callus on 4, 5 and 6 weeks after planting. Dark condition was the best conditions for callus induction of stevia. The texture of callus was compact on all treatments and the callus dominant color produced is golden brown

  14. MRI and CT features of hyperplastic callus in osteogenesis imperfecta tarda

    International Nuclear Information System (INIS)

    Dobrocky, I.; Seidl, G.; Grill, F.

    1999-01-01

    We describe the MRI and CT findings of hyperplastic callus formation simulating a tumour of pelvis in patient with osteogenesis imperfecta tarda. Possible differential diagnoses and the impact of different imaging techniques on the correct diagnosis are discussed. (orig.)

  15. Tissue culture of Cecropia glaziovii Sneth (urticaceae: vegetative micropropagation and plant regeneration from callus Cultura de tecidos de Cecropia glaziovii Sneth (Urticaceae: micropropagação vegetativa e regeneração de plantas via calos

    Directory of Open Access Journals (Sweden)

    Marcos Nopper Alves

    2010-10-01

    Full Text Available Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS medium supplemented with 6-benzylaminopurine alone (BAP (1.0, 5.0 or 10.0 mg L-1 or combined with -naphthalene acetic acid (NAA (1.0 or 2.0 mg L-1, after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D (5.0 mg L-1 combined with BAP (1.0 mg L-1. Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA (0.1 mg L-1 alone or combined with 2,4-D (1.0 or 5.0 mg L-1 was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA (1.0 mg L-1. Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.Cecropia glaziovii é uma planta lenhosa, popularmente usada no Brasil como medicinal. Métodos que visem a sua propagação clonal podem ser de grande utilidade na preservação de seus genótipos de elite. Foram examinados efeitos de diferentes reguladores de crescimento e explantes na forma

  16. Anti-aging activities of Pyrus pyrifolia var culta plant callus extract

    African Journals Online (AJOL)

    saline (PBS, pH 7.4) at 95 °C for 2 h and filtering through .... callus extract also exhibited dose-dependent .... toxic effects on the skin. ... copper-containing enzyme that catalyzes two .... herbal extract: implication for the mechanism underlying.

  17. Are bone turnover markers capable of predicting callus consolidation during bone healing?

    Science.gov (United States)

    Klein, P; Bail, H J; Schell, H; Michel, R; Amthauer, H; Bragulla, H; Duda, G N

    2004-07-01

    The aim of this study was to determine the ability of the following bone turnover markers to monitor the course of callus consolidation during bone healing: Carboxy-terminal propeptide of procollagen type I (PICP), skeletal alkaline phosphatase (sALP), and amino-terminal propeptide of type III procollagen (PIlINP). Since interfragmentary movements have been proven to possess the ability to document the progression of bone healing in experimental studies, correlations between bone turnover markers and interfragmentary movements in vivo were investigated. Therefore, two different types of osteosyntheses representing different mechanical situations at the fracture site were compared in an ovine osteotomy model. Blood samples were taken preoperatively and postoperatively in weekly intervals over a nine-week healing period. At the same intervals, interfragmentary movements were measured in all sheep. After nine weeks, animals were sacrificed and the tibiae were evaluated both mechanically and histologically. Wide interindividual ranges were observed for all bone turnover markers. The systemic PICP level did not increase with callus consolidation. The bone-healing model seemed to influence the systemic level of PIIINP and sALP but no general correlation between bone turnover markers and interfragmentary movements could be detected. No differences between the different types of osteosyntheses and thus the different mechanical situations were observed. All analyzed markers failed as general predictors for the course of callus consolidation during bone healing.

  18. Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers.

    Science.gov (United States)

    Seguí-Simarro, José M; Nuez, Fernando

    2007-01-01

    In this work, some of the different in vitro developmental pathways into which tomato microspores or microsporocytes can be deviated experimentally were explored. The two principal ones are direct embryogenesis from isolated microspores and callus formation from meiocyte-containing anthers. By means of light and electron microscopy, the process of early embryogenesis from isolated microspores and the disruption of normal meiotic development and change of developmental fate towards callus proliferation, morphogenesis, and plant regeneration have been shown. From microspores isolated at the vacuolate stage, embryos can be directly induced, thus avoiding non-androgenic products. In contrast, several different morphogenic events can be triggered in cultures of microsporocyte-containing anthers under adequate conditions, including indirect embryogenesis, adventitious organogenesis, and plant regeneration. Both callus and regenerated plants may be haploid, diploid, and mostly mixoploid. The results demonstrate that both gametophytic and sporophytic calli occur in cultured tomato anthers, and point to an in vitro-induced disturbance of cytokinesis and subsequent fusion of daughter nuclei as a putative cause for mixoploidy and genome doubling during both tetrad compartmentalization and callus proliferation. The potential implications of the different alternative pathways are discussed in the context of their application to the production of doubled-haploid plants in tomato, which is still very poorly developed.

  19. The effects of cultural practice methods on fruit orchard rehabilitation after flooding crisis in Songkhla province

    Directory of Open Access Journals (Sweden)

    Chanaweerawan, S.

    2004-01-01

    Full Text Available To rehabilitate the fruit orchards exposed to flooding crisis that occurred in year 2000 in Songkhla province, an investigation of the effects of cultural practice methods was done in the farmers’ orchards at 2 experimental sites (at Tumbol Kho Hong and Tumbol Kuan Lang, Amphur Hat Yai. The first site, at Tumbol Kho Hong, was mixed cropping (durian, longkong and mangosteen. The second site, at Tumbol Kuan Lang, was a monocrop of pummelo orchard. The experiment was comprised a stratified sampling method with 3 treatments: 1 control, 2 fertilization (15-15-15, 8-24-24 and 13-13-21 and soil improvement with humic acid (S and 3 foliar application (glucose was applied with 16-12-0+micronutrients+extracted seaweed and 7-13-34+12.5Zn+extracted seaweed+Ca-B spraying with fertilization and soil improvement (F+S. The results from the both experimental sites showed that the F+S treatment exhibited the best result. This promoted the plant growth and yield of fruit trees. In addition, the other orchards surrounding the experimental sites were surveyed. It was noticeable that fruit trees grown in raising-beds could recover and exhibit normal fruit bearing. This pointed out that the impact of flooding on fruit orchards would be possibly alleviated by a drainage system.

  20. MRI and CT features of hyperplastic callus in osteogenesis imperfecta tarda

    Energy Technology Data Exchange (ETDEWEB)

    Dobrocky, I. [Diagnostic Center Meidling, Vienna (Austria); Seidl, G. [Diagnostic Center Meidling, Vienna (Austria)]|[Universitaetsklinik fuer Radiodiagnostik, Vienna (Austria); Grill, F. [Orthopaedisches Spital Wien Speising, Vienna (Austria)

    1999-05-01

    We describe the MRI and CT findings of hyperplastic callus formation simulating a tumour of pelvis in patient with osteogenesis imperfecta tarda. Possible differential diagnoses and the impact of different imaging techniques on the correct diagnosis are discussed. (orig.) With 3 figs., 5 refs.

  1. callus culture

    African Journals Online (AJOL)

    steve

    1Harran University, Faculty of Agriculture, Department of Food Engineering, ... conduct the study with the independent variables being: percent depletion of KH2PO4 or NH4NO3 and ... phenolic compounds for prevention of disorders related to.

  2. Effects of plant growth regulators on callus induction from Cananga ...

    African Journals Online (AJOL)

    The Cananga odorata callus was initiated from petals of the C. odorata flowers on MS medium and B5 vitamins containing 30 g/L sugar and 3 g/L agar. The medium was also supplemented with different concentrations of 1-naphtalene acetic acid (NAA), and combinations of NAA with 6-benzylaminpurine (BAP) as the plant ...

  3. In-vitro regeneration of castor bean (ricinus communis, L) and physic nut (jatropha curcas, L.)

    International Nuclear Information System (INIS)

    Afful, N. T.

    2008-06-01

    Investigations were conducted on in vitro regeneration of physic nut (Jatropha curcas, 1.) and castor bean (Ricinus communis, 1.) using zygotic embryo culture, shoot tip and meristem culture as well as somatic embryogenesis. Zygotic embryos from different stages of fruit maturity cultured on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), kinetin and 2-isopentyl (2iP) produced vigorous seedlings with roots. However the optimal concentration of cytokinin for seedling development depended on the type cytokinin as well as the maturity stage of the fruit. Embryos from yellow fruits of Jatropha had the highest germination (100%) on an MS medium supplemented with 0.5 mg/L kinetin or 1 mg/L BAP while in Ricinus embryos from black fruits had the highest percentage germination (97%) on an MS medium modified with I mg/L 2iP or 0.5 or I mg/L BAP. Decrease in embryo viability especially in Jatropha as the seed matured may be due to high oil content in matured (black) fruit. Oil content in both Jatropha and Ricinus increased as the fruit matured from green to black and this may have effect on germination of zygotic embryos. In vitro culture response of shoot tip or meristem was significantly influenced by the concentration of growth regulators in the culture medium as well as the plant species. In both plant species, 2iP resulted in highest percentage shoot regeneration. The optimal concentration of 2iP was 0.5 mg/L for Ricinus shoot tip and 1.5 or 2.0 mg/L for Jatropha, indicating genotypic difference between the species. However excessive callus formation and browning in Ricinus led to the loss of all the regenerants. Shoot regeneration from meristem explants was also influenced by growth regulators with 2iP again being the best. Cultures of cotyledon and leaf lobe explants on MS medium supplemented with 2, 4-D and picloram resulted in callus induction with the exception of cotyledons obtained from black fruits. However transfer of calli onto fresh

  4. Plant regeneration from organ culture in white Guinea Yam

    Energy Technology Data Exchange (ETDEWEB)

    Nwachukwu, E C; Mbanaso, E N.A. [National Root Crops Research Inst., Umudike, Umuahia, Abia State (Nigeria); Sonnino, A [Centro Recerche Energia, ENEA, Rome (Italy)

    1997-07-01

    Explants from leaves, leaf segments, petioles and internodal stem of in vitro grown seedlings of white guinea yam, Dioscorea rotundata Poir, cv. `Obiaoturugo` were cultured on defined media. NAA at concentrations of 0.5-1.0 mg/1 induced shoot regeneration from petiolar and inter-nodal stem pieces, and rooting occurred with little or no callusing from whole leaves or leaf segments. With concentration of 3.0-10.0 m/1 NAA, explants from petioles, inter-nodal stem, whole leaves and leaf segments formed callus which produced roots. These explants developed plantlets when subcultured on MS medium supplemented with 2.0 mg/1 BAP and 0.1 mg/1 NAA. (author). 11 refs, 1 tab.

  5. Plant regeneration from organ culture in white Guinea Yam

    International Nuclear Information System (INIS)

    Nwachukwu, E.C.; Mbanaso, E.N.A.; Sonnino, A.

    1997-01-01

    Explants from leaves, leaf segments, petioles and internodal stem of in vitro grown seedlings of white guinea yam, Dioscorea rotundata Poir, cv. 'Obiaoturugo' were cultured on defined media. NAA at concentrations of 0.5-1.0 mg/1 induced shoot regeneration from petiolar and inter-nodal stem pieces, and rooting occurred with little or no callusing from whole leaves or leaf segments. With concentration of 3.0-10.0 m/1 NAA, explants from petioles, inter-nodal stem, whole leaves and leaf segments formed callus which produced roots. These explants developed plantlets when subcultured on MS medium supplemented with 2.0 mg/1 BAP and 0.1 mg/1 NAA. (author). 11 refs, 1 tab

  6. High capacity of plant regeneration from callus of interspecific hybrids with cultivated barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Bagger Jørgensen, Rikke; Jensen, C. J.; Andersen, B.

    1986-01-01

    Callus was induced from hybrids between cultivated barley (Hordeum vulgare L. ssp. vulgare) and ten species of wild barley (Hordeum L.) as well as from one backcross line ((H. lechleri .times. H. vulgare) .times. H. vulgare). Successful callus induction and regeneration of plants were achieved from...... explants of young spikes on the barley medium J 25-8. The capacity for plant regeneration was dependent on the wild parental species. In particular, combinations with four related wild species, viz. H. jubatum, H. roshevitzii, H. lechleri, and H. procerum, regenerated high numbers of plants from calli....

  7. Protein and Glycoprotein Patterns Related to Morphogenesis in Mammillaria gracillis Pfeiff. Tissue Culture

    Directory of Open Access Journals (Sweden)

    Biljana Balen

    2002-01-01

    Full Text Available As plants with Crassulacean Acid Metabolism (CAM, cacti are highly affected by artificial environmental conditions in tissue culture. Plants of Mammillaria gracillis Pfeiff. (Cactaceae propagated in vitro produced callus spontaneously. This habituated callus regenerated normal and hyperhydric shoots without the addition of growth regulators. In order to compare habituated callus with the tumorous one, cactus cells were transformed with two strains of Agrobacterium tumefaciens: the wild strain B6S3 (tumour line TW and the rooty mutant GV3101 (tumour line TR. Gene expression in cactus plants, habituated callus, regenerated shoots and two tumour lines was analysed at the level of cellular and extracellular protein and glycoprotein profiles. Proteins were separated by SDS-polyacrylamide gel electrophoresis and 2-D PAGE electrophoresis and silver stained. Concavalin A-peroxidase staining detected glycoproteins with D-manose in their glycan component on protein blots. Developmentally specific protein patterns of Mammillaria gracillis tissue lines were detected. The 2-D PAGE electrophoresis revealed some tissue specific protein groups. The cellular glycoprotein of 42 kDa detected by ConA was highly expressed in undifferentiated tissues (habituated callus, TW and TR tumours and in hyperhydric regenerants. Tumours produced extracellular proteins of 33, 23 and 22 kDa. The N glycosylation of cellular and extracellular proteins was related to specific developmental stage of cactus tissue.

  8. Efficiency of anther culture technique in the production of wheat double haploids

    Directory of Open Access Journals (Sweden)

    Kondić-Špika Ankica Đ.

    2008-01-01

    Full Text Available The objective of the study was to investigate efficiency of anther culture in the production of spontaneous double haploids from randomly selected heterozygous genotypes of wheat (Triticum aestivum L.. Anthers of 20 F1 wheat combinations were grown in vitro on a modified Potato-2 medium. All of the examined genotypes have shown the ability to produce pollen calluses as well as to regenerate green plants. On average for the whole experiment material, 47.2 calluses were produced per 100 cultured anthers. The green plant regeneration ranged from 0.8 to 13.4 green plants per spike, with an overall mean of 5.8. From the total of 582 regenerated green plants, 47.9% (279 were spontaneous double haploids. The final average yield from the study was 2.8 double haploids per spike.

  9. Analysis of soybean tissue culture protein dynamics using difference gel electrophoresis

    Science.gov (United States)

    Excised hypocotyls from developing soybean (Glycine max (L.) merr. cv. Jack) were cultivated on agar-solidified medium until callus formed. The calli were then propagated in liquid medium until stable, relatively uniform, finely-divided suspension cultures were obtained. Cells were typically transfe...

  10. Efficient production of a bioactive Bevacizumab monoclonal antibody using the 2A self-cleavage peptide in transgenic rice callus

    Directory of Open Access Journals (Sweden)

    Lei Chen

    2016-08-01

    Full Text Available Bevacizumab, a humanized monoclonal antibody (mAb targeting to the vascular endothelial growth factor (VEGF, has been widely used in clinical practice for the treatment of multiple cancers. Bevacizumab was mostly produced by the mammalian cell expression system. We here reported the first plant-derived Bevacizumab by using transgenic rice callus as an alternative gene expression system. Codon-optimized Bevacizumab light chain (BLC and heavy chain (BHC genes were designed, synthesized as a polyprotein with a 2A self-cleavage linker peptide from the Foot-and-mouth disease virus (FMDV, cloned into a plant binary vector under a constitutive maize ubiquitin promoter, and transformed into rice nuclear genome through Agrobacterium-mediated transformation. Southern blot and western blot analyses confirmed the integration and expression of BLC and BHC genes in transgenic rice callus. Enzyme linked immunosorbent assay (ELISA analysis indicated that the rice-derived Bevacizumab mAb was biologically active and the recombinant mAb was expressed at high levels (160.7-242.8 mg kg-1FW in transgenic rice callus. The mAb was purified by using protein A affinity chromatography and the purified antibody was tested for its binding affinity with its target hVEGF antigen by ELISA. Rice callus produced Bevacizumab and a commercial Bevacizumab (Avastin were shown to have similar binding affinity to hVEGF. These results indicated that rice callus produced Bevacizumab could have similar biological activity and might potentially be used as a cost-effective biosimilar molecule in future cancer treatment.

  11. The Effect of Plant Growth Regulators and Different Explants on the Response of Tissue Culture and Cell Suspension Cultures of German Chamomile (Matricaria chamomilla L.

    Directory of Open Access Journals (Sweden)

    L. Koohi,

    2014-07-01

    Full Text Available German chamomile (Matricaria chamomilla L. is one of the most important medicinal plants that its essential oils used in different medicinal industries. In this study which was carried out in 2013 growing season at the Faculty of Agricultural Sciences of the University of Mohaghegh Ardabili, the in vitro response of leaf and hypocotyl explants of German Chamomile in B5 medium supplemented with different levels of plant growth regulators including 2,4-D, naphthalene acetic acid (NAA, kinetin and 6-benzylaminopurine (BAP were investigated in a factorial experiment based on completely randomized design (CRD.In addition, cell suspension cultures were established and characterized. Hypocotyl and leaf explants exhibited cell proliferation and produced callus within 1-2 weeks. The highest fresh weight of the callus (264.1 mg was produced by leaf explants in the medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l BAP. However, the leaf explants cultured on medium containing 1.5 mg/l 2,4-D showed the lowest cell proliferation and callus yield (40.42 mg. The highest percentage of root induction from leaf explants (58.73% was observed on the medium containing 4 mg/l 2,4-D and 1 mg/l Kin, and from hypocotyl explants (48.61% was observed on medium supplemented with 1.5 mg/l NAA. The 42.22% of calli derived from hypocotyl explants on B5 medium supplemented with 4 mg/l NAA and 3 mg/l BAP, were friable. Cell suspension cultures of German chamomile were established by transferring of hypocotyl-derived friable calli into the MS medium supplemented with 1.5 mg/l 2,4-D and 1 mg/l kinetin. The growth curve of cell proliferations started 4 days after culture and continued to grow until day 13th, where the cells entered stationary phase.

  12. Genomics of biotrophic, plant-infecting plasmodiophorids using in vitro dual cultures.

    Science.gov (United States)

    Bulman, Simon; Candy, Judith M; Fiers, Mark; Lister, Ros; Conner, Anthony J; Eady, Colin C

    2011-07-01

    The plasmodiophorids are a phylogenetically distinct group of parasitic protists that infect plants and stramenopiles, causing several important agricultural diseases. Because of the obligate intracellular part of their lifecycle, none of the plasmodiophorids has been axenically cultured. Further, the molecular biology of the plasmodiophorids is poorly understood because pure cultures are not available from any species. We report on an in-vitro dual culture system of the plasmodiophorids Plasmodiophora brassicae and Spongospora subterranea with their respective plant hosts, Brassica rapa and Solanum tuberosum. We show that these plasmodiophorids are capable of initiating and maintaining stable, long-term plant cell callus cultures in the absence of exogenous plant growth regulators. We show that callus cultures harbouring S. subterranea provide an excellent starting material for gene discovery from this organism by constructing a pilot-scale DNA library. Bioinformatic analysis of the sequences established that almost all of the DNA clones from this library were from S. subterranea rather than the plant host. The Spongospora genome was found to be rich in retrotransposable elements, and Spongospora protein-coding genes were shown to contain introns. The sequence of a near full-length non-LTR retrotransposon was obtained, the first transposable element reported from a cercozoan protist. Copyright © 2010 Elsevier GmbH. All rights reserved.

  13. The effect of plant growth regulators on callus initiation in wormwood ...

    African Journals Online (AJOL)

    Studies were carried out in the Biotechnology laboratory of Plant Science Department of Ahmadu Bello University Zaria, Nigeria to study the effect of some plant growth regulators on the in vitro initiation of callus using the leaves of Chiyong variety of Artemisia annua. The explants were sterilized and incubated on Murashige ...

  14. Effect of 60Co γ irradiation with seed and shoot-tip of Brassica campestris L. var on its culture in vitro

    International Nuclear Information System (INIS)

    Liao Feixiong; Yu Rangcai; Pan Ruichi

    2003-01-01

    The survival rate in vitro of shoot-tips of Brassica campestris L. var from seeds irradiated by 60 Co γ-rays decreased with the increase of dose. Irradiation inhibited proliferation of shoot-tip, induction of callus from cotyledons and differentiation of the callus. The age of explant contributed to the effect of irradiation in the culture. Irradiation stimulated the proliferation of shoot-tip with dose less than 50 Gy. Based on the effect of irradiation in the tissue culture, the effective dose recommended was about 200 Gy for seeds, 50-100 Gy for pre-soaked germinating seeds and 40-70 Gy for shoot-tips in vitro, respectively

  15. Genetic Transformation of Metroxylon sagu (Rottb.) Cultures via Agrobacterium-Mediated and Particle Bombardment

    Science.gov (United States)

    Ibrahim, Evra Raunie

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance. PMID:25295258

  16. Genetic Transformation of Metroxylon sagu (Rottb. Cultures via Agrobacterium-Mediated and Particle Bombardment

    Directory of Open Access Journals (Sweden)

    Evra Raunie Ibrahim

    2014-01-01

    Full Text Available Sago palm (Metroxylon sagu is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L. Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

  17. Preliminary results on in vitro mutagenesis studies on Lansium domesticum Corr

    International Nuclear Information System (INIS)

    Mohd Nazir Basiran; Shuhaimi Shamsuddin; Sakinah Ariffin

    2000-01-01

    Dokong (Lansium domesticum Corr.) which belongs to the family Meliaceae is an important fruit trees for the Malaysian fruit industry. Despite the various types that have been identified, the genetic variability is still too narrow for meaningful breeding efforts. Dokong also has long juvenility period and the fruits are pathenocarpically developed. The fruits are often not uniform in size, the tree is prone to bark borers, and the tree architecture needs a lot of pruning for better fruit formation and facilitate easier harvesting. A lot of breeding efforts is needed to improve the genetic characteristics of this species before it can really have an industrial impact. Induced mutation and in vitro culture are two approaches which may be more efficient for genetic improvement. Results of radiosensitivity studies showed that irradiation doses between 50 and 70 Gy is effective enough to induced mutations in seeds. Initial attempts to develop in vitro cultures of Lansium showed that shoot-tips and axillary buds can be cultured to produce plantlets in a medium containing 3 mg/L kinetin and 4 mg/L indoleacetic acid. The procedures can be optimised to develop an efficient micropropagation system. However, attempts to initiate callus cultures have not been successful

  18. Meios de cultura, reguladores de crescimento e fontes de nitrogênio na regulação da calogênese do pau-brasil (Caesalpinia echinata Lam. Culture media, growth regulators and nitrogen sources in callus formation regulation of Brazilwood (Caesalpinia echinata Lam.

    Directory of Open Access Journals (Sweden)

    Elias Terra Werner

    2010-12-01

    Full Text Available A regulação da calogênese é o primeiro passo para a embriogênese somática indireta in vitro. Nesse sentido, foram investigados os efeitos de diferentes meios de cultura, reguladores de crescimento e fontes nitrogenadas no desenvolvimento de calos em tecidos foliares de Caesalpinia echinata (pau-brasil. Explantes foliares foram cultivados em meios de cultura MS, B5, White e WPM. Testou-se, também, o efeito dos compostos nitrogenados (NH4NO3, KNO3 e glutamina e a interação entre auxinas (2,4-D, AIA e AIB e citocininas (BAP e KIN na desdiferenciação celular indicado pela formação de calos. Os meios MS, B5 e White não foram significantemente diferentes e proporcionaram melhores resultados. Em meio MS contendo as fontes nitrogenadas 2,4 g L-1 NH4NO3 e 1,35 g L-1 glutamina + 4,11 g L-1 de KNO3 na presença de 1,0 mg L-1 2,4-D e 5,0 mg L-1 BAP ocorreu maior proliferação celular indicada pela massa fresca dos calos. As combinações dos reguladores 0,5 mg L-1 2,4-D + 5 mg L-1 BAP e 0,5 mg L-1 AIB + 5,0 mg L-1 BAP suplementado com 2,4 g L-1 NH4NO3 em meio MS estimularam o crescimento dos calos. De maneira geral, os calos apresentaram aspectos friáveis, não embriogênicos, com acúmulo de fenóis e presença de áreas meristemáticas (meristemóides.The regulation of callus formation is the first step to indirect somatic embryogenesis in vitro. Therefore, we investigated hormonal and nutrient factors on development of callus in leaf tissues of Caesalpinia echinata (Brazilwood. Explants were cultivated in culture media MS, B5, WPM, and White. The effect of nitrogen sources (NH4NO3, KNO3 and glutamine and the interaction between auxins (2,4-D, IAA and IBA and cytokinins (BAP and KIN were also evaluated. Media MS, B5 and White were not significantly different and provided better results. MS medium containing 2.4 g L-1 NH4NO3, 1.35 g L-1 glutamine, and 4.11 g L-1 of KNO3 in the presence of 1 mg.L-1 2,4-D and 5 mg.L-1 BAP increased cellular

  19. A Whole-Genome Microarray Study of Arabidopsis thaliana Semisolid Callus Cultures Exposed to Microgravity and Nonmicrogravity Related Spaceflight Conditions for 5 Days on Board of Shenzhou 8

    Directory of Open Access Journals (Sweden)

    Svenja Fengler

    2015-01-01

    Full Text Available The Simbox mission was the first joint space project between Germany and China in November 2011. Eleven-day-old Arabidopsis thaliana wild type semisolid callus cultures were integrated into fully automated plant cultivation containers and exposed to spaceflight conditions within the Simbox hardware on board of the spacecraft Shenzhou 8. The related ground experiment was conducted under similar conditions. The use of an in-flight centrifuge provided a 1 g gravitational field in space. The cells were metabolically quenched after 5 days via RNAlater injection. The impact on the Arabidopsis transcriptome was investigated by means of whole-genome gene expression analysis. The results show a major impact of nonmicrogravity related spaceflight conditions. Genes that were significantly altered in transcript abundance are mainly involved in protein phosphorylation and MAPK cascade-related signaling processes, as well as in the cellular defense and stress responses. In contrast to short-term effects of microgravity (seconds, minutes, this mission identified only minor changes after 5 days of microgravity. These concerned genes coding for proteins involved in the plastid-associated translation machinery, mitochondrial electron transport, and energy production.

  20. Production of solid mutants in citrus, utilizing new approaches and techniques

    International Nuclear Information System (INIS)

    Spiegel-Roy, P.; Kochba, J.

    1975-01-01

    Conditions for embryoid differentiation in Shamouti orange ovular callus were studied. Lines differing in embryogenic capacity were established. Ageing of callus prior to subculture enhanced embryoid development. A pronounced habituation effect has been found in most cultures. Protoplasts have been obtained from callus after treatment with cellolytic enzymes. Irradiation of any embryogenic line of callus with 12-20 kR seemed to promote embryoid formation, after a time lag in their appearance. Transferrence of embryoids into agar+sucrose and GA 3 and a further transfer into agar+sucrose, GA 3 and adenine sulphate gave best results as to rooting of embryoids and plant survival. The technique of using decapitated nucellar seedlings for mutagenic treatment was developed further with Shamouti orange and Marsh grapefruit. Irradiation 48-72 hours after decapitation with 2 kR resulted in shoot neoformation and survival not much below that of control, while 6 kR impeded de novo shoot formation in Marsh grapefruit. A tendency towards mutants with earlier fruit maturity was found in mV 2 plants from material originating from irradiation with the 8-kR dose. (author)

  1. Improvement of efficient in vitro regeneration potential of mature callus induced from Malaysian upland rice seed (Oryza sativa cv. Panderas).

    Science.gov (United States)

    Mohd Din, Abd Rahman Jabir; Iliyas Ahmad, Fauziah; Wagiran, Alina; Abd Samad, Azman; Rahmat, Zaidah; Sarmidi, Mohamad Roji

    2016-01-01

    A new and rapid protocol for optimum callus production and complete plant regeneration has been assessed in Malaysian upland rice (Oryza sativa) cv. Panderas. The effect of plant growth regulator (PGR) on the regeneration frequency of Malaysian upland rice (cv. Panderas) was investigated. Mature seeds were used as a starting material for callus induction experiment using various concentrations of 2,4-D and NAA. Optimal callus induction frequency at 90% was obtained on MS media containing 2,4-D (3 mg L(-1)) and NAA (2 mg L(-1)) after 6 weeks while no significant difference was seen on tryptophan and glutamine parameters. Embryogenic callus was recorded as compact, globular and light yellowish in color. The embryogenic callus morphology was further confirmed with scanning electron microscopy (SEM) analysis. For regeneration, induced calli were treated with various concentrations of Kin (0.5-1.5 mg L(-1)), BAP, NAA and 0.5 mg L(-1) of TDZ. The result showed that the maximum regeneration frequency (100%) was achieved on MS medium containing BAP (0.5 mg L(-1)), Kin (1.5 mg L(-1)), NAA (0.5 mg L(-1)) and TDZ (0.5 mg L(-1)) within four weeks. Developed shoots were successfully rooted on half strength MS free hormone medium and later transferred into a pot containing soil for acclimatization. This cutting-edge finding is unique over the other existing publishable data due to the good regeneration response by producing a large number of shoots.

  2. Relationship between endogenous hormonal content and somatic organogenesis in callus of peach (Prunus persica L. Batsch) cultivars and Prunus persica×Prunus dulcis rootstocks.

    Science.gov (United States)

    Pérez-Jiménez, Margarita; Cantero-Navarro, Elena; Pérez-Alfocea, Francisco; Le-Disquet, Isabel; Guivarc'h, Anne; Cos-Terrer, José

    2014-05-01

    The relationship between endogenous hormones content and the induction of somatic peach plant was studied. To induce multiple shoots from callus derived from the base of stem explants of the scion cultivars 'UFO-3', 'Flariba' and 'Alice Bigi', and the peach×almond rootstocks 'Garnem' and 'GF677', propagated plants were cultured on Murashige and Skoog salts augmented with 0.1mgL(-1) of indolebutyric acid, 1mgL(-1) of 6-benzylaminopurine and 3% sucrose. The highest regeneration rate was obtained with the peach×almond rootstocks. Endogenous levels of abscisic acid (ABA), indole-3-acetic acid (IAA), zeatin (Z), zeatin riboside (ZR), ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), salicylic acid (SA), and jasmonic acid (JA) were analyzed in the organogenic callus. Lower levels of several hormones, namely Z, ZR, ABA, and ACC were found in the peach×almond rootstock compared to peach cultivars, while IAA and SA presented inconclusive returns. These results suggest that the difference in somatic organogenesis capacity observed in peach and peach×almond hybrids is markedly affected by the endogenous hormonal content of the studied genotypes. Copyright © 2014 Elsevier GmbH. All rights reserved.

  3. Endochondral fracture healing with external fixation in the Sost knockout mouse results in earlier fibrocartilage callus removal and increased bone volume fraction and strength.

    Science.gov (United States)

    Morse, A; Yu, N Y C; Peacock, L; Mikulec, K; Kramer, I; Kneissel, M; McDonald, M M; Little, D G

    2015-02-01

    Sclerostin deficiency, via genetic knockout or anti-Sclerostin antibody treatment, has been shown to cause increased bone volume, density and strength of calluses following endochondral bone healing. However, there is limited data on the effect of Sclerostin deficiency on the formative early stage of fibrocartilage (non-bony tissue) formation and removal. In this study we extensively investigate the early fibrocartilage callus. Closed tibial fractures were performed on Sost(-/-) mice and age-matched wild type (C57Bl/6J) controls and assessed at multiple early time points (7, 10 and 14days), as well as at 28days post-fracture after bony union. External fixation was utilized, avoiding internal pinning and minimizing differences in stability stiffness, a variable that has confounded previous research in this area. Normal endochondral ossification progressed in wild type and Sost(-/-) mice with equivalent volumes of fibrocartilage formed at early day 7 and day 10 time points, and bony union in both genotypes by day 28. There were no significant differences in rate of bony union; however there were significant increases in fibrocartilage removal from the Sost(-/-) fracture calluses at day 14 suggesting earlier progression of endochondral healing. Earlier bone formation was seen in Sost(-/-) calluses over wild type with greater bone volume at day 10 (221%, p<0.01). The resultant Sost(-/-) united bony calluses at day 28 had increased bone volume fraction compared to wild type calluses (24%, p<0.05), and the strength of the fractured Sost(-/-) tibiae was greater than that that of wild type fractured tibiae. In summary, bony union was not altered by Sclerostin deficiency in externally-fixed closed tibial fractures, but fibrocartilage removal was enhanced and the resultant united bony calluses had increased bone fraction and increased strength. Crown Copyright © 2014. Published by Elsevier Inc. All rights reserved.

  4. Calogênese em Cissus sicyoides L. a partir de segmentos foliares visando à produção de metabólitos in vitro Calluses from Cissus sicyoides L. leaves

    Directory of Open Access Journals (Sweden)

    F.R. Rodrigues

    2010-09-01

    faz-se necessário a adição de 6,0 mg L-1 de BAP ao meio de cultivo. Identificou-se a presença de heterosídeos cardiotônicos em calos de Cissus sicyoides L.Secondary metabolites are essentially produced and extracted from plants grown in the field under influence of seasonal variations. The use of biotechnological techniques is an alternative resource for drug production. Among these techniques, tissue culture through callus genesis is highlighted, since callus growth is desirable to induce somaclonal variation and physiological studies, especially when the presence of secondary metabolites can be related to cell growth. The aim of this work was to establish a protocol for Cissus sicyoides L. callus genesis from leaf segments in order to produce metabolites in vitro. Thus, leaf segments removed from adult plants grown in the field were used as explants. After disinfestation, the material was inoculated into MT medium + 1.0 mg L-1 NAA and kept in a BOD chamber, with controlled temperature and luminosity. After 30 days, the percentage of surviving explants and the percentage of contamination were evaluated. For culture, MT medium + 1.0 mg L-1 NAA was used, varying BAP concentrations: 2.0, 4.0, 6.0 and 12.0 mg L-1. In the cultivation, the number of compact and friable calluses was counted. For the first and second subculture, the material was introduced into MT medium + 1.0 mg L-1 NAA, varying the same BAP concentrations; the number of friable calluses formed and the size of callus mass were described. The number of replicates formed during subcultures, and fresh and dry matter (g were also obtained. Then, phytochemical tests were done in order to identify some compounds. The adopted time and concentration of sodium hypochlorite proved to be inefficient for disinfestation. For Cissus sicyoides L. callus genesis from leaf segments, the addition of 6.0 mg L-1 BAP to the culture medium is needed. Cardiotonic heterosides were detected in Cissus sicyoides L. calluses.

  5. Impact of application of zinc oxide nanoparticles on callus induction, plant regeneration, element content and antioxidant enzyme activity in tomato (Solanum lycopersicum Mill. under salt stress

    Directory of Open Access Journals (Sweden)

    Alharby Hesham F.

    2016-01-01

    Full Text Available The properties of nanomaterials and their potential applications have been given considerable attention by researchers in various fields, especially agricultural biotechnology. However, not much has been done to evaluate the role or effect of zinc oxide nanoparticles (ZnO-NP in regulating physiological and biochemical processes in response to salt-induced stress. For this purpose, some callus growth traits, plant regeneration rate, mineral element (sodium, potassium, phosphorous and nitrogen contents and changes in the activity of superoxide dismutase (SOD and glutathione peroxidase (GPX in tissues of five tomato cultivars were investigated in a callus culture exposed to elevated concentrations of salt (3.0 and 6.0 g L-1NaCl, and in the presence of zinc oxide nanoparticles (15 and 30 mg L-1. The relative callus growth rate was inhibited by 3.0 g L-1 NaCl; this was increased dramatically at 6.0 g L-1. Increasing exposure to NaCl was associated with a significantly higher sodium content and SOD and GPX activities. Zinc oxide nanoparticles mitigated the effects of NaCl, and in this application of lower concentrations (15 mg L-1 was more effective than a higher concentration (30 mg L-1. This finding indicates that zinc oxide nanoparticles should be investigated further as a potential anti-stress agent in crop production. Different tomato cultivars showed different degrees of tolerance to salinity in the presence of ZnO-NP. The cultivars Edkawy, followed by Sandpoint, were less affected by salt stress than the cultivar Anna Aasa.

  6. Using Haworthia Cultured Cells as an Aid in Teaching Botany

    Science.gov (United States)

    Majumdar, Shyamal K.; Castellano, John M.

    1977-01-01

    Callus induction from species of Haworthia can be done quickly in the laboratory with minimal equipment to study tissue dedifferentiation and cellular redifferentiation. It is shown that the cultured cell can also be used to study and evaluate the effects of various mutagens, carcinogens, and pesticides in controlled environments. (Author/MA)

  7. Apple MdACS6 Regulates Ethylene Biosynthesis During Fruit Development Involving Ethylene-Responsive Factor.

    Science.gov (United States)

    Li, Tong; Tan, Dongmei; Liu, Zhi; Jiang, Zhongyu; Wei, Yun; Zhang, Lichao; Li, Xinyue; Yuan, Hui; Wang, Aide

    2015-10-01

    Ethylene biosynthesis in plants involves different 1-aminocyclopropane-1-carboxylic acid synthase (ACS) genes. The regulation of each ACS gene during fruit development is unclear. Here, we characterized another apple (Malus×domestica) ACS gene, MdACS6. The transcript of MdACS6 was observed not only in fruits but also in other tissues. During fruit development, MdACS6 was initiated at a much earlier stage, whereas MdACS3a and MdACS1 began to be expressed at 35 d before harvest and immediateley after harvest, respectively. Moreover, the enzyme activity of MdACS6 was significantly lower than that of MdACS3a and MdACS1, accounting for the low ethylene biosynthesis in young fruits. Overexpression of MdACS6 (MdACS6-OE) by transient assay in apple showed enhanced ethylene production, and MdACS3a was induced in MdACS6-OE fruits but not in control fruits. In MdACS6 apple fruits silenced by the virus-induced gene silencing (VIGS) system (MdACS6-AN), neither ethylene production nor MdACS3a transcript was detectable. In order to explore the mechanism through which MdACS3a was induced in MdACS6-OE fruits, we investigated the expression of apple ethylene-responsive factor (ERF) genes. The results showed that the expression of MdERF2 was induced in MdACS6-OE fruits and inhibited in MdACS6-AN fruits. Yeast one-hybrid assay showed that MdERF2 protein could bind to the promoter of MdACS3a. Moreover, down-regulation of MdERF2 in apple flesh callus led to a decrease of MdACS3a expression, demonstrating the regulation of MdERF2 on MdACS3a. The mechanism through which MdACS6 regulates the action of MdACS3a was discussed. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  8. Plant regeneration from petiole segments of some species in tissue culture

    Directory of Open Access Journals (Sweden)

    Krystyna Klimaszewska

    2013-12-01

    Full Text Available The regeneration ability of 21 plant species belonging to 14 families was tested. The method of tissue culture in vitro was applied, on basic MS medium with an addition of growth regulators from the auxin and cytokinin groups. From among the investigated plant groups Peperomia scandens and Caladium × hortulanum were capable of plant regeneration, Passiilora coerulea regenerated shoots, Hedera helix, Begonia glabra, Coleus blumei, Fuchsia hybrida, Passiflora suberosa and Peperomia eburnea formed callus and roots, Kalanchoe blossfeldiana, Pelargonium grandiflorum, P. peltatum, P. radula, Coleus shirensis and Magnolia soulangeana produced callus, Philodendron scandens, Rhododendron smirnovii, Hibiscus rosa-sinensis, Coprosma baueri, Cestrum purpureum and Solanum rantonnetii did not exhibit any regeneration reactions.

  9. Chemical characterization and anti-inflammatory effect of rauvolfian, a pectic polysaccharide of Rauvolfia callus.

    Science.gov (United States)

    Popov, S V; Vinter, V G; Patova, O A; Markov, P A; Nikitina, I R; Ovodova, R G; Popova, G Yu; Shashkov, A S; Ovodov, Yu S

    2007-07-01

    The pectic polysaccharide named rauvolfian RS was obtained from the dried callus of Rauvolfia serpentina L. by extraction with 0.7% aqueous ammonium oxalate. Crude rauvolfian RS was purified using membrane ultrafiltration to yield the purified rauvolfian RSP in addition to glucan as admixture from the callus, with molecular weights 300 and 100-300 kD, respectively. A peroral pretreatment of mice with the crude and purified samples of rauvolfian (RS and RSP) was found to decrease colonic macroscopic scores, the total area of damage, and tissue myeloperoxidase activity in colons as compared with a colitis group. RS and RSP were shown to stimulate production of mucus by colons of the colitis mice. RSP appeared to be an active constituent of the parent RS. The glucan failed to possess anti-inflammatory activity.

  10. Flavonoid Production, Growth and Differentiation of Stelechocarpus burahol (Bl.) Hook. F. and Th. Cell Suspension Culture.

    Science.gov (United States)

    Aini Habibah, Noor; Moeljopawiro, Sukarti; Dewi, Kumala; Indrianto, Ari

    2017-01-01

    Stelechocarpus burahol is a plant containing flavonoid compounds that have the potential for use as an antihyperuricemic for gout medication. This study was performed to assess flavonoid production, growth and cell differentiation of S. burahol in cell suspension culture. Mesocarp was planted in Murashige and Skoog (MS) medium supplemented with 7.5 mg L-1 picloram for the induction of callus. Non-embryonic callus obtained was used in the formation of cell suspension cultures. Growth of cells was determined by fresh and dry weights. During the culturing, the fresh weight, dry weight and flavonoid content were determined as a result of culture status. The growth of the S. burahol cell suspension was slow, the stationary phase occurred at 30 days. The production of flavonoids was not in line with the growth of cells and the maximum production occurred on the 15th day of the log phase. The globular-shaped cells dominated the cell suspension culture at all ages. Fluorescein diacetate (FDA) staining of cells derived from cell cultures aged for 36 days showed that some cells were still viable. The results show that flavonoid production, growth and cell differentiation of a S. burahol cell suspension culture differed according to the culture age.

  11. In vitro propagation and elemental analysis of phyllanthus nirui L.: an anti-plasmodial plant

    International Nuclear Information System (INIS)

    Adusei-Fosu, K.

    2010-12-01

    Three accessions of Phyllanthus niruri from three different localities were assessed for their fruit or seed germination in vivo and in vitro as well as embryogenic calli and subsequent embryo induction. Accessions collected from Kwabenya, Kasoa or Aburi did not germinate when nursed. However, seeds from 3, 5 or 7 days dehisced fruits germinated with 7 days having the highest percentage (68.8%) of germination when nursed in the same substrate suggesting that there was fruit wall imposed dormancy. To improve percentage germination, seeds were cultured on MS medium supplemented with 0-1.2 mg/l BAP or kinetin. At this treatment, seed cultured on MS medium supplemented with 1.2 mg/l had the highest percentage (61.1% ) of germination. Alternate regeneration via nodal cuttings cultured on MS basal medium supplemented with BAP with NAA or IBA resulted in multiple shoot induction which augurs well for rapid clonal multiplication. Additionally, regeneration via somatic embryogenesis using leaf lobe explants cultured on MS, OK W, Schenk basal medium with vitamins supplemented with 2,4-0 or Picloram resulted in only callus induction. Callus induction was high when concentrations of (0.2-1.0 mg/l) 2,4-0 were used with concentrations of (2.0-3.0 mg/l) Picloram was able to induce calli. The transfer of calli to MS medium supplemented with BAP resulted in multiple shoot regeneration. Calli also grew in size and were weighed after three series of subcultures indicating that they could be used for extraction of antiplasmodial active ingredients. Instrumental Neutron Activation Analysis (lNAA) revealed the presence of five macro and four micro nutrients but no toxic element making P. niruri useful as a medicinal plant. (au)

  12. Effect Of Explant Source And Different Medium Culture On Friable Embryogenic Callus Induction Of Four Cultivars Of Cassava Manihot Esculenta Crantz

    Directory of Open Access Journals (Sweden)

    Simplice Prosper Yandia

    2015-08-01

    Full Text Available In order to obtain Friable Embryogenic Callus FEC for protoplast isolation we have evaluated in this research the competance for Friable Embryogenic Callus FEC of four cassava cultivars M61033 Rendre Yalipe and Six-mois in media containing MS supplemented with 8mgl 24-D MS supplemented with 10 mgl BAP and GD supplemented with 12mgl picloram using apical bud AB and immature leaves lobes ILL as explants. In general in the medium GD12mgl picloram the highest efficiencies of FEC ranged from 58 to 87 and the highest score of FEC ranged from 4.2 to 5.4 with explants AB however we have observed with explants ILL the efficiencies of somatic embryos ranged form 41 to 75 and the score ranged from 4.1 to 4.4. The mediums MS28 mgl 24-D have induced with explants AB the efficiencies of FEC ranged from 43 to 57 and the score ranged from 3.1 to 3.8 however with ILL explants the efficiencies of FEC ranged from 39 to 49 and the score ranged from 2.9 to 3.7. The least FEC were observed in the medium MS210 mgl BAP with BA explants however the efficiencies ranged from 6 to 11 and the score ranged from 1.1 to 1.8. Whereas the efficiencies of FEC with ILL explants ranged from 4 to 7 and the score ranged from 0.5 to 0.8. All of four cultivars showed capability of producing FEC although their efficiency varied according to gonotype donors explants and medium taking into acount. Abbreviations GD Gressoff and Doy MS Murashige and Skoog 24-dichloro phenoxyacetic acid BAP Benzylamino-purin-Acid AB Apical Bud ILL Immature Leaves lobes

  13. Reprogramming of H3K27me3 is critical for acquisition of pluripotency from cultured Arabidopsis tissues.

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    Chongsheng He

    2012-08-01

    Full Text Available In plants, multiple detached tissues are capable of forming a pluripotent cell mass, termed callus, when cultured on media containing appropriate plant hormones. Recent studies demonstrated that callus resembles the root-tip meristem, even if it is derived from aerial organs. This finding improves our understanding of the regeneration process of plant cells; however, the molecular mechanism that guides cells of different tissue types to form a callus still remains elusive. Here, we show that genome-wide reprogramming of histone H3 lysine 27 trimethylation (H3K27me3 is a critical step in the leaf-to-callus transition. The Polycomb Repressive Complex 2 (PRC2 is known to function in establishing H3K27me3. By analyzing callus formation of mutants corresponding to different histone modification pathways, we found that leaf blades and/or cotyledons of the PRC2 mutants curly leaf swinger (clf swn and embryonic flower2 (emf2 were defective in callus formation. We identified the H3K27me3-covered loci in leaves and calli by a ChIP-chip assay, and we found that in the callus H3K27me3 levels decreased first at certain auxin-pathway genes. The levels were then increased at specific leaf genes but decreased at a number of root-regulatory genes. Changes in H3K27me3 levels were negatively correlated with expression levels of the corresponding genes. One possible role of PRC2-mediated H3K27me3 in the leaf-to-callus transition might relate to elimination of leaf features by silencing leaf-regulatory genes, as most leaf-preferentially expressed regulatory genes could not be silenced in the leaf explants of clf swn. In contrast to the leaf explants, the root explants of both clf swn and emf2 formed calli normally, possibly because the root-to-callus transition bypasses the leaf gene silencing process. Furthermore, our data show that PRC2-mediated H3K27me3 and H3K27 demethylation act in parallel in the reprogramming of H3K27me3 during the leaf-to-callus transition

  14. Progress in selection for sodium chloride, 2,4-D dichlorophenoxy acetic acid (2,4-D) and streptomycin tolerance in Citrus sinensis ovular callus lines

    International Nuclear Information System (INIS)

    Kochba, J.; Spiegel-Roy, P.

    1982-01-01

    Citrus sinensis (cultivar Shamouti) nucellar embryogenic callus lines with greatly increased tolerance to salinity (NaCl), 2,4-D and streptomycin were selected. Selected lines were found stable after removal of selection pressure. Gamma irradiation at 8-16 kR was also employed and found to speed up selections. Embryos from NaCl and 2,4-D tolerant lines also showed increased tolerance. Embryogenesis in selected lines, suppressed during selection procedures, was regained by growing cultures in the presence of galactose or lactose as the sole carbon source. A schedule was worked out furthering development of embryos into plantlets. Conditions for adventive shoot formation from embryonic shoot segments were established, thus allowing cloning of embryos. A procedure was worked out for suspension culture and agar plating of cell groups. (author)

  15. Rice improvement, involving altered flower structure more suitable to cross-pollination, using in vitro culture in combination with mutagenesis

    International Nuclear Information System (INIS)

    Min, S.K.

    1998-01-01

    Anther and somatic tissue culture in combination with mutagenesis were carried out to evaluate the efficiency of different mutagenic treatments of various in vitro culture materials, and to obtain some promising variants for rice improvement. Results indicated that in japonica rice radiation treatment of dry seeds and young panicles influenced the percentage of green plantlets regeneration from anther culture. Both treatments increased significantly the percentage of regenerated green plantlets in comparison with the control. Irradiation with 30 Gy of rice callus increased also the percentage of regenerated green plantlets. For indica rice, the combination of the suitable dose of gamma rays irradiation on seeds and an improved medium, increased the percentage of callus induction. This approach made it possible to use anther culture in indica rice breeding. Somatic tissue cultures combined with radiation-induced mutagenesis led to the development of a number of promising mutants including some new cytoplasm-nucleus interacting male-sterile lines with almost 100% stigma exertion. Their development would be of practical significance for increasing the genetic diversity for production of hybrid rice. (author)

  16. Gravitational and magnetic field variations synergize to cause subtle variations in the global transcriptional state of Arabidopsis in vitro callus cultures

    Directory of Open Access Journals (Sweden)

    Manzano Ana I

    2012-03-01

    Full Text Available Abstract Background Biological systems respond to changes in both the Earth's magnetic and gravitational fields, but as experiments in space are expensive and infrequent, Earth-based simulation techniques are required. A high gradient magnetic field can be used to levitate biological material, thereby simulating microgravity and can also create environments with a reduced or an enhanced level of gravity (g, although special attention should be paid to the possible effects of the magnetic field (B itself. Results Using diamagnetic levitation, we exposed Arabidopsis thaliana in vitro callus cultures to five environments with different levels of effective gravity and magnetic field strengths. The environments included levitation, i.e. simulated μg* (close to 0 g* at B = 10.1 T, intermediate g* (0.1 g* at B = 14.7 T and enhanced gravity levels (1.9 g* at B = 14.7 T and 2 g* at B = 10.1 T plus an internal 1 g* control (B = 16.5 T. The asterisk denotes the presence of the background magnetic field, as opposed to the effective gravity environments in the absence of an applied magnetic field, created using a Random Position Machine (simulated μg and a Large Diameter Centrifuge (2 g. Microarray analysis indicates that changes in the overall gene expression of cultured cells exposed to these unusual environments barely reach significance using an FDR algorithm. However, it was found that gravitational and magnetic fields produce synergistic variations in the steady state of the transcriptional profile of plants. Transcriptomic results confirm that high gradient magnetic fields (i.e. to create μg* and 2 g* conditions have a significant effect, mainly on structural, abiotic stress genes and secondary metabolism genes, but these subtle gravitational effects are only observable using clustering methodologies. Conclusions A detailed microarray dataset analysis, based on clustering of similarly expressed genes (GEDI software, can detect underlying global

  17. Production of Gymnemic Acid Depends on Medium, Explants, PGRs, Color Lights, Temperature, Photoperiod, and Sucrose Sources in Batch Culture of Gymnema sylvestre

    Directory of Open Access Journals (Sweden)

    A. Bakrudeen Ali Ahmed

    2012-01-01

    Full Text Available Gymnema sylvestre (R.Br. is an important diabetic medicinal plant which yields pharmaceutically active compounds called gymnemic acid (GA. The present study describes callus induction and the subsequent batch culture optimization and GA quantification determined by linearity, precision, accuracy, and recovery. Best callus induction of GA was noticed in MS medium combined with 2,4-D (1.5 mg/L and KN (0.5 mg/L. Evaluation and isolation of GA from the calluses derived from different plant parts, namely, leaf, stem and petioles have been done in the present case for the first time. Factors such as light, temperature, sucrose, and photoperiod were studied to observe their effect on GA production. Temperature conditions completely inhibited GA production. Out of the different sucrose concentrations tested, the highest yield (35.4 mg/g d.w was found at 5% sucrose followed by 12 h photoperiod (26.86 mg/g d.w. Maximum GA production (58.28 mg/g d.w was observed in blue light. The results showed that physical and chemical factors greatly influence the production of GA in callus cultures of G. sylvestre. The factors optimized for in vitro production of GA during the present study can successfully be employed for their large-scale production in bioreactors.

  18. Development of an irrigation control device based on solar radiation and its adaptability for cultivation of high soluble solid tomato fruit in root zone restriction culture

    International Nuclear Information System (INIS)

    Nitta, M.; Shibuya, K.; Kubai, K.; Komatsu, H.; Hosokawa, T.; Nakamura, K.

    2009-01-01

    An irrigation control device based on solar radiation was developed to allow automatic irrigation management for high soluble solid tomato fruit production in root zone restriction culture. Its adaptability for long-term cultivation (planting carried out in early September and harvesting ending in late June) of high soluble solid tomato fruit in root zone restriction culture was examined. The following results were obtained: 1. The control device was composed of generally available electronic parts. A change of setting was possible for the irrigation starting point, the irrigation time period, and the once amount of irrigation. For the first irrigation of the day, one of two irrigation control modes can be chosen; the first determines irrigation dependent on the solar radiation after the irrigated time of the previous day. The second mode irrigates at a set time. 2. The correlation between the total integrated solar radiation and the evapotranspiration rate of tomato plants were investigated. Positive correlations were observed for each month from October to June. Moreover, total integrated solar radiation per unit evapotranspiration was different for each month. 3. In long-term cultivation of tomato fruit using this device, the marketable yield of high soluble solid tomato fruit (more than Brix 8%) was 9.7t/10a. 4. This device exhibited the necessary adaptability for use in long-term cultivation of high soluble solid tomato fruit in root zone restriction culture, by changing the set value of the irrigation starting point and the irrigation time period in accordance with the growth period

  19. Mechanical stimulation enhanced estrogen receptor expression and callus formation in diaphyseal long bone fracture healing in ovariectomy-induced osteoporotic rats.

    Science.gov (United States)

    Chow, S K H; Leung, K S; Qin, J; Guo, A; Sun, M; Qin, L; Cheung, W H

    2016-10-01

    Estrogen receptor (ER) in ovariectomy-induced osteoporotic fracture was reported to exhibit delayed expression. Mechanical stimulation enhanced ER-α expression in osteoporotic fracture callus at the tissue level. ER was also found to be required for the effectiveness of vibrational mechanical stimulation treatment in osteoporotic fracture healing. Estrogen receptor(ER) is involved in mechanical signal transduction in bone metabolism. Its expression was reported to be delayed in osteoporotic fracture healing. The purpose of this study was to investigate the roles played by ER during osteoporotic fracture healing enhanced with mechanical stimulation. Ovariectomy-induced osteoporotic SD rats that received closed femoral fractures were divided into five groups, (i) SHAM, (ii) SHAM-VT, (iii) OVX, (iv) OVX-VT, and (v) OVX-VT-ICI, where VT stands for whole-body vibration treatment and ICI for ER antagonization by ICI 182,780. Callus formation and gene expression were assessed at 2, 4, and 8 weeks postfracture. In vitro osteoblastic differentiation, mineralization, and ER-α expression were assessed. The delayed ER expression was found to be enhanced by vibration treatment. Callus formation enhancement was shown by callus morphometry and micro-CT analysis. Enhancement effects by vibration were partially abolished when ER was modulated by ICI 182,780, in terms of callus formation capacity at 2-4 weeks and ER gene and protein expression at all time points. In vitro, ER expression in osteoblasts was not enhanced by VT treatment, but osteoblastic differentiation and mineralization were enhanced under estrogen-deprived condition. When osteoblastic cells were modulated by ICI 182,780, enhancement effects of VT were eliminated. Vibration was able to enhance ER expression in ovariectomy-induced osteoporotic fracture healing. ER was essential in mechanical signal transduction and enhancement in callus formation effects during osteoporotic fracture healing enhanced by vibration

  20. Mutation breeding in Philippine fruits

    International Nuclear Information System (INIS)

    Espino, R.R.C.

    1987-09-01

    Studies were made to establish standard conditions for mutation induction by gamma-irradiation to be performed in combination with in-vitro culture for banana and citrus spp. Besides this, radio-sensitivity of seeds and/or plantlets of mango, sugar apple, soursop, lanzones and Jack fruit was investigated and primary observation on the occurrence of mutation was made. For the mutagenesis of banana shoot tip cultures, radio-sensitivity of plantlets derived from the culture as well as fresh-cultured shoots was examined and phenotypes indicative of mutation, such as chlorophyl streaking, slow growth, pigmentation and varied bunch orientation were recorded. Isozyme analysis for mutated protein structure was not conclusive. In the in-vitro culture of Citrus spp., seeds placed on fresh media as well as germinating seeds and two-leaf stage seedlings in test tubes were examined for their radio-sensitivity. Irradiated materials were propagated for further observation. In these two crops, basic methodology for mutation induction with combined use of in-vitro culture and gamma-irradiation was established. In mango, sugar apple, soursop, lanzones and Jack fruit, basic data on radiosensitivity were obtained. In mango, leaf abnormalities were observed after the treatment of scions

  1. Isolation of lactic acid bacteria with potential protective culture characteristics from fruits

    Science.gov (United States)

    Hashim, Nurul Huda; Sani, Norrakiah Abdullah

    2015-09-01

    Lactic acid bacteria are also known as beneficial microorganisms abundantly found in fermented food products. In this study, lactic acid bacteria were isolated from fresh cut fruits obtained from local markets. Throughout the isolation process from 11 samples of fruits, 225 presumptive lactic acid bacteria were isolated on MRS agar medium. After catalase and oxidase tests, 149 resulted to fit the characteristics of lactic acid bacteria. Further identification using Gram staining was conducted to identify the Gram positive bacteria. After this confirmation, the fermentation characteristics of these isolates were identified. It was found that 87 (58.4%) isolates were heterofermentative, while the rest of 62 (41.6%) are homofermentative lactic acid bacteria. Later, all these isolates were investigated for the ability to inhibit growth of Staphylococcus aureus using agar spot assay method. Seven (4.7%) isolates showed strong antagonistic capacity, while 127 (85.2%) and 8 (5.4%) isolates have medium and weak antagonistic capacity, respectively. The other 7 (4.7%) isolates indicated to have no antagonistic effect on S. aureus. Results support the potential of LAB isolated in this study which showed strong antagonistic activity against S. aureus may be manipulated to become protective cultures in food products. While the homofermentative or heterofermentative LAB can be utilized in fermentation of food and non-food products depending on the by-products required during the fermentation.

  2. Morphological, biochemical and genetic influence of mutagen treatments on medicinal plant tissue cultures

    International Nuclear Information System (INIS)

    Onisei, T.; Toth, E.; Tesio, B.; Floria, F.

    1994-01-01

    Gamma rays and/or alkylant agents have been applied on callus tissue, young regenerants and cell suspension in order to establish their effect on morphogenesis, regeneration ability and biosynthetic potential. Growth dynamics, morpho-anatomic variables, secondary metabolite production, cell cytogenetics, enzyme specific activities, isoperoxidase and isoesterase patterns were analyzed in relation to the morphogenetic response of Atropa belladonna, Datura innoxia, Lavandula angustifolia, Chamomilla recutita, Digitalis lanata and Vinca minor tissue cultures. The effects of gamma-ray doses varied from one species to another; 10 to 20 Gy were generally able to stimulate growth and plant regeneration (via organogenesis and somatic embryogenesis), while 10 to 50 Gy enhanced secondary metabolite biosynthesis both in callus and cell suspension culture. Semnificative increase of secondary metabolite production was obtained when treatments with EMS (0.1-0.2%) have been applied to young regenerants. Many differences in biological features and biochemical behaviour were registered 20 days and one year, respectively, after treatment. (author)

  3. Nuclear morphology, polyploidy, and chromatin elimination in tissue culture of Allium fistulosum L.

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    Andrzej Joachimiak

    2011-01-01

    Full Text Available The morphology of cell nuclei in callus obtained from root-tip meristems of Allium fistulosum L. (Monocotyledoneae, Alliaceae was analysed. The most interesting phenomena observed in long-term callus culture were the different mechanisms of cell polyploidization, enlargement of telomeric segments of heterochromatin, and extensive chromatin elimination, associated with instability of nuclei size and DNA content. Protruding heterochromatin "spikes" were observed on the surface of some di- and polyploid nuclei. The presence of these spikes was connected with the formation of small heterochromatic micronuclei frequently found in the cytoplasm. It is suggested that these micronuclei are produced by direct elimination of heterochromatin from the interphase nuclei. Polyploid cells accumulated with each successive cell collection. The ploidy level attained by highly polyploid cells was 15C-220C. The shape of the nuclei and heterochromatin distribution suggest that polyploid nuclei in A. fistulosum tissue culture are produced by endoreduplication and by restitution cycles.

  4. Cell-wall polysaccharide composition and glycanase activity of Silene vulgaris callus transformed with rolB and rolC genes.

    Science.gov (United States)

    Günter, Elena A; Shkryl, Yury N; Popeyko, Oxana V; Veremeichik, Galina N; Bulgakov, Victor P

    2015-03-15

    The aim of this research is to investigate the effects of the Agrobacterium rhizogenes rol genes on the composition of cell-wall polysaccharides and glycanase activity in the campion callus. The expression of the rolC gene reduces the yield of campion pectin, while the expression of the rolB or rolC gene inhibits the volumetric production of both pectin and intracellular arabinogalactan. The rol genes are involved in regulating the activity of glycanases and esterases, thereby contributing to the modification of polysaccharide structures, their molecular weight (Mw) and the degree of pectin methyl esterification (DE). The increase in pectin arabinose residue appears to be connected to a decrease in intracellular and extracellular α-l-arabinofuranosidase activity in transgenic campion calluses. In transgenic calluses expressing the rolB and rolC genes, the increase in pectin galactose residue is likely due to a decrease in β-galactosidase activity. The decrease in the Mw of pectin and its d-galacturonic acid content appears to be connected to an increase in extracellular polygalacturonase activity. Finally, the increase in pectinesterase activity causes a decrease in the DE of pectin. Thus, the expression of rolB and rolC genes in campion callus has a considerable effect on pectin's sugar composition, DE and Mw, while it appears to have an insignificant influence on intracellular and extracellular arabinogalactans. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Thidiazuron-enhanced biosynthesis and antimicrobial efficacy of silver nanoparticles via improving phytochemical reducing potential in callus culture of Linum usitatissimum L.

    Science.gov (United States)

    Anjum, Sumaira; Abbasi, Bilal Haider

    2016-01-01

    Green synthesis of silver nanoparticles (AgNPs) by using plants is an emerging class of nanobiotechnology. It revolutionizes all domains of medical sciences by synthesizing chemical-free AgNPs for various biomedical applications. In this report, AgNPs were successfully synthesized by using whole plant extract (WPE) and thidiazuron-induced callus extract (CE) of Linum usitatissimum. The phytochemical analysis revealed that the total phenolic and flavonoid contents were higher in CE than that in WPE. Ultraviolet-visible spectroscopy of synthesized AgNPs showed a characteristic surface plasmon band in the range of 410-426 nm. Bioreduction of CE-mediated AgNPs was completed in a shorter time than that of WPE-mediated AgNPs. Scanning electron microscopy showed that both types of synthesized AgNPs were spherical in shape, but CE-mediated AgNPs were smaller in size (19-24 nm) and more scattered in distribution than that of WPE-mediated AgNPs (49-54 nm). X-ray diffraction analysis confirmed crystalline nature (face-centered cubic) of both types of AgNPs. Fourier-transform infrared spectroscopy revealed that the polyphenols and flavonoids were mainly responsible for reduction and capping of synthesized AgNPs. Energy dispersive X-ray analysis further confirmed the successful synthesis of AgNPs. Moreover, the synthesized AgNPs were found to be stable over months with no change in the surface plasmon bands. More importantly, CE-mediated AgNPs displayed significantly higher bactericidal activity against multiple drug-resistant human pathogens than WPE-mediated AgNPs. The present work highlighted the potent role of thidiazuron in in vitro-derived cultures for enhanced biosynthesis of chemical-free AgNPs, which can be used as nanomedicines in many biomedical applications.

  6. Mutation induction and isolation in potato through true seed and tuber mutagenesis and use of tissue culture

    International Nuclear Information System (INIS)

    Upadhya, M.D.; Abraham, M.J.; Dass, B.; Chandra, R.

    1982-01-01

    Advance MV generation clones from hydrazine-sulphate-treated 'O.T' cultures have been field evaluated and 12 cultures have been selected for yield trials. One culture, DN-31-3, has been found to be day-neutral in its tuberization behaviour. Four JL/RA clones have been selected after a large-scale field trial. These clones are the selections from gamma-irradiated self seeds of Kufri Lauvkar (A-7416) and hybrid A-2235. Similarly 15 MV 3 clones have been selected from the populations raised from EMS- and DES-treated self seeds of A-2235. Day-neutral mutants have been selected from the fourth to seventh sprouts taken from EMS-treated tuber halves of Kufri Jyoti. From the sixth sprout harvest from EMS-treated Kufri Jyoti halves, one mutant, BCN-6-2, has been isolated which showed less than 30 cysts of G. rostochiensis in the MV 2 generation. This clone has been multiplied and made disease-free through apical meristem culture. Through the use of a new medium, PM-32, the plating efficiency of mechanically isolated single callus cells of dihaploid PH-258 is 30-35%. Nitsch's medium has been modified to formulate a new medium for direct embryogenesis in single callus cells of Phulwa. All stages up to the globular stage could be observed after five to six weeks of culture. Efforts were continued for the enzymatic isolation of single-leaf mesophyll cells from dihaploid PH-258. A new cell separation medium has been formulated which gives 80% viable cells. The LD 50 and LD 100 doses of EMS for the single callus cells of dihaploid PH-255 were found to be 500 ppm and 1000 ppm respectively. (author)

  7. Biotechnological characteristics of callusogenesis in maize immature embryo culture under the influence of abscisic acid and 6-benzylaminopurine

    Directory of Open Access Journals (Sweden)

    O. E. Abraimova

    2010-02-01

    Full Text Available The effect of abscisic acid and 6-benzylaminopurine on the induction of callus tissue in maize immature embryo culture was studied. For the majority of investigated genotypes abscisic acid stimulated, but 6-benzylaminopurine inhibited the formation of morphogenic calli in induction medium. It was noted that genotype appeared to be an important factor that determined the character of the influence of phytohormonal composition of the medium on the induction of the specific types of calli. Using of 0.04-0.10 mg/l abscisic acid is recommended for biotechnological production of morphogenic callus tissue in dependence of donor plant genotype.

  8. Lodgepole pine: the first evidence of seed-based somatic embryogenesis and the expression of embryogenesis marker genes in shoot bud cultures of adult trees.

    Science.gov (United States)

    Park, So-Young; Klimaszewska, Krystyna; Park, Ji-Young; Mansfield, Shawn D

    2010-11-01

    Of the various alternatives for cloning elite conifers, somatic embryogenesis (SE) appears to be the best option. In recent years, significant areas of lodgepole pine (Pinus contorta) forest have been devastated by the mountain pine beetle (MPB) in Western Canada. In an attempt to establish an SE propagation system for MPB-resistant lodgepole pine, several families displaying varying levels of resistance were selected for experimentation involving shoot bud and immature seed explants. In bud cultures, eight embryogenic lines were induced from 2 of 15 genotypes following various treatments. Genotype had an important influence on embryogenic culture initiation, and this effect was consistent over time. These lines were identified by microscopic observation and genetic markers. Despite the abundance of early somatic embryos, the cultures have yet to develop into mature embryos. In contrast, immature zygotic embryos (ZEs) cultured from megagametophytes initiated SE at an early dominance stage via nodule-type callus in 1 of 10 genotypes. As part of the study, putative embryogenesis-specific genes, WOX2 (WUSCHELL homeobox 2) and HAP3A, were analyzed in cultures of both shoot bud explants and ZEs. On the basis of these analyses, we postulate that PcHAP3A was expressed mainly in callus and may be involved in cell division, whereas WOX2 was expressed mainly in embryonal mass (EM)-like tissues. The findings from this study, based on molecular assessment, suggest that the cell lines derived from bud cultures were truly EM. Moreover, these experimental observations suggest that PcWOX2 could be used as an early genetic marker to discriminate embryogenic cultures from callus.

  9. Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni Bertoni = Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni Bertoni

    Directory of Open Access Journals (Sweden)

    Ana Paula Patrão

    2007-07-01

    Full Text Available As folhas de Stevia rebaudiana (Bertoni Bertoni (Asteraceae contêm glicosídeos diterpenóides (GDS, que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS, suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D, e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM, suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN. Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDS.The leaves from Stevia rebaudiana (Bertoni Bertoni (Asteraceae contain diterpenoid glycosides (GDS, which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D; MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP and other developed in our laboratory the Woody Plant Medium (WPM with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN. The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an

  10. Regeneration of roots from callus reveals stability of the developmental program for determinate root growth in Sonoran Desert Cactaceae.

    Science.gov (United States)

    Shishkova, Svetlana; García-Mendoza, Edith; Castillo-Díaz, Vicente; Moreno, Norma E; Arellano, Jesús; Dubrovsky, Joseph G

    2007-05-01

    In some Sonoran Desert Cactaceae the primary root has a determinate root growth: the cells of the root apical meristem undergo only a few cell division cycles and then differentiate. The determinate growth of primary roots in Cactaceae was found in plants cultivated under various growth conditions, and could not be reverted by any treatment tested. The mechanisms involved in root meristem maintenance and determinate root growth in plants remain poorly understood. In this study, we have shown that roots regenerated from the callus of two Cactaceae species, Stenocereus gummosus and Ferocactus peninsulae, have a determinate growth pattern, similar to that of the primary root. To demonstrate this, a protocol for root regeneration from callus was established. The determinate growth pattern of roots regenerated from callus suggests that the program of root development is very stable in these species. These findings will permit future analysis of the role of certain Cactaceae genes in the determinate pattern of root growth via the regeneration of transgenic roots from transformed calli.

  11. Studies on the reaction in tissue culture of tomato genotypes under biotic stress

    Directory of Open Access Journals (Sweden)

    Ewa Hanus-Fajerska

    2014-01-01

    Full Text Available Plant regeneration in vitro from virus-infected somatic tomato (Lycopersicon sp. tissue was performed. Regeneration experiments were started after the determination of virus presence, using enzyme-linked immunosorbent assay, in leaves used as a source of explants. Leaf explants infected with selected strains of tomato mosaic Tobamovirus or cucumber mosaic Cucumovirus respectively, were cultured on a standarised MS agar medium to induce adventitious shoots, which were afterwards excised, rooted in vitro and cultured to plants. Explants were also screened for their ability to produce callus. Diverse effects of viral infection, ranging from stimulation to inhibition of callus formation and of morphogenesis rate, were observed. The health condition of the tissue proved to affect regeneration potential of Lycopersicon esculentum, whereas wild accesions did not react in that case so distinctly. In cultivated tomato was encountered the decline in competence to reproduce shoots adventitiously in infected tissue. There was also relationship between donor plant health condition and adventitious root formation in regenerated shoots. Experiments with short-term cultures of L. esculenum reveled also that a certain number of shoots regenerated from diseased tissue can be virus-free.

  12. Methods for suspension culture, protoplast extraction, and transformation of high-biomass yielding perennial grass Arundo donax.

    Science.gov (United States)

    Pigna, Gaia; Dhillon, Taniya; Dlugosz, Elizabeth M; Yuan, Joshua S; Gorman, Connor; Morandini, Piero; Lenaghan, Scott C; Stewart, C Neal

    2016-12-01

    Arundo donax L. is a promising biofuel feedstock in the Mediterranean region. Despite considerable interest in its genetic improvement, Arundo tissue culture and transformation remains arduous. The authors developed methodologies for cell- and tissue culture and genetic engineering in Arundo. A media screen was conducted, and a suspension culture was established using callus induced from stem axillary bud explants. DBAP medium, containing 9 µM 2,4-D and 4.4 µM BAP, was found to be the most effective medium among those tested for inducing cell suspension cultures, which resulted in a five-fold increase in tissue mass over 14 days. In contrast, CIM medium containing 13 µM 2,4-D, resulted in just a 1.4-fold increase in mass over the same period. Optimized suspension cultures were superior to previously-described solidified medium-based callus culture methods for tissue mass increase. Suspension cultures proved to be very effective for subsequent protoplast isolation. Protoplast electroporation resulted in a 3.3 ± 1.5% transformation efficiency. A dual fluorescent reporter gene vector enabled the direct comparison of the CAMV 35S promoter with the switchgrass ubi2 promoter in single cells of Arundo. The switchgrass ubi2 promoter resulted in noticeably higher reporter gene expression compared with that conferred by the 35S promoter in Arundo. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Swelling and Degradation of Calcium-Pectic Gel Particles Made of Pectins of Silene vulgaris and Lemna minor Сallus Cultures at Different Concentrations of Pectinase in an Artificial Colon Environment

    Directory of Open Access Journals (Sweden)

    Anatoly A. Shubakov

    2018-03-01

    Full Text Available Spherical calcium-pectic gel particles (CaPGPs were obtained from pectins of callus cultures (CC of campion Silene vulgaris (silenan and duckweed Lemna minor (lemnan, as well as from commercial apple and citrus pectins by the method of ionotropic gelation. We studied the morphological characteristics of the obtained gel particles and found that the largest gel particles were formed from citrus pectin (CP, and the densest gel particles were formed from lemnan pectin (LP. The swelling and degradation of CaPGPs were comparatively evaluated by incubation in a simulated gastrointestinal environment. The swelling and degradation of CaPGPs formed from pectins of the silenan and lemnan callus cultures were studied at different concentrations of pectinase (0.4, 0.8 and 1.7 mg/ml in the simulated fluid of the colon. It has been established that the CaPGPs obtained from lemnan are more resistant to degradation in the simulated colon fluid than CaPGP obtained from silenan. It was shown that the concentration of pectinase in the simulated fluid of the colon and the type of pectin affect the degradability of CaPGPs formed on the basis of pectins of callus cultures.

  14. Somatic embryogenesis in cell cultures of Glycine species.

    Science.gov (United States)

    Gamborg, O L; Davis, B P; Stahlhut, R W

    1983-08-01

    This report describes the development of procedures for the production of somatic embryos in cell cultures of Glycine species including soybean. The conditions for callus induction and initiation of rapidly growing cell suspension cultures were defined. Methods for inducing embryogenesis were tested on 16 lines of several Glycine species and cultivars of soybean. The SB-26 Culture of a G. soja gave the best results and was used in the experiments. Embryogenesis required the presence of picloram or 2,4-D. AMO 1618, CCC, PP-333 and Ancymidol enhanced the embryogenesis frequency. Plants of the G. soja (SB-26) were grown to maturity from seed-derived shoot tips. Characteristics of the plants are discussed.

  15. [18S-25S rDNA variation in tissue culture of some Gentiana L. species].

    Science.gov (United States)

    Mel'nyk, V M; Andrieiev, I O; Spiridonova, K V; Strashniuk, N M; Kunakh, V A

    2007-01-01

    18S-25S rDNA of intact plants and tissue cultures of G. acaulis, G. punctata and G. lutea have been investigated by using blot-hybridization. The decrease of rDNA amount was found in the callus cultures as compared with the plants. In contrast to other species, G. lutea showed intragenome heterogeneity of rRNA genes as well as qualitative rDNA changes in tissue culture, in particular appearance of altered repeats. The relationship between the peculiarities of rRNA gene structure and their rearrangements in in vitro culture was suggested.

  16. In vitro neoplastic transformation of plant callus tissue by γ-radiation

    International Nuclear Information System (INIS)

    Pandey, K.N.; Sabharwal, P.S.

    1979-01-01

    Tumours have been induced by γ-radiation in callus tissue derived from a monocotyledonous flowering plant, Haworthia mirabilis Haw. The transformed tissue exhibited compact texture, excessive cell proliferation and loss of capacity for organogenesis. Tumors were characterized by their ability to undergo continuous autonomous growth on minimal media in the subsequent 4 generations of subculture. In contrast, the nonirradiated control tissue grew with friable texture, required inositol or growth hormones and showed prolific differentiation of vegetative buds. (Auth.)

  17. Comparative study of antibacterial and antifugal activity of callus culture and adult plants extracts from Alternanthera maritima (Amaranthaceae Estudo comparativo da atividade antibacteriana e antifúngica de extratos obtidos da cultura de calos e da planta adulta de Alternanthera maritima (Amaranthaceae

    Directory of Open Access Journals (Sweden)

    Marcos J. Salvador

    2004-06-01

    Full Text Available The aim of this study was to evaluate the antibacterial and antifungal activity of callus culture (two different hormonal combination culture medium and adult plants (two collect extracts from Alternanthera maritima (Amaranthaceae investigating the maintenance of antimicrobial activity in vivo and in vitro. The antibacterial and antifungal activity was determined by the agar-well diffusion method against thirty strains of microorganisms including Gram-positive and Gram-negative bacteria, yeasts and dermatophytes. All the organic crude extracts studied were bioactive. Extracts of aerial parts and roots of adult plants collected during the same period of years of 1995 and 1998 (Restinga de Maricá (RJ, collect 1 and 2 inhibited the growth of several microorganisms (bacteria, yeasts and dermatophytes with inhibition halo between 6 and 20 mm. Plant cell callus culture extracts obtained from two culture conditions were also bioactive. Thus, the positive results suggest that the A. maritima extracts should be further studied to determine the bioactive chemical compounds as well as to understand the possible mechanisms of action and evaluate their toxicity looking toward a pharmaceutical employment.Neste estudo procedeu-se a avaliação da atividade antibacteriana e antifúngica dos extratos brutos de Alternanthera maritima (Amaranthaceae planta in natura de duas coletas distintas e obtidos por cultura de células buscando-se averiguar a manutenção da atividade antimicrobiana dos extratos obtidos da planta in vivo e in vitro. A ação antibacteriana e antifúngica foi determinada pelo método de difusão em ágar (técnica do poço utilizando-se trinta cepas de microrganismos indicadores (bactérias Gram-positivas e Gram-negativas, leveduras e dermatófitos. Todos os extratos obtidos com solventes orgânicos avaliados apresentaram-se bioativos com halos de inibição de 6 a 20 mm. Os extratos da planta in natura das duas coletas (Restinga de Marica

  18. A continuous culture system of direct somatic embryogenesis in microspore-derived embryos of Brassica juncea.

    Science.gov (United States)

    Prabhudesai, V; Bhaskaran, S

    1993-03-01

    An efficient culture system has been developed for repeated cycles of somatic embryogenesis in microspore-derived embryos of Brassica juncea without a callus phase. Haploid embryos produced through anther culture showed a high propensity for direct production of somatic embryos in response to 2 mgL(-1) BA and 0.1 mgL(-1) NAA. The embryogenic cultures which comprised the elongated embryonal axis of microspore-derived embryos when explanted and grown on the medium of same composition produced a large number of secondary embryos. These somatic embryos in turn underwent axis elongation and produced more somatic embryos when explanted and cultured. This cycle of repetitive somatic embryogenesis continued with undiminished vigour passage after passage and was monitored for more than a year. Somatic embryos from any passage when isolated at cotyledonary stage and grown on auxin-free medium for 5 days and then on a medium containing NAA (0.1 mgL(-1)), developed into complete plants with a profuse root system and were easily established in the soil. The cytology of the root tips of these plants confirmed their haploid nature. The total absence of callus phase makes the system ideal for continuous cloning of androgenic lines, Agrobacterium-mediated transformation and mutation induction studies.

  19. Somatic embryogenesis, scanning electron microscopy, histology and biochemical analysis at different developing stages of embryogenesis in six date palm (Phoenix dactylifera L.) cultivars.

    Science.gov (United States)

    Aslam, Junaid; Khan, Saeed Ahmad; Cheruth, Abdul Jaleel; Mujib, Abdul; Sharma, Maheshwar Pershad; Srivastava, Prem Shanker

    2011-10-01

    An efficient somatic embryogenesis system has been established in six date palm (Phoenix dactylifera L.) cultivars (Barhee, Zardai, Khalasah, Muzati, Shishi and Zart). Somatic embryogenesis (SE) was growth regulators and cultivars dependent. Friable embryogenic callus was induced from excised shoot tips on MS medium supplemented with various auxins particularly 2,4-dichlorophenoxyacetic acid (2,4-D, 1.5 mg 1(-l)). Suspension culture increased embryogenesis potentiality. Only a-naphthaleneacetic acid (NAA, 0.5 mg 1(-1)) produced somatic embryos in culture. Somatic embryos germinated and converted into plantlets in N(6)-benzyladenine (BAP, 0.75 mg 1(-l)) added medium following a treatment with thidiazuron (TDZ, 1.0 mg 1(-l)) for maturation. Scanning electron microscopy showed early stages of somatic embryo particularly, globular types, and was in masses. Different developing stages of embryogenesis (heart, torpedo and cotyledonary) were observed under histological preparation of embryogenic callus. Biochemical screening at various stages of somatic embryogenesis (embryogenic callus, somatic embryos, matured, germinated embryos and converted plantlets) of date palm cultivars has been conducted and discussed in detail. The result discussed in this paper indicates that somatic embryos were produced in numbers and converted plantlets can be used as a good source of alternative propagation. Genetic modification to the embryo precursor cell may improve the fruit quality and yield further.

  20. Medicinal mushroom Ganoderma lucidum as a potent elicitor in production of t-resveratrol and t-piceatannol in peanut calluses.

    Science.gov (United States)

    Yang, Ming-Hua; Lin, Yi-Ju; Kuo, Chang-Hsin; Ku, Kuo-Lung

    2010-09-08

    Phytoalexins t-resveratrol and t-piceatannol, the well-known health-promoting active components in plants, are secondary metabolites generated upon biotic or abiotic stresses. We have reported UV-irradiated peanut callus is a potent means to produce these compounds (J. Agric. Food Chem. 2005, 53, 3877). In this work, the effects of fungi and chemical elicitors on induction of t-resveratrol and t-piceatannol were examined. Results showed the investigated fungi Botryodiplodia theobromae and Reishi Ganoderma lucidum were generally more effective than chemical stress methyl jasmonate, salicylic acid, and sucrose. As high as 15.46+/-9.85 microg of t-resveratrol and 6.93+/-2.03 microg of t-piceatannol could be elicited in each gram of callus by sterilized G. lucidum mycelium (80 mg). Although much more sterilized G. ludicum mycelia was required to induce similar level of t-resveratrol and t-piceatannol in comparison to the sterilized B. theobromae mycelia (1 mg), uptake of the G. ludicum mycelium may provide a variety of health-promoting effects. Our findings suggest G. ludicum mycelium-treated peanut callus is a good source of bioactive components.

  1. Biosynthesis of Silver Nanoparticles Using Taxus yunnanensis Callus and Their Antibacterial Activity and Cytotoxicity in Human Cancer Cells

    Directory of Open Access Journals (Sweden)

    Qian Hua Xia

    2016-09-01

    Full Text Available Plant constituents could act as chelating/reducing or capping agents for synthesis of silver nanoparticles (AgNPs. The green synthesis of AgNPs has been considered as an environmental friendly and cost-effective alternative to other fabrication methods. The present work described the biosynthesis of AgNPs using callus extracts from Taxus yunnanensis and evaluated their antibacterial activities in vitro and potential cytotoxicity in cancer cells. Callus extracts were able to reduce silver nitrate at 1 mM in 10 min. Transmission electron microscope (TEM indicated the synthesized AgNPs were spherical with the size range from 6.4 to 27.2 nm. X-ray diffraction (XRD confirmed the AgNPs were in the form of nanocrystals. Fourier transform infrared spectroscopy (FTIR suggested phytochemicals in callus extracts were possible reducing and capping agents. The AgNPs exhibited effective inhibitory activity against all tested human pathogen bacteria and the inhibition against Gram-positive bacteria was stronger than that of Gram-negative bacteria. Furthermore, they exhibited stronger cytotoxic activity against human hepatoma SMMC-7721 cells and induced noticeable apoptosis in SMMC-7721 cells, but showed lower cytotoxic against normal human liver cells (HL-7702. Our results suggested that biosynthesized AgNPs could be an alternative measure in the field of antibacterial and anticancer therapeutics.

  2. Sustainable production of azadirachtin from differentiated in vitro cell lines of neem (Azadirachta indica)

    Science.gov (United States)

    Singh, Mithilesh; Chaturvedi, Rakhi

    2013-01-01

    Azadirachtin has high industrial demand due to its immediate application as an ecofriendly, biodegradable biopesticide and also due to its various other significant bioactivities. To date, the only commercially feasible way to produce azadirachtin is extraction from seeds, but their availability is very limited as the tree flowers only once a year and only one-third of the fruits are collected due to operational problems. Further, due to the strict out-breeding nature of the plant, the seeds are highly heterozygous, resulting in inconsistent metabolite production. Therefore, in the present study, to achieve sustainable production of azadirachtin, dedifferentiated and redifferentiated calli derived from various explants of neem—zygotic embryo, leaf and ovary—were investigated for their potential to biosynthesize azadirachtin. High-performance liquid chromatography analysis of the in vitro cell lines showed the presence of azadirachtin in all the samples tested, the content of which in cultured cells varied with explant source and cell differentiation response. The presence of azadirachtin in samples was further confirmed by positive electrospray ionization mass spectroscopy. The zygotic embryo cultures of neem accumulated much higher amounts of azadirachtin than leaf and ovary cultures. Furthermore, organized in vitro callus cultures (redifferentiated) supported higher azadirachtin biosynthesis, while unorganized callus cultures (dedifferentiated) supported the least. The maximum azadirachtin content of 2.33 mg g−1 dry weight was obtained from redifferentiated immature zygotic embryo cultures.

  3. Flavors of the city: access to regional fruit and fruit consumption in the State of Acre, Brazil

    Directory of Open Access Journals (Sweden)

    Ana Carolina Barbosa de Lima

    Full Text Available In 2010 more than 70% of the population in the Brazilian Amazon was living in urban centers. This article looks at the effect of urbanization on market availability and consumption of regional fruits in the state of Acre. The east and west region were used as proxies for urbanization, and quantitative and qualitative methods were combined in interviews with regional fruit vendors and consumers. Open markets in large cities provided a greater variety of regional fruits for purchase, yet fruit consumption was more diverse in the less urbanized west, than in the east. This pattern reveals the importance of fruit tree diversity in home gardens and urban forested fragments, as well as of non-monetary exchanges of goods as promoters of variety in fruit consumption. Findings suggest that children may be benefiting the most from this consumption. Also, certain regional fruits have gained a 'cultural marker' status and are widely consumed regardless of the urbanization rates. Nevertheless, this article demonstrates how urbanization affects the diversity of fruit consumption in different social groups, and how this process is mediated by access, income level, and health concerns.

  4. Preliminary Screening of Antioxidant and Antibacterial Activities and Establishment of an Efficient Callus Induction in Curculigo latifolia Dryand (Lemba

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    Reza Farzinebrahimi

    2016-01-01

    Full Text Available Leaf, seed, and tuber explants of C. latifolia were inoculated on MS medium supplemented with various concentrations of BAP and IBA, alone or in combinations, to achieve in vitro plant regeneration. Subsequently, antioxidant and antibacterial activities were determined from in vitro and in vivo plant developed. No response was observed from seed culture on MS media with various concentrations of PGRs. The highest percentage of callus was observed on tuber explants (94% and leaf explants (89% when cultured on MS media supplemented with IBA in combination with BAP. A maximum of 88% shoots per tuber explant, with a mean number of shoots (8.8±1.0, were obtained on MS medium supplemented with combinations of BAP and IBA (2.5 mg L−1. The best root induction (92% and mean number (7.6±0.5 from tuber explants were recorded on 2.5 mg L−1 IBA alone supplemented to MS medium. The higher antioxidant content (80% was observed from in vivo tuber. However, tuber part from the intact plant showed higher inhibition zone in antibacterial activity compared to other in vitro and in vivo tested parts.

  5. Improvement of rice anther culture and application of the technique in mutation breeding

    International Nuclear Information System (INIS)

    Chen Qiufang; Wang Cailian; Lu Yimei; Jin Wei

    2001-01-01

    The ability of callus formation and green plant regeneration was very different for different rice type and varieties in anther culture. The differentiation and regeneration of green plants were obviously improved when the rice anthers at about 30 d after culture on induction medium were irradiated with 20 Gy of γ-rays and calli were cultured on the differentiation medium containing 30 mg/L colchicines. The stimulation effect of γ-irradiation combined with colchicines was much better than that of their single use. Mutation frequency and selective efficiency in M 2 were obviously increased by application of the technique

  6. Induksi Embriogenesis Somatik dari Jaringan Endosperma Jeruk Siam (Citrus nobilis Lour. cv Simadu

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    Mia Kosmiatin

    2014-07-01

    Full Text Available ABSTRACTTriploid plants can be obtained from endosperm tissues through somatic embryogenesis regeneration. This research aimed to obtain somatic embryogenesis regeneration technique of tangerine endosperm. There were 3 experiments conducted in this research: 1 Embryogenic callus induction of tangerine endosperm. Endosperms isolated from fruits that were harvested from mother plants 11-13 weeks after anthesis and cultured on Murashige and Skoog (MS medium by modified vitamin Morel and Wetmore (MW which treated by 0.1 mg L-1 biotin, 500 mg L-1 malt extract (ME, 500 mg L-1 Casein hydrolisate (CH, 500 mg L-1 ME + 0.1 mg L-1 biotin, and 500 mg L-1CH + 0.1 mg L-1 biotin, 2 Maturation and germination of somatic embryos conducted by embryogenic callus cultured on MS medium by vitamin MW modified with addition of ABA, glutamine, and biotin, and 3 Plantlet elongation conducted on MS medium modified by MW vitamin with addition of GA3 and Kinetin. The best induction medium for embryogenic callus was modified MS enriched with 3 mg L-1 BA and 500 L-1 CH or ME, in succession 84.0 and 80.0%. The best medium for somatic embryos maturation with normal morphological plantlets (54.8% was modified MS medium without plant growth regulator with higher rate of solidified agent (from 2.5 to 3 g L-1 Phytagel. Plantlets elongation was highly (0.9 cm occurred on modified MS with enriched of 2.5 mg L-1 GA3. Keywords: Citrus nobilis (Lour., endosperm culture, in vitro, Simadu tangerine

  7. Effect of the genotype and gamma irradiation on the anther cultures of a 10x10 diallel cross of wheat

    International Nuclear Information System (INIS)

    Molina, L.; Aldana, F.

    2001-01-01

    Anther culture responsiveness, irradiation effect and reciprocal effect were evaluated on ten genotypes (V1-V10) and a 101x0 diallel cross. Gamma irradiation dose of 100 Gy was applied to seeds of parents and F 1 cross from which the donor plants were grown. Non-irradiated donor plants were also used for comparison. Anthers were plated on potato-2 callus induction medium and calli formed were transferred to MS medium supplemented with sucrose (3%), indolacetic acid (1.0 mg/L), kinetin (1.0 mg/L), inositol (100 mg/L) and solidified with agar (0.7%). Genotypes showed big differences for callus induction, plant regeneration and anther culturability rate. The most responsive materials were V2, V10 and V5 with 76.0, 27.4 and 10.8 green plants per 100 anthers respectively. No irradiation effect was found for the parents nor the F 1 crosses on the pooled data. Mean anther culture response of specific genotypes showed that irradiation significantly increased anther culturability rate of V3 from 0.1 to 27.6 green plants per 100 anthers. No reciprocal effect was observed. (author)

  8. Overexpression of a repressor MdMYB15L negatively regulates anthocyanin and cold tolerance in red-fleshed callus.

    Science.gov (United States)

    Xu, Haifeng; Yang, Guanxian; Zhang, Jing; Wang, Yicheng; Zhang, Tianliang; Wang, Nan; Jiang, Shenghui; Zhang, Zongying; Chen, Xuesen

    2018-04-14

    The cold-induced metabolic pathway and anthocyanin biosynthesis play important roles in plant growth. In this study, we identified a bHLH binding motif in the MdMYB15L protein using protein sequence analyses. Yeast two-hybrid and pull-down assays showed that MdMYB15L could interact with MdbHLH33. Overexpressing MdMYB15L in red-fleshed callus inhibited the expression of MdCBF2 and resulted in reduced cold tolerance but did not affect anthocyanin levels. Chip-PCR and EMSA analysis showed that MdMYB15L could bind the type II cis-acting element found in the MdCBF2 promoter. Overexpressing MdMYB15L in red-fleshed callus overexpressing MdbHLH33 also reduced cold tolerance and reduced MdbHLH33-induced anthocyanin biosynthesis. Knocking out the bHLH binding sequence of MdMYB15L (LBSMdMYB15L) prevented LBSMdMYB15L from interacting with MdbHLH33. Overexpressing LBSMdMYB15L in red-fleshed callus overexpressing MdbHLH33 also reduced cold tolerance and reduced MdbHLH33-induced anthocyanin biosynthesis. Together, these results suggested that an apple repressor MdMYB15L might play a key role in the cold signaling and anthocyanin metabolic pathways. Copyright © 2018 Elsevier Inc. All rights reserved.

  9. Plant germination and production of callus from the yellow hornpoppy (Glaucium flavum): the first stage of micropropagation.

    Science.gov (United States)

    Mohamed, M E; Arafa, A M; Soliman, S S; Eldahmy, S I

    2014-09-01

    The yellow hornpoppy, Glaucium flavum Cr. (Fam. Papaveraceae) is a perennial herb, distributed in the Mediterranean region, including Egypt. The plant contains many benzyl isoquinoline alkaloids from the aporphine type such as glaucine, isoboldine, 1-chelidonine, 1-norchelidonine and 3-O-methylarterenol, making it to display various medicinal activities including antitussive, anticancer, antioxidant, antimicrobial, antiviral, hypoglycemic, analgesic, antipyretic, bronchodilator and anti-inflammatory effects. The plant is now rare and endangered in the Egyptian flora due to urban sprawl. The present study looks into Glaucium flavum seeds' in vitro germination as well as the ability of the explants taken from the growing seedlings to form stable callus lines in order to enable micropropagation as a way to save the rare plant. The study also scans the production of different medicinally valuable alkaloids, particularly glaucine, in produced callus.

  10. Induction of mutations in garlic by combined use of gamma-rays and tissue culture

    International Nuclear Information System (INIS)

    Zhen Hai Rou

    1997-01-01

    Callus cultures were initiated from leaf explants of garlic on MS medium supplemented with 1.0 mg/l KIN + 1.0 mg/l IAA + 2.0 mg/l 2,4-D. Plantlets were induced from leaf calli on MS medium with 2.0 mg/l BA + 2.0 mg/l IAA. Bulblets were induced from plantlets on MS medium containing 3.0 mg/l ITA. Callus growth and plantlets induction were remarkably inhibited with irradiation doses of 8 and 10 Gy. It was found that doses of 3 and 5 Gy were suitable to induce variation. Somatic embryos were induced on MS medium supplemented with 2.0 mg/l 2,4-D + 500 mg/l casein, 1000 mg/l yeast extract and 3 to 5% sucrose. (author). 11 tabs

  11. Induction of mutations in garlic by combined use of gamma-rays and tissue culture

    Energy Technology Data Exchange (ETDEWEB)

    Rou, Zhen Hai [Horticultural Research Inst., Shanghai Academy of Agricultural Sciences, Shanghai (China)

    1997-07-01

    Callus cultures were initiated from leaf explants of garlic on MS medium supplemented with 1.0 mg/l KIN + 1.0 mg/l IAA + 2.0 mg/l 2,4-D. Plantlets were induced from leaf calli on MS medium with 2.0 mg/l BA + 2.0 mg/l IAA. Bulblets were induced from plantlets on MS medium containing 3.0 mg/l ITA. Callus growth and plantlets induction were remarkably inhibited with irradiation doses of 8 and 10 Gy. It was found that doses of 3 and 5 Gy were suitable to induce variation. Somatic embryos were induced on MS medium supplemented with 2.0 mg/l 2,4-D + 500 mg/l casein, 1000 mg/l yeast extract and 3 to 5% sucrose. (author). 11 tabs.

  12. Regeneração de plantas híbridas entre Lycopersicon esculentum e L. peruvianum a partir de calos com dois anos de cultura in vitro Regeneration of Lycopersicon esculentum X L. peruvianum hybrid plants from two year old callus culture

    Directory of Open Access Journals (Sweden)

    Walter José Siqueira

    1988-01-01

    Full Text Available Calos obtidos da cultura in vitro de embrião imaturo do cruzamento interespecífico L. esculentum x L. peruvianum, praticamente perderam a capacidade morfogenética, após dois anos de subcultura. Na tentativa de recuperação do processo de organogênese desses calos, realizaram-se dois experimentos, utilizando-se os fitorreguladores ácido indolacético (IAA e 6-benziladenina (6-BA, cujas concentrações foram combinadas em dialélicos de 5 x 5 e 3 x 3. A composição de sais minerais e vitaminas baseou-se no meio de Murashige e Skoog, adicionando-se sacarose a 3% e ágar a 0,8%, e ajustando-se o pH final dos meios de cultura para 5,5. As condições para o dialélico 5 x 5 foram fotoperíodo de 16 horas de luz a 600 lux e temperatura de 25 ± 3°C. No dialélico 3 x 3, os tratamentos foram mantidos em câmara de crescimento a 2.000 lux, sob a mesma variação de temperatura e fotoperíodo. Em cada frasco, inoculou-se um calo com cerca de 1 cm³, totalizando quinze repetições. Avaliaram-se o desenvolvimento de calos, atribuindo-se uma escala de notas de 1 a 5, e a presença de plantas (organogênese após 30 dias de cultura. Observou-se o número total de plantas por tratamento, bem como o desenvolvimento das plantas em centímetro. No dialélico 5 x 5, a organogênese foi apenas incipiente em três tratamentos, porém as melhores combinações para o desenvolvimento dos calos foram de 0,5, 2,5 e 5,0µM de IAA com 2,5µM de 6-BA. No dialélico 3 x 3, houve a indução de plantas em sete tratamentos, sendo mais eficientes 25 e 50µM de 6-BA, sem auxina. O tratamento de 0,5 e 10,0µM de IAA e 6-BA, respectivamente, permitiu simultaneamente o crescimento de calos e a regeneração de plantas. Nota-se a influência das condições ambientais de manutenção das culturas, principalmente da intensidade de luz.Callus from Lycopersicon esculentum x L. peruvianum interespecific hybrids cultured in vitro lost their morphogenetic abilities after two

  13. Tolerance of wheat genotypes to lead in in vitro culture

    Directory of Open Access Journals (Sweden)

    Šesek Stanislav

    2000-01-01

    Full Text Available We studied the effects of five lead concentrations (10-7, 10-6, 10-5, 10-4 and 10-3 M on callus growth and dry matter content in the callus tissue. Two winter wheat (Triticum aestivum L. cultivars, Balkan and Proteinka, were used to isolate mature embryos. The embryos were grown on a modified MS (Murashige and Skoog, 1962 nutrient medium to which lead in the form of Pb(NO32 was added. Calluses from the control group were grown on a lead-free medium. During cultivation, the growth of callus tissue was observed until, 30 days alter isolation, fresh callus weight and dry matter content were measured The results showed that there were significant differences between the genotypes with regard to their response to certain lead concentrations. The highest concentration (10-3 M significantly decreased fresh callus weight relative to the control- by 43% in Balkan and 22% in Proteinka. At the same lead concentration, the callus tissue dry matter content of Balkan increased significantly (23% relative to the control, while the increase of the same parameter in Proteinka was not significant (8.6% relative to the control. The lower lead concentrations had a lees pronounced effect, although the 10-6 M dose had a stimulatory effect on callus tissue growth in Balkan, while the 10-7 M one had the same effect in Proteinka. .

  14. Effects of gamma-radiation treatment in somatic cell culture of indica rice Basmati 370 selection

    Energy Technology Data Exchange (ETDEWEB)

    Shaokai, Min; Xiufang, Qi; Zhenmin, Xiong; Chengzhang, Zhao [China National Rice Research Institute, Hangzhou (China)

    1990-01-01

    Full text: Radiation treatment was applied at different doses to germinating embryo, callus, callus with green spots and callus with developing shoots. Results indicate: (1) induction rate of the callus was reduced with increased radiation dose applied to germinating embryos; (2) radiation effect on redifferentiation percentage and/or green plantlets percentage of the callus varied with different treatment dates in the order of callus treatment > treatment of callus with green spots > treatment of callus with developing shoots > treatment of germinating embryo; (3) in vitro irradiation reduced the fertility of regenerated plants, but was able to increase their exerted stigma rate and the frequency of early maturing variation in their progenies; (4) a number of promising mutants including 2 male sterile plants were obtained from the progenies of in vitro irradiation. The radiation treatment with 2.5-5.OkR to callus with green spots or shoots is a method worthy of adoption in rice improvement. (author)

  15. The Mexican Fruit Fly Eradication Programme

    Energy Technology Data Exchange (ETDEWEB)

    Reyes F, Jesus; Santiago M, Guillermo; Hernandez M, Porfirio [Comision Nacional de Sanidad Agropecuaria (Mexico)

    2000-07-01

    The goal of the Mexican Fruit Fly Eradication Programme is to control, suppress or eradicate from Mexico four species of fruit flies of economic and quarantine importance (Anastrepha ludens Loew, A. obliqua Macquart, A. serpentina Wied. and A. striata Schiner). These pests cause damage amounting to US$710 million per year. In addition to this cost, there are other expenses from pest control actions and the loss of international markets, because fruit importing countries have established stringent quarantine measures to restrict the entry of these pests. For purposes of the programme's implementation, Mexico was divided into three working zones, defined by agro-ecological characteristics, the number of fruit fly species present and the size of fruit growing regions. In addition, a cost:benefit analysis was carried out which indicated that the rate of return, in a 12-year time frame, might be as much as 33:1 in Northern Mexico, and 17:1 in the rest of the country, for an area over 100,000 hectares. Eradication technology involves: 1) surveys of pest populations by trapping and host fruit harvesting to monitor the presence and density of fruit flies, 2) reduction of pest populations applying cultural practices and using selective bait sprays, 3) mass release of sterile flies and augmentative release of parasitoids to eliminate populations and, 4) enforcement of quarantine measures to protect fruit fly free areas.

  16. The Mexican Fruit Fly Eradication Programme

    International Nuclear Information System (INIS)

    Reyes F, Jesus; Santiago M, Guillermo; Hernandez M, Porfirio

    2000-01-01

    The goal of the Mexican Fruit Fly Eradication Programme is to control, suppress or eradicate from Mexico four species of fruit flies of economic and quarantine importance (Anastrepha ludens Loew, A. obliqua Macquart, A. serpentina Wied. and A. striata Schiner). These pests cause damage amounting to US$710 million per year. In addition to this cost, there are other expenses from pest control actions and the loss of international markets, because fruit importing countries have established stringent quarantine measures to restrict the entry of these pests. For purposes of the programme's implementation, Mexico was divided into three working zones, defined by agro-ecological characteristics, the number of fruit fly species present and the size of fruit growing regions. In addition, a cost:benefit analysis was carried out which indicated that the rate of return, in a 12-year time frame, might be as much as 33:1 in Northern Mexico, and 17:1 in the rest of the country, for an area over 100,000 hectares. Eradication technology involves: 1) surveys of pest populations by trapping and host fruit harvesting to monitor the presence and density of fruit flies, 2) reduction of pest populations applying cultural practices and using selective bait sprays, 3) mass release of sterile flies and augmentative release of parasitoids to eliminate populations and, 4) enforcement of quarantine measures to protect fruit fly free areas

  17. Plant Explants Grown on Medium Supplemented with Fe3O4 Nanoparticles Have a Significant Increase in Embryogenesis

    Directory of Open Access Journals (Sweden)

    Inese Kokina

    2017-01-01

    Full Text Available Development of nanotechnology leads to the increasing release of nanoparticles in the environment that results in accumulation of different NPs in living organisms including plants. This can lead to serious changes in plant cultures which leads to genotoxicity. The aims of the present study were to detect if iron oxide NPs pass through the flax cell wall, to compare callus morphology, and to estimate the genotoxicity in Linum usitatissimum L. callus cultures induced by different concentrations of Fe3O4 nanoparticles. Two parallel experiments were performed: experiment A, where flax explants were grown on medium supplemented with 0.5 mg/l, 1 mg/l, and 1.5 mg/l Fe3O4 NPs for callus culture obtaining, and experiment B, where calluses obtained from basal MS medium were transported into medium supplemented with concentrations of NPs identical to experiment A. Obtained results demonstrate similarly in both experiments that 25 nm Fe3O4 NPs pass into callus cells and induce low toxicity level in the callus cultures. Nevertheless, calluses from experiment A showed 100% embryogenesis in comparison with experiment B where 100% rhizogenesis was noticed. It could be associated with different stress levels and adaptation time for explants and calluses that were transported into medium with Fe3O4 NPs supplementation.

  18. Efforts towards domestication of wild tomato (solanum pimpinellifolium L.) using mutation and in vitro culture techniques

    International Nuclear Information System (INIS)

    Quartey, E. K.

    2010-06-01

    antioxidant properties (90% DPPH inhibition at 75 and 100 ml volumes) compared to the control (84.4% DPPH inhibition at 100 ml). Fruits of variant line C-2*1(4) (yellow fruits) recorded higher total solids of 17.9% and the lowest pH value of 4.17 compared to 12.8% and 4.36 for fruits from un-irradiated plants. The highest amounts of total soluble solids (7%) were contained in fruits harvested from variant lines G-1 *4(4) (light red fruits) and P-2*3(29) (deep red fruits) as against 5.6% for fruits from control plants. Similarly, fruits of line X- 1*4(9) (deep red fruits) contained highest amounts of potassium (7.31 g/kg in pericarp; 4.80 g/kg in pulp + seeds), sodium (106.3 mg/kg in pericarp; 120.6 mg/kg in pulp + seeds) and manganese (10.03 mg/kg in pericarp; 39.21 mg/kg in pulp + seeds). Fruits of lines G-1*4(4), SA-2*1(27) (deep red fruits) and C-2*1(4) contained highest amounts of magnesium (0.17 g/kg in pericarp; 0.34 g/kg in pulp + seeds), calcium (0.22 g/kg in pericarp and pulp + seeds) and vanadium (0.28 mg/kg in pericarp; 0.35 mg/kg in pulp + seeds) respectively. Soft vines propagated in vivo recorded optimum performance on a substrate of 3:1:1 of topsoil, cow dung and coconut fibre respectively (91.7% shoot regeneration). Hormone-free MS basal medium supported full plantlet regeneration (100%) from nodal cuttings under in vitro culture conditions. Hypocotyls proved to be better explants compared to cotyledons for callus induction using same MS medium supplemented with phytohormones at concentrations of 1.0 mg/L BAP + 1.5 mg/L 2, 4-D under 16/8-hr light/dark after three (3) weeks in culture. Significant differences (p≤0.05) were recorded for most of the traits studied among the selected variants. These include lycopene content, total solids and total soluble solids. There were also improvements in the macro and micro elemental composition of fruits of the selected variants. Additionally lower pH value, which is a more acceptable characteristic for processing of

  19. Effect of gamma-radiations on haploid cultured cells of Datura innoxia

    International Nuclear Information System (INIS)

    Jain, R.K.; Maherchandani, N.; Sharma, D.R.; Chowdhury, V.K.

    1981-01-01

    The effects of gamma-radiations were studied in haploid cultured cells of Datura innoxia. Growth of callus cultures and shoot differentiation were stimulated at low doses (0.2 and 1.0 kR), while the higher dose (5.0 kR) was inhibitory. Root differentiation was observed only in cultures exposed to 1.0 kR dose. Enzyme activities of alpha-amylase, peroxidase, malate dehydrogenase and phosphatases, and the amounts of buffer extractable proteins were stimulated at 0.2 and 1.0 kR and inhibited at 5.0 kR. Mitotic index too decreased at 5.0 kR. Mean nuclear volume increased with increase in radiation dose, probably due to increased ploidy. (author)

  20. Induksi Ginogenesis melalui Kultur Multi Ovule Slice dan Kultur Ovary Slice Dianthus chinensis

    Directory of Open Access Journals (Sweden)

    Suskandari Kartikaningrum

    2013-10-01

    Full Text Available Callus induction was studied in five genotypes of Dianthus chinensis using 2.4 D and NAA. Calluses can be obtainedfrom unfertilized ovule culture and ovary culture. The aim of the research was to study gynogenic potential and responseof Dianthus chinensis through ovule slice and ovary slice culture for obtaining haploid plants. Five genotypes of Dianthuschinensis and five media were used in ovule slice culture and two genotypes and three medium were used in ovary culture.Flower buds in the 7th stage were incubated for the purpose of dark pre-treatment at 4 oC for one day. Ovules and ovaries wereisolated and cultured in induction medium. Cultures were incubated for the purpose of dark pre-treatment at 4 oC for seven days, followed by 25 oC light incubation. The result showed that 2.4D was better than NAA in inducing callus. Percentage of regenerated calluses were produced in V11, V13 and V15 genotypes in M7 medium (MS + 2 mg L-1 2.4D + 1 mg L-1 BAP + 30 g L-1 sucrose and M10 medium (MS + 1 mg L-1 2.4D + 1 mg L-1 BAP + 20 g L-1 sucrose. All calluses originated from ovule and ovary cultures flowered prematurely. Double haploid (V11-34 were obtained from ovule slice culture based on PER (peroksidase and EST (esterase isoenzym marker.Keywords: ovule slice culture, ovary slice culture, callus, Dianthus sp., haploid

  1. Cultivation and fruit body production of Lentinus squarrosulus Mont ...

    African Journals Online (AJOL)

    The mycelia growth of Lentinus squarrosulus culture on the leaves and bark of common fruit trees were investigated. The effect of supplementing these fruit trees with 25% each of rice bran, horse dung, poultry droppings, cow dung, fresh cassava flour and oil palm waste fiber on the mycelia growth of this fungus was also ...

  2. Delayed fracture healing and increased callus adiposity in a C57BL/6J murine model of obesity-associated type 2 diabetes mellitus.

    Directory of Open Access Journals (Sweden)

    Matthew L Brown

    Full Text Available INTRODUCTION: Impaired healing and non-union of skeletal fractures is a major public health problem, with morbidity exacerbated in patients with diabetes mellitus (DM. DM is prevalent worldwide and affects approximately 25.8 million US adults, with >90% having obesity-related type 2 DM (T2DM. While fracture healing in type 1 DM (T1DM has been studied using animal models, an investigation into delayed healing in an animal model of T2DM has not yet been performed. METHODS: Male C57BL/6J mice at 5 weeks of age were placed on either a control lean diet or an experimental high-fat diet (HFD for 12 weeks. A mid-diaphyseal open tibia fracture was induced at 17 weeks of age and a spinal needle was used for intra-medullary fixation. Mice were sacrificed at days 7, 10, 14, 21, 28, and 35 for micro-computed tomography (μCT, histology-based histomorphometry and molecular analyses, and biomechanical testing. RESULTS: HFD-fed mice displayed increased body weight and impaired glucose tolerance, both characteristic of T2DM. Compared to control mice, HFD-fed mice with tibia fractures showed significantly (p<0.001 decreased woven bone at day 28 by histomorphometry and significantly (p<0.01 decreased callus bone volume at day 21 by μCT. Interestingly, fracture calluses contained markedly increased adiposity in HFD-fed mice at days 21, 28, and 35. HFD-fed mice also showed increased PPARγ immunohistochemical staining at day 14. Finally, calluses from HFD-fed mice at day 35 showed significantly (p<0.01 reduced torsional rigidity compared to controls. DISCUSSION: Our murine model of T2DM demonstrated delayed fracture healing and weakened biomechanical properties, and was distinctly characterized by increased callus adiposity. This suggests altered mesenchymal stem cell fate determination with a shift to the adipocyte lineage at the expense of the osteoblast lineage. The up-regulation of PPARγ in fracture calluses of HFD-fed mice is likely involved in the proposed

  3. Enhanced production of vanillin flavour metabolites by precursor feeding in cell suspension cultures of Decalepis hamiltonii Wight & Arn., in shake flask culture.

    Science.gov (United States)

    Matam, Pradeep; Parvatam, Giridhar; Shetty, Nandini P

    2017-12-01

    The flavour rich tuberous roots of Decalepis hamiltonii are known for its edible and medicinal use and have become endangered due to commercial over-exploitation. Besides 2-Hydroxy-4-methoxy benzaldehyde (2H4MB), other flavour metabolites in tuberous roots include vanillin, 4-Methoxy Cinnamic acid derivatives, aromatic alcohols etc. So far, there are no reports on the pathway of 2H4MB biosynthesis nor there is an organized work on biotransformation using normal and cell suspension cultures for obtaining these metabolites using precursors. The main aim of the study is to develop a method for enhanced production of flavour attributing metabolites through ferulic acid (FA) feeding to the D. hamiltonii callus culture medium. Biomass of D. hamiltonii cell suspension cultures was maximum (200.38 ± 1.56 g/l) by 4th week. Maximum production of 2H4MB was recorded on 4th week (0.08 ± 0.01 mg/100 g dry weight) as quantified by HPLC. Addition of 0.1-1.5 mM ferulic acid as precursor in the culture medium showed significant ( p  vanillin, 2H4MB, vanillic acid, ferulic acid were of 0.1 ± 0.02 mg/100 g, 0.44 ± 0.01 mg/100 g, 0.52 ± 0.04 mg/100 g, 0.18 ± 0.02 mg/100 g DW respectively in 4 weeks of cultured cells supplemented with 1 mM ferulic acid as a precursor. The results indicate that, substantial increase in the levels of flavour metabolites in D. hamiltonii callus suspension culture was achieved. This would be having implications in biosynthesis of respective vanilla flavour attributing metabolites at very high levels for their large scale production.

  4. Mutation breeding in sugar-cane (Saccharum sp. hybrid) by gamma irradiation of cuttings and tissue cultures

    International Nuclear Information System (INIS)

    Siddiqui, S.H.; Javed, M.

    1982-01-01

    The sugar-cane variety Co 547, which is highly susceptible to smut disease (Ustilago scitaminea Syd.) and is also late maturing, was exposed to different doses of gamma radiation to study its radiosensitivity and to induce smut disease resistance and early maturity. The radiosensitivity of the variety showed that an optimum dose (LD 50 ) was 2.0 kR and the working dose range was found to be 1.5-3.0 kR, whereas doses higher than 4.0 kR drastically affected the growth and germination. A broad spectrum of variability in reaction to disease resistance was observed after radiation exposure. This facilitated the isolation of disease-resistant mutants. Twenty-three mutants showing varied reaction to smut under field infection conditions were tested for two years by artificial inoculation using the dip method. From these studies 15 stable mutants were isolated. Of 15 mutants, seven showed promising performance in cane yield and sucrose contents. The tissue culture technique was used to determine the potential of different commercial clones for callusing. In test explants callusing was achieved readily and proliferation of callus was fairly good in all the clones. (author)

  5. Parameters necessary for in vitro hydroponic pea plantlet flowering and fruiting

    Science.gov (United States)

    To date, the development of culture systems to stimulate the reproductive and fruiting processes in vitro has not been earnestly exploited. Nonetheless, the ability to promote sterile flowering and fruiting would have commercial applications in the agricultural, food and medical industries. Non-ir...

  6. Penicillium digitatum metabolites on synthetic media and citrus fruits

    DEFF Research Database (Denmark)

    Ariza, M.R.; Larsen, Thomas Ostenfeld; Petersen, Bent O.

    2002-01-01

    Penicillium digitatum has been cultured on citrus fruits and yeast extract sucrose agar media (YES).Cultivation of fungal cultures on solid medium allowed the isolation of two novel tryptoquivaline-like metabolites, tryptoquialanine A (1) and tryptoquialanine B (2), also biosynthesized on citrus...

  7. Induction and Analysis of the Alkaloid Mitragynine Content of a Mitragyna speciosa Suspension Culture System upon Elicitation and Precursor Feeding

    Directory of Open Access Journals (Sweden)

    Nor Nahazima Mohamad Zuldin

    2013-01-01

    Full Text Available This study aimed to determine the effects of different concentrations and combinations of the phytohormones 2,4-dichlorophenoxy acetic acid (2,4-D, kinetin, 6-benzylaminopurine (BAP, and 1-naphthaleneacetic acid (NAA on callus induction and to demonstrate the role of elicitors and exogenous precursors on the production of mitragynine in a Mitragyna speciosa suspension culture. The best callus induction was achieved from petiole explants cultured on WPM that was supplemented with 4 mg L−1 2, 4-D (70.83%. Calli were transferred to liquid media and agitated on rotary shakers to establish Mitragyna speciosa cell suspension cultures. The optimum settled cell volume was achieved in the presence of WPM that contained 3 mg L−1 2,4-D and 3% sucrose (9.47±0.4667 mL. The treatment of cultures with different concentrations of yeast extract and salicylic acid for different inoculation periods revealed that the highest mitragynine content as determined by HPLC was achieved from the culture treated with 250 mg L−1 yeast extract (9.275±0.082 mg L−1 that was harvested on day 6 of culturing; salicylic acid showed low mitragynine content in all concentrations used. Tryptophan and loganin were used as exogenous precursors; the highest level of mitragynine production was achieved in cultures treated with 3 μM tryptophan and harvested at 6 days (13.226±1.98 mg L−1.

  8. Synthesis and absolute configurations of the cytotoxic polyacetylenes isolated from the callus of Panax ginseng.

    Science.gov (United States)

    Fujimoto, Y; Satoh, M; Takeuchi, N; Kirisawa, M

    1990-06-01

    Panaxacol (1) and dihydropanaxacol (2), cytotoxic polyacetylenes isolated from the callus of Panax ginseng, were synthesized starting from D-(-)-diethyl tartrate. The absolute configuration of 1 was determined to be 9R, 10R and the absolute configuration at C-3 of 2 was tentatively assigned as 3S by the application of the R(+)-alpha-methoxy-alpha-(trifluoro methyl)phenylacetyl (MTPA) method.

  9. FY 1998 industrial technology R and D project. Report on the results of the development of utilization technology of biological resource such bioconsortia system (Development of production technology of biological use petroleum substituting fuels); 1998 nendo sangyo gijutsu kenkyu kaihatsu jigyo. Fukugo seibutsukeinado seibutsu shigen riyo gijutsu kaihatsu (seibutsu riyo sekiyu daitai nenryo seizo gijutsu no kaihatsu)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1996-03-01

    Technology development was conducted for production of useful substances using the bioconsortia system. In FY 1998, the development of element technology was studied. As to the utilization technology of gut symbiotic microorganisms such as termite and longhorn beetle, clone types of gut microorganism complex system were analyzed to find out the diversification. In the decay of wood, co-culture of two species of mold fungus was increased in efficiency than single culture. More than 90% was classified/identified of the mold fungi accumulated (in wood piece)/separated for the utilization of plant symbiotic microorganisms. For the production of petroleum substituting useful resource, conditions were established of callus induction from immature embryos of tropical oil crops, especially oil palm, and of regeneration of a lot of small plants from the callus. To establish the Agrobacterium-mediated transformation system of oil palm, the binary vector harboring the reporter gene and selectable marker gene was constructed. Using the vector, oil palm tissues are inoculated and infected with agrobacteria. To heighten the function of palm oil, 10 particular clones were selected from the complementary DNA library obtained from oil palm fruit tissues. The genetic study of germs was also made. (NEDO)

  10. Effect of different in vitro culture extracts of black pepper (Piper nigrum L.) on toxic metabolites-producing strains.

    Science.gov (United States)

    Ahmad, Nisar; Abbasi, Bilal Haider; Fazal, Hina

    2016-03-01

    In the present study, the effect of different in vitro cultures (callus, in vitro shoots) and commercially available peppercorn extract was investigated for its activity against toxic metabolite-producing strains (Escherichia coli, Pseudomonas aeroginosa, Salmonella typhi, Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Candida albicans). These in vitro cultures were extracted with ethanol, hexane, and chloroform, and the antipathogenic activity was determined by well-diffusion method. Hexane extract of callus showed 22 mm zone of inhibition against B. cereus, 23 mm against S. aureus, while regenerated shoots and seeds have shown 24.3 and 26 mm zones of inhibition. The ethanolic extracts of regenerated Piper shoots have shown 25 mm activity against S. aureus, 21 mm against B. cereus, and 16 mm in the case of C. albicans in comparison with standard antibiotics. Peppercorn extracts in chloroform and ethanol had shown activities against B. cereus (23.6 mm) and B. subtilis (23.5 mm). During in vitro organogenesis and morphogenesis, cells and tissues produced a comparable phytochemicals profile like mother plant. Morphogenesis is critically controlled by the application of exogenous plant-growth regulators. Such addition alters the hormonal transduction pathways, and cells under in vitro conditions regenerate tissues, which are dependant on the physiological state of cells, and finally enhance the production of secondary metabolites. To the best of our knowledge, this is the first report to compare the antimicrobial potential of in vitro regenerated tissues and peppercorn with standard antibiotics. In conclusion, most of the extracts showed pronounced activities against all the pathogenic microbes. This is a preliminary work, and the minimum inhibitory concentration values needs to be further explored. Regenerated tissues of P. nigrum are a good source of biologically active metabolites for antimicrobial activities, and callus culture presented itself as

  11. Constitutive, Institutive and Up-Regulation of Carotenogenesis Regulatory Mechanism via In Vitro Culture Model System and Elicitors

    International Nuclear Information System (INIS)

    Rashidi Othman; Fatimah Azzahra Mohd Zaifuddin; Norazian Mohd Hassan

    2015-01-01

    Phyto hormone abscisic acid (ABA) plays a regulatory role in many physiological processes in plants and is regulated and controlled by specific key factors or genes. Different environmental stress conditions such as water, drought, cold, light, and temperature result in increased amounts of ABA. The action of ABA involves modification of gene expression and analysis of in vitro callus model system cultures revealed several potential of constitutive, institutive and up-regulation acting regulatory mechanisms. Therefore, this study was aimed at establishing in vitro cultures as potential research tools to study the regulatory mechanisms of the carotenoid biosynthesis in selected plant species through a controlled environment. The presence and absence of zeaxanthin and neoxanthin in callus cultures and intact plants could be explained by changes in gene expression in response to stress. Abiotic stress can alter gene expression and trigger cellular metabolism in plants. This study suggested that the key factors which involved in regulatory mechanisms of individual carotenoid biosynthesis in a particular biology system of plants can be either be silenced or activated. Therefore, based on the results in this study environmental stress is made possible for enhancement or enrichment of certain carotenoid of interest in food crops without altering the genes. (author)

  12. Radiation effect in another culture rice (Oryza Sativa L.) variety Krispo-38

    International Nuclear Information System (INIS)

    Montepeque, R.; Molina, L.G.; Lopez, J.J.; Pazos, W.; Ramirez, J.

    1993-01-01

    Seeds of the rice variety Krispo-38 were irradiated with 0, 100, 200, 300 and 400 Gray (Gy) and sown in the greenhouse. From each treatment, others containing uninucleate pollen grains were collected and cultured on a N6 agar medium with 5% sucrose concentration and supplemented with mg/l naphthalene acetic acid (NAA), 1 mg/l kinetin and 1 mg/l 2,4 dichlorophenoxyacetic acid (2,4-D). The pollen grains were induced to develop callus. The percentage of others that produced calli varied from 0.8 for the 400-Gy treatment to 3.3% for the control. The calli were transferred to N6 medium with 3% sucrose concentration, supplemented with 0.5 mg/l NAA and 1 mg/l kinetin. The percentage of calli that produced green plants varied from 1.9 for the 300-Gy treatment to 10.5 for the 200 Gy treatment. Plants developed in 2-5 weeks after callus transplant. A total of 101 green plants was obtained

  13. p Effe prolife ect of s eration cultur season and s res of H nal coll ...

    African Journals Online (AJOL)

    SAM

    2014-08-07

    Aug 7, 2014 ... of rubber tree occurred both in summer and rainy season. In case of mangosteen, .... of flowering ( and 43.75%. ected from d collected in 3 allus (Figure from the first d white in colo nal collectio om culturing oliferate on M. 0.2 mg.L-1 N callus from a ..... rare and endangered oak species of Greece. Plant Cell ...

  14. Plant regeneration in wheat mature embryo culture

    African Journals Online (AJOL)

    Kamil Haliloğlu

    2011-11-09

    Nov 9, 2011 ... Success in genetic engineering of cereals depends on the callus formation and efficient plant regeneration system. Callus formation and plant regeneration of wheat mature embryos ... compiled by modification of methods previously mentioned in ..... of more and readily available nutrition than artificial cul-.

  15. Terminology of pollination biology applied to fruit culture.

    Directory of Open Access Journals (Sweden)

    Douglas de Almeida Pereira

    2009-03-01

    Full Text Available The aim of this work was to familiarize a target audience with the terminology of scientific communication in reproductive biology with emphasis on pollination in fruit trees. This is fundamental to the professional who aims to develop and publish technical and scientific writings in this important area of research. To this end, a glossary of the usual terms employed in scientific journals and research institutes is presented.

  16. The effects of cellulase on capsaicin production in freely suspended cells and immobilized cell cultures of capsicum annuum

    International Nuclear Information System (INIS)

    Islek, C.

    2014-01-01

    The effect of different concentrations of cellulase on the production of capsaicin in freely suspended cell and immobilized cell cultures of Kahramanmara pepper seeds (Capsicum annuum L.) were studied. Calluses were obtained from in vitro germinated hypocotyl explants of pepper seedlings and cell suspensions were prepared from these calluses. Immobilized cell suspension cultures with calcium alginate and free cell suspension cultures were obtained by using cell suspensions. Elicitor such as cellulase (5-30 micro g/ml), was applied both for the free and immobilized cell suspensions and control group without elicitor was prepared. The concentration of capsaicin in freely suspended cells, immobilized cells and their filtrates were identified by HPLC after extraction with ethyl acetate. It was found that the immobilization process had an increasing effect on the capsaicin accumulation. The concentration of capsaicin in the immobilized cells for both control groups and elicitor added samples was higher than the free cells. In general, capsaicin concentration in the filtrate for free cells was higher than the immobilized cells. When all the cellulase and the sampling hours were compared, the highest capsaicin concentration for the immobilized cells was determined as 362,91 micro g/ml f.w. at the 24th hour for 30 micro g/ml cellulase applied samples. (author)

  17. Isolation and culture of protoplast from leaves of Lactuca sativa

    Directory of Open Access Journals (Sweden)

    Witool Chaipakdee

    2007-07-01

    Full Text Available Protoplasts were isolated from leaves of lettuce (Lactuca sativa L. seedlings after in vitro germination for 25, 30, 40 and 50 days. The leaves were stripped and incubated in various combinations of cellulase and pectinase. Protoplasts were cultured on MS medium containing various kinds and concentrations of plant growth regulators in different culture systems including liquid media, hanging, drop culture and solid media. Results revealed that the highest number of viable protoplasts, 14.1x105 cells per gram of fresh weight, was obtained from 30 day-old leaves of lettuce seedlings and isolated by using 2% cellulase in combination with 1% pectinase. Liquid MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA promoted the highest cell division up to 17.67%. First division of protoplasts was observed at 4 days after culture and microcolony formation occurred at the 4th week after culturing. Unfortunately, neither callus formation nor plantlet regeneration were obtained.

  18. Study of the agroindustrial alterations induced by the irradiated tissue culture in sugar cane, variety NA 56-79

    International Nuclear Information System (INIS)

    Figueiredo Junior, O.

    1991-01-01

    The use of plant tissue culture and the application of gamma radiation as mutation inducing agents, in the sugar cane plant, variety NA 5679, are studied. The variation in the contents of brix, pol, fiber, purity, extraction, phosphorus, nitrogen, reducing sugars as well as the morphological characteristics are analysed. The 'callus' obtained by the tissue culture were irradiated with 20, 40, and 60 Gy doses. The statistical analysis indicated that the method of tissue culture may, eventually, increase the contents of the technological parameters and the dosages of gamma radiation were not efficient for such purpose. (M.A.C.)

  19. Cultural contrast

    Institute of Scientific and Technical Information of China (English)

    周志

    2016-01-01

    Chinese cultural contains a great number of styles;culture differentiation does not depend on region differentiation.This research would interpret what difference between Hong Kong and Shenzhen.1.Food culture in china Traditional Chinese medicine suggests eating local seasonal fruit and vegetables,as they are most suitable for the body during a particular season.It is also divided food into 3 characteristics:cooling foods,warming foods and balance or

  20. Anther development stage and gamma radiation effects on tomato anther-derived callus formation; Efeitos do estadio de desenvolvimento da antera e da radiacao gama na formacao de calos derivados de anteras de tomate

    Energy Technology Data Exchange (ETDEWEB)

    Brasileiro, Ana Christina R.; Willadino, Lilia [Universidade Federal Rural de Pernambuco, Recife, PE (Brazil). Lab. de Cultura de Tecidos Vegetais. E-mail: lilia@truenet.com.br; Guerra, Marcelo [Pernambuco Univ., Recife, PE (Brazil). Dept. de Botanica. Lab. de Citogenetica Vegetal; Colaco, Waldeciro [Pernambuco Univ., Recife, PE (Brazil). Dept. de Energia Nuclear. Lab. de Radioagronomia; Meunier, Isabelle [Universidade Federal Rural de Pernambuco, Recife, PE (Brazil). Dept. de Engenharia Florestal; Camara, Terezinha R. [Universidade Federal Rural de Pernambuco, Recife, PE (Brazil). Dept. de Quimica. Lab. de Cultura de Tecidos Vegetais

    1999-12-01

    Two experiments were carried (I) to determine tomato anther development stage influence on callus production; and (II) to investigate gamma radiation effects on anther culture. In the first experiment, anthers of a tomato hybrid (IPA 5 x Rotam 4-F{sub 1}) were grown on three media. Although calli were induced at all stages of anther development, varying from prophase I to mono nucleate microspore, callus frequency decreased as anther development progressed and calli induction were not significantly affected by all media tested. Anthers containing prophase I meiocytes produced the highest calli frequency. Anther and flower bud length both were significantly correlated with anther development stage. In the second experiment, seed and floral buds of tomato hybrids IPA 5 x Rotam 4 (F{sub 2}), IPA 6 x Rotam 4 (F{sub 2}) and IPA 8 x 217.1 (F{sub 2}) were submitted to gamma-ray and anthers were plated on two media described by Gresshoff and Doy (1972) supplemented with 2.0 mg L{sup -1} NAA + 5.0 mg L{sup -1} KIN and 2.0 mg L{sup -1} NAA + 1.0 mg L{sup -1} KIN. No significant differences for genotype and dosage testes were found for calli formation. (author)

  1. Isolation, Culture and Characterization of Hirsutella sinensis Mycelium from Caterpillar Fungus Fruiting Body.

    Science.gov (United States)

    Ko, Yun-Fei; Liau, Jian-Ching; Lee, Chien-Sheng; Chiu, Chen-Yaw; Martel, Jan; Lin, Chuan-Sheng; Tseng, Shun-Fu; Ojcius, David M; Lu, Chia-Chen; Lai, Hsin-Chih; Young, John D

    2017-01-01

    The caterpillar fungus Ophiocordyceps sinensis (previously called Cordyceps sinensis) has been used for centuries in Asia as a tonic to improve health and longevity. Recent studies show that O. sinensis produces a wide range of biological effects on cells, laboratory animals and humans, including anti-fatigue, anti-infection, anti-inflammatory, antioxidant, and anti-tumor activities. In view of the rarity of O. sinensis fruiting bodies in nature, cultivation of its anamorph mycelium represents a useful alternative for large-scale production. However, O. sinensis fruiting bodies harvested in nature harbor several fungal contaminants, a phenomenon that led to the isolation and characterization of a large number of incorrect mycelium strains. We report here the isolation of a mycelium from a fruiting body of O. sinensis and we identify the isolate as O. sinensis' anamorph (also called Hirsutella sinensis) based on multi-locus sequence typing of several fungal genes (ITS, nrSSU, nrLSU, RPB1, RPB2, MCM7, β-tubulin, TEF-1α, and ATP6). The main characteristics of the isolated mycelium, including its optimal growth at low temperature (16°C) and its biochemical composition, are similar to that of O. sinensis fruiting bodies, indicating that the mycelium strain characterized here may be used as a substitute for the rare and expensive O. sinensis fruiting bodies found in nature.

  2. Isolation, Culture and Characterization of Hirsutella sinensis Mycelium from Caterpillar Fungus Fruiting Body.

    Directory of Open Access Journals (Sweden)

    Yun-Fei Ko

    Full Text Available The caterpillar fungus Ophiocordyceps sinensis (previously called Cordyceps sinensis has been used for centuries in Asia as a tonic to improve health and longevity. Recent studies show that O. sinensis produces a wide range of biological effects on cells, laboratory animals and humans, including anti-fatigue, anti-infection, anti-inflammatory, antioxidant, and anti-tumor activities. In view of the rarity of O. sinensis fruiting bodies in nature, cultivation of its anamorph mycelium represents a useful alternative for large-scale production. However, O. sinensis fruiting bodies harvested in nature harbor several fungal contaminants, a phenomenon that led to the isolation and characterization of a large number of incorrect mycelium strains. We report here the isolation of a mycelium from a fruiting body of O. sinensis and we identify the isolate as O. sinensis' anamorph (also called Hirsutella sinensis based on multi-locus sequence typing of several fungal genes (ITS, nrSSU, nrLSU, RPB1, RPB2, MCM7, β-tubulin, TEF-1α, and ATP6. The main characteristics of the isolated mycelium, including its optimal growth at low temperature (16°C and its biochemical composition, are similar to that of O. sinensis fruiting bodies, indicating that the mycelium strain characterized here may be used as a substitute for the rare and expensive O. sinensis fruiting bodies found in nature.

  3. Oxygen regulation of uricase and sucrose synthase synthesis in soybean callus tissue is exerted at the mRNA level

    DEFF Research Database (Denmark)

    Xue, Z T; Larsen, K; Jochimsen, B U

    1991-01-01

    The effect of lowering oxygen concentration on the expression of nodulin genes in soybean callus tissue devoid of the microsymbiont has been examined. Poly(A)+ RNA was isolated from tissue cultivated in 4% oxygen and in normal atmosphere. Quantitative mRNA hybridization experiments using nodule...

  4. Influence of auxins combinations on accumulation of reserpine in the callus of Rauvolfia tetraphylla L.

    Science.gov (United States)

    Anitha, S; Kumari, B D Ranjitha

    2007-11-01

    Reserpine is a monoterpene indole alkaloid used to treat hypertension because of its hypotensive property and psychiatric disorders because of its tranquilizing effect. Protocol has been standardized to enhance the synthesis of reserpine in leaf derived calli of Rauvolfia tetraphylla L. by adjusting the auxins combinations in the medium consisting of MS nutrient salts and B5 vitamins. Auxins such as naphthalene acetic acid (NAA), indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) were used in 1-5 microM concentration along with 9 microM concentration of 2,4 dichlorophenoxy acetic acid (2,4-D), which was found suitable for callus induction. The combination of (2,4-D) with NAA had been proved to accumulate maximum amount of reserpine followed by 2,4-D with IBA. The IAA with 2,4-D combination yielded very less amount of reserpine than the other combinations and 9 microM 2,4-D alone. The results suggest that there may be synergetic effect of NAA with 2,4-D and IBA with 2,4-D for increase in the biomass and reserpine accumulation and antagonistic effect of IAA with 2,4-D for the above said factors in the callus.

  5. Influence of silicon on production and fruit quality of strawberry

    Directory of Open Access Journals (Sweden)

    Maria Ligia Souza Silva

    2013-12-01

    Full Text Available In Brazil, the culture of strawberries is an important activity for small and medium farmers. Among the important factors for culture is fertilization, which can interfere in production and fruit quality. Silicon (Si is one of the little studied in fertilization, which demonstrated some beneficial to several different cultures. The objective of this study was to evaluate the influence of Si on production and concentration of anthocyanin and titratable acid in the fruit and reading SPAD (chlorophyll in the leaves of the strawberry. The experiment was conducted in a greenhouse using pots containing 14.5 kg of limed soil, fertilized and three plants (cultivar Milsei Tudla were transplanted to pots. Treatments consisted of application of Si at 0, 12.5, 25, 50 and 100 mg kg-1 rates, via soil or foliar. During this experiment the reading of SPAD (chlorophyll and the end of the experiment, the shoots was collected, dried, weighed, ground and analyzed by content and accumulation Si. The fruits were weighed and analyzed the concentration of anthocyanin, titratable acidity and Si concentration. The fertilization with Si, via soil or foliar applied, contributed to the increase in fruit production, especially with the application via soil. Applying foliar or soil promoted an increase in the values of titratable acidity and anthocyanins. Rates of Si via soil or foliar applied influenced the reading SPAD (chlorophyll, in the leaves. The Si independent of the form of application promoted improvements in strawberry crop.

  6. Evaluation of Posterolateral Lumbar Fusion in Sheep Using Mineral Scaffolds Seeded with Cultured Bone Marrow Cells

    Directory of Open Access Journals (Sweden)

    María D. Cuenca-López

    2014-12-01

    Full Text Available The objective of this study is to investigate the efficacy of hybrid constructs in comparison to bone grafts (autograft and allograft for posterolateral lumbar fusion (PLF in sheep, instrumented with transpedicular screws and bars. Hybrid constructs using cultured bone marrow (BM mesenchymal stem cells (MSCs have shown promising results in several bone healing models. In particular, hybrid constructs made by calcium phosphate-enriched cells have had similar fusion rates to bone autografts in posterolateral lumbar fusion in sheep. In our study, four experimental spinal fusions in two animal groups were compared in sheep: autograft and allograft (reference group, hydroxyapatite scaffold, and hydroxyapatite scaffold seeded with cultured and osteoinduced bone marrow MSCs (hybrid construct. During the last three days of culture, dexamethasone (dex and beta-glycerophosphate (β-GP were added to potentiate osteoinduction. The two experimental situations of each group were tested in the same spinal segment (L4–L5. Spinal fusion and bone formation were studied by clinical observation, X-ray, computed tomography (CT, histology, and histomorphometry. Lumbar fusion rates assessed by CT scan and histology were higher for autograft and allograft (70% than for mineral scaffold alone (22% and hybrid constructs (35%. The quantity of new bone formation was also higher for the reference group, quite similar in both (autograft and allograft. Although the hybrid scaffold group had a better fusion rate than the non-hybrid scaffold group, the histological analysis revealed no significant differences between them in terms of quantity of bone formation. The histology results suggested that mineral scaffolds were partly resorbed in an early phase, and included in callus tissues. Far from the callus area the hydroxyapatite alone did not generate bone around it, but the hybrid scaffold did. In nude mice, labeled cells were induced to differentiate in vivo and monitored

  7. EVALUATING THE ACCUMULATION TREND OF L-DOPA IN DARK-GERMINATED SEEDS AND SUSPENSION CULTURES OF Phaseolus vulgaris L. BY AN EFFICIENT UV-SPECTROPHOTOMETRIC METHOD

    Directory of Open Access Journals (Sweden)

    Samira Rahmani-Nezhad

    Full Text Available Seed germination and plant cell cultures provide an alternative mean for producing secondary metabolites. The present study is an attempt to evaluate the effect of seed dark germination and some elicitors and precursors on the production of L-DOPA in Phaseolus vulgaris L. Callus cultured on Murashige and Skoog medium supplemented with various concentrations of different plant growth regulators. L-DOPA produced was quantified by UV-spectrophotometric method. In this study, a user-friendly, quick, and economical UV-spectrophotometric method was described to determine L-DOPA content in extracts from 33 biotypes of Phaseolus vulgaris L. The method is based on the nitrosation of L-DOPA to form a yellow solution and then formation of a red solution by adding base which is measurable at 470 nm. According to our statistical studies, this method showed high efficiency and selectivity for quantitative determination of L-DOPA in herbal extracts from dried plant seeds, dark-germinated seeds and callus cultures. L-DOPA content in dark-germinated seeds and suspension cultures increased significantly to approximately several-fold compared to the control. The implication from this study is that elicitor treatment and precursor feeding of Phaseolus vulgaris L. can significantly improve the parkinson’s relevant L-DOPA content.

  8. Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

    NARCIS (Netherlands)

    Wang, Yue; Kronenburg-van de Ven, van B.C.E.; Menzel, T.R.; Maliepaard, C.A.; Shen, X.; Krens, F.A.

    2012-01-01

    To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in

  9. Influence of electric conductivity management on greenhouse tomato yield and fruit quality

    OpenAIRE

    Dorai , Martine; Papadopoulos , Athanasios; Gosselin , André

    2001-01-01

    International audience; The osmotic and ionic effects of the electrical conductivity (EC) of the nutrient solution and its interactions with climatic factors and cultural practices on tomato yield and fruit quality are reviewed. Adjusting the salinity of the nutrient solution allows growers to modify water availability to the crop and hence improve fruit quality. At some point, however, increases in salinity limit marketable yield. Under high ECs, fruit size is inversely related to EC while t...

  10. Biochemical markers of embryogenesis in tissue cultures of the maize inbred B73

    International Nuclear Information System (INIS)

    Everett, N.P.; Wach, M.J.; Ashworth, D.J.

    1985-01-01

    Stable embryogenic, organogenic and undifferentiated cell lines of the maize (Zea mays L.) inbred B73 were used to assess the value of using isozyme analyses and the composition of secreted polysaccharides to identify embryogenic cells. Esterase, glutamate dehydrogenase, alcohol dehydrogenase and β-glucosidase all possessed developmentally regulated isozymes but only esterase and glutamate dehydrogenase could be used to distinguish between embryogenic and shoot-forming cultures. Embryogenic callus and suspension cultures secreted a mucilagenous polysaccharide whose production was stimulated by 2, 4-dichlorophenozyacetic acid (2, 4-D). The polysaccharide was different from root slime and corn hull gum and may be related to the 'cementing layer' in maize kernels (author)

  11. Growing Arabidopsis in vitro: cell suspensions, in vitro culture, and regeneration.

    Science.gov (United States)

    Barkla, Bronwyn J; Vera-Estrella, Rosario; Pantoja, Omar

    2014-01-01

    An understanding of basic methods in Arabidopsis tissue culture is beneficial for any laboratory working on this model plant. Tissue culture refers to the aseptic growth of cells, organs, or plants in a controlled environment, in which physical, nutrient, and hormonal conditions can all be easily manipulated and monitored. The methodology facilitates the production of a large number of plants that are genetically identical over a relatively short growth period. Techniques, including callus production, cell suspension cultures, and plant regeneration, are all indispensable tools for the study of cellular biochemical and molecular processes. Plant regeneration is a key technology for successful stable plant transformation, while cell suspension cultures can be exploited for metabolite profiling and mining. In this chapter we report methods for the successful and highly efficient in vitro regeneration of plants and production of stable cell suspension lines from leaf explants of both Arabidopsis thaliana and Arabidopsis halleri.

  12. Mutagenic treatments towards increasing the frequency of day-neutral mutations and standardization of procedures for tissue culture, in potato

    International Nuclear Information System (INIS)

    Upadhya, M.D.; Chandra, R.; Abraham, M.J.

    1976-01-01

    Various chemical mutagens and gamma radiation have been used on single dormant eyes and true seeds with a view to finding effective mutagenic treatment for the induction of day-length neutral mutants in potato using an effective screening technique for the isolation of day-length neutral mutants. Sodium meta bisulphite (SMS) was found to be an efficient mutagen in inducing mutations for this trait in true seeds although the same concentrations, when used for treating the single tuber eyes proved lethal. Pre-soaking the seeds for 24 hrs prior to treatment with 0.0025M SMS gave highest frequency of the mutants followed by 48 hrs presoaking, indicating a sensitive stage during the cell cycle in true seeds. Other mutagen treatments gave different frequencies of mutations. The highest frequency of day-length neutral mutants was observed when seeds irradiated with 40 Kr of gamma radiation were treated with 0.05M hydrazinium dichloride solution. Screening procedures have also been standardised with the development of synethetic media for the isolation of biochemical mutants at the true seed level. Initial efforts have yielded mutants resistant to LD 100 doses of ethionine. Another aspect of the study was to develop a proper potato callus culture technique. A medium has been developed to produce and maintain callus from potato leaf strips. Efforts on the regeneration of shoot and roots from callus, have so far lead to differentiation of callus to form roots. The ultimate aim of these studies is to develop plantlets from single cell which would form the units of mutation induction and isolation. (author)

  13. Factors influencing Agrobacterium-mediated embryogenic callus transformation of Valencia sweet orange (Citrus sinensis) containing the pTA29-barnase gene.

    Science.gov (United States)

    Li, D D; Shi, W; Deng, X X

    2003-12-01

    Valencia sweet orange (Citrus sinensis (L.) Osbeck) calluses were used as explants to develop a new transformation system for citrus mediated by Agrobacterium tumefaciens. Factors affecting Agrobacterium-mediated transformation efficiency included mode of pre-cultivation, temperature of cocultivation and presence of acetosyringone (AS). The highest transformation efficiency was obtained with a 4-day pre-cultivation period in liquid medium. Transformation efficiency was higher when cocultivation was performed for 3 days at 19 degrees C than at 23 or 28 degrees C. Almost no resistant callus was obtained if the cocultivation medium lacked AS. The transformation procedure yielded transgenic Valencia plants containing the pTA29-barnase gene, as verified by PCR amplification and confirmed by Southern blotting. Because male sterility is a common factor leading to seedlessness in citrus cultivars with parthenocarpic characteristics, production of seedless citrus genotypes by Agrobacterium-mediated genetic transformation is a promising alternative to conventional breeding methods.

  14. Organ and plantlet regeneration of Menyanthes trifoliata through tissue culture

    Directory of Open Access Journals (Sweden)

    Urszula Adamczyk-Rogozińska

    2014-01-01

    Full Text Available The conditions for the regeneration of plants through organogenesis from callus tissues of Menyanthes trifoliata are described. The shoot multiplication rate was affected by basal culture media, the type and concentration of cytokinin and subculture number. The best response was obtained when caulogenic calli were cultured on the modified Schenk and Hildebrandt medium (SH-M containing indole-3-acetic acid (IAA 0,5 mg/l and 6-benzyladenine (BA 1 mg/l or zeatin (2 mg/l. Under these conditions ca 7 shoots (mostly 1 cm or more in length per culture in the 5th and 6th passages could be developed. In older cultures (after 11-12 passages there was a trend for more numerous but shorter shoot formation. All regenerated shoots could be rooted on the SH-M medium supplemented with 0.5 mg/l IAA within 6 weeks; 80% of in vitro rooted plantlets survived their transfer to soil.

  15. [Induction and in vitro culture of hairy roots of Dianthus caryophyllus and its plant regeneration].

    Science.gov (United States)

    Shi, Heping; Zhu, Yuanfeng; Wang, Bei; Sun, Jiangbing; Huang, Shengqin

    2014-11-01

    To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.

  16. Nonsterol Triterpenoids as Major Constituents of Olea europaea

    Science.gov (United States)

    Stiti, Naïm; Hartmann, Marie-Andrée

    2012-01-01

    Plant triterpenoids represent a large and structurally diverse class of natural products. A growing interest has been focused on triterpenoids over the past decade due to their beneficial effects on human health. We show here that these bioactive compounds are major constituents of several aerial parts (floral bud, leaf bud, stem, and leaf) of olive tree, a crop exploited so far almost exclusively for its fruit and oil. O. europaea callus cultures were analyzed as well. Twenty sterols and twenty-nine nonsteroidal tetra- and pentacyclic triterpenoids belonging to seven types of carbon skeletons (oleanane, ursane, lupane, taraxerane, taraxastane, euphane, and lanostane) were identified and quantified by GC and GC-MS as free and esterified compounds. The oleanane-type compounds, oleanolic acid and maslinic acid, were largely predominant in all the organs tested, whereas they are practically absent in olive oil. In floral buds, they represented as much as 2.7% of dry matter. In callus cultures, lanostane-type compounds were the most abundant triterpenoids. In all the tissues analyzed, free and esterified triterpene alcohols exhibited different distribution patterns of their carbon skeletons. Taken together, these data provide new insights into largely unknown triterpene secondary metabolism of Olea europaea. PMID:22523691

  17. Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni Bertoni - DOI: 10.4025/actascihealthsci.1090 Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni Bertoni - DOI: 10.4025/actascihealthsci.1090

    Directory of Open Access Journals (Sweden)

    Regina Aparecida Correia Gonçalves

    2008-03-01

    Full Text Available As folhas de Stevia rebaudiana (Bertoni Bertoni (Asteraceae contêm glicosídeos diterpenóides (GDS, que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS, suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D, e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM, suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN. Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDSThe leaves from Stevia rebaudiana (Bertoni Bertoni (Asteraceae contain diterpenoid glycosides (GDS, which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D; MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP and other developed in our laboratory the Woody Plant Medium (WPM with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN. The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an

  18. Effect of γ-ray irradiation on in vitro culture and plant regeneration of alfalfa

    International Nuclear Information System (INIS)

    Zhang Xiaodong; Lin Tingan

    1992-01-01

    60 Co γ-ray irradiation ranged 0-16 kR was used to treat the cotyledons and hypocotyls of 5 cultivars of a alfalfa (Medicago sativa L.). The effects of irradiation on the frequency of callus, fresh weight of callus, the frequency of somatic embryo induction and plantlet regeneration were studied. The results showed as follows: the radiosensitivities of cotyledon was significantly higher than that of hypocotyl. Exposure of 2 kR could improve the growth of callus, embryogenesis and plantlet regeneration at various levels. Exposure of 12 kR completely inhibited the callus formation from explants of cotyledon. Callus from two cultivars, England 648 and Jining alfalfa, were irradiated with 0-8 kR. The results showed that exposure below 2 kR could promote the growth of callus and the formation of somatic embryo at different levels. The exposure of 8 kR had the effect of lethal. The optimum exposure for genetic improvement of alfalfa in vitro by inducing mutation was considered to be 4-6 kR

  19. Effect of gamma irradiation and ethyl methane sulphonate on flavour formation in garlic (Allium sativum L. ) cultures

    International Nuclear Information System (INIS)

    Malpathak, N.P.; David, S.B.

    1990-01-01

    In callus cultures both gamma irradiation and EMS treatments produced stimul ation of flavour formation at lower as well as at higher dosages. Different results were obtained when the plantlets differentiated from both the treatments were analysed. There was a gradual increase in flavour formation with increasing dosages, which declined at a higher dosages. Cytological observations showed an increase in polyploidy with increasing dosages. (author). 5 refs., 5 tabs

  20. A new approach for in vitro regeneration of tomato plants devoid of exogenous plant growth hormones.

    Science.gov (United States)

    Plana, Dagmara; Fuentes, Alejandro; Alvarez, Marta; Lara, Regla M; Alvarez, Félix; Pujol, Merardo

    2006-10-01

    Many available methodologies for in vitro regeneration of commercial tomato varieties promote not only the production of normal shoots but also individual leaves, shoots without apical meristems and vitrified structures. All these abnormal formations influence and diminish the regeneration efficiency. At the basis of this phenomenon lies callus development. We optimized an alternative procedure by which the regeneration occurs without abnormal shoot formation. The portion including the proximal part of hypocotyls and the radicle was cultured on medium consisting of Murashige and Skoog salts, 4 mg/L thiamine, 100 mg/L mio-inositol and 3% sucrose. After two-three weeks, 60% explants showed adventitious shoot formation. No changes in the morphological characteristics of regenerated plants and fruits were observed as compared with parents. Karyotypic analysis of regenerated plants showed no variations in chromosome number. The optimized procedure offers the advantage of tomato plant regeneration avoiding callus formation, which enables normal plant recovery with an efficiency ranging from 1.45 +/- 0.05 to 2.57 +/- 0.06 shoots per explant in Campbell-28, Amalia, Lignon, and Floradel cultivars.

  1. Protocols for Callus and Somatic Embryo Initiation for Hibiscus sabdariffa L. (Malvaceae): Influence of Explant Type, Sugar, and Plant Growth Regulators

    Science.gov (United States)

    A significant work on callus induction and somatic embryogenesis was realized for Hibiscus sabdariffa. Two genotypes (Hibiscus sabdariffa and Hibiscus sabdariffa var. altissima) two sugars (sucrose and glucose) and three concentrations (1 %, 2%, 3%) of each sugar, 3 explant types (root, hypocotyl, c...

  2. Morphological and Molecular Identification of Colletotrichum acutatum from Tomato Fruit

    Directory of Open Access Journals (Sweden)

    Svetlana Živković

    2010-01-01

    Full Text Available Colletotrichum gloeosporioides, Colletotrichum acutatum, Colletotrichum coccodes, and Colletotrichum dematium are the four main species of Colletotrichum that cause tomato anthracnose. In Serbia, the occurrence of anthracnose on tomato fruit has been recorded during the last several years. Typical fruit symptoms include dark, sunken, and circular lesion with orange conidial masses. Pathogen isolates were obtained from a diseased tomato fruits, on PDA medium forming a white to gray colonies. The cultures developed black acervuli around the center of the colony. Conidia were hyaline, aseptate, and fusiform or rarely cylindrical. Appressoria were smooth, simple, clavate to ovate, and variedfrom light to dark brown. Pathogenicity tests with representative isolates were conducted on symptomless, detached tomato fruits. All tested isolates caused anthracnose lesions on tomato fruit after 7 days of incubation. Koch’s postulates were fulfilled by reisolationfrom inoculated tomato fruits. PCR analysis (using species-specific primer pair, CaInt2/ITS4 of genomic DNA from tomato isolates resulted in an amplification product of 490 bp, specific for C. acutatum, further confirming the identity of the pathogen. Based onmorphological and molecular characteristics, the isolates from tomato fruit were determined as C. acutatum.

  3. Cloning higher plants from aseptically cultured tissues and cells

    Science.gov (United States)

    Krikorian, A. D.

    1982-01-01

    A review of aseptic culture methods for higher plants is presented, which focuses on the existing problems that limit or prevent the full realization of cloning plants from free cells. It is shown that substantial progress in clonal multiplication has been made with explanted stem tips or lateral buds which can be stimulated to produce numerous precocious axillary branches. These branches can then be separated or subdivided and induced to root in order to yield populations of genetically and phenotypically uniorm plantlets. Similarly, undifferentiated calluses can sometimes be induced to form shoots and/or roots adventitiously. Although the cell culture techniques required to produce somatic embryos are presently rudimentary, steady advances are being made in learning how to stimulate formation of somatic or adventive embryos from totipotent cells grown in suspension cultures. It is concluded that many problems exist in the producing and growing of totipotent or morphogenetically competent cell suspensions, but the potential benefits are great.

  4. Low-magnitude high-frequency vibration enhances gene expression related to callus formation, mineralization and remodeling during osteoporotic fracture healing in rats.

    Science.gov (United States)

    Chung, Shu-Lu; Leung, Kwok-Sui; Cheung, Wing-Hoi

    2014-12-01

    Low magnitude high frequency vibration (LMHFV) has been shown to improve anabolic and osteogenic responses in osteoporotic intact bones and during osteoporotic fracture healing; however, the molecular response of LMHFV during osteoporotic fracture healing has not been investigated. It was hypothesized that LMHFV could enhance osteoporotic fracture healing by regulating the expression of genes related to chondrogenesis (Col-2), osteogenesis (Col-1) and remodeling (receptor activator for nuclear factor- κ B ligand (RANKL) and osteoproteger (OPG)). In this study, the effects of LMHFV on both osteoporotic and normal bone fracture healing were assessed by endpoint gene expressions, weekly radiographs, and histomorphometry at weeks 2, 4 and 8 post-treatment. LMHFV enhanced osteoporotic fracture healing by up-regulating the expression of chondrogenesis-, osteogenesis- and remodeling-related genes (Col-2 at week 4 (p=0.008), Col-1 at week 2 and 8 (p<0.001 and p=0.008) and RANKL/OPG at week 8 (p=0.045)). Osteoporotic bone had a higher response to LMHFV than normal bone and showed significantly better results as reflected by increased expression of Col-2 and Col-1 at week 2 (p<0.001 for all), larger callus width at week 2 (p=0.001), callus area at week 1 and 5(p<0.05 for all) and greater relative area of osseous tissue (p=0.002) at week 8. This study helps to understand how LMHFV regulates gene expression of callus formation, mineralization and remodeling during osteoporotic fracture healing. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  5. SPECTRAL QUALITY AFFECTS MORPHOGENESIS ON ANTHURIUM PLANTLET DURING IN VITRO CULTURE

    Directory of Open Access Journals (Sweden)

    Kurniawan Budiarto

    2010-10-01

    Full Text Available This paper elucidates the effects of LEDs spectral on callus induction, proliferation and shoot development of anthurium plantlet derived from leaf explants. The research was conducted at the Ornamental Research Station, Fukuyama, Japan from January to August 2008. Three experimental series were designed to determine the effects of LED-based spectral compositions i.e. 100% red, 75% red + 25% blue, 50% red + 50% blue, 25% red + 75% blue and 100% blue LEDs on morphogenetic process of callus formation derived from leaf explants up to plantlet formation on two anthurium cultivars, Violeta and Pink Lady. The results showed no differences among cultivars tested but interaction of factors studied were found in all parameters observed. LEDs spectral gave significant influence on the morphogenetic processes from callus induction to complete plantlet formation. Progressive initial callus was promoted with the decrease of blue LEDs portion. Conversely, to proliferate globose to torpedo callus formation, more blue light was required than red LEDs. During shoot induction and formation, hastened shoot initiation and number of shoots were achieved in higher blue LEDs portions, but not in root formations.

  6. Quantitative transient GUS expression in J-104 rice calli through manipulation of in vitro culture conditions.

    Directory of Open Access Journals (Sweden)

    Maylin Pérez Bernal

    2009-10-01

    This paper purposes suitable conditions for callus induction and co-cultivation with Agrobacterium tumefaciens of J-104 rice cultivar. It was evaluated the effect of different concentrations of 2.4-D and agar, and the inclusion of L-proline and L-glutamine in callus culture medium. The use of 2.5 mg/L 2.4-D and 0.8% agar allowed the highest percentage of embryogenic calli. Callus formation was improved considerably with 500 mg/L of L-proline and L-glutamine in the culture medium. Different factors were studied throughout co-cultivation of calli with A. tumefaciens: inoculation time, co-cultivation temperature, concentration of acetosyringone and co-cultivation period. Transient GUS expression was quantified by fluorometry in all co-cultivated calli. The best results were obtained with the following conditions: 10 min as inoculation time, 100µM acetosyringone in co-cultivation medium, temperature of 20ºC, and 3 days as co-cultivation period. Key words: Agar; callus; co-cultivation; fluorometric GUS activity. Resumen Se describen las condiciones óptimas para la callogénesis y cocultivo de callos con Agrobacterium tume-faciens de la variedad de arroz J-104. Se determinó el efecto de diferentes concentraciones de 2.4-D, agar y de L-prolina y L-glutamina en el medio de cultivo de callos. El uso de 2,5 mg/L de 2.4-D y 0,8% de agar permitió lograr el porcentaje más alto de callos embriogénicos. La formación de callos fue mejorada considerablemente con la adición de 500 mg/L de L-prolina e igual concentración de L-glutamina en el medio de cultivo. Se estudiaron diferentes factores en el cocultivo de los callos con A. tumefaciens: tiempo de inoculación, concentración de acetosiringona, temperatura y tiempo de cocultivo. Para comparar el efecto de cada factor sobre la expresión GUS se cuantificó la actividad transitoria mediante fluorimetría. Los valores más altos de actividad fluorimétrica fueron obtenidos con las siguientes condiciones: 10 min de

  7. Successful selling of packed fresh fruit and vegetable

    Directory of Open Access Journals (Sweden)

    Radmila Presová

    2009-01-01

    Full Text Available The paper deals with the marketability of packaged fruit and vegetables, as a new trend of satisfying consumer needs, in connection with selling-culture of stall selling and self-stores sales. It emphasize fact that food have to be wrapped into the quality, health packages, under the conditions of the Act. 477/2001 Coll.Specific data for the analytical part of this paper was provided by Hortim International Ltd. This company has operate on the Czech market since 1995. Analysis of the economic results confirmes the legitimacy of the sale of packaged fresh fruits and vegetables, but there will be neccesary longer lasting promotion. This fact also confirmed the research of consumer demand, that was made by authors. Determinant for the success sales are these factors: consumer confidence that the packaged products of fruit and vegetables are high quality.

  8. The use of Citrus tissue culture for mutation breeding. Effects of plant growth substances and gamma irradiation on embryogenesis

    International Nuclear Information System (INIS)

    Kochba, J.; Spiegel-Roy, P.

    1976-01-01

    An embryogenic callus subcultured from unfertilized ovules of the 'Shamouti' orange (Citrus sinensis) was established and is used for mutation-breeding. The callus is habituated and lines of differing embryogenic potential were established. The effect of growth substances and of gamma-irradiation on embryogenesis were studied. Auxins and cytokinins inhibited embryogenesis while inhibitors of auxin synthesis and a cytokinin antagonist significantly stimulate embryogenesis in an embryogenic line. A non-embryogenic callus line did not respond to these treatments. Stimulation of embryogenesis was observed when callus but not when the medium was irradiated. Age of callus prior to subculture and irradiation intensities modify irradiation induced embryogenesis by changing optimal dose range and radiosensitivity of the callus. Addition of IAA to unirradiated medium resulted in increased embryogenesis and greatly stimulated plantlet development in certain combinations of irradiation dose and IAA concentration. (author)

  9. In vitro regeneration of 'Feizixiao' litchi (Litchi chinensis Sonn.)

    African Journals Online (AJOL)

    Tuoyo Aghomotsegin

    Different developmental stages of SEs were obtained from the callus on MS ... methods. Key words: litchi (Litchi chinensis Sonn.), anther culture, callus, regeneration, somatic embryogenesis. ...... Induction and maintenance of the embryogenic.

  10. Bacterial cellulose production by Gluconacetobacter xylinus by employing alternative culture media.

    Science.gov (United States)

    Jozala, Angela Faustino; Pértile, Renata Aparecida Nedel; dos Santos, Carolina Alves; de Carvalho Santos-Ebinuma, Valéria; Seckler, Marcelo Martins; Gama, Francisco Miguel; Pessoa, Adalberto

    2015-02-01

    Bacterial cellulose (BC) is used in different fields as a biological material due to its unique properties. Despite there being many BC applications, there still remain many problems associated with bioprocess technology, such as increasing productivity and decreasing production cost. New technologies that use waste from the food industry as raw materials for culture media promote economic advantages because they reduce environmental pollution and stimulate new research for science sustainability. For this reason, BC production requires optimized conditions to increase its application. The main objective of this study was to evaluate BC production by Gluconacetobacter xylinus using industry waste, namely, rotten fruits and milk whey, as culture media. Furthermore, the structure of BC produced at different conditions was also determined. The culture media employed in this study were composed of rotten fruit collected from the disposal of free markets, milk whey from a local industrial disposal, and their combination, and Hestrin and Schramm media was used as standard culture media. Although all culture media studied produced BC, the highest BC yield-60 mg/mL-was achieved with the rotten fruit culture. Thus, the results showed that rotten fruit can be used for BC production. This culture media can be considered as a profitable alternative to generate high-value products. In addition, it combines environmental concern with sustainable processes that can promote also the reduction of production cost.

  11. PHYTOCHEMICAL STUDY OF CELL CULTURE JATROPHA CURCAS

    Directory of Open Access Journals (Sweden)

    KOMAR RUSLAN

    2011-01-01

    Full Text Available Jatropha curcas belongs to the Euphorbiaceae family which has potential economically. This plant has been reported to contain toxic compounds such as curcin and phorbol ester and its derivatives. These compounds may become a problem if J. curcas will be explored as a source of biofuel. In order to provide safety plants, the research on the study of phytochemical and initiation of cell and organ culture have been carried out. J curcas which has been collected from different regions in Indonesia showed to contain relatively the same profile of chemical contents. Dominant compounds that were detected by GCMS are hidrocarbon such as 2-heptenal, decadienal, hexsadecane, pentadecane, cyclooctane etc, fatty acid such as oktadecanoate acid, etthyl linoleate, ethyl stearate, heksadecanoate acid and steroid such as stigmasterol, fucosterol, sitosterol. No phorbol ester and its derivatives have been detected yet by the GCMS method. Callus and suspension cultures of J. curcas have been established to be used for further investigation.

  12. Iodine uptake and distribution in horticultural and fruit tree species

    Directory of Open Access Journals (Sweden)

    Alessandra Caffagni

    2012-07-01

    Full Text Available Iodine is an essential microelement for humans and iodine deficiency disorder (IDD is one of the most widespread nutrient-deficiency diseases in the world. Iodine biofortification of plants provides an attractive opportunity to increase iodine intake in humans and to prevent and control IDD. This study was conducted to investigate the iodine uptake and accumulation in edible portion of two fruit trees: plum and nectarine, and two horticultural crops: tomato and potato. Two type of iodine treatments (soil and foliar spray application, and, for fresh market tomato, two production systems (open field and greenhouse hydroponic culture were tested. The distribution of iodine in potato stem and leaves, and in plum tree fruits, leaves, and branches was investigated. Iodine content of potato tubers after postharvest storage and processing (cooking, and iodine content of nectarine fruits after postharvest storage and processing (peeling were also determined. Differences in iodine accumulation were observed among the four crops, between applications, and between production systems. In open field, the maximum iodine content ranged from 9.5 and 14.3 μg 100 g−1 for plum and nectarine fruit, to 89.4 and 144.0 μg 100 g−1 for potato tuber and tomato fruit, respectively. These results showed that nectarine and plum tree accumulated significantly lower amounts of iodine in their edible tissues, in comparison with potato and tomato. The experiments also indicated hydroponic culture as the most efficient system for iodine uptake in tomato, since its fresh fruits accumulated up to 2423 μg 100 g−1 of iodine. Iodine was stored mainly in the leaves, in all species investigated. Only a small portion of iodine was moved to plum tree branches and fruits, and to potato stems and tubers. No differences in iodine content after fruit peeling was observed. A significant increase in iodine content of potato was observed after baking, whereas a significant decrease was

  13. Obtaining unique large kernel rice using chemical mutagenesis in tissue culture

    International Nuclear Information System (INIS)

    Alyoshin, N.E.; Avakyan, E.R.; Alyoshin, E.P.

    2001-01-01

    Full text: Lines with improved characters have been received by chemical mutagenesis in rice tissue culture. The japonica rice (Oryza sativa L.) varieties 'Krasnodarskii 424', 'Dubovskii 129', 'Slavyanetz', 'Liman', 'Lomello', 'VNIIR 2471' were used for mutation induction. Nnitrozo-N-methylurea (MNH) has been used as a mutagen. Two approaches were applied: 1. Development mutants by mutagenic treatment of seeds 2. Development regenerants from somatic tissue culture. In the first case, dry seeds with removed covering glumes have been treated with a solution of NMH (exposure 24 hours, tested concentrations 0.05%; 0.1%; 0.2%). After treatment seeds have been rinsed and planted into the soil in vessels. The effect of mutagen was very much genotype dependant. The highest frequency of mutants were observed in the following concentrations of MNH: for variety VNIIR 2471 - 0.05-0.1%, for variety Slavyanetz - 0.1%; for Lomello - 0.2%; for Linman - 0.05% and 0.2%. The mutant N 95, which has been selected from variety Liman after treatment with 0.2% concentration of mutagen, had the following improved characters: vegetation period 103 days (110 days for the parent variety); plant height 93.2 cm (98.2 cm - parent variety); length of the main panicle 17.2 cm; 1000 grain mass 44.9 g (39.2 g - parent variety). Mutant line N 101 selected from the same variety Liman after treatment with 0.05% concentration of mutagen mutated also in many characters: vegetation period 103 days; plant height 106 cm; 1000 grain mass was 47.0 g. In the second experiment, a somatic callus of the 2nd passage from varieties Kransnodarskii 424, Dubovskii 129, Slavyanetz, Liman were treated with the solution of mutagen NMH (concentration: 0.05%; 0.1%; 0.2% + 0.1% PABA by 40 minutes at Certomat shaking machine (100 rev./min). The treated callus has been cultivated at MS regeneration media (4 mg 2.4 D + 20 mg /l of sucrose) and MS intermediate media (non-hormonal + PABA) to obtain regenerants. Plant

  14. Somatic embryogenesis and plant regeneration from cell suspension cultures of Cucumis sativus L.

    Science.gov (United States)

    Chee, P P; Tricoli, D M

    1988-06-01

    A procedure for the regeneration of whole cucumber plants (Cucumis sativus L. cv. Poinsett 76) by embryogenesis from cell suspension cultures is described. Embryogenic callus was initiated from the primary leaves of 14-17 day old plants. Suspension cultures of embryogenic cells were grown in liquid Murashige and Skoog basal medium containing 5 uM 2,4,5-trichlorophenoxyacetic acid and 4 uM 6-benzylaminopurine. Suspension cultures were composed of a population of cells that were densely cytoplasmic and potentially embryogenic. Differentiation of embryos was enhanced by washing the suspension culture cells with MS basal medium containing 0.5% activated charcoal and twice with MS basal medium followed by liquid shake cultures in MS basal medium. Sixty to 70 percent of the embryos prewashed with activated charcoal germinated into plantlets with normal morphology. Embryos obtained from suspension cultured cells without prewashing with activated charcoal organized into plantlets with abnormal primary leaves. Morphologically normal plantlets were obtained by excising the shoot tips and transferring them to fresh medium.

  15. The effect of plant growth regulators on optimization of tissue culture ...

    African Journals Online (AJOL)

    Mature seeds of four upland rice cultivars namely Kusan, Lamsan, Selasi and Siam were assessed for callus induction and plant regeneration on different concentrations and combinations of plant growth regulators, incorporated into MS (Murashige and Skoog) basal medium. Callus induction frequency was significantly ...

  16. Semi-wild chimpanzees open hard-shelled fruits differently across communities.

    Science.gov (United States)

    Rawlings, Bruce; Davila-Ross, Marina; Boysen, Sarah T

    2014-07-01

    Researchers investigating the evolutionary roots of human culture have turned to comparing behaviours across nonhuman primate communities, with tool-based foraging in particular receiving much attention. This study examined whether natural extractive foraging behaviours other than tool selection differed across nonhuman primate colonies that had the same foods available. Specifically, the behaviours applied to open the hard-shelled fruits of Strychnos spp. were examined in three socially separate, semi-wild colonies of chimpanzees (Pan troglodytes) that lived under shared ecological conditions at Chimfunshi Wildlife Orphanage, and were comparable in their genetic makeup. The chimpanzees (N=56) consistently applied six techniques to open these fruits. GLMM results revealed differences in the number of combined technique types to open fruits across the colonies. They also showed colony differences in the application of three specific techniques. Two techniques (full biting and fruit cracking) were entirely absent in some colonies. This study provides empirical evidence that natural hard-shelled fruit-opening behaviours are distinct across chimpanzee colonies, differences that most likely have not resulted from ecological and genetic reasons.

  17. [Fresh fruit and occultism as ways to salvation: conversions in Leipzig's alternative culture at around 1900].

    Science.gov (United States)

    Bigalke, Bernadett

    2008-01-01

    During the time of the Wilhelmine Empire, there were multiple interdependencies between adherents of the life reform movement (vegetarians, naturopathists, nudists, etc.) and new religious movements such as esoteric groups like the theosophists in the alternative cultural milieu around 1900. These networks became visible in the form of double memberships in associations. However, there were also ambiguous affiliations, migration between groups and syncretistic beliefs without institutional belonging. The similarity between patterns of argumentation for this specific lifestyle and the congruence of chosen goals, ways and goods of salvation become particularly clear in this context. These forms of "methodical lifestyle" may lead to the development of a specific ethos or habitus (Max Weber). To illustrate these processes, this article analyses the report of a Leipzig lady who ate raw fruits and vegetables only, and examines her broader social context. Thereby the analysis will employ sociological theories of conversion to explain the case of Hedwig Bresch.

  18. Integrated Management of Causal Agents of Postharvest Fruit Rot of Apple

    Directory of Open Access Journals (Sweden)

    Mila Grahovac

    2011-01-01

    Full Text Available One of the major causes of poor quality and fruit loss (during storage and transportare diseases caused by phytopathogenic fungi. Economic losses which are the consequenceof the phytopathogenic fungus activity after harvest exceed the losses in the field.Themost important postharvest fungal pathogens of apple fruits are: Botrytis cinerea Pers. exFr., Penicillium expansum (Lk. Thom., Cryptosporiopsis curvispora (Peck. Grem., Colletotrichumgloeosporioides (Penz. Sacc., Monilinia sp., Gloeosporium album Osterw, Alternaria alternata(Fr. Keissler, Cladosporium herbarium Link., Cylindrocarpon mali (Alles. Wollenw., Stemphyliumbotryosum Wallr. The use of available protection technologies can significantly reducelosses caused by pathogens in storage. The concept of integrated pest management (IPMin apple fruits i.e. sustainable approach in control of causal agents of postharvest fruit rot,using cultural, physical, biological and chemical measures, to minimize economic, healthand risks to consumers and environment, is presented in the paper.

  19. The Influence of Coconut Water and Activated Charcoal in MS Medium on In Vitro Callus Regeneration of Dendrobium sp. Cultivar Bertha Chong Orchids

    Directory of Open Access Journals (Sweden)

    Dessi Novita Sari

    2015-09-01

    Full Text Available Dendrobium is one of the most commercial orchids. In Vitro technique is one of solution to fulfill the market demand of Dendrobium. Organic matters, such as coconut water, and activated charcoal are often given to in vitro medium to regenerate orchids callus. The addition of activated charcoal is not only adsorbing toxic substances but also organic matters. The aimof this researchistofindthe best combination for callus regeneration medium. The research was conducted at the Biological Cell and Molecular Laboratory, Mathematics and Natural Science Faculty of Syiah Kuala University, Darussalam, Banda Aceh since March to November 2013. The method used is experimental with Completely Randomized Factorial Design with two factor; treatments of coconut water and activated charcoal. The result showed that the combinationof 150mL/Lcoconut waterand2,0g/Lactivated charcoal is the best resultbecauseit is the onlytreatment that have capability in producingplantletswithin60days.

  20. Avocado fruit (Persea americana Mill) exhibits chemo-protective potentiality against cyclophosphamide induced genotoxicity in human lymphocyte culture.

    Science.gov (United States)

    Paul, Rajkumar; Kulkarni, Paresh; Ganesh, Narayan

    2011-01-01

    Diets rich in fruits and vegetables have been associated with reduced risks for many types of cancers. Avocado (Persea americana Mill.) is a widely consumed fruit containing many cancer preventing nutrients, vitamins and phytochemicals. Studies have shown that phytochemicals extracted from the avocado fruit selectively induce cell cycle arrest, inhibit growth, and induce apoptosis in precancerous and cancer cell lines. Our recent studies indicate that phytochemicals extracted with 50% Methanol from avocado fruits help in proliferation of human lymphocyte cells and decrease chromosomal aberrations induced by cyclophosphamide. Among three concentrations (100 mg, 150 mg and 200 mg per Kg Body Weight), the most effective conc. of extract was 200 mg/Kg Body Wt. It decreased significant level of numerical and structural aberrations (breaks, premature centromeric division etc. up to 88%, p avocado fruit can be utilized for making active chemoprotective ingredient for lowering the side effect of chemotherapy like cyclophosphamide in cancer therapy.

  1. Frozen fruit skin prick test for the diagnosis of fruit allergy.

    Science.gov (United States)

    Garriga, Teresa; Guilarte, Mar; Luengo, Olga; Guillén, Mercé; Labrador-Horrillo, Moisés; Fadeeva, Tatiana; Sala, Anna; Cardona, Victória

    2010-12-01

    Diagnosis of fruit sensitisation by skin prick test (SPT) is fast and easy to perform. Nevertheless, some fruit is not available throughout the year. Freezing aliquots of these fresh fruits to be defrosted would be a good solution to perform SPT at any time. To compare the reproducibility of SPT with Rosaceae and Cucurbitaceae frozen fruit with fresh and commercial fruit extracts. SPT with the following fruit were performed: apricot, cherry, strawberry, nectarine, Japanese medlar, peach, (peel and pulp), yellow and red plum, melon and watermelon. We compared fresh fruit, commercial extract and fruit which had been frozen at -18 degrees C. Results were read by planimetry (Inmunotek prick-film) after 15 minutes. The study group comprised 48 patients (9 males, 39 females) with a mean age of 31, 6 +/- 2.0 years. Concordance of positive and negative results was extremely high and significant in all cases. Correlation between frozen fruit and commercial extract, frozen fruit and fresh and commercial extract and fresh fruit was statistically significant in all cases except for strawberry. The use of frozen fruit is a valid method, as the performance of the SPT is similar to that of fresh fruit. This enables diagnostic procedures with seasonal fruit at any time of the year.

  2. Global analysis of differentially expressed genes and proteins in the wheat callus infected by Agrobacterium tumefaciens.

    Directory of Open Access Journals (Sweden)

    Xiaohong Zhou

    Full Text Available Agrobacterium-mediated plant transformation is an extremely complex and evolved process involving genetic determinants of both the bacteria and the host plant cells. However, the mechanism of the determinants remains obscure, especially in some cereal crops such as wheat, which is recalcitrant for Agrobacterium-mediated transformation. In this study, differentially expressed genes (DEGs and differentially expressed proteins (DEPs were analyzed in wheat callus cells co-cultured with Agrobacterium by using RNA sequencing (RNA-seq and two-dimensional electrophoresis (2-DE in conjunction with mass spectrometry (MS. A set of 4,889 DEGs and 90 DEPs were identified, respectively. Most of them are related to metabolism, chromatin assembly or disassembly and immune defense. After comparative analysis, 24 of the 90 DEPs were detected in RNA-seq and proteomics datasets simultaneously. In addition, real-time RT-PCR experiments were performed to check the differential expression of the 24 genes, and the results were consistent with the RNA-seq data. According to gene ontology (GO analysis, we found that a big part of these differentially expressed genes were related to the process of stress or immunity response. Several putative determinants and candidate effectors responsive to Agrobacterium mediated transformation of wheat cells were discussed. We speculate that some of these genes are possibly related to Agrobacterium infection. Our results will help to understand the interaction between Agrobacterium and host cells, and may facilitate developing efficient transformation strategies in cereal crops.

  3. Global Analysis of Differentially Expressed Genes and Proteins in the Wheat Callus Infected by Agrobacterium tumefaciens

    Science.gov (United States)

    Zhou, Xiaohong; Wang, Ke; Lv, Dongwen; Wu, Chengjun; Li, Jiarui; Zhao, Pei; Lin, Zhishan; Du, Lipu; Yan, Yueming; Ye, Xingguo

    2013-01-01

    Agrobacterium-mediated plant transformation is an extremely complex and evolved process involving genetic determinants of both the bacteria and the host plant cells. However, the mechanism of the determinants remains obscure, especially in some cereal crops such as wheat, which is recalcitrant for Agrobacterium-mediated transformation. In this study, differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were analyzed in wheat callus cells co-cultured with Agrobacterium by using RNA sequencing (RNA-seq) and two-dimensional electrophoresis (2-DE) in conjunction with mass spectrometry (MS). A set of 4,889 DEGs and 90 DEPs were identified, respectively. Most of them are related to metabolism, chromatin assembly or disassembly and immune defense. After comparative analysis, 24 of the 90 DEPs were detected in RNA-seq and proteomics datasets simultaneously. In addition, real-time RT-PCR experiments were performed to check the differential expression of the 24 genes, and the results were consistent with the RNA-seq data. According to gene ontology (GO) analysis, we found that a big part of these differentially expressed genes were related to the process of stress or immunity response. Several putative determinants and candidate effectors responsive to Agrobacterium mediated transformation of wheat cells were discussed. We speculate that some of these genes are possibly related to Agrobacterium infection. Our results will help to understand the interaction between Agrobacterium and host cells, and may facilitate developing efficient transformation strategies in cereal crops. PMID:24278131

  4. Proteins Play Important Role in Intercellular Adhesion Affecting on Fruit Textural Quality

    DEFF Research Database (Denmark)

    Bahadur Adhikari, Khem; Shomer, Ilan

    2012-01-01

    Fruit textural quality is becoming a major quality parameter for export, postharvest preservation, handling and processing. The main determinant of textural quality is intercellular adhesion (ICA) as attributed by the cell wall (CW) and its components. The importance of CW protein in ICA strength......Fruit textural quality is becoming a major quality parameter for export, postharvest preservation, handling and processing. The main determinant of textural quality is intercellular adhesion (ICA) as attributed by the cell wall (CW) and its components. The importance of CW protein in ICA...... strengthening was exempli ed in Medjoul date (Phoenix dactylifera L.) fruit, as a model. Fruit mesocarp sensitively responded to culture environment which was assayed in vitro at pH 3.5( pKa) in presence of organic acid molecules. The max penetration force, as a measure of ICA strength, of p...

  5. Produção de frutanos em calos e plântulas clonadas in vitro de Viguiera discolor Baker (Asteraceae Fructan production in callus and in vitro cloned seedlings of Viguiera discolor Baker (Asteraceae

    Directory of Open Access Journals (Sweden)

    Nair Massumi Itaya

    2005-09-01

    the previous culture medium, supplemented with 0.5 mg L-1 NAA, which allowed the production of uniform plants, and formation of non thickened roots, tuberous roots and callus-like structures (callus type 1. Analysis of these materials showed the presence of inulin-type fructans. Incubation of stem nodes in the presence of 2,4-D induced growth of friable callus (callus type 2, in which fructans and their synthesis enzymes sucrose: sucrose 1-fructosyltransferase (SST and fructan: fructan 1fructosyltransferase (FFT were detected. Although in lower concentrations, the ratio SST/FFT activities and their fructans were similar to values found in plants cultivated under natural conditions. Stem nodes incubation on hormone-free medium resulted in regeneration of 50% plantlets. In vitro propagation, of V. discolor, may allow large-scale multiplication and conservation of this species, as well as fructan production under this condition.

  6. Heterogeneity in pineapple fruit quality results from plant heterogeneity at flower induction.

    Science.gov (United States)

    Fassinou Hotegni, V Nicodème; Lommen, Willemien J M; Agbossou, Euloge K; Struik, Paul C

    2014-01-01

    Heterogeneity in fruit quality constitutes a major constraint in agri-food chains. In this paper the sources of the heterogeneity in pineapple in the field were studied in four experiments in commercial pineapple fields. The aims were to determine (a) whether differences in pineapple fruit quality among individual fruits are associated with differences in vigor of the individual plants within the crop at the time of artificial flower induction; and (b) whether the side shoots produced by the plant during the generative phase account for the fruit quality heterogeneity. Two pineapple cultivars were considered: cv. Sugarloaf and cv. Smooth Cayenne. Plant vigor at the time of artificial flower induction was measured by three variates: the number of functional leaves, the D-leaf length and their cross product. Fruit quality attributes measured at harvest time included external attributes (weight and height of fruit, infructescence and crown) and internal quality attributes [total soluble solids (TSS), pH, translucent flesh]. Results showed that the heterogeneity in fruit weight was a consequence of the heterogeneity in vigor of the plants at the moment of flower induction; that effect was mainly on the infructescence weight and less or not on the crown weight. The associations between plant vigor variates at flower induction and the internal quality attributes of the fruit were poor and/or not consistent across experiments. The weight of the slips (side shoots) explained part of the heterogeneity in fruit weight, infructescence weight and fruit height in cv. Sugarloaf. Possibilities for reducing the variation in fruit quality by precise cultural practices are discussed.

  7. Heterogeneity in pineapple fruit quality results from plant heterogeneity at flower induction

    Directory of Open Access Journals (Sweden)

    V. Nicodeme eFassinou Hotegni

    2014-12-01

    Full Text Available Heterogeneity in fruit quality constitutes a major constraint in agri-food chains. In this paper the sources of the heterogeneity in pineapple in the field were studied in four experiments in commercial pineapple fields. The aims were to determine (a whether differences in pineapple fruit quality among individual fruits are associated with differences in vigor of the individual plants within the crop at the time of artificial flower induction; and (b whether the side shoots produced by the plant during the generative phase account for the fruit quality heterogeneity. Two pineapple cultivars were considered: cv. Sugarloaf and cv. Smooth Cayenne. Plant vigor at the time of artificial flower induction was measured by three variates: the number of functional leaves, the D-leaf length and their cross product. Fruit quality attributes measured at harvest time included external attributes (weight and height of fruit, infructescence and crown and internal quality attributes (total soluble solids, pH, translucent flesh. Results showed that the heterogeneity in fruit weight was a consequence of the heterogeneity in vigor of the plants at the moment of flower induction; that effect was mainly on the infructescence weight and less or not on the crown weight. The association between plant vigor variates at flower induction and the internal quality attributes of the fruit were poor and/or not consistent across experiments. The weight of the slips (side shoots, explained part of the heterogeneity in fruit weight, infructescence weight and fruit height in cv. Sugarloaf. Possibilities for reducing the variation in fruit quality by precise cultural practices are discussed.

  8. Social attraction mediated by fruit flies' microbiome.

    Science.gov (United States)

    Venu, Isvarya; Durisko, Zachary; Xu, Jianping; Dukas, Reuven

    2014-04-15

    Larval and adult fruit flies are attracted to volatiles emanating from food substrates that have been occupied by larvae. We tested whether such volatiles are emitted by the larval gut bacteria by conducting tests under bacteria-free (axenic) conditions. We also tested attraction to two bacteria species, Lactobacillus brevis, which we cultured from larvae in our lab, and L. plantarum, a common constituent of fruit flies' microbiome in other laboratory populations and in wild fruit flies. Neither larvae nor adults showed attraction to axenic food that had been occupied by axenic larvae, but both showed the previously reported attraction to standard food that had been occupied by larvae with an intact microbiome. Larvae also showed significant attraction to volatiles from axenic food and larvae to which we added only either L. brevis or L. plantarum, and volatiles from L. brevis reared on its optimal growth medium. Controlled learning experiments indicated that larvae experienced with both standard and axenic used food do not perceive either as superior, while focal larvae experienced with simulated used food, which contains burrows, perceive it as superior to unused food. Our results suggest that flies rely on microbiome-derived volatiles for long-distance attraction to suitable food patches. Under natural settings, fruits often contain harmful fungi and bacteria, and both L. brevis and L. plantarum produce compounds that suppress the growth of some antagonistic fungi and bacteria. The larval microbiome volatiles may therefore lead prospective fruit flies towards substrates with a hospitable microbial environment.

  9. Aplicações da cultura de tecidos vegetais em fruteiras do Cerrado Applications of tissue culture techniques in Brazilian Cerrado fruit trees

    Directory of Open Access Journals (Sweden)

    Hernane Fernandes Pinhal

    2011-07-01

    Full Text Available Atualmente, percebe-se uma preocupação em relação às plantas do cerrado, com grande enfoque nas fruteiras em função de suas características e usos. Apesar de ser uma área ainda pouco explorada, é crescente o número de estudos dessas espécies nativas, dentre eles, os que abrangem as técnicas de cultura de tecidos. Isso se deve uma vez que essa ferramenta biotecnológica permite a propagação de espécies com dificuldade de germinação, minimiza o problema de sementes recalcitrantes, promove a produção de mudas em larga escala, complementa bancos de germoplasma e facilita as trocas de materiais genéticos. Dessa maneira, esta revisão visa a sumarizar o histórico e panorama atual das aplicações da cultura de tecidos em fruteiras do cerrado, proporcionando sustentação para novos estudos.Currently, it's been given a huge concern to the cerrado plants, focusing on fruit trees due to their characteristics and uses. Despite being a fairly unexplored area, the number of studies on these native species has increased, especially those involving tissue culture techniques. That's because this biotechnological tool provides the propagation of species with germination difficulty, reduces problems of recalcitrant seeds, promotes large scale seedling production, complements germplasm banks and facilitates the exchange of genetic materials. Therefore, this review summarizes the history and current situation of tissue culture techniques applied to Brazilian Cerrado fruit trees, providing support to further studies.

  10. Occurrence of Anthracnose on Fruits of Asian Pear Tree Caused by Colletotrichum acutatum

    OpenAIRE

    Kim, Wan Gyu; Hong, Sung Kee; Park, Yeong Seob

    2007-01-01

    Anthracnose symptoms often occurred on fruits of Asian pear trees grown in Anseong, Naju, Seonghwan and Pyeongtaek in Korea during the harvesting period from 2000 to 2005. A total of 28 isolates of Colletotrichum sp. were obtained from the anthracnose symptoms. All the isolates were identified as Colletotrichum acutatum based on their morphological and cultural characteristics. Four isolates of the fungus were tested for pathogenicity to fruits of Asian pear tree by artificial inoculation. Al...

  11. Isolation and culture of Celosia cristata L cell suspension protoplasts

    Directory of Open Access Journals (Sweden)

    Retno Mastuti

    2003-06-01

    Full Text Available Developmental competence of Celosia cristata L. cell suspension-derived protoplasts was investigated. The protoplasts were isolatedfrom 3- to 9-d old cultures in enzyme solution containing 2% (w/v Cellulase YC and 0.5% (w/v Macerozyme R-10 which was dissolvedin washing solution (0.4 M mannitol and 10 mM CaCl2 at pH 5.6 for 3 hours. The highest number of viable protoplasts was releasedfrom 5-d old culture of a homogenous cell suspension. Subsequently, three kinds of protoplast culture media were simultaneously examinedwith four kinds of concentration of gelling agent. Culturing the protoplasts on KM8p medium solidified with 1.2% agarose significantlyenhanced plating efficiency as well as microcolony formation. Afterwards, the microcalli actively proliferated into friable watery calluswhen they were subcultured on MS medium supplemented with 0.3 mg/l 2,4-D and 1.0 mg/l kinetin. Although the plant regenerationfrom the protoplasts-derived calli has not yet been obtained, the reproducible developmental step from protoplasts to callus in thisstudy may facilitate the establishment of somatic hybridization using C. cristata as one parent.

  12. Irradiation effect on in vitro organogenesis, callus growth and plantlet development of Gerbera jamesonii Efeito da irradiação na organogênese in vitro, crescimento de calos e desenvolvimento de plântulas de gerbera

    Directory of Open Access Journals (Sweden)

    Nor A Hasbullah

    2012-06-01

    Full Text Available The present work was carried out to study the effects of gamma irradiation on in vitro growth of explants, callus and the formation of shoots and plantlets. Irradiation is known to exhibit or inhibit the differentiation of cells and growth of plants in vitro, which helps in producing new plant varieties. Gamma irradiation is one of the physical mutagens that are widely used for mutation breeding. A gradual decline was observed in the number of shoots regenerated from irradiated petiole explants compared to control. Numbers of shoots regenerated from irradiated petiole explant cultured on Murashige & Skoog medium supplemented with 2.0 mg L-1 BAP and 0.5 mg L-1 NAA was reduced to 6.6±0.9 from 7.5±0.4 (control when explants were exposed to 20 Gray of irradiation dose. Similar observation was reported on effects of gamma irradiation on in vitro propagated plantlets. Gradual decline was observed based on plant height as the dose of gamma irradiation increased. A significant decline was observed in the fresh weight of irradiated callus compared to control. In this case, growth responses of callus were strongly influenced by the radiation dose. The fresh weight of callus was reduced to 76.4±2.2% compared to 89.7±0.5% of control when callus tissues were exposed to 20 Gy.O presente trabalho foi realizado para estudar os efeitos da radiação gama no crescimento in vitro de explantes de calos, e a formação de brotos e mudas. A irradiação é conhecida por induzir ou inibir a diferenciação de células e o crescimento das plantas in vitro, o que ajuda na produção de novas variedades vegetais. Radiação gama é um dos agentes mutagenicos que são amplamente utilizados para o melhoramento através da mutação. Um declínio gradual foi observado no número de brotos regenerados a partir de explantes de pecíolos irradiados comparado com o controle. O número de brotações regeneradas de explantes de pecíolos irradiados, cultivados em meio

  13. An inventory of recent innovations in fruit and fruit products

    NARCIS (Netherlands)

    Zajac, J.; Lans, van der I.A.

    2009-01-01

    The goals of this study were to make an inventory of recent and ongoing fruit and fruit product innovations, to assess what novelty or improvement they offer, and whether consumers could identify and/or recognise them. Researchers from 11 European countries submitted 386 examples of fruit and fruit

  14. Efeito da concentração de sais e fitorreguladores na indução de calos em carqueja Callus induction in "carqueja" as affected by salt concentrations and growth regulators

    Directory of Open Access Journals (Sweden)

    Fabiano Guimarães Silva

    2003-06-01

    Full Text Available Realizou-se este trabalho com o objetivo de estudar condições nutricionais e hormonais para maximizar a produção de calos friáveis de carqueja [Baccharis trimera (Less. DC]. Foi verificado que a iniciação de calo é dependente de fitorreguladores e da concentração do meio. A melhor indução de calo ocorreu em meio MS contendo 50% da concentração de sais, inositol e vitaminas, suplementado com 15,0 mM ANA. Proliferação de brotos foi obtida pelo uso de TDZ.The influence of various growth regulators and medium concentrations, in different quantities, on the in vitro callus induction of carqueja [Baccharis trimera (Less. DC.] was evaluated. It was found that the callus initiation was dependent on both, the growth regulator and medium concentration. The highest callus induction and development were obtained by using 15.0 mM 1-napthaleneacetic acid (NAA as growth regulators and half strength of salts, vitamins, and myo-inositol of Murashige and Skoog medium. In vitro shoot proliferation was obtained by using thidiazuron.

  15. AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION OF SORGHUM USING TISSUE CULTURE-BASED AND POLLEN-MEDIATED APPROACHES

    Directory of Open Access Journals (Sweden)

    Elkonin L.A.

    2012-08-01

    Full Text Available Genetic transformation is a powerful tool for genetic improvement of arable crops. Genetic engineering approaches are especially important for modification of starch and protein contents, vitamin and micronutrient concentration, improvement of nutritive value of protein fractions, and increase tolerance to environmental stresses. Application of transgenic technologies for genetic improvement of sorghum, a highly productive heat tolerant and drought resistant crop, is extremely important since climate aridization in many regions all over the globe hampers sustainable production of traditional cereals, such as wheat, maize and barley. However, sorghum, in spite of great number of investigations, is one of the most recalcitrant crop species to genetic modification. The most frequently reported problems are a low frequency of transformation and silencing of transgenes. Using the A. tumefaciens strain AGL0/p35SGIB with the bar and gus-intron genes under the nos and CaMV35S promoters, respectively, we studied different methods of Agrobacterium-mediated genetic transformation of the grain sorghum: in vitro culture-based techniques, by inoculation of immature embryos or embryo-derived calli, and pollen-mediated approach, by inoculation of flowering panicles. Four lines of grain sorghum – Milo-10, [9E] Milo-10 (CMS-line, KVV-114, and KVV-45 – were used. In both approaches, for activation of vir-genes agrobacterial cell suspension was grown in the AB or modified AB media with acetosyringone at room temperature. In vitro culture approach was effective for obtaining transgenic plants in the lines Milo-10 and KVV-45, which were able to produce embryogenic callus from immature embryos after their co-cultivation with agrobacterial cell suspension. Callus cultures tolerant to glufosinate ammonium (GA and capable to plant regeneration were obtained. The frequency of immature embryos producing PCR-positive transgenic plants varied in different experiments

  16. Effect of growing conditions at greenhouse on vitamin E content in sweet pepper (Capsicum annuum L. fruits

    Directory of Open Access Journals (Sweden)

    Marcin Horbowicz

    2013-12-01

    Full Text Available The studies on effect of growing conditions on vitamin E content in fruits of two cultivars of sweet pepper (Bendigo F1 and Lamuyo F1 were carried out. The influence of growing substrate type (sphagnum peat or rockwool, Flormin, Poland, season of fruits harvesting (Summer or Autumn and dosages of mineral Iiquid nutrition were investigated. It was found that fruits of sweet pepper grown in rockwool contained more vitamin E than those from sphagnum peat substrate (ring culture, during Summer harvesting„ ln Autumn differences appeared too. but most of them were not statistically proved. The sweet pepper fruits harvested at Summer contained more vitamin E than fruits from Autumn. There were not significant differences among vitamin E level in fruits of the plants cultivated in rockwool at different dosages of fertilizers.

  17. Electrolytic treatment and biosurfactants applied to the conservation of Eugenia uniflora fruit

    Directory of Open Access Journals (Sweden)

    Guilherme DILARRI

    2016-01-01

    Full Text Available Abstract Microorganisms are the primary responsible for food poisoning and food spoilage. The purpose of this study was to evaluate different fruit washing methods with tap water, electrolyzed water and rhamnolipids solution produced by Pseudomonas aeruginosa LBI, in order to inhibit microbial growth. The tested organism was Eugenia uniflora. The fruits were washed and periodically inoculated into culture media to evaluate and count the colonies on the fruit surface. It was also observed the deterioration level of the fruits after each treatment. The results showed that treatment with rhamnolipids were the most efficient, inhibiting the growth of fungi and bacteria. The electrolyzed water proved to be very efficient in bacterial inhibition at the initial time, but in the final time it did not present any inhibitory effect. The electrolyzed water was also not effective in eliminating fungus. Washing with tap water was the less efficient treatment of all. The only treatment that showed an increased durability has been with rhamnolipids, increasing shelf life by up to two days. Thus rhamnolipids are the most recommended method for fruits sanitation.

  18. Comparison of the nutrient content of fresh fruit juices vs commercial fruit juices.

    Science.gov (United States)

    Densupsoontorn, Narumon; Jirapinyo, Pipop; Thamonsiri, Nuchnoi; Wongarn, Renu; Phosuya, Panarat; Tritiprat, Amornrat; Patraarat, Siriphan; Pidatcha, Pannee; Suwannthol, Lerson

    2002-08-01

    To compare the types and quantities of carbohydrate, electrolytes, pH and osmolarity of fresh fruit juices and commercial fruit juices. Forty kinds of fresh fruits available in Thai markets were analyzed for types and quantities of carbohydrate, electrolyte, pH and osmolarity and compared with previously obtained data for commercial fruit juices. Most fresh fruit juices did not contain sucrose, whereas, commercial fruit juices mostly have sucrose in the range of 3-112 g/L. Although both fruit juices were acidic (pH varied from 3.6-6.7 and 3.2-5.8 of fresh juice and commercial juice), fresh fruit juices had a more neutral pH than commercial fruit juices. Apple, guava, orange, pear, and pineapple juices from commercial fruit juices had a high osmolarity compared with fresh fruit juices. All types of fresh fruit juices contained less sodium than commercial ones, whereas, most fresh fruit juices contained more potassium, phosphorus, and magnesium than commercial fluids. The nutrient content of fresh fruit juices and commercial fruit juices from the same kinds of fruits are not the same, possibly due to the manufacturing process. Therefore, physicians should know the composition of fruit juices in order to advise patients properly.

  19. The role of fruit colour in avian fruit selection: an objective approach

    OpenAIRE

    Schmidt, Veronika

    2002-01-01

    To explain the prevalence of red and black fruits in fruit colour patterns, the following hypotheses were addressed, using reflectance spectra of fruits as colour assessment: 1. Birds prefer red and black fruits, or these hues are cues for food recognition in migrants or fledglings. 2. Fruit colours correlate with chemical compounds. 3. Fruit colours serve as advertisement for ripe fruits. Reflectance spectra are the most objective colour assessment currently possible. Birds show no colour pr...

  20. Somatic embryogenesis and plant regeneration in tissue cultures of sweet potato (Ipomea batatas Poir.).

    Science.gov (United States)

    Liu, J R; Cantliffe, D J

    1984-06-01

    Leaf, shoot-tip, stem, and root explants of sweet potato (Ipomea batatas Poir.) gave rise to two kinds of callus on nutrient agar medium containing 0.5 to 2.0 mg/l 2,4-D. One callus, bright- to pale-yellow, was compact and organized, while the other was dull-yellow and friable. The former callus gave rise to numerous globular and heart-shaped embryoids. When transferred onto hormone-free medium, the embryoids readily developed into a torpedo-shape before germination. The plantlets were transplanted to soil where they flowered and formed storage roots at maturity.

  1. Consumer Preference Towards Fruit Leather Attributes of Madurese Exotic Tropical Fruits

    Directory of Open Access Journals (Sweden)

    Elys Fauziyah

    2018-01-01

    Full Text Available Madura island has high potential for producing tropical fruits, but it still not being well managed especially concerning with the value added Fruit leather is a product created by using various fruits and simple technology application. Fruit leather is categorized as new product on the market, therefore it is important to know consumer preference towards fruit leather attributes so that producer can design an acceptable product in the market. The research investigated attributes within the levels that become consumer preference in purchasing fruit leather product. There were 60 samples respondents taken accidentally at Bangkalan Plaza Shopping area. Method being used was conjoint analysis. Result showed that fruit leather being chosen by consumers as preference are gummy, mixed fruit taste, yellow color small roll shape, at 100 grams and in a plastic tube package.

  2. Eating fruits and vegetables. An ethnographic study of American and French family dinners.

    Science.gov (United States)

    Kremer-Sadlik, Tamar; Morgenstern, Aliyah; Peters, Chloe; Beaupoil, Pauline; Caët, Stéphanie; Debras, Camille; le Mené, Marine

    2015-06-01

    The French eat more fruits and vegetables than Americans and have lower rates of childhood obesity. This ethnographic study compares various aspects of meal environment in sixteen households in LA, California and Paris, France, and offers insights on the relationship between local practices and preferences and children's consumption of fruits and vegetables. Our analysis of video-recorded naturalist data reveals that the consumption of fruits and vegetables is linked to the cultural organization of dinner--what, when and how food is served--and to local beliefs about children's eating practices. We also found that the French model for dinnertime prioritizes the eating of fruits and vegetables more than the American model does. We propose that local eating models should be taken into account in research on childhood obesity and in prevention programs. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Focus on Fruits: 10 Tips to Eat More Fruits

    Science.gov (United States)

    ... lunch, pack a tangerine, banana, or grapes to eat or choose fruits from a salad bar. Individual containers of fruits like peaches or applesauce are easy to carry and convenient for lunch. 7 Enjoy fruit at dinner, too At dinner, add crushed pineapple to coleslaw ...

  4. In vitro flowering and fruiting in culture of dendrobium officinate kimura et migo (orchidaceae)

    International Nuclear Information System (INIS)

    Qian, X.; Wang, C.; Ouyang, T.; Tian, M.

    2014-01-01

    Dendrobium officinate Kimura et Migo. is a rare and endangered perennial orchid, which is valued for its attractive flowers and medicinal uses. Its three-to-five-year breeding cycle makes propagation difficult, so we investigated In vitro production of flowers and seeds for this species. Calluses were induced from shoot-tip explants of D. officinate on Murashige and Skoog (MS) medium with 0.2 mg L-1 benzyladenine (BA) and 0.05 mg L-1 naphthalene acetic acid (NAA). Multiple shoots were regenerated after protocorm-like bodies (PLBs) were transferred onto the same basal medium with 1.0 mg L-1 NAA. The plantlets, 2-4 cm in height, maintained In vitro were induced to flower, and the highest rates of inflorescence (83.2%) and normal flowers (73.6%) were produced on the MS medium with 15% (v/v) coconut water and 0.1 mg L-1 thidiazuron (TDZ) within 9 weeks. Histological analysis showed the origin of the floral primordial and normal morphologies of the pollen and female organs. The normal flowers were artificially pollinated and capsules developed. Viable seeds were produced and most of them germinated on the MS medium. This reproduction system could be used to study the molecular basis of flowering or to improve plant breeding programs. (author)

  5. Prediction of the time course of callus stiffness as a function of mechanical parameters in experimental rat fracture healing studies--a numerical study.

    Directory of Open Access Journals (Sweden)

    Tim Wehner

    Full Text Available Numerous experimental fracture healing studies are performed on rats, in which different experimental, mechanical parameters are applied, thereby prohibiting direct comparison between each other. Numerical fracture healing simulation models are able to predict courses of fracture healing and offer support for pre-planning animal experiments and for post-hoc comparison between outcomes of different in vivo studies. The aims of this study are to adapt a pre-existing fracture healing simulation algorithm for sheep and humans to the rat, to corroborate it using the data of numerous different rat experiments, and to provide healing predictions for future rat experiments. First, material properties of different tissue types involved were adjusted by comparing experimentally measured callus stiffness to respective simulated values obtained in three finite element (FE models. This yielded values for Young's moduli of cortical bone, woven bone, cartilage, and connective tissue of 15,750 MPa, 1,000 MPa, 5 MPa, and 1 MPa, respectively. Next, thresholds in the underlying mechanoregulatory tissue differentiation rules were calibrated by modifying model parameters so that predicted fracture callus stiffness matched experimental data from a study that used rigid and flexible fixators. This resulted in strain thresholds at higher magnitudes than in models for sheep and humans. The resulting numerical model was then used to simulate numerous fracture healing scenarios from literature, showing a considerable mismatch in only 6 of 21 cases. Based on this corroborated model, a fit curve function was derived which predicts the increase of callus stiffness dependent on bodyweight, fixation stiffness, and fracture gap size. By mathematically predicting the time course of the healing process prior to the animal studies, the data presented in this work provides support for planning new fracture healing experiments in rats. Furthermore, it allows one to transfer and

  6. Developmental gene regulation during tomato fruit ripening and in-vitro sepal morphogenesis

    Directory of Open Access Journals (Sweden)

    Ishida Betty K

    2003-08-01

    Full Text Available Abstract Background Red ripe tomatoes are the result of numerous physiological changes controlled by hormonal and developmental signals, causing maturation or differentiation of various fruit tissues simultaneously. These physiological changes affect visual, textural, flavor, and aroma characteristics, making the fruit more appealing to potential consumers for seed dispersal. Developmental regulation of tomato fruit ripening has, until recently, been lacking in rigorous investigation. We previously indicated the presence of up-regulated transcription factors in ripening tomato fruit by data mining in TIGR Tomato Gene Index. In our in-vitro system, green tomato sepals cultured at 16 to 22°C turn red and swell like ripening tomato fruit while those at 28°C remain green. Results Here, we have further examined regulation of putative developmental genes possibly involved in tomato fruit ripening and development. Using molecular biological methods, we have determined the relative abundance of various transcripts of genes during in vitro sepal ripening and in tomato fruit pericarp at three stages of development. A number of transcripts show similar expression in fruits to RIN and PSY1, ripening-associated genes, and others show quite different expression. Conclusions Our investigation has resulted in confirmation of some of our previous database mining results and has revealed differences in gene expression that may be important for tomato cultivar variation. We present new and intriguing information on genes that should now be studied in a more focused fashion.

  7. Association Study Reveals Novel Genes Related to Yield and Quality of Fruit in Cape Gooseberry (Physalis peruviana L.).

    Science.gov (United States)

    García-Arias, Francy L; Osorio-Guarín, Jaime A; Núñez Zarantes, Victor M

    2018-01-01

    Association mapping has been proposed as an efficient approach to assist plant breeding programs to investigate the genetic basis of agronomic traits. In this study, we evaluated 18 traits related to yield, (FWP, NF, FWI, and FWII), fruit size-shape (FP, FA, MW, WMH, MH, HMW, DI, FSI, FSII, OVO, OBO), and fruit quality (FIR, CF, and SST), in a diverse collection of 100 accessions of Physalis peruviana including wild, landraces, and anther culture derived lines. We identified seven accessions with suitable traits: fruit weight per plant (FWP) > 7,000 g/plant and cracked fruits (CF) peruviana .

  8. Biotechnological approaches to enhance salidroside, rosin and its derivatives production in selected Rhodiola spp. in vitro cultures.

    Science.gov (United States)

    Grech-Baran, Marta; Sykłowska-Baranek, Katarzyna; Pietrosiuk, Agnieszka

    Rhodiola (Crassulaceae) an arctic-alpine plant, is extensively used in traditional folk medicine in Asian and European countries. A number of investigations have demonstrated that Rhodiola preparations exhibit adaptogenic, neuroprotective, anti-tumour, cardioprotective, and anti-depressant effects. The main compounds responsible for these activities are believed to be salidroside, rosin and its derivatives which became the target of biotechnological investigations. This review summarizes the results of the diverse biotechnological approaches undertaken to enhance the production of salidroside, rosin and its derivatives in callus, cell suspension and organ in vitro cultures of selected Rhodiola species.

  9. Pineapple Fruit Collapse: Newly Emerging Disease of Pineapple Fruit in Lampung, Indonesia

    OpenAIRE

    Joko Prasetyo; Titik Nur Aeny

    2014-01-01

    ABSTRACT Pineapple fruit collapse: newly emerging disease of pineapple fruit in Lampung, Indonesia Recently, a new disease on pineapple fruit has occurred in Lampung. Symptoms of the disease are complex. Fruits rotted and exuded copious liquid from the inter- fruitlet tissues accompanied by gas bubbles. Open spaces were formed inside the rotten fruit. Dissection of diseased fruit showed many cavities within its sceletal fibres and bad odour was exerted from the rotten tissues. A bacterial...

  10. Phenolic compounds in cultures of tissues of tea plants and the effect of light on their synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Koretskaya, T.F.; Zaprometov, M.N.

    1975-01-01

    Stem and leaf calluses of tea plants (Camellia sinensis) retain the capacity for synthesis of phenolic compounds. The content of phenolic compounds comprises 2 to 5 percent of dry weight, the main share (80 to 95 percent) belonging to catechins and leucoanthocyans, including their polymeric forms. The following compounds were detected in callus tissue: (--)-epicatechin, (+)-catechin, two leucoanthocyans, and several unidentified phenolic compounds that fluoresce in UV. (--)-Epicatechin is predominant. In contrast to tissues of an intact plant, the callus does not contain gallocatechins or free gallic acid under the given cultivation conditions. The content of phenolic compounds changes in proportion to callus growth, their greatest amount being formed during the phase of intensive growth. Light stimulates synthesis of phenolic compounds, including the most reduced group of flavonoids, viz., leucoanthocyans and catechins.

  11. Studies on trigonelline from Moringa oleifera and its in vitro regulation by feeding precursor in cell cultures

    Directory of Open Access Journals (Sweden)

    Manas Mathur

    2012-04-01

    Full Text Available Trigonelline (N-methylnicotinate biosynthesized from nicotinate is one of the metabolically active pyridine alkaloid, widely distributed in plant kingdom. In the present study trigonelline has been isolated from various plant parts and callus cultures of Moringa oleifera Lam., Moringaceae, and was identified using TLC, GLC, GC-MS, which was comparable to that of the standard trigonelline. The trigonelline recovery was found to be maximum in the pods and minimum in flowers. In order to enhance the production of trigonelline in vitro grown cultures, different treatment doses of nicotinic acid (250, 500 and 750 mg L-1 were supplemented in the medium as precursor. Maximum increase (up to 1.10 fold was observed in the treatment dose of 500 mg L-1 of nicotinic acid.

  12. Consumer Preference Towards Fruit Leather Attributes of Madurese Exotic Tropical Fruits

    OpenAIRE

    Elys Fauziyah

    2018-01-01

    Madura island has high potential for producing tropical fruits, but it still not being well managed especially concerning with the value added Fruit leather is a product created by using various fruits and simple technology application. Fruit leather is categorized as new product on the market, therefore it is important to know consumer preference towards fruit leather attributes so that producer can design an acceptable product in the market. The research investigated attributes within the l...

  13. [Spectral navigation technology and its application in positioning the fruits of fruit trees].

    Science.gov (United States)

    Yu, Xiao-Lei; Zhao, Zhi-Min

    2010-03-01

    An innovative technology of spectral navigation is presented in the present paper. This new method adopts reflectance spectra of fruits, leaves and branches as one of the key navigation parameters and positions the fruits of fruit trees relying on the diversity of spectral characteristics. The research results show that the distinct smoothness as effect is available in the spectrum of leaves of fruit trees. On the other hand, gradual increasing as the trend is an important feature in the spectrum of branches of fruit trees while the spectrum of fruit fluctuates. In addition, the peak diversity of reflectance rate between fruits and leaves of fruit trees is reached at 850 nm of wavelength. So the limit value can be designed at this wavelength in order to distinguish fruits and leaves. The method introduced here can not only quickly distinguish fruits, leaves and branches, but also avoid the effects of surroundings. Compared with the traditional navigation systems based on machine vision, there are still some special and unique features in the field of positioning the fruits of fruit trees using spectral navigation technology.

  14. Shoot organogenesis in oleaster (Elaeagnus angustifolia L.)

    African Journals Online (AJOL)

    STORAGESEVER

    2009-02-04

    Feb 4, 2009 ... Regenerated plantlets were acclimatized and successfully transplanted to soil. Key words: shoot organogenesis, callus, ... saline and alkaline soils (Economou and Maloupa, 1995). Its fruits have been used as a .... cally inert compounds as reported by Kaminek (1992). Direct shoot organogenesis without ...

  15. Pineapple Fruit Collapse: Newly Emerging Disease of Pineapple Fruit in Lampung, Indonesia

    Directory of Open Access Journals (Sweden)

    Joko Prasetyo

    2014-03-01

    Full Text Available ABSTRACT Pineapple fruit collapse: newly emerging disease of pineapple fruit in Lampung, Indonesia Recently, a new disease on pineapple fruit has occurred in Lampung. Symptoms of the disease are complex. Fruits rotted and exuded copious liquid from the inter- fruitlet tissues accompanied by gas bubbles. Open spaces were formed inside the rotten fruit. Dissection of diseased fruit showed many cavities within its sceletal fibres and bad odour was exerted from the rotten tissues. A bacterial entity was isolated  from the diseased materials. In a pathogenicity test, the isolated bacteria caused the same symptom as mentioned. In the growing-on test the crown of the heavily infected fruit  showed  heart rot symptom.  Those  indicated that the disease was pineapple fruit collapse. Both symptoms were known related to the same causal agent, Erwinia chrysanthemi (pineapple strain Dickeya sp.. In our opinion, this is the first report of pineapple fruit collapse in Indonesia.

  16. Enhancement effect of shikonin in cell suspension culture and transfermanant culture by radiation application

    International Nuclear Information System (INIS)

    Kim, Jae Sung; Lee, Young Keun; Chung, Byung Yeoup; Lee, Young Bok; Hwang Hye Yeon

    2004-10-01

    The cell lines 679, 679-29 and 622-46 of L. erythrorhizon could be selected on LS agar medium for the production shikonin in cell suspension culture. The shikonin was increased moderately in suspension culture of cell line 622-46 in LS liquid medium containing BA 2 mg·L -1 and IAA 0.2 mg·L -1 in the dark, and was increased by adding 1 μM Cu 2+ and 100 μM methyl jasmonate The accumulation of shikonin in the liquid medium was increased significantly by 2 Gy irradiation to callus of cell line 622-46 and culture in LS liquid medium containing BA 2 mg·L -1 and IAA 0.2 mg·L -1 in the dark and shikonin in cell debris was higher by 16 Gy irradiation. The activity of p-hydroxybenzoate geranyltransferase was increased by irradiation of 2 Gy and 16 Gy of γ radiation. Seedling hypocotyles of L. erythrorhizon were infected with Agrogacterium rhizogenes strain 15834 harboring a binary vector with an intron bearing the GUS (β-glucuronidase) gene driven by cauliflower mosaic virus (CaMV) 35S promotor as well as the HPT (hygromycin phosphotransferase) gene as the selection marker. Hairy roots isolated were hygromycin resistant and had integrated GUS gene in DNA. The root tip grown on M-9 medium showed normal pigment production pattern in border cells and root hairs

  17. Control of postharvest Botrytis fruit rot of strawberry by volatile organic compounds of Candida intermedia.

    Science.gov (United States)

    Huang, R; Li, G Q; Zhang, J; Yang, L; Che, H J; Jiang, D H; Huang, H C

    2011-07-01

    A study was conducted to identify volatile organic compounds or volatiles produced by Candida intermedia strain C410 using gas chromatography-mass spectrometry, and to determine efficacy of the volatiles of C. intermedia in suppression of conidial germination and mycelial growth of Botrytis cinerea and control of Botrytis fruit rot of strawberry. Results showed that, among 49 volatiles (esters, alcohols, alkenes, alkanes, alkynes, organic acids, ketones, and aldehydes) identified from C. intermedia cultures on yeast extract peptone dextrose agar, two compounds, 1,3,5,7-cyclooctatetraene and 3-methyl-1-butanol, were the most abundant. Synthetic chemicals of 1,3,5,7-cyclooctatetraene; 3-methyl-1-butanol; 2-nonanone; pentanoic acid, 4-methyl-, ethyl ester; 3-methyl-1-butanol, acetate; acetic acid, pentyl ester; and hexanoic acid, ethyl ester were highly inhibitory to conidial germination and mycelial growth of B. cinerea. Inhibition of conidial germination and mycelial growth of B. cinerea by volatiles of C. intermedia was also observed. Meanwhile, results showed that incidence and severity of Botrytis fruit rot of strawberry was significantly (P intermedia cultures or C. intermedia-infested strawberry fruit. These results suggest that the volatiles of C. intermedia C410 are promising biofumigants for control of Botrytis fruit rot of strawberry.

  18. Effects of Selected Physicochemical Parameters on Zerumbone Production of Zingiber zerumbet Smith Cell Suspension Culture.

    Science.gov (United States)

    Jalil, Mahanom; Annuar, Mohamad Suffian Mohamad; Tan, Boon Chin; Khalid, Norzulaani

    2015-01-01

    Zingiber zerumbet Smith is an important herb that contains bioactive phytomedicinal compound, zerumbone. To enhance cell growth and production of this useful compound, we investigated the growth conditions of cell suspension culture. Embryogenic callus generated from shoot bud was used to initiate cell suspension culture. The highest specific growth rate of cells was recorded when it was cultured in liquid Murashige and Skoog basal medium containing 3% sucrose with pH 5.7 and incubated under continuous shaking condition of 70 rpm for 16 h light and 8 h dark cycle at 24°C. Our results also revealed that the type of carbohydrate substrate, light regime, agitation speed, and incubation temperature could affect the production of zerumbone. Although the zerumbone produced in this study was not abundant compared to rhizome of Z. zerumbet, the possibility of producing zerumbone during early stage could serve as a model for subsequent improvement.

  19. Effects of Selected Physicochemical Parameters on Zerumbone Production of Zingiber zerumbet Smith Cell Suspension Culture

    Directory of Open Access Journals (Sweden)

    Mahanom Jalil

    2015-01-01

    Full Text Available Zingiber zerumbet Smith is an important herb that contains bioactive phytomedicinal compound, zerumbone. To enhance cell growth and production of this useful compound, we investigated the growth conditions of cell suspension culture. Embryogenic callus generated from shoot bud was used to initiate cell suspension culture. The highest specific growth rate of cells was recorded when it was cultured in liquid Murashige and Skoog basal medium containing 3% sucrose with pH 5.7 and incubated under continuous shaking condition of 70 rpm for 16 h light and 8 h dark cycle at 24°C. Our results also revealed that the type of carbohydrate substrate, light regime, agitation speed, and incubation temperature could affect the production of zerumbone. Although the zerumbone produced in this study was not abundant compared to rhizome of Z. zerumbet, the possibility of producing zerumbone during early stage could serve as a model for subsequent improvement.

  20. Occurrence of Anthracnose on Fruits of Asian Pear Tree Caused by Colletotrichum acutatum.

    Science.gov (United States)

    Kim, Wan Gyu; Hong, Sung Kee; Park, Yeong Seob

    2007-12-01

    Anthracnose symptoms often occurred on fruits of Asian pear trees grown in Anseong, Naju, Seonghwan and Pyeongtaek in Korea during the harvesting period from 2000 to 2005. A total of 28 isolates of Colletotrichum sp. were obtained from the anthracnose symptoms. All the isolates were identified as Colletotrichum acutatum based on their morphological and cultural characteristics. Four isolates of the fungus were tested for pathogenicity to fruits of Asian pear tree by artificial inoculation. All the isolates induced anthracnose symptoms on the fruits by wound inoculation but not by unwound inoculation. The anthracnose symptoms induced by artificial inoculation were similar to those observed in the orchard. This is the first report of anthracnose of Asian pear tree caused by Colletotrichum acutatum.