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1.
S. Arumugam F. H. Chu S. Y. Wang S. T. Chang 《Journal of plant biochemistry and biotechnology.》2009,18(2):197-201
An efficient plant regeneration system was established from immature leaflet-derived callus of Acacia confusa Merr, through organogenesis. Under optimized culture conditions, the high rate of callus induction and proliferation was obtained in 35 days on MMS medium supplemented with 2,4-D (3 mg l?1) + NAA (0.01 mg l?1) + Kin (0.05 mg l?1). The highest percentage of shoot regeneration response (95%) and greatest number of shoots (52.9) were obtained after the 46-day transfer of green nodular calli onto the shoot regeneration medium (WPM) supplemented with the BA 3 mg l?1 + NAA 0.05 mg l?1 + Zeatin 0.1 mg l?1 + AdSO4 5 mg l?1 combination. Efficient shoot elongation was achieved by transferring the clusters of adventitious shoot buds to medium (half-strength MS) containing GA, (1 mg l?1) and BA (0.05 mg l?1), within 30 days. The elongated shoots were rooted on half-strength MS medium supplemented with 4 mg l?1 IBA and 0.05 mg l?1 Kin in the 42-day culture. Rooted plantlets were hardened and successfully established in soil. The field-established plants were morphologically normal and fertile. 相似文献
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3.
The influence of plant growth substances, medium and potting mixture on protocorm development, differentiation, growth and establishment of Zygopetalum intermedium was assessed. Embryo from mature green but unripe capsule cultured on half strength Murashige and Skoog medium containing 1.5 g l?1 AC with 0.25 mg l?1 PBZ and 0.1 mg l?1 of NAA swollen in 59.7 days, followed by formation of globular bodies in 64 days and protocorm development in 70.7 days. Nitsch medium in combination with 0.5 mg l?1 of BAP and 0.25 mg l?1 of Triacontanol resulted in shoot and root differentiation and maximum plant growth in vitro. Plantlets with 4–5 well-developed leaves with roots pre hardened in medium supplemented with 0.25 mg l?1 each of PBZ and Triacontanol transferred to community pots filled with potting mixture of coco peat and tree fern (1:1) resulted in 72.3% survival ex vitro. 相似文献
4.
Plant regeneration from protoplast culture of Crocus cancellatus was investigated using regenerable embryogenic calli obtained from shoot meristem culture on LS (Linsmaier and Skoog, 1965)
medium containing 4 mg l−1 kinetin and 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Protoplasts were isolated directly from embryogenic calli. The best protoplast growth
was found on those embedded in Ca-alginate beads and cultured with nurse cells in MS (Murashige and Skoog, 1962) medium supplemented
with 2 mg l−1 kinetin, 1 mg l−1 2,4-D and 100 mg l−1 ascorbic acid at 25 °C in darkness. After 4–5 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads,
but the protoplasts without immobilization in Ca-alginate beads showed very low cell division. Growth of the microcalli in
the medium with nurse cells was much better than in the medium without nurse cells. Transferring beads onto half strength
MS medium supplemented with 0.2 mg l−1 kinetin and 0.1 mg l−1 2,4-D, increased the growth of embryogenic calli. Somatic embryo development was observed either on half strength MS medium
growth regulator free or with 1 mg l−1 abscisic acid. Matured embryos germinated on half strength MS medium containing 25 mg l−1 of gibberelic acid. Plantlet formation was obtained on half strength MS medium containing 1 mg l−1 6-benzyladenine and 1 mg l−1 α-naphthaleneacetic acid at 20 °C in a 16/8 h light/dark cycle.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
5.
Protoplasts were isolated from cell suspensions derived from cotyledon and hypocotyl Gentiana kurroo (Royle). Cell walls were digested with an enzyme cocktail containing cellulase, macerozyme, driselase, hemicellulase and
pectolyase in CPW solution. Protoplast viability ranged from 88 to 96%. Three techniques of culture and six media were evaluated
in terms of their efficiency in producing viable cultures and regenerating whole plants. With liquid culture, cell division
occurred in only a low number of the protoplasts isolated, and no plant regeneration was successful. Cell division occurred
within 2 or 3 days in case of agarose solidified media. After 10 days of culture, the number of dividing cells was the highest
with modified MS medium in which NH4NO3 was replaced with 3.0 g l−1 glutamine. The best results were obtained with agarose bead cultures: plating efficiency was 68.7% and 58.1% for protoplasts
isolated from cotyledon and hypocotyl derived suspensions, respectively. The results were achieved with using medium containing
0.5 mg l−1 2,4-D + 1.0 mg l−1 kinetin or 2.0 mg l−1 BAP + 1.0 mg l−1 dicamba + 0.1 mg l−1 NAA + 80 mg l−1 adenine sulfate. Protocalluses transferred on the following composition of plant growth regulators: 0.5 mg l−1 2,4-D + 1.0 mg l−1 kinetin or 1.0 mg l−1 kinetin + 0.5 mg l−1 GA3 + 80.0 mg l−1 adenine sulfate developed in embryogenic cultures. However, the best embryo production occurred with the first one. Later
embryos were transferred to half-strength MS mineral salts to promote plants formation. Flow cytometry studies revealed increased
amounts of DNA in about one third of the regenerants. 相似文献
6.
《Enzyme and microbial technology》1987,9(3):153-155
In a study of the control of metabolite formation, prodigiosin production by Serratia marcescens was used as a model. Specific production rates of prodigiosin formation were determined using batch culture technique. Sucrose as carbon source and NH4NO3 as nitrogen source resulted in a specific production rate of 0.476 mg prodigiosin (g cell dry weight)−1 h−1. Prodigiosin formation and productivity was inversely correlated to growth rate when the bacterium was grown under carbon limitation on a defined medium in a chemostat culture. The maximum specific growth rate (μmax) was 0.54 h−1 and prodigiosin was formed in amounts over 1 mg l−1 up to a growth rate (μ) of 0.3 h−1 at steady state conditions. At a dilution rate of 0.1 h−1 growth at steady state with carbon and phosphate limitation supported prodigiosin formation giving a similar specific yield [1.17 mg prodigiosin (g cell dry weight)−1 and 0.94 mg g−1, respectively], however, cells grown with nitrogen limitation [(NH4)2SO4] did not form prodigiosin. Productivity in batch culture was 1.33 mg l−1 h−1 as compared to 0.57 mg l−1 h−1 in the chemostat. 相似文献
7.
Samart Taikhao Suwannee Junyapoon Aran Incharoensakdi Saranya Phunpruch 《Journal of applied phycology》2013,25(2):575-585
The effects of several physiological parameters on H2 production rate in the unicellular halotolerant cyanobacterium Aphanothece halophytica were investigated. Under nitrogen deprivation, the growth of cells was inhibited, but H2 production rate was enhanced approximately fourfold. Interestingly, cells grown under sulfur deprivation exhibited a decrease in cell growth, H2 production rate, and bidirectional hydrogenase activity. Glucose was the preferred sugar source for H2 production by A. halophytica, but H2 production decreased at high glucose concentrations. H2 production rate was optimum when cells were grown in the presence of 0.75 M?NaCl, or 0.4 μM?Fe3+, or 1 μM?Ni2+. The optimum light intensity and temperature for H2 production were 30 μmol photons m?2?s?1 and 35 °C, respectively. A two-stage culture of A. halophytica was performed in order to overcome the reduction of cell growth in N-free medium. In the first stage, cells were grown in normal medium to accumulate biomass, and in the second stage, H2 production by the obtained biomass was induced by growing cells in N-free medium supplemented with various chemicals for 24 h. A. halophytica grown in N-free medium containing various MgSO4 concentrations had a high H2 production rate between 11.432 and 12.767 μmol H2 mg?chlorophyll a (chl a)?1?h?1, a 30-fold increase compared to cells grown in normal medium. The highest rate of 13.804 μmol H2 mg?chl a ?1?h?1 was obtained when the N-free growth medium contained 0.4 μM Fe3+. These results suggested the possibility of using A. halophytica and some other halotolerant cyanobacteria thriving under extreme environmental conditions in the sea as potential sources for H2 production in the future. 相似文献
8.
Haifeng Yu 《Journal of applied phycology》2012,24(4):669-673
The effects of organic carbon sources on cell growth and exopolysaccharide (EPS) production of dissociated Nostoc flagelliforme cells under mixotrophic batch culture were investigated. After 7?days of cultivation, glycerol, acetate, sucrose, and glucose increased the final cell density and final EPS concentrations, and mixotrophic growth achieved higher biomass concentrations. The increase in cell growth was particularly high when glucose was added as the sole carbon source. On the other hand, EPS production per dry cell weight was significantly enhanced by adding acetate. For more effective EPS production, the effects of the mixture of glucose and acetate were investigated. Increasing the ratio of glucose to acetate resulted in higher growth rate with BG-11 medium and higher EPS productivity with BG-110 medium (without NaNO3). When the medium was supplemented with a mixture of glucose (4.0?g?L?1) and acetate (2.0?g?L?1), 1.79?g?L?1 biomass with BG-11 medium and 879.6?mg?L?1 of EPS production with BG-110 medium were achieved. Adopting this optimal ratio of glucose to acetate established in flask culture, the culture was also conducted in a 20-L photobioreactor with BG-11 medium for 7?days. A maximum biomass of 2.32?g?L?1 was achieved, and the EPS production was 634.6?mg?L?1. 相似文献
9.
Cornelius W. Sullivan 《Journal of phycology》1979,15(2):210-216
Silicic acid taken up from the growth medium by Navicula pelliculosa (Bréb.) Hilse was shown to enter at least two compartments: i) soluble pools; ii) insoluble fraction comprised predominantly of the silica frustule. Soluble Si pools were extracted by a variety of agents from cells uniformly labeled for ten generations in medium containing 68Ge-Si(OH)4. 100 C water soluble and 0 C perchloric acid (PCA) soluble Si pools of 680 mM Si·l?1 and 490 mM Si·l?1 cell water represented 13 and 9%, respectively, of total, cell Si in exponential growth phase cells. Uniformly labeled cells synchronized by the combined synchronization technique accumulate at the cell cycle stage where silica frustule development is initiated. These cells contain water and PCA soluble pools of 10 nmol Si·106 cells?1 and-8.8 nmol Si·106 cells?1, respectively. On addition of Si(OH)4, a rapid uptake ensues allowing the Si pool to expand 2.5-fold, apparently to provide precursors of the silica frustule. 相似文献
10.
The optimisation of submerged culture conditions and nutritional requirements was studied for the production of exopolysaccharide (EPS) fromPleurotus nebrodensis. The optimal temperature and initial pH for both mycelial growth and EPS production in shake flask cultures were 25 °C and 8.0, respectively. Maltose was found the most suitable carbon source for both mycelial biomass and EPS production. Yeast extract was favourable nitrogen source for both mycelial biomass and EPS production. Optimum concentration of each medium component was determined using the orthogonal matrix method. The optimal combination of the media constituents for mycelial growth and EPS production was as follows: 200 g l?1 bran, 25 g l?1 maltose, 3 g l?1 yeast extract, 1 g l?1 KH2PO4, 1 g l?1 MgSO4 7H2O. Under the optimal conditions, the mycelial biomass (4.13 g l?1) and EPS content (2.40 g l?1) ofPleurotus nebrodensis was 2.3 and 3.6 times compared to the control with basal medium respectively. 相似文献
11.
Simon H. T. Raharjo Witjaksono Miguel A. Gomez-Lim Guillermo Padilla Richard E. Litz 《In vitro cellular & developmental biology. Plant》2008,44(4):254-262
Embryogenic avocado cultures derived from ‘Hass’ protoplasts were genetically transformed with the plant defensin gene (pdf1.2) driven by the CaMV 35S promoter in pGPTV with uidA as a reporter gene and bar, the gene for resistance to phosphinothricin, the active ingredient of the herbicide Finale® (Basta) (Bayer Environmental Science, Research Triangle Park, Durham, NC ). Transformation was mediated by Agrobacterium tumefaciens strain EHA105. Transformed cultures were selected in the presence of 3.0 mg l?1 phosphinothricin in liquid maintenance medium for 3–4 mo. Liquid maintenance medium consisted of modified MS medium containing (per liter) 12 mg NH4NO3 and 30.3 mg KNO3 and supplemented with 0.1 mg l?1 thiamine HCl, 100 mg l?1 myo-inositol, 30 g l?1 sucrose, 3.0 mg l?1 phosphinothricin, and 0.41 μM picloram. Somatic embryo development from transformed cultures was initiated on MS medium supplemented with 45 g l?1 sucrose, 4 mg l?1 thiamine HCl, 100 mg l?1 myo-inositol, 10% (v/v) filter-sterilized coconut water, 3.0 mg l?1 phosphinothricin, and 6.0 g l?1 gellan gum. Limited plant recovery occurred from somatic embryos on semi-solid MS medium supplemented with 3.0 mg l?1 phosphinothricin, 4.44 μM 6-benzylaminopurine (BA), and 2.89 μM GA3; transformed shoots were micrografted on in vitro-grown seedling rootstocks. Approximately 1 yr after acclimatization in the greenhouse, transformed shoots were air-layered to recover transformed roots. Genetic transformation of embryogenic cultures, somatic embryos, and regenerated plants was confirmed by polymerase chain reaction (PCR), Southern blot hybridization, the XGLUC reaction for uidA, and application of the herbicide Finale® to regenerated plants. 相似文献
12.
Nodal explants of in vivo plants and in vitro seedlings of Wattakaka volubilis were cultured on Murashige and Skoog medium fortified with various concentrations of cytokinins — BA (0.5–5 mg l?1), KN (0.5–10 mg l?1),TDZ (0.05–1 mg l?1) either singly or in combination with NAA (0.1 mg l?1). KN proved best for inducing healthy shoots in both in vitro and in vivo derived explants. Maximum number of shoots (14.1±0.84) with 80% regeneration frequency was obtained from nodal explants of seedlings cultured on 5 mg 1?1 KN + 0.1 mg l?1 NAA. In vivo nodal explants produced a maximum of 4.2 shoots on MS medium fortified with 2 mg l?1 BA+0.1 mg l?1 NAA. The differentiated shoots from both could be rooted with 85% frequency on 1/2 strength MS medium (1% sucrose) with 0.6% agar + 1 mg l?1 IBA + 0.2 mg l?1 KN. Rooted shoots were transplanted to vermiculite-soil (3:1) mixture in polyethylene covered pots with 45% transplantation success. Peroxidase isozymes (native PAGE) analysis helped to verify the variation in regenerated plants. 相似文献
13.
Bo Zhao Foster A. Agblevor John G. Jelesko 《Plant Cell, Tissue and Organ Culture》2014,117(2):157-165
Previously, increased partitioning of the natural product nicotine from tobacco hairy roots into the culture media was achieved by altering the expression of the nicotine uptake permease gene. The present study demonstrated that further increases in nicotine yield in the media were attained by using surfactant-stabilized microbubbles. Compared to other non-ionic surfactants (Tween 20 and Tween 80) and the ionic surfactant SDS, Triton X-100 (TX100) both increased total nicotine production and exudation into the hairy root culture media. In comparison to surfactant-free medium, TX100 at 10, 25, and 50 mg l?1 did not show strong inhibition of hairy root growth. At 4,000 rpm shear speed, microbubbles stabilized by 10, 25, and 50 mg l?1 TX100 had k L a of 22.3, 36.2, and 44.1 h?1 in Gamborg’s B5 medium, respectively, in comparison to 16.4 h?1 with conventional air sparging. In a 1-l bioreactor, microbubbles stabilized by TX100 were applied to hairy roots after the inoculated root tips were self-immobilized by branching. With microbubble dispersion, dissolved oxygen rapidly increased from 60 to 85 %, and hairy root growth rate increased. Nicotine accumulation in culture medium with microbubbles reached 146 mg l?1 after 30 days cultivation. These results show that combining genetic modification with surfactant-stabilized microbubble dispersion can substantially increase levels of nicotine in the media of hairy root cultures. 相似文献
14.
Morphogenesis was induced in Eucalyptus globulus seeds, cotyledons, hypocotyls and leaves from in vitro clonal plantlets. Globular structures were observed after 2 weeks induction on B5 culture medium supplemented with 10% coconut water, 0.05–0.5 mg l?1 6-benzylaminopurine (BAP) and 0.5 mg l?1 indole-3-butyric acid (IBA). These continued to proliferate under dark conditions until the 2nd to 3rd subculture. Following transfer to a photoperiod of 16 h light, shoots evolved from these globular structures and developed further to plantlets. The influence of several factors, including culture medium composition, sucrose concentration, the type, concentration and combination of growth regulators and the presence of coconut water was studied. The percentage of explants showing globular structure formation and the number of globular structures per explant were evaluated. Macroscopic, histological and scanning electron microscopic studies revealed that the morphogenic process involved mainly organogenic nodules with fewer globular somatic embryos. The nodules gave rise to shoots and subsequently complete plants following incubation on B5 Gamborg medium containing 0.5 mg l?1 IBA and 30 g l?1 sucrose, which promoted root formation. 相似文献
15.
In order to determine the most suitable in vitro tissue culture and plant regeneration conditions for the small flowered willow herb (Epilobium parviflorum Schreb), various explants were cultured on semi-solid MS media containing factorial combinations of plant growth regulators. Callus induction from hypocotyl, cotyledon, petiole and leaf explants was achieved on media containing 2,4-dichlorophenoxy acetic acid (2,4-D) and kinetin (KIN). All other growth regulator combinations [□-naphtaleneacetic acid (NAA) ± benzylaminopurine (BAP), NAA ± thidiazuron (TDZ), indol acetic acid (IAA) ± Zeatin (ZEA)] tested failed to respond. The best results with cotyledon- and petiole- derived callus were obtained from MS medium supplemented with 1.0 mg l?1 2,4-D + 0.1 mg l?1 KIN and 2.0 mg l?1 2,4-D + 0.2 mg l?1 KIN. It was observed that B5 basal medium was more effective than MS basal medium for producing seedling and the most effective seed sterilizing solution was 25 % (v/v) sodium hypochlorite (NaOCl). No plant regeneration was observed in either callus induction or during the subculturing stage. This is the first report on in vitro tissue culture study within the genus Epilobium. 相似文献
16.
Regis Le-Feuvre Claudia Triviño Ana M. Sabja Michèle Bernier-Cardou Michael R. Moynihan Krystyna Klimaszewska 《In vitro cellular & developmental biology. Plant》2013,49(1):30-40
Repeated attempts to genetically transform Pinus radiata embryonal masses through cocultivation with Agrobacterium tumefaciens on MSG medium were unproductive due to Agrobacterium overgrowth. Timentin at either 200 or 400 mg?l?1 was ineffective in inhibiting bacterial growth after cocultivation. In this study, the causes of the abundant bacterial growth were investigated by comparing MSG medium with two other media (mLV and DCR) commonly used in conifer somatic embryogenesis. Statistical analysis of the growth data (optical density and number of cell-forming units) showed that bacterium grew significantly more on MSG than on mLV or DCR during the 48-h cocultivation. This enhanced growth was attributed to the higher concentration of L-glutamine in MSG. Lowering the concentration of L-glutamine in MSG to 0.5 g?l?1 resulted in similar growth of Agrobacterium compared with the other two media. MSG was also superior for the growth of radiata pine cells, with a statistically significant difference after 14 d of culture. Hence, to avoid bacterial overgrowth during and after cocultivation, a two-medium protocol was developed in which cocultivation was carried out on mLV, followed by 5 d on mLV with 400 mg?l?1 Timentin. Selection for transformed cells and further control of bacterial growth was then performed using MSG with Timentin and Geneticin. By sequential application of these two media, 2,096 cell colonies were selected; of these, 94 were analyzed and 49 were transgenic. These results highlight yet another factor that might be critical for the success of transformation experiments but has not been sufficiently studied until now: the growth dynamics and ability to eliminate A. tumefaciens on various plant tissue culture media. 相似文献
17.
Kåre Lindström 《Hydrobiologia》1983,104(1):325-328
Keratella cochlearis (Gosse) was cultured non-axenically in Carefoot medium diluted with Erken water at 5 °C, 15 °C and 20 °C with Rhodomonas minuta (Skuja) as a food alga. The rotifer reached ca. 120 ind. ml?1, having generation times of 2–7 days, a Q10-value of ca. 2, and at the lowest temperature >20% longer posterior spines. When co-cultured with Chlorella sp., at 0–30 mg Ca l?1 and 1.6 meq NaHCO3 l?1 in medium L 11 at 20 °C, the maximum generation time and individual numbers were 3–4 days and up to 100 ind. ml?1, respectively. Animal numbers increased in relation to nutrient multiples, up to two multiples, of the culture medium L 16. Growth and length were reduced, although the width increased above two multiples of this culture medium. The trace metal tolerance was broad and increased additions of a metal mixture (L 11) slightly increased the length of the rotifers. No major changes in the length were observed when HCO3 or Ca were varied in the culture medium (L 11), although a decrease in the length was noted in old cultures. 相似文献
18.
The industrially-important thermostable lipase, TliA, was extracellularly produced in the recombinant Pseudomonas fluorescens by the homologous expression of TliA and its cognate ABC protein exporter, TliDEF. To increase the secretory production of TliA, we optimized the growth temperature and the culture medium of P. fluorescens. The total amount and the specific productivity of lipase was highest at 25 °C of cell growth temperature, although maximal cell growth was observed at 30 °C. Using the culture medium composed of 20 g dextrin l?1, 40 g Tween 80 l?1 and 30 g peptone l?1, TliA was produced at a level of 2,200 U ml?1 in a flask culture. The TliA production increased about 3.8-fold (8,450 U ml?1) in batch fermentation using a 2.5 l fermentor, which was about 7.7-fold higher than that of previously reported TliA production. 相似文献
19.
Possibilities of adventitious buds induction on the cotyledons obtained from sterile seedlings ofAbies concolor xAbies grandis hybrid were investigated. The following variables influencing bud induction and their further development were studied: the effect of expiant age, the effect of different growth regulators and their concentrations and duration of their application. The most suitable expiants proved to be the cotyledons of 7 d old seedlings. The most efficient cytokinin was benzylaminopurine (S mg l?1) in combination with napthaleneacetic acid (0.01 mg l?1). The most optimal duration of treatment was 17 to 21 d culture of explants on induction medium. Shoot growth was achieved on basal medium to which 14 mg 1?1 spermidine was added. 相似文献
20.
Sangeeta Nath Alak K. Buragohain 《Journal of plant biochemistry and biotechnology.》2003,12(2):167-169
A rapid clonal propagation system has been developed for the medicinally important herb Centella asiatica (L) Urban by shoot tip (2–3 cm long) culture. The shoot tips isolated from mature plants were inoculated on MS medium incorporated with BA alone or in combination with NAA and Kn. The optimum number of shoots (3.38) with optimum number of leaves per shoot (4.25) were attained on MS medium supplemented with 4.0 mg l?1 BA and 0.1 mg l?1 NAA. On transferring the microshoots on full strength MS medium supplemented with various concentrations of IBA (1.0-3.0 mg l?1) and NAA (0.5-2.0 mg l?1), profuse rooting (46.8 per shoot) was obtained in MS basal medium with 2.0 mg l?1 IBA with root length of 19.7 cm. Well rooted plantlets were acclimatized successfully by adjusting the temperature and humidity for 3–4 weeks after transfer to pots filled with sterilized vermiculite soil: sand (1:1)mixture. This micropropgation protocol could be useful for raising a stock of genetically homogenous material for field cultivation within a very short period. 相似文献