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1.
Hiroshi Yonemitsu Koichiro Shimomura Motoyoshi Satake Shunji Mochida Masahiko Tanaka Thoru Endo Akira Kaji 《Plant cell reports》1990,9(6):307-310
Hairy roots were obtained following inoculation of the stems of Lobelia inflata L. with Agrobacterium rhizogenes strain ATCC 15834. These hairy roots contained agropine and mannopine. In addition, lobeline was detected by HPLC and confirmed by mass spectrometry. Various media were tested for the growth of hairy roots as well as for the content of lobeline in hairy roots. The growth rate of hairy roots cultured in Nitsch and Nitsch's medium was approximately one third of those cultured in other media. The lobeline content of hairy roots (18–54 g/g dry weight) cultured in these media was the same order of magnitude compared with that of roots of L. inflata (24 g/g dry weight) cultivated in pots. The hairy roots cultured in Nitsch and Nitsch's medium were morphologically different from those cultured in other media.Abbreviations MS medium
Murashige and Skoog's medium
- 1/2 MS medium
one-half strength of the standard Murashige and Skoog's medium
- B5 medium
Gamborg's B5 medium
- NN medium
Nitsch and Nitsch's medium
- FW
fresh weight
- DW
dry weight 相似文献
2.
Hypocotyl explants of three leguminous forest tree species, Albizia amara, A. lucida and A. richardiana, have differentiated shoot buds on B5 basal medium. Maximum number of shoots per explant developed on basal medium augmented with 2,4-D (0.1 M) in A. amara (2) and BA (10 M) for both A. lucida (2) and A. richardiana (1.6). Higher concentrations of auxins in the medium, in general, enhanced rooting and callusing but cytokinins promoted the growth of green calli. BA enchanced the differentiation of shoots in the three species. The in vitro grown shoots of A. amara and A. richardiana, after subculturing on B5+1 M IAA developed roots (up to 30–40%). These plants have been successfully transferred to the field.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
6-benzylaminopurine
- BM
Gamborg's B5 medium with 0.9% agar+3% sucrose
- IAA
Indole-3-acetic acid
- IBA
Indole-3-butyric acid
- Kn
Kinetin
- NAA
-naphthaleneacetic acid 相似文献
3.
Summary Hairy root cultures of Solanum aculeatissimum were established by trans-formation using Agrobacterium rhizogenes strain 15834. Root growth and production of steroidal saponin were investigated under various culture conditions. Transformed roots grew better in Gamborg's B5 medium containing 3 % sucrose under continuous light than in the dark. Also, the roots turned light green when cultured under continuous light. Green hairy roots produced aculeatiside A (6.71mg ·) L–1 and aculeatiside B (6.39mg · L–1) after 8 weeks of culture, while no steroidal saponin was detected in hairy roots cultured in the dark. Of the three culture media tested, Gamborg's B5 medium was superior for growth and steroidal saponin production. Growth and steroidal saponin production were enhanced when 100g · L–1 auxin except for 2,4-D was added to the medium. The addition of 2,4-D inhibited growth. Production of steroidal saponin was highest with NAA. Transformed roots used in this experiment were confirmed that hairy roots examined contain both TL-DNA and TR-DNA region of Ri plasmid by PCR amplification analysis of DNA.Abbreviations MS medium
Murashige and Skoog's medium (1962)
- B5 medium
Gamborg's B5 medium (1968)
- LS medium
Linsmaier and Skoog's medium(1965)
- HPLC
High performance liquid chromatography
- NAA
-Naphthaleneacetic acid
- IAA
Indole-3-acetic acid
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- PCR
polymerase chain reaction 相似文献
4.
Roots of marigold (Tagetes patula L.) accumulate thiophenes, heterocyclic sulfurous compounds with strong biocidal activity. In detached roots cultured in vitro, the thiophene content was 5 mol·(g fresh weight)-1 which is 25-times higher than in roots attached to the plant. In roots derived from tissues transformed by Agrobacterium tumefaciens and A. rhizogenes, the morphology and thiophene content varied with the bacterial strain used. Transformation stimulated the elongation of the root tips and the formation of lateral roots but lowered the thiophene level to 20–50% relative to the concentration in untransformed detached roots. A negative correlation was found between the number of laterals in a root system and the thiophene content. Extensive branching and a decrease in thiophene accumulation was evoked in untransformed roots by indole-3-acetic acid (1–10 mol·l-1) added to the medium. Within the roots, the highest thiophene concentrations were found in the tips. The results indicate that auxin directly or indirectly plays a role in the regulation of the thiophene level in root tips.Abbreviations B5
Gamborg's B5 medium
- IAA
Indole-3-acetic acid 相似文献
5.
Multiple shoots have developed from axillary buds excised from in vitro grown seedlings of Acacia auriculiformis on Gamborg's (B5) basal medium supplemented with coconut milk (5 or 10%) and benzylaminopurine (10-6M). These shoots, if transferred individually to indole-3-acetic acid (10-7M) or naphthaleneacetic acid (10-6 or 10-7M) augmented B5 medium, produced roots at their base.Abbreviations B5
Gamborg's basal medium (BM)
- CM
coconut milk
- BA
6-benzylaminopurine
- Kn
kinetin
- IAA
indole-3-acetic acid
- NAA
-naphthaleneacetic acid 相似文献
6.
Protoplasts were isolated from cotyledons and foliage leaves of cotton (Gossypium hirsutum and G. barbadense). Cotyledon protoplasts were larger and responded to culture better than leaf protoplasts. Cotyledon derived protoplasts regenerated cell walls and formed microcolonies of 2–3 cells in G. hirsutum and 5–8 cells in G. barbadense. However, the microcolonies did not grow beyond this stage. Protoplast yield and viability, cell wall regeneration and cell division were influenced by several factors, e.g., genotype, age, tissue and growth condition of donor plant, enzyme mixture and concentration, preplasmolysis period, incubation period, and culture medium.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- GA3
gibberellic acid
- p CPA
p-chlorophenoxyacetic acid
- MES
2[N-morpholino]ethanesulfonic acid 相似文献
7.
D. A. Godbole M. N. Kunachgi U. A. Potdar K. V. Krishnamurthy A. F. Mascarenhas 《Plant cell reports》1984,3(2):75-78
Plantlets regenerated from shoot apices, cotyledons and callus cultures in Moth bean, Vigna aconitifolia (JACQ) Marechal, a drought resistant legume and pulse crop, were rooted and transferred to soil. Explants for these studies were derived from seedlings pre-conditioned by germination of seeds on B5BA and WMB (control).Abbreviations MS
Murashige and Skoog (1962)
- B5
B5 basal medium (Gamborg et al 1968)
- B5BA
B5 basal medium containing BA (2.25 mg/l)
- WMB
Modified White's medium (Mascarenhas et al 1976)
- BA
6-benzyladenine
- IAA
indole-3-acetic acid
- NAA
1-napthaleneaceticacid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IBA
indolebutyric acid
- 2iP
N(–2 isopentyl) adenine
- CM
coconut milk
NCL Communication No. 3375 相似文献
8.
A new approach to direct somatic embryogenesis in Medicago 总被引:2,自引:0,他引:2
A highly efficient system for direct somatic embryogenesis is described. Leaf sections originating from young trifoliate leaves of Medicago falcata line 47/1–5 and Medicago sativa line No2/9R, directly produced embryos after cultivation in liquid B5IV induction medium. In comparison with indirect somatic embryogenesis the system omits the callus stage and thus allows shortening of the process of somatic embryogenesis in alfalfa by 35–40 days. It permits the avoidance of secondary changes occurring during the process of dedifferentiation. A modified B5/3H medium containing Polyethylene Glycol 6000 promoted embryo development from globular up to torpedo stage. It was clearly shown that 2.5% Polyethylene Glycol stimulated this process for both H. falcata 47/1–5 and M. sativa No 2/9R. Maturation of torpedo stage embryos was carried out on solidified or liquid abscisic acidcontaining medium. A 30M abscisic acid concentration was optimal in allowing one embryo to yield one plant. Somatic embryo conversion to plants and plant regeneration was performed on Murashige and Skoog medium. Regenerated plants showed a normal morphology.Abbreviations ABA
Abscisic acid
- B5
Medium of Gamborg et al.(1968)
- COT
Cotyledone stage embryos
- 2,4-D
2,4-dichlorphenoxyacetic acid
- FW
Fresh weight
- GA3
Gibberellin A3
- MS
Medium of Murashige and Skoog (1962)
- PEG
Polyethylene Glycol
- POLY
Polyembryos 相似文献
9.
Neera Bhalla-Sarin Suman Bagga Sudhir K. Sopory Sipra Guha-Mukherjee 《Plant cell reports》1986,5(5):322-324
Calli from young embryos of Cocos nucifera L. were induced on B5 medium supplemented with IAA-conjugates (IAA-asp or IAA-ala) at a concentration of 2.0 mg/1 and callusing was increased by about 10% if both IAA-conjugates, IAA-asp and IAA-ala were added together. Differentiation of shoots and roots was achieved by transferring calli to B5 medium supplemented with either IAA-asp (2.0 mg/1)+Kn(2.0 mg/1) or NAA (2.0 mg/1). Complete plantlets were obtained on B5 medium supplemented with NAA (0.5 mg/1)+BAP (2.0 mg/1)+PVP (1.0 g/1).Abbreviations IAA
Indole-3-acetic acid
- IAA-ala
Indole acetyl-L-alanine
- IAA-asp
Indole acetyl-L-aspartic acid
- Kn
Kinetin
- BAP
N6-benzylaminopurine
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- NAA
-naphthalene acetic acid
- PVP
Polyvinylpyrolidone 相似文献
10.
Establishment of hairy root cultures of Psoralea species 总被引:1,自引:0,他引:1
Christophe Nguyen Frédéric Bourgaud Paul Forlot Armand Guckert 《Plant cell reports》1992,11(8):424-427
Eight Psoralea species (Leguminosae) were inoculated with Agrobacterium rhizogenes, strains 8196 and 9402. Hairy roots were only induced by strain 9402. Attention was focussed on Psoralea lachnostachys. Transformed roots grew very rapidly in Gamborg B5 liquid medium with a doubling time of the culture of 38 hours. Whatever the culture conditions, the two furanocoumarins usually found in roots of Psoralea plants, psoralen and angelicin, were not detected in cultured transformed and non transformed roots even when some chitosan was added to the medium. However, 669 g.g–1 dry matter of psoralen and 215 g.g–1 dry matter of angelicin were found in roots from soil grown plants. A possible translocation of these compounds from the aerial parts to the roots is suggested.Abbreviations B5 medium
Gamborg's medium (Flow laboratories's formulation)
- NAA
Naphthaleneacetic acid 相似文献
11.
Somatic embryogenesis and regeneration of plants in the bamboo Dendrocalamus strictus 总被引:1,自引:0,他引:1
Somatic embryogenesis leading to plant regeneration has been achieved in the bamboo, Dendrocalamus strictus, by culturing seeds (caryopses) on B5 basal medium supplemented with 2,4-dichlorophenoxyacetic acid. Callus cultures obtained from the embryonal end of the seeds differentiated chlorophyllous embryoids. On transfer to a germination medium (B5 liquid, sucrose, indolebutyric acid, and -naphthaleneacetic acid) 40% of the embryoids developed into plantlets. Further development of the plantlets occured on B5 liquid medium (half strength) + sucrose (1%) + IBA (5 × 10–7M) + NAA (10–7M).Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- IBA
Indolebutyric acid
- NAA
-naphthaleneacetic acid 相似文献
12.
Somatic embryogenesis and plant regeneration in diploid Allium fistulosum × A. cepa F1 hybrid onions
Procedures were developed for disinfestation of non-dormant basal plate tissue excised from field grown basal plate tissue of diploid Allium fistulosum × A. cepa F1 hybrid onions. Contamination levels varied with the season and vegetative development of plant material. Callus initiated from basal plate tissue and immature inflorescences of the F1 hybrids was maintained on a BDS-based medium containing 0.75 mg/l picloram and 2.0 mg/l BA. When this medium was supplemented with vitamins and glycine, and with proline at 2.5 gm/1, somatic embryos began to form. Their development continued on a BDS-based shoot promotion medium containing 0.03 mg/l picloram and 0.32 mg/l 2iP supplemented with vitamins, glycine and proline. Genotypes differed significantly in the numbers of structures regenerated. Plantlets from somatic embryos were rooted into BDS or half-strength BDS medium without growth substances and were successfully transferred to sterilized potting mix in plastic commercial corsage boxes.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid
- 2iP
N6-(2-isopentenyl)adenine
- NAA
1-naphthylacetic acid
- BDS
Gamborg's B5 medium modified by Dunstan and Short (1977a) 相似文献
13.
Coronilla varia L. (crownvetch) plants were regenerated from callus cultures through somatic embryogenesis. Callus cultures were initiated using hypocotyls excised from sterile seedlings. Cultures were then transferred from a modified Gamborg's B5 medium containing 2,4-D to a medium containing no plant growth regulators (basal B5). Formation of embryos was evident in 12 of 32 callus lines after transfer of callus to BOi2Y (modified Blayde medium supplemented with 100 mg inositol and 2 g yeast extract/L). Basal B5 supplemented with 10 mM asparagine or 20 mM NH4Cl could be substituted for BOi2Y. Embryos subsequently transferred to basal B5 developed roots and shoots. Plants thus formed were first transferred to vermiculite and then to soil.Contribution No. 8219 of the U.S. Regional Pasture Reasearch Laboratory, USDA-ARS, University Park, PA, U.S.A. 相似文献
14.
Summary Rapidly proliferating and highly regenerable suspension cultures of somatic embryos of Carica papaya x C. cauliflora were used for protoplast isolation. On average, protoplast yield was 1.5×106/g fresh weight of somatic embryos. Protoplasts were first cultured in liquid KM8P-S medium for 2 weeks and then plated in the same medium solidified with 1% agarose. About 1.4% of the protoplasts developed directly into somatic embryos. Protoplast-derived somatic embryos proliferated rapidly through direct embryogenesis on modified MS medium supplemented with 1 mg/1 ABA, and developed into plantlets upon transfer to MS medium devoid of plant growth regulators. The plantlets were successfully transplanted to soil.Abbreviations MS
Murashige and Skoog medium (1962)
- KM8P
Kao and Michayluk medium (1975)
- NAA
-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- ABA
abscisic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- CPW
Frearson et al. medium (1973) 相似文献
15.
The studies for production of saikosaponins by tissue culture of Bupleurum falcatum L. were carried out to produce saikosaponins with several kinds of media and plant hormones. Among the media and plant hormones studied, Gamborg's B-5 [23] medium containing 0.5 ppm kinetin (k) and 1.0 ppm 3-indolebutyric acid (IBA) was the most effective medium and hormone for production of saikosaponins. The highest content of saikosaponin-d in the dried cells was 0.26%, which was similar to a concentration of Bupleuri Radix.Abbreviations
MS
medium used by Murashige and Skoog [22]
-
G
medium used by Gamborg (B 5) [23]
-
W
medium used by White [24]
-
NN
medium used by Nitsch and Nitsch [25]
-
k
Kinetin
-
BAP
6-Benzylaminopurine
-
2,4 D
2,4-Dichlorophenoxyacetic acid
-
NAA
-Naphtylacetic acid
-
IBA
3-Indolebutyric acid
-
IAA
3-Indoleacetic acid
-
ssd
saikosaponin-d
-
PM
Production medium
- dw
Dry weight 相似文献
16.
Summary Some 19 strains ofAspergillus niger,A. oryzae, andPaccilomyces spp. are tested for their ability to grow on the supernatant remaining after the expressed juice from sugar beet tops and meadow grass has been heat-treated to precipitate crude leaf protein, and supplemented as required by glucose or ammonium sulphate. With effective strains ofA. niger,A. oryzae,P. elegans orP. variotii and an optimized carbohydrate/nitrogen ratio, over 70% of the organic content of the supernatant is rapidly converted into mycelial biomass of high protein content. 相似文献
17.
Six sugarbeet (Beta vulgaris L.) lines (GWI-248, SPB-11, MonoHy 55, SMS-1, EL45 and FC607) were tested for regeneration. Shoot cultures were initiated in vitro from naked, sterilized embryos obtained from mature seed. Excised petioles from cultured shoots were plated on Gamborg's B5 medium and four modified Murashige and Skoog (MS) media. A medium containing MS inorganic salts supplemented with 0.4 mg/1 N6-benzyladenine, 0.1 mg/1 indole-3-butyric acid, ten vitamins and six amino acids, termed RV, was superior for both adventitious shoot and callus formation. Callus was observed only on RV medium and only on petioles that did not develop adventitious buds directly. Rooting of regenerated shoots and development of complete plants was accomplished by transfer to Gamborg's B5 medium with 5 mg/l indole-3-butyric acid as the sole phytohormone. The complete process of regeneration through adventitious shoot production took from 4 to 6 weeks from explants to rooted plants. The callus that formed on nonorganogenic petioles was regenerative when transferred to fresh RV medium. Regeneration from callus occurred mainly by shoot organogenesis but also by somatic embryogenesis at a low frequency.Abbreviations BA
N6-benzyladenine
- IBA
indole-3-butyric acid
Contribution from Missouri Agricultural Experiment Station. Journal Series No. 10394. University of Missouri, Columbia, MO 63873, USA Mention of trade or company name does not constitute a guarantee or warranty of the product by University of Missouri-Columbia or U.S.D.A. Agricultural Research Service and does not imply their approval to the exclusion of other products that may be suitable. 相似文献
18.
The toxicity of kanamycin, hygromycin B, geneticin, methotrexate and cefotaxime on zygotic embryos ofPicea glauca was studied. Embryos placed on bud induction medium produced approximately 20 adventitious buds per embryo under control conditions. Addition of antibiotics reduced the number of bud-forming embryos. Using the percentage of embryos with buds as an indication of antibiotic toxicity, two-day-old explants were found to be more sensitive than nine-day-old. Kanamycin toxicity was enhanced by cefotaxime and this effect increased with increases in concentration of either antibiotic. Although no morphological difference was observed after 21 days, embryos growing on medium containing 20 g ml–1 kanamycin showed a decrease of 73% in dry weight and 23% in protein content per embryo when compared to control embryos. Similarly, a decrease of 38% in dry weight and 40% in protein content per embryo was found in embryos on medium containing 300 g ml–1 cefotaxime.Abbreviations BA
6 benzyl-aminopurine
- EDTA
ethylenediamine-tetraacetic acid
- PVP 10
polyvinylpyrrolidone (MW 10,000)
- Tris-HCl
Tris [hydroxymethyl] amino hydrochloride
NRCC No. 30262 相似文献
19.
An efficient system for gene transfer into plants of Brassica juncea var. India Mustard, mediated by Agrobacterium tumefaciens. was developed through the manipulation of the culture medium and the use of the appropriate Agrobacterium strain. High frequency shoot regeneration (90–100%) was obtained from hypocotyl explants grown on medium containing 0.9% agarose, 3.3 mg/L AgNO3 and 0.5–2 mg/L BA in combination with 0.01–0.05 mg/L 2,4-D or 0.1–1 mg/L NAA. Of all the Agrobacterium strains tested, A. tumefaciens A208-SE, carrying the disarmed Ti plasmid and a binary vector pROA93, was the most effective for B. juncea transformation. pROA93 carries the coding sequences of the NPTII and the GUS genes, both driven by a common CaMV 35S promoter in two divergent directions. Inoculated explants grown on the selection medium in the presence of 0.5 mg/L BA and 0.1 mg/L NAA gave rise to transgenic shoots at the highest frequency (9%). All Ro transgenic plants were phenotypically normal, but variation in expression patterns of the GUS gene occurred among the transgenic plants in an organ- and tissue-specific manner. Both the NPTII and the GUS genes were transmitted to the R1 seed progeny and showed co-segregation.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- NPTII
neomycin phosphotransferase type II
- GUS
-glucuronidase
- CaMV
cauliflower mosaic virus
- MS
Murashige and Skoog
- X-Gluc
5-bromo-4-chloro-3-indolyl-D--glucuronic acid
- IBA
indolebutyric acid
- SDS
sodium dodecyl sulfate 相似文献
20.
The partitioning of carbon between reserve polysaccharide and alkaloid secondary products was investigated in batch cultures of transformed roots of Datura stramonium grown in media in which the carbon substrate concentration was held constant and the level of mineral nutrients was varied. The growth and accumulation of starch and hyoscyamine was examined in roots grown at temperatures of 20°C, 25°C or 30°C in media containing 5% sucrose and levels of mineral nutrients varying from 1/4 to twice the standard level of Gamborg's B5 salts. The dry matter content was highest (up to 15% w/w) in roots grown at either 20°C or 25°C in medium of the lowest ionic strenth (1/4 B5 salts) and decreased as the ionic strength was raised (down to 7% w/w with 2 B5 salts). Up to half of this decrease could be accounted for by loss of starch from the roots. At 20°C and 25°C, the starch content of the roots grown in medium of the lowest ionic strength (1/4 B5) was 40 mg g-1 and 22 mg g-1 fresh weight respectively but decreased to less than 1 mg g-1 weight at either temperature when the ionic strength of the medium was raised to 2 B5. At 30°C, starch accumulation was severely inhibited in all media. In contrast, varying either the temperature or the ionic strength of the medium had only a small effect on hyoscyamine accumulation which remained at between 0.4–0.6 mg g-1 fresh weight. Although increases in the level of mineral salts had little effect on the hyoscyamine content of the roots, total yields however, increased due to stimulation of growth. Time course experiments showed that cultures grown at either 20°C or 25°C continued to accumulate both starch and hyoscyamine into late stationary phase. 相似文献