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1.
Factors affecting Agrobacterium tumefaciens-mediated transformation in several black poplar clones 总被引:4,自引:0,他引:4
M. Confalonieri A. Balestrazzi S. Bisoffi R. Cella 《Plant Cell, Tissue and Organ Culture》1995,43(3):215-222
Transient expression of the uidA reporter gene was used in preliminary experiments with two oncogenic and two disarmed Agrobacterium tumefaciens strains in order to test the efficiency of T-DNA transfer to N084 x Populus nigra and N107 x P. nigra clones. The oncogenic strain A281 pKIWI105 produced the highest average number of GUS spots per leaf disc. In order to optimize the production of transgenic plantlets from different P. nigra clones (San Giorgio, Jean Pourtet, N084 x P. nigra and N107 x P. nigra, respectively), two A. tumefaciens strains (GV2260 p35S GUS, A281pKIWI105) and bacterial concentrations (7×108; 1.2×09 bacteria ml-1) were used. Following co-cultivation with A281 pKIWI105, the frequency of leaf discs producing kanamycin-resistant calli was not significantly different between the clones and bacteria concentrations used. Transformed shoots were regenerated from all clones, except for Jean Pourtet. Co-cultivation of leaf discs with GV2260 p35S GUS produced very few calli which died when transferred to selective regeneration medium. In addition, the effects of acetosyringone and leaf wounding were evaluated for the San Giorgio and Jean Pourtet clones, using the same strains. Factors which significantly affected the transformation efficiency of leaf explants were the P. nigra clone, the A. tumefaciens strain, and the presence of acetosyringone. Genetic transformation of calli and regenerated plantlets was confirmed by their ability to grow and root on Woody Plant Medium containing kanamycin, by histochemical -glucuronidase assays, and Southern blot hybridization analyses.Abbreviations BA
benzyladenine
- GUS
-glucuronidase
- IBA
indolebutyric acid
- MS
Murashige and Skoog
- NAA
-naphthaleneacetic acid
-
nptII
neomycin phosphotransferase II gene
-
uidA
-glucuronidase gene
- WPM
Woody Plant Medium 相似文献
2.
Summary Twenty-three independent kanamycin resistant lines were obtained after cocultivation of longterm embryogenic cultures of three Asparagus officinalis L. genotypes with an Agrobacterium tumefaciens strain harboring ß-glucuronidase and neomycin phosphotransferase II genes. All the lines showed ß-glucuronidase activity by histological staining. DNA analysis by Southern blots of the kanamycin resistant embryogenic lines and of a plant regenerated from one of them confirmed the integration of the T-DNA.Abbreviations GUS
ß-glucuronidase
- X-Gluc
5-bromo-4-chloro-3indolyl ß-D-glucuronic acid
- NPT II
neomycin phosphotransferase II 相似文献
3.
Agrobacterium-mediated transformation of sweet orange and regeneration of transgenic plants 总被引:1,自引:6,他引:1
Leandro Peña Magdalena Cervera José Juárez Antonio Navarro José A. Pina Nuria Durán-Vila Luis Navarro 《Plant cell reports》1995,14(10):616-619
Summary Transgenic sweet orange (Citrus sinensis L. Osbeck) plants have been obtained by Agrobacterium tumefaciens-mediated gene transfer. An hypervirulent A. tumefaciens strain harboring a binary vector that contains the chimeric neomycin phosphotransferase II (NPT II) and ß-glucuronidase (GUS) genes was cocultivated with stem segments from in vivo grown seedlings. Shoots regenerated under kanamycin selection were harvested from the stem segments within 12 weeks. Shoot basal portions were assayed for GUS activity and the remaining portions were shoot tip grafted in vitro for production of plants. Integration of the GUS gene was confirmed by Southern analysis. This transformation procedure showed the highest transgenic plant production efficiency reported for Citrus.Abbreviations BA
benzyladenine
- CaMV
cauliflowermosaic virus
- GUS
ß-glucuronidase
- LB
Luria Broth
- MS
Murashige and Skoog
- NAA
naphthalenacetic acid
- NOS
nopaline synthase
- NPT II
neomycin phosphotransferase II
- PEG
polyethylene glycol
- RM
rooting medium
- SRM
shoot regeneration medium 相似文献
4.
An efficient system for Agrobacterium tumefaciens-mediated transformation of Solanum gilo was established. The marker genes for kanamycin resistance and ß-glucuronidase expression were introduced. A comparison between cotyledon and hypocotyl explants showed that while regeneration was better from hypocotyl explants, cotyledon explants gave better transformation efficiency (46% vs. 32%). Four levels of kanamycin selection (100, 150, 200 and 250 mg/l) were tested for effect on transformation efficiency with each type of explant. Lower levels of kanamycin worked better using cotyledon explants, while higher levels of kanamycin worked better for hypocotyl explants. All nine t0 plants tested for expression of the kan
r
gene were positive. The progeny of three of these plants showed a pattern of classical Mendelian inheritance (3 to 1) for both the kan
r
and the ß-glucuronidase genes.Abbreviations MS
Murashige and Skoog (1962) medium
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- NPTII
neomycin phosphotransferase
- GUS
ß-glucuronidase 相似文献
5.
Monique F. van Wordragen Jan de Jong Hans B. M. Huitema Hans J. M. Dons 《Plant cell reports》1991,9(9):505-508
Summary To develop an Agrobacterium mediated transformation protocol for chrysanthemum we studied the transformation efficiency of commonly used A.tumefaciens strains on 14 genotypes by comparing tumour size and frequency. One genotype was analyzed in detail using 14 strains of both A.tumefaciens and A.rhizogenes. Only a few genotype/strain combinations resulted in significant tumour formation. Especially 0-type strains were highly efficient. An 0-type strain was used to transfer genes for neomycine phosphotransferase (NPT II) and ß-glucuronidase (GUS) to a susceptible cultivar. Transfer of the GUS gene was confirmed by using the Polymerase Chain Reaction (PCR). 相似文献
6.
Jovanka Miljuš-Djukić Mirjana Nešković Slavica Ninković Radomir Crkvenjakov 《Plant Cell, Tissue and Organ Culture》1992,29(2):101-108
Genetic transformation of buckwheat (Fagopyrum esculentum Moench.) and regeneration of transgenic plants were obtained by using Agrobacterium tumefaciens strains as vectors. Buckwheat cotyledons were excised from imbibed seeds, co-cultivated with A. tumefaciens and subjected to previously reported protocols for callus and shoot regeneration. The transformation with oncogenic strains was confirmed by opine and DNA analyses of tumour tissue extracts. Plants were regenerated on cotyledon fragments incubated with strain A281, harboring pGA472, which carries the neomycin phosphotransferase II gene for kanamycin resistance. The transformation of resistant shoot clones was confirmed by NPTII enzyme assay and DNA hybridization. A large number of transformed shoots were rooted and fertile plantlets were raised in the greenhouse. Transgenic plants comprised pin and thrum clones, which were allowed to cross-pollinate. In about 180 R2 seeds tested for kanamycin resistance, the ratio of resistant to sensitive seedlings was roughly 3:1.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
dichloro-phenoxyacetic acid
- 2iP
6-(, ,-dimethylallyl-amino)-purine
- IBA
indole-3-butyric acid
- IAA
indole-3-acetic acid
- Km
kanamycin
- NPTII
neomycin phosphotransferase II 相似文献
7.
Kazuhito Akama Hideaki Shiraishi Shozo Ohta Kenzo Nakamura Kiyotaka Okada Yoshiro Shimura 《Plant cell reports》1992,12(1):7-11
Summary The efficiency of Agrobacterium-mediated transformation of Arabidopsis thaliana was compared with different organs, Arabidopsis ecotypes, and Agrobacterium strains. Efficiency of shoot regeneration was examined using hypocotyl, cotyledon and root explants prepared from young seedlings. Hypocotyl expiants had the highest regeneration efficiency in all of the four Arabidopsis ecotypes tested, when based on a tissue culture system of callus-inducing medium (CIM: Valvekens et al. 1988) and shoot-inducing medium (SIM: Feldmann and Marks 1986). Histochemical analysis using the ß-glucuronidase (GUS) reporter gene showed that the gusA gene expression increased as the period of preincubation on CIM was extended, suggesting that dividing cells are susceptible to Agrobacterium infection. In order to obtain transgenic shoots, hypocotyl explants preincubated for 7 or 8 days on CIM were infected with Agrobacterium containing a binary vector which carries two drug-resistant genes as selection markers, and transferred to SIM for selection of transformed shoots. Of four Arabidopsis ecotypes and of three Agrobacterium strains examined, Wassilewskija ecotype and EHA101 strain showed the highest efficiency of regeneration of transformed shoots. By combining the most efficient factors of preincubation period, Arabidopsis ecotype, tissue, and bacterial strain, we obtained a transformation efficiency of about 80–90%. Southern analysis of 124 transgenic plants showed that 44% had one copy of inserted T-DNA while the others had more than one copy.Abbreviations AIM
Agrobacterium infection medium
- CIM
callus-inducing medium
- CTAB
cetyltrimethylammonium bromide
- 2,4-D
2,4-dichlorophenoxy-acetic acid
- GUS
ß-glucuronidase
- hph
hygromycin phosphotransferase
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- 2ip
N -(2-isopentenyl) adenine
- NPTII
neomycin phosphotransferase II
- RIM
root-inducing medium
- 35S
cauliflower mosaic virus 35S promoter
- SIM
shoot-inducing medium 相似文献
8.
Summary TransgenicPelargonium X hortorum have been producedvia Agrobacterium tumefaciens-mediated transformation. The regeneration protocol used provided a regeneration frequency approximately to 95 percent. Clumps of regenerants, from cotyledons and hypocotyls ofPelargonium X hortorum seedlings, were inoculated with the disarmed strain EHA101 ofAgrobacterium tumefaciens. This strain contains a binary vector carrying neomycin phosphotransferase II, hygromycin B phosphotransferase and ß-glucuronidase genes. Selection on the regeneration medium supplemented with hygromycin allowed production of transgenic plants in up to 20% of the inoculated explants. The insertion of foreign DNA was demonstrated by Southern and polymerase chain reaction analysis: these experiments indicated that the inserted T-DNA is not full length for most of the plants. All RO transgenic plants exhibited a normal phenotype and are fertile.Abbreviations GUS
ß-glucuronidase coding sequence
- PCR
polymerase chain reaction
- CaMV
cauliflower mosaic virus
- NPTII
neomycin phosphotransferase coding sequence
- NOS
nopaline synthase gene promoter and terminator
- HPH
hygromycin B phosphotransferase coding sequence
- SDS
sodium dodecyl sulphate
- EDTA
(ethylenedinitro trilo)tetra-acetic acid disodium salt 相似文献
9.
Summary
Agrobacterium-mediated gene transformation of Populus tremuloides Michx was accomplished by co-cultivation of leaf disks excised from greenhouse plants with Agrobacterium tumefaciens containing a binary Ti-plasmid vector harboring chimeric neomycin phosphotransferase (NPT II) and ß-glucuronidase (GUS) genes. Shoot regeneration in the presence of kanamycin was achieved when thidiazuron (TDZ) was used as a plant growth regulator. Transformation was verified by amplification of NPT II and GUS gene fragments from genomic DNA of transgenic plants with polymerase chain reaction (PCR) and integration of these genes into nuclear genome of transgenic plants was confirmed by genomic Southern hybridization analysis. Histochemical assay revealed the expression of GUS gene in leaf, stem and root tissues of transgenic plants, further confirming the integration and expression of T-DNA in these plants. This protocol allows effective transformation and regeneration of quaking aspen using greenhouse-grown materials as an explant source. Whole plant regeneration from cuttings of fieldgrown mature quaking aspen and hybrid poplar (P. alba x P. grandidentata) was also readily achieved by using this protocol, which represents a potential system for producing transgenic quaking aspen and hybrid poplar of valuable genotypes.Abbreviations AMV RNA4
Alfalfa mosaic virus RNA4
- BA
6-benzyladenine
- CaMV
cauliflower mosaic virus
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EDTA
ethylenediaminetetraacetic acid
- FAA
formalin-acetic acid-alcohol
- GUS
ß-glucuronidase
- NAA
1-naphthylacetic acid
- NPT II
neomycin phosphotransferase II
- PCR
polymerase chain reaction
- SDS
sodium dodecyl sulphate
- TE
Tris-Cl/EDTA
- TDZ
N-phenyl-N-1,2,3-thiadiazol-5-yl-urea (thidiazuron)
- WPM
woody plant medium (Lloyd and McCown 1980)
- X-GLUC
5-bromo-4-chloro-3-indolyl-ß-glucuronic acid 相似文献
10.
Leaf pieces of in vitro-cultured plantlets of the wild potato species Solanum brevidens Phil. were cocultivated with Agrobacterium tumefaciens that contained nptII and uidA genes on the disarmed plasmid pBI121. Independent transgenic shoots were regenerated from solidified and liquid medium that contained 50 mg l–1 kanamycin. Two Agrobacterium strains were investigated for transformation efficiency. GV2260, which contained p35SGUSINT, resulted in a 11% transformation frequency, compared with 1% using LBA4404. Transformation rates were 7% in liquid culture and 3% on solidified medium. All kanamycinresistant, putatively transformed plantlets were confirmed positive by histochemical GUS assays. GUS activity in 22 independently transformed plants was quantified by fluorometric assay. Southern analysis of randomly selected transgenic plants showed that each transgenic plant contained at least one copy of the uidA gene.Abbreviations GUS
ß-glucuronidase
- MS
Murashige-Skoog medium
- BA
6-benzylaminopurine
- 2ip
6-(, -dimethylallylamino)purine
- IAA
indole-3-acetic acid
- GA3
gibberellic acid
-
npt II
neomycin phosphotransferase II
- NOS
nopaline synthase
- MUG
4-methyl umbelliferyl glucuronide
- MU
7-hydroxy-4-methylcoumarin
- X-gluc
5-bromo-4-chloro-3-indolyl ß-D-glucuronic acid 相似文献
11.
Antonella Furini Csaba Koncz Francesco Salamini Dorothea Bartels 《Plant cell reports》1994,14(2-3):102-106
Summary An efficient procedure for Agrobacterium tumefaciens- mediated transformation of the desiccation-tolerant plant Craterostigma plantagineum has been developed. Leaf explants were inoculated with A. tumefaciens strain GV3101 carrying the gene for kanamycin- or hygromycin-resistance and the ßglucuronidase reporter gene. Parameters which affected the transformation efficiency were the age of the explant, the degree of wounding and the presence of an antioxidant in the medium. Under optimal conditions, calli originated in more than 80% of leaf explants. Transformed plants were obtained from more than 50% of the cultured calli during regeneration in the presence of a suitable antibiotic. The stable integration of T-DNA was confirmed by Southern blot analysis and its expression by assays for ß-glucuronidase activity.Abbreviations GUS
ß-glucuronidase
- MUG
4-methyl-umbelliferyl ß-D-glucuronide
- ABA
abscisic acid
- NPTII
neomycin phosphotransferase II
- CaMV
cauliflower mosaic virus
- MSAR
modified MS medium
- MS
Murashige and Skoog 相似文献
12.
Genetic transformation of alfalfa somatic embryos and their clonal propagation through repetitive somatic embryogenesis 总被引:3,自引:0,他引:3
Slavica Ninković Jovanka Miljuš-Djukić Mirjana Nešković 《Plant Cell, Tissue and Organ Culture》1995,42(3):255-260
Genetically transformed alfalfa (Medicago sativa L., cv. Zajearska 83) plantlets were obtained by inoculating somatic embryos with Agrobacterium tumefaciens strains A281/pGA472 and LBA4404/pBI121. Single somatic embryos, 5–7 mm long, were released from a repetitively embryogenic culture, wounded, and cocultivated with the bacteria. The agar-solidified culture medium contained mineral salts, vitamins, 40 g l–1 sucrose, 1 g l–1 yeast extract and 0.05 mg l–1 BA. Five clones, transformed with A281/pGA472, and 4 clones transformed with LBA4404/pBI121, were selected for proliferation by repetitive somatic embryogenesis, on media containing 100 mg l–1 of kanamycin. The transformation of kanamycin-resistant clones was confirmed by assaying the activity of neomycin phosphotransferase II and/or -glucuronidase enzymes, and by the Southern blot analysis. It is suggested that the transformation/regeneration system based on somatic embryogenesis may be suitable for establishing transgenic alfalfa lines. The relatively low frequency of embryo transformation is compensated for by abundant proliferation in secondary somatic embryogenesis.Abbreviations BA
6-benzyladenine
- GUS
-glucuronidase
- Km
kanamycin
- NPTII
neomycin phosphotransferase II
- X-gluc
5-bromo-4-chloro-3-indolyl--glucuronic acid
- BM
basal medium 相似文献
13.
Summary Using an Agrobacterium tumefaciens binary vector (pAL4404, pBI131), we have demonstrated the transfer of the -glucuronidase gene into the flax (Linum usitatissimum L.) cultivar Glenelg after selection for kanamycin resistance. The transformed lines were obtained by inoculation and subsequent regeneration of hypocotyl segments. The callus that formed on the cut surfaces of the hypocotyl segments was isolated three weeks after infection and was subsequently subcultured to yield shoots. This procedure generated a large number of transgenic shoots over a relatively short period of time. The transformation efficiencies obtained were the highest reported so far for this plant species.Abbreviations 2,4-D, 2,4
dichlorophenoxyacetic acid
- GUS
glucuronidase
- MS
Murasbige and Skoog (1962) medium
- MU
4-methyl-umbelliferone
- MUG
4-methylumbelliferyl-glucuronide
- NPTII
neomycin phosphotransferase II
- PCR
polymerase chain reaction 相似文献
14.
Summary A procedure for the regeneration of fertile transgenic white mustard (Sinapis alba L.) is presented. The protocol is based on infection of stem explants of 7–9 day old plants with an Agrobacterium tumefaciens strain harboring a disarmed binary vector with chimeric genes encoding neomycin phosphotransferase and -glucuronidase. Shoots are regenerated from callus-forming explants within 3–4 weeks. Under selection, 10% of the explants with transgenic embryonic callus develop into fertile transgenic plants. Rooting shoots transferred to soil yield seeds within 14–16 weeks following transformation. Integration and expression of the T-DNA encoded marker genes was confirmed by histochemical glucuronidase assays and Southern-DNA hybridization using primary transformants and S1-progeny. The analysis showed stable integration and Mendelian inheritance of trans-genes in transformed Sinapis lines.Abbreviations BAP
6-benzylaminopurine
- CaMV
cauliflower mosaic virus
- GUS
-glucuronidase
- IBA
indole-3-butyric acid
- IM
infection medium
- NAA
1-naphthalene acetic acid
-
neo
gene encoding NPTII
- NPTII
neomycin phosphotransferase
- RIM
root-inducing medium
- SEM
shoot-elongation medium
- SIM
shoot-inducing medium
- t-nos
polyadenylation site of the nopaline synthase gene
-
uidA
gene encoding GUS
- WM
wash medium
- X-Gluc
5-bromo-4-chloro-3-indolyl -D-glucuronide 相似文献
15.
Mehmet Babaoglu Matthew S. McCabe J. Brian Power Michael R. Davey 《Acta Physiologiae Plantarum》2000,22(2):111-119
A procedure for regenerating plants of Lupinus mutabilis from shoot apices, from which the leaf primordia and initial cell layer(s) of the apical meristem were removed, has been
used to generate transgenic plants following Agrobacterium tumefaciens-mediated gene delivery. Transformation competent cells, from which buds developed, were located at the periphery of the apical
meristem. Kanamycin resistant plants were obtained which expressed β-glucuronidase activity. Integration of the neomycin phosphotransferase
II and β-glucuronidase genes into the genomes of transgenic plants was confirmed by non-radioactive DNA-DNA hybridisation.
This is the first report of the generation of transgenic plants in L. mutabilis. 相似文献
16.
A highly efficient transformation procedure was developed for Lobelia erinus. Leaf or cotyledon discs were inoculated with Agrobacterium tumefaciens strain EHA105 harboring the binary vector plasmid pIG121Hm, which contains a -glucuronidase gene with an intron as a reporter gene and both the neomycin phosphotransferase II and hygromycin phosphotransferase genes as selectable markers. The hygromycin-resistant calli produced on the selection medium were transferred to MS medium supplemented with 0.5 mg/l benzyladenine and 0.2 mg/l indole-3-acetic acid for regeneration of adventitious shoots. Transgenic plants were obtained as a result of the high regeneration rate of the transformed calli, which was as high as 83%. In contrast, no transgenic plant was obtained by the procedure of direct shoot formation following inoculation with A. tumefaciens. Transgenic plants flowered 3–4 months after transformation. Integration of the transgenes was detected using PCR and Southern blot analysis, which revealed that one to several copies were integrated into the genomes of the host plants. The transformation frequency at the stage of whole plants was very high—45% per inoculated disc.Abbreviations BA: 6-Benzyladenine - 2,4-D: 2,4-Dichlorophenoxyacetic acid - GUS: -Glucuronidase - IAA: Indole-3-acetic acidCommunicated by G.C. Phillips 相似文献
17.
A protocol for Agrobacterium-mediated transformation of leaf pieces from two almond cultivars has been developed. This protocol gave a transformation percentage of 48.6 for the cultivar MN51 and 27.9 for the cultivar Supernova. The Agrobacterium tumefaciens strain LBA 4404 carrying a binary vector plasmid pBinGUSint has been used. A critical step for optimal transformation was the pre-culture period of leaf pieces prior selection on medium containing kanamycin. Both cultivars showed an almost doubled transformation percentage when the selection was started at day 6 compared to day 0. The presence of neomycin phosphotransferase II and ß-glucuronidase genes carried by pBinGUSint was confirmed in transformed expiants both by Southern blotting analysis and enzyme activity assays.Abbreviations CaMV
cauliflower mosaic virus
- GUS
ß-glucuronidase
- NPTII
neomycin phosphotransferase II
- -NAA
-naphthaleneacetic acid
- BAP
benzylaminopurine
- IAA
indole-3-acetic acid
- Kin
kinetin
- kn
kanamycin
- cx
cefotaxime
- cb
carbenicillin
- Tris
trishydroxymethylaminomethane
- PVPP
insoluble polyvinylpyrrolidone-40
- X-gluc
5-bromo-4-chloro-3-indolyl-ß-D-glucuronic acid
- EDTA
ethylenediaminetetraacetic acid
- SDS
sodium dodecil sulfate
- SSC
sodium chloride (3 M) sodium citrate (0.3 M) 相似文献
18.
The US Department of Energy recently released a 6.8X draft of the genome sequence for Nisqually-1, a genotype of black cottonwood
(Populus trichocarpa). To improve its utility for functional genomics research, having an efficient means for transformation and regeneration
is necessary. To examine several parameters known to affect the transformation rate, we cocultivated leaf disc and stem explants
with a strain ofAgrobacterium tumefaciens harboring a binary plasmid vector containing genes for both neomycin phosphotransferase (NPTII) and β-glucuronidase (GUS). Shoot regeneration from stem explants was observed in the presence of kanamycin when thidiazuron was incorporated in the
selection medium. Transformation efficiency was influenced by the level of thidiazuron to which explants were exposed during
the early stages of shoot induction. Histochemical assays revealed expression of theGUS gene in leaf, stem, and root tissues of transgenic plants. Polymerase chain reaction confirmed the presence of both selectable
marker and reporter genes in all lines that stained positive for β-glucuronidase activity. By use of our modified protocol,
transgenic plants were recovered within 6 mo at an efficiency of 6%, adequate to produce a large number of transgenic events
with modest effort. 相似文献
19.
Three different regeneration systems, viz. direct regeneration of adventitious shoot buds from explant, regeneration through
callus cultures and somatic embryos were compared to see their effect on transfer of neomycin phosphotransferase (nptII) and β-glucuronidase (GUS) reporter gene (gus) to Morus alba clone M5, through Agrobacterium tumefaciens mediated transformation. Pre-conditioning and co-cultivation durations had a marked effect on transformation frequency. The
highest transformation frequency of 18.6% was obtained using direct induction of adventitious shoot buds. Expression and presence
of transgene were assayed histochemically and through polymerase chain reaction. Southern analysis of GUS and PCR positive
transformants confirmed stable integration of transgenes with two to four copy numbers. The selected transformants showed
normal phenotype under in vitro and field conditions. 相似文献
20.
Anin vitro transformation method has been developed for stem explants of fast-growing willow clones (Salix spp.) usingAgrobacterium tumefaciens as a vector. Transformants obtained with the strains C58 and GV3101 (pGV3851::pLD1) were selected on hormone-free medium and on medium containing kanamycin, respectively. Transformation was confirmed by Southern blot analysis and nopaline assay. Inoculation of green-house grown plants with nopaline and octopine wildtype strains and shoot or root inducing mutant strains caused undifferentiated tumors at a frequency of 0 to 80%, depending on theSalix genotype and the bacterial strain used.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- Km
kanamycin
- NPT
neomycin phosphotransferase 相似文献