共查询到20条相似文献,搜索用时 23 毫秒
1.
Willem J. Stiekema Freek Heidekamp Jeanine D. Louwerse Harrie A. Verhoeven Paul Dijkhuis 《Plant cell reports》1988,7(1):47-50
Tuber discs of Solanum tuberosum cv Bintje and Désirée were cocultivated with an Agrobacterium tumefaciens binary vector, carrying both the neomycine phosphotransferase and the E. coli -glucuronidase gene fused to resp. the nopaline synthase and Cauliflower mosaic virus 35S promotor.Inoculated tuber discs produce transgenic shoots in selective media containing kanamycin. The transgenic plants are phenotypically normal and contain the euploid number of chromosomes. Both the neomycin phosphotransferase as well as the -glucuronidase gene are expressed conferring resp. kanamycin resistance and -glucuronidase activity to the plants.Abbreviations GUS
-glucuronidase
- NPT
neomycin phosphotransferase
- CaMV
Cauliflower Mosaic Virus
- BAP
6-benzylaminopurine
- GA3
gibberellic acid
- NAA
naphthalineacetic acid
- LB
Luria Broth
- MU
methylumbelliferone 相似文献
2.
Agrobacterium-mediated transformation of white clover using direct shoot organogenesis 总被引:1,自引:0,他引:1
Christine R. Voisey Derek W. R. White Brigitta Dudas Ruth D. Appleby Paul M. Ealing Alicia G. Scott 《Plant cell reports》1994,13(6):309-314
Summary White clover (Trifolium repens L.) plants from the cultivars Grasslands Huia and Grasslands Tahora have been transformed using Agrobacterium-mediated T-DNA transfer. Transgenic plants regenerated directly from cells of the cotyledonary axil. To transform white clover, shoot tips from 3 day old seedlings were co-cultivated with A. tumefaciens strain LBA4404 carrying the plasmid vector pPE64. This vector contains the neomycin phosphotransferase II gene (nptII) and -glucuronidase reporter gene (gus) both under the control of the CaMV 35S promoter. Kanamycin-resistant plants regenerated within 42 days after transfer onto selective media. Integration of the nptII and gus genes into the white clover genome was confirmed using Southern blotting, and histochemical analysis indicated that the gus gene was expressed in a variety of tissues. In reciprocal crosses between a primary transformant and a non-transformed plant the introduced gus gene segregated as a single dominant Mendelian trait.Abbreviations BAP
6-benzylaminopurine
- NAA
-naphthaleneacetic acid
- MS
Murashige and Skoog
- GUS
-glucuronidase
- X-GLUc
5-bromo-4-chloro-3-indolyl--D-glucuronide
- MUG
methylumbelliferyl--D-glucuronide
- CaMV
Cauliflower Mosaic Virus
- NPTII
neomycin phosphotransferase II
- OCS
octopine synthase
- 4-MU
4-methyl umbelliferone 相似文献
3.
Regeneration of transgenic plants of Prunus armeniaca containing the coat protein gene of Plum Pox Virus 总被引:1,自引:0,他引:1
Margit Laimer da Câmara Machado Artur da Câmara Machado Veronika Hanzer Hans Weiss Ferdinand Regner Herta Steinkellner Diethard Mattanovich Regina Plail Elisabeth Knapp Birgit Kalthoff Hermann Katinger 《Plant cell reports》1992,11(1):25-29
Summary A system was developed which allows the transfer of foreign genes into apricot cultivars. We report the transformation and regeneration of Prunus armeniaca plants with Agrobacterium tumefaciens strain LBA 4404 containing various binary plasmids, pBinGUSint, carrying the marker gene ß-glucuronidase (GUS) and pBinPPVm, carrying the coat protein gene of Plum Pox Virus (PPV). The marker gene GUS was used for optical evaluation of the efficiency of the transformation system. The coat protein gene of PPV was used to introduce coat protein mediated resistance against one of the most important pathogens of stone fruit trees in Europe and the whole Mediterranean area. This is the first report of the successful integration of a viral coat protein gene into a fruit tree species, opening a new perspective on the control of the disease.Abbreviations GUS
ß-glucuronidase
- PPV
Plum Pox Virus
- BA
6-benzylaminopurine
- NPTII
neomycin phosphotransferase II
- CP
coat protein
- CaMV
Cauliflower Mosaic Virus
- P35S 35S
promoter
- MS
Murashige and Skoog
- PCR
polymerase chain reaction
- P/C/I
phenol/chloroform/isoamylalcohol
- RNase
ribonuclease
- dNTP
deoxyribonucleosidetriphosphate
- DMSO
dimethyl sulfoxide 相似文献
4.
Julie R. Kikkert Dominique Hébert-Soulé Patricia G. Wallace Michael J. Striem Bruce I. Reisch 《Plant cell reports》1996,15(5):311-316
Transgenic plantlets of Chancellor grapevine (Vitis L. complex interspecific hybrid) were produced via biolistic transformation. Embryogenic cell suspensions were bombarded with 1 m tungsten particles coated with pBI426 which encodes a fusion peptide between -glucuronidase (GUS) and neomycin phosphotransferase II (NPTII). The fusion peptide is under the control of a double 35S Cauliflower Mosaic Virus promoter and a leader sequence from Alfalfa Mosaic Virus. The cells were placed on kanamycin-containing media (10, 25 or 50 mg/l) 2 d after bombardment. Activated charcoal reduced cell browning. Embryos were first observed on selective media 14–29 weeks after bombardment. More than 1600 clusters of embryos were germinated and/or assayed for GUS. Of 621 embryos assayed for GUS expression, 182 (29.3%) were positive. PCR confirmed the presence of the NPTII gene in all 5 GUS-positive and 2 GUS-negative (bombarded) embryos tested. In germination experiments, 15% of the embryo clusters produced at least one plant with normal shoot growth. Of 164 normal plants assayed for GUS expression, 37 (22.6%) were positive. The NPTII gene was amplified by PCR in 1 (of 1) GUS-positive and 4 (of 5) GUS-negative bombarded plants, but not in non-bombarded control plants. Southern blotting confirmed integration of the NPTII gene in all 3 of the GUS and PCR-NPTII positive plants tested. Biolistics is an efficient method for transformation of Chancellor and should be applicable to other important grape cultivars.Abbreviations AC
activated charcoal
- GUS
-glucuronidase
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
6-benzylaminopurine
- NAA
-naphthalene acetic acid
- TDZ
thidiazuron
- NPTII
neomycin phosphotransferase II
- Km
kanamycin
- MS
Murashige and Skoog (1962) medium
- WPM
Woody Plant Medium of Lloyd and McCown (1980) 相似文献
5.
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
6.
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
7.
Two clones of Populus nigra L. were tested in vivo and in vitro for their susceptibility to three different Agrobacterium tumefaciens wild-type strains evaluating number and size of resulting calluses. Strain C58 proved to be the most virulent.Various parameters affecting Agrobacterium-mediated transformation of P. nigra clones were further analyzed using ß-glucuronidase gene transient expression. The clone Jean Pourtet proved to be more susceptible than the clone San Giorgio. A. tumefaciens strain A281 pKIWI105 proved to be the most virulent. The optimal procedure involved dipping of leaf discs into a bacterial suspension (7×108 cells/ml) for 20 min, followed by a 48 h co-cultivation period on semi-solid regeneration medium.Leaf explants were co-cultivated with two disarmed A. tumefaciens strains. Plantlets of San Giorgio were regenerated, tested for ß-glucuronidase activity and rooted on selective medium containing kanamycin. Polymerase chain reaction analysis and Southern blot hybridization confirmed the integration of the neomycin phosphotransferase II gene into the poplar genome.Abbreviations BAP
6-benzyl-aminopurine
- CaMV
Cauliflower Mosaic Virus
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GUS and gus
ß-glucuronidase
- hpt
hygromycin phosphotransferase
- IBA
indole-3-butyric acid
- KIN
kinetin
- LB
Luria Bertani
- MS
Murashige and Skoog
- NAA
ßnaphthaleneacetic acid
- NOS
Nopaline synthase
- NPTII and nptII
neomycin phosphotransferase II
- PCR
Polymerase chain reaction
- PVC
poly-vinyl-cloride
- SDS
sodium dodecyl sulfate
- SSC
sodium cloride-sodium citrate
- Tris
tris(hydroxymethyl)amino-methane
- WPM
Woody Plant Medium 相似文献
8.
Induced expression of a chimeric gene construct in transgenic lettuce plants using tobacco pathogenesis-related protein gene promoter region 总被引:3,自引:0,他引:3
The expression of a stress- and salicylic acidinducible protein gene from tobacco, PR1a protein gene, was determined after its Introduction to lettuce (Lactuca sativa L.) plants. The 5 flanking 2.4 Kb fragment from PR1a gene was joined to the bacterial -glucuronidase (GUS) gene (PR-GUS) and introduced into lettuce cotyledons by Agrobacterium-mediated gene transfer using a binary vector containing a kanamycin-resistance gene as a selectable marker. As a control with constitutive expression, the chimeric gene consisting of CaMV 35S RNA promoter and GUS gene (35S-GUS) was used. An improved method for shoot formation directly from lettuce cotyledons was used effectively for transformation, shortening the time for regeneration. In 70% or more of kanamycin-resistant regenerated lettuce plants, into which PR-GUS or 35S-GUS was introduced, high GUS activity and integration of the chimeric gene into the lettuce genome were detected. By treatment with salicylic acid, GUS activity increased 3- to 50-fold in PR-GUS transformants, however, no increase was detected in 35S-GUS plants. These results showed that the promoter of the stress-inducible tobacco PR1a protein gene was introduced into lettuce plants, and the introduced chimeric gene was expressed normally under the regulated control of the PRla promoter.Abbreviations BA
N6-benzyladenine
- GUS
-glucuronidase
- NAA
-naphthaleneacetic acid
- Km
kanamycin
- Kms
kanamycin resistant
- Km0
kanamycin sensitive
- NPT- II
neomycin phosphotransferase II
- PR
pathogenesis-related
- SA
salicylic acid
- MS
Murashige and Skoog medium
- NOS
nopaline synthase 相似文献
9.
Stable transformation of papaya via microprojectile bombardment 总被引:27,自引:0,他引:27
Maureen M. M. Fitch Richard M. Manshardt Dennis Gonsalves Jerry L. Slightom John C. Sanford 《Plant cell reports》1990,9(4):189-194
Summary Stable transformation of papaya (Carica papaya L.) has been achieved following DNA delivery via high velocity microprojectiles. Three types of embryogenic tissues, including immature zygotic embryos, freshly explanted hypocotyl sections, and somatic embryos derived from both, were bombarded with tungsten particles carrying chimeric NPTII and GUS genes. All tissue types were cultured prior to and following bombardment on half-strength MS medium supplemented with 10 mg 1–1 2,4-D, 400 mg 1–1 glutamine, and 6% sucrose. Upon transfer to 2,4-D-free medium containing 150 mg 1–1 kanamycin sulfate, ten putative transgenic isolates produced somatic embryos and five regenerated leafy shoots. Leafy shoots were produced six to nine months following bombardment. Tissues from 13 of these isolates were assayed for NPTII activity, and 10 were positive. Six out of 15 isolates assayed for GUS expression were positive. Three isolates were positive for both NPTII and GUS,Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- GUS
-glucuronidase
- X-gluc
5-Br-4-Cl-3-indolyl--D-glucuronic acid
- CaMV
cauliflower mosaic virus
- NOS
nopaline synthase
- NPTII
neomycin phosphotransferase II
Journal Series no. 3448 of the Hawaii Institute of Tropical Agriculture and Human Resources 相似文献
10.
Internode explants ofin vitro plants ofForsythia x intermedia Spring Glory were transformed with thegus andnpt II genes after inoculation with theA. tumefaciens strain EHA 101 harbouring the plasmid pFAJ3000. Shoot organogenesis took place from callused edges of explants. The first transformed buds were detected 4 to 6 weeks after transfer on regeneration medium, containing 25 mg/l kanamycin as selective agent. An average of 1% of explants regenerated transgenic shoots.-glucuronidase assays and culture on kanamycin-containing medium provided the first indication of integration and expression of introduced genes in transformants. Southern blot and polymerase chain reaction amplification analyses gave molecular confirmation of genetic transformation. Transgenic plants were acclimatized in the greenhouse. Enzymatic assays on several organs of mature plants still showed -glucuronidase activity, thus confirming stable integration of T-DNA in the plant genome.Abbreviations BAP
6-benzyl-aminopurine
- CaMV
Cauliflower Mosaic Virus
- GUS andgus
-glucuronidase
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige and Skoog
- NOS
nopaline synthase
- NPT II andnpt II
neomycin phosphotransferase II
- PCR
polymerase chain reaction
- SDS
sodium dodecyl sulphate
- SSC
sodium chloride-sodium citrate
- X-Gluc
5-bromo-4-cbloro-3-indolyl glucuronide 相似文献
11.
High frequency shoot regeneration and Agrobacterium-mediated DNA transfer in Canola (Brassica napus)
Muhammad Ramzan Khan Hamid Rashid Muhammad Ansar Zubeda Chaudry 《Plant Cell, Tissue and Organ Culture》2003,75(3):223-231
Five different varieties of Brassica napus (Cyclone, Dunkled, Oscar, Rainbow and KS75) were tested for their regeneration response. Cyclone showed a very high frequency of regeneration (92%). The use of silver nitrate was a pre-requisite for efficient shoot regeneration. Hypocotyls were selected as the starting material for transformation experiments on the basis of high transient GUS expression. Explants were co-cultivated with Agrobacterium strain EHA101 harboring a binary vector pIG121Hm containing neomycin phosphotransferase II (NPTII) gene, conferring resistance to kanamycin, hygromycin phosphotransferase (HPT) gene, conferring resistance to hygromycin as selectable markers and -glucuronidase (GUS) gene as a reporter. Acetosyringone promoted the transformation but was not an absolute requirement. A pre-selection period of 7 days after co-cultivation was essential for successful transformation. Kanamycin was efficient selective agent for selection and maximum transformation efficiency was 24%. GUS activity was evident in leaf tissues. All the transgenic plants have an expected band of 0.43 kb fragment by PCR analysis confirming the presence of foreign DNA into plant genome. 相似文献
12.
Transgenic peanut plants were produced using Agrobacterium mediated gene transfer. Primary leaf explants of peanut were co-cultivated with Agrobacterium tumefaciens LBA 4404 harbouring the binary plasmid pBI 121 (conferring -glucuronidase activity and resistance to kanamycin) and cultured on regeneration medium supplemented with kanamycin to select putatively transformed shoots. They were rooted and plants were transferred to soil. Stable integration and expression of the transgenes were confirmed by NPT II assay, Southern blot hybridization and GUS assay.Abbreviations BA
6-benzyladenine
- GUS
-glucuronidase
- IAA
indole-3-acetic acid
- NAA
-naphthaleneacetic acid
- NOS
nopaline synthase
- NPT II
neomycin phosphotransferase II
- SDS
Lauryl sulfate 相似文献
13.
Summary A rapid regeneration system was used for studies ofAgrobacterium-mediated transformation inPisum sativum L. Cotyledonary node explants were inoculated withAgrobacterium tumefaciens strains containing binary vectors carrying genes for nopaline synthase (NOS),β-glucuronidase (GUS), and neomycin phosphotransferase (NPTII) and placed on selection medium containing either 75 or 150 mg/liter
kanamycin. A GUS encoding gene (uidA) containing an intron was used to monitor gene expression from 6 to 21 days postinoculation. GUS activity could be observed
6 days after inoculation in the area of the explant in which regeneration-occurred. Regenerating tissue containing transformed
cells was observed in explants on selection medium 21 days postinoculation. Using this system, a single transgenic plant was
obtained. Progeny of this plant, which contained two T-DNA inserts, demonstrated segregation for the inserts and for expression
of the NOS gene in the selfed R1 progeny. NPTII activity was observed in the R2 generation, indicating inheritance and expression of the foreign DNA over at least two generations. Attempts to repeat this
procedure were unsuccessful. 相似文献
14.
Summary A simple and efficient gene transfer system of trifoliate orange (Poncirus trifoliata Raf.) was developed using epicotyl segments. The segments were infected with Agrobacterium harboring the binary vector pBI121 or pBI101-O12-p1. Both vectors contained the neomycin phosphotransferase II (NPTII) and the -glucuronidase (GUS) genes. In the plasmid pBI101-O12-p1, the GUS gene was directed to the promoter region of ORF12 (rolC) of the Ri plasmid. On a selection medium containing 100 or 200 g/ml kanamycin, adventitious shoots were formed from 21.7–44.6% of the segments. Histochemical GUS assay showed that 55.4–87.7% of the shoots expressed the GUS gene. The stable integration of this gene was also confirmed by polymerase chain reaction (PCR) analysis and by Southern blot analysis. When the pBI101-O12-p1 plasmid was used, the GUS activity was found to be located in phloem cells of leaf, stem and root. More than 100 transformed plants were obtained using this method within 2–3 months. 相似文献
15.
Shigeto Kiyokawa Yasuhiro Kikuchi Hiroshi Kamada Hiroshi Harada 《Plant cell reports》1996,15(8):606-609
We have developed an Agrobacterium -mediated transformation system for commercial Begonia species. The leaf explants of Begonia semperflorens, Begonia x hiemalis and B. tuberhybrida were inoculated with Agrobacterium tumefaciens LBA4404 harboring a binary vector pBI121 which contains rolA, B and C genes of an agropine type Ri plasmid (pRiA4b). Kanamycin resistant shoots of B. tuberhybrida were obtained on MS agar medium supplemented with 0.1 mg/l NAA, 0.5 mg/l BA, 500 mg/l claforan and 100 mg/l kanamycin. These shoots exhibited GUS activity and Southern analysis showed a single copy insertion into the genome. When the transgenic plants were transferred to soil, they displayed the phenotype specific to the transgenic plants by A. rhizogenes such as dwarfness, delay of flowering, and wrinkled leaves and petals.Abbreviations NAA
1-naphthaleneacetic acid
- BA
benzyladenine
- MS
Murashige and Skoog (1962)
- GUS
-glucuronidase (EC 3.2.1.31)
- NPT II
neomycin phosphotransferase (EC 2.7.1.95)
- NOS
nopaline synthase
- CaMV
Cauliflower Mosaic Virus
- CTAB
cetyl triethylammonium bromide
- X-GLUC
5-bromo-4-chloro-3-indolyl--D-glucuronide
- PCR
polymerase chain reaction 相似文献
16.
John C. Thomas Mark J. Guiltinan Silvia Bustos Terry Thomas Craig Nessler 《Plant cell reports》1989,8(6):354-357
Summary
Daucus carota hypocotyl sections were transformed withAgrobacterium tumefaciens LBA4404 containing CaMV 35S promoter, -glucuronidase coding sequence and the nopaline synthase (Nos) poly adenylation sequences in Bin 19. Sliced sterile seedling hypocotyl segments were preincubated for 2 days, co-cultivated withAgrobacterium for an additional 2 days, and then transferred to medium containing 100ug/ml of kanamycin and 400ug/ml carbenicillin. In 6 weeks kanamycin resistant calli were obtained in 5.8% of the explants from one variety. Calli were subcultured on solid medium, and in 4 weeks introduced into suspension culture. NPTII and Southern blot analysis confirmed that three selected lines were transformed with 1–3 copies of the GUSII construction. GUS activity in transformants was 5 to 250 fold over background.Abbreviations NPT II
neomycin phosphotransferase II
- Nos
nopaline synthase
- GUS
-glucuronidase
- CaMV
cauliflower mosaic virus
- 2,4-D
2,4 dichlorophenoxyacetic acid
- PMSF
phenylmethylsulfonyl fluoride 相似文献
17.
Agrobacterium-mediated transformation of sweet orange and regeneration of transgenic plants 总被引:7,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
18.
Jean Charles Leple Ana Cristina Miranda Brasileiro Marie France Michel Francis Delmotte Lise Jouanin 《Plant cell reports》1992,11(3):137-141
Summary Leaf or stem explants of a hybrid poplar clone (Populus tremula X Populus alba), sensitive to Agrobacterium tumefaciens, were co-cultivated either by an octopine or a nopaline disarmed A. tumefaciens modified strain. Transformed poplar shoots were readily regenerated from explants. The protocol was improved using the nopaline disarmed strain C58/pMP90 with the binary vector pBI121. This protocol was then used to test three other vectors. The first one, possessing a nptII gene fused to the CaMV 19S promoter, permitted regeneration of transformed shoots in presence of 50 to 100 mg/l kanamycin. The two other vectors carried an additional nptII gene under the control of the CaMV 35S or CaMV 35S promoter with a double enhancer sequence (CaMV 70). CaMV 70 promoter provided consistently higher level of gene expression than the other promoters in both callus and leaf tissues.Abbreviations CaMV
Cauliflower Mosaïc Virus
- 2iP
2-isopentenyladenine
- GUS and gus
ß-glucuronidase
- NAA
1-naphthaleneacetic acid
- NPTII and nptII
neomycin phosphotransferase II
- NOS
Nopaline synthase, X-Gluc: 5-bromo-4-chloro-3-indolyl ß-D glucuronide
- Ap
ampicillin
- Gn
gentamycin
- Km
kanamycin
- Rf
rifampicin
- St
streptomycin
This work is dedicated to the late Marie France Michel who initiated the poplar biotechnology project at INRA. 相似文献
19.
Elke Kemper Christoph Grevelding Jeff Schell Robert Masterson 《Plant cell reports》1992,11(3):118-121
Summary A modified root explant transformation method has been developed that is effective in producing transgenic Arabidopsis thaliana plants which are methotrexate resistant due to the integration of T-DNA vectors containing a chimeric dihydrofolate reductase gene. Molecular analysis shows that transformed methotrexate resistant plants contain the expected T-DNA construct with the chimeric gene. This transformation method also works well with other plant selectable markers, including hygromycin phosphotransferase and neomycin phosphotransferase II.Abbreviations DHFR
(dihydrofolate reductase)
- HPT
(hygromycin phosphotransferase)
- NPTII
(neomycin phosphotransferase)
- CaMV
(Cauliflower Mosaic Virus)
- MES
(2-[N-morpholino]ethanesulfonic acid)
- BAP
(N6-benzylaminopurine)
- NAA
(naphthaleneacetic acid)
- 2,4-D
(2,4 dichlorophenoxyacetic acid)
- 2iP
(6-(dimethylallylamino)-purine)
- 2iPAde
(6-(dimethylallylamino)-ade-nine)
- IAA
(indole-3-acetic acid)
- IBA
(indole-3-butyric acid) 相似文献
20.
Summary Direct delivery of DNA into embryogenic pollen was used to produce transgenic plants in tobacco. A plasmid bearing the ß-glucuronidase (GUS) marker gene in fusion with the 35S-promoter was introduced by microprojectile bombardment into mid-binucleate pollen of Nicotiana tabacum that had been induced to form embryos by a starvation treatment. In cytochemical expression assays, 5 out of 104 pollen grains were GUS+. Visual selection by staining with a non-lethal substrate for GUS was used to manually isolate transformed embryos. From the initial population of embryogenic GUS+ pollen, 1–5% developed into multicellular structures and 0.02% formed regenerable embryos. Two haploid transformants were regenerated. GUS expression was detected in different parts of the plants, and Southern analysis confirmed stable integration of the foreign DNA. Diploidisation was induced by injection of colchicine into the stem near adventitious buds. Offspring from selfings and backcrosses of one transformant were tested for GUS expression and by Southern blots. All F1-plants were transgenic, in accordance with Mendelian inheritance.Abbreviations GUS
ß-glucuronidase
- CaMV
Cauliflower Mosaic Virus
- MCS
multicellular structure
- NPTII
neomycin phosphotransferase
- PEG
polyethylene glycol
- X-gluc
5-bromo-4-chloro-3-indolyl glucuronide
- DAPI
4,6-diamidino-2-phenylindole
- Tris
Tris(hydroxymethyl)aminomethane hydrochloride
- EDTA
ethylenedinitrilo tetraacetic acid, disodium salt dihydrate 相似文献