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  • 1
    Electronic Resource
    Electronic Resource
    Transgenic plant production mediated by Agrobacterium in Indica rice (1996)
    Springer
    Plant cell reports 15 (1996), S. 727-730 
    Details
    ISSN: 1432-203X
    Keywords: Acetosyringone ; Agrobacterium ; Indica rice ; Oryza sativa L. ; Transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A reproducible system has been developed for the production of transgenic plants in indica rice using Agrobacterium-mediated gene transfer. Three-week-old scutella calli served as an excellent starting material. These were infected with an Agrobacterium tumefaciens strain EHA101 carrying a plasmid pIG121Hm containing genes for β-glucuronidase (GUS) and hygromycin resistnace (HygR). Hygromycin (50 mg/l) was used as a selectable agent. Inclusion of acetosyringone (50μM) in the Agrobacterium suspension and co-culture media proved to be indispensable for successful transformation. Transformation efficiency of Basmati 370 was 22% which was as high as reported in japonica rice and dicots. A large number of morphologically normal, fertile transgenic plants were obtained. Integration of foreign genes into the genome of transgenic plants was confirmed by Southern blot analysis. GUS and HygR genes were inherited and expressed in R1 progeny. Mendelian segregation was observed in some R1 progeny.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232216
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  • 2
    Electronic Resource
    Electronic Resource
    Transient expression of uidA constructs in in vitro onion (Allium cepa L.) cultures following particle bombardment and Agrobacterium-mediated DNA delivery (1996)
    Springer
    Plant cell reports 15 (1996), S. 958-962 
    Details
    ISSN: 1432-203X
    Keywords: Transformation ; particle bombardment ; Agrobacterium ; Allium cepa.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Particle bombardment and Agrobacterium-mediated DNA delivery into immature embryos and microbulbs were used to investigate the expression of the uidA gene in in vitro onion cultures. Both methods were successful in delivering DNA and subsequent uidA expression was observed. Optimal transient β-glucuronidase activity was observed in immature embryos that had been pre-cultured for three days and bombarded at a distance of 3 cm from the stopping plate, under 25 in Hg vacuum, using 900–1300 psi rupture discs. The CaMV35S-uidA gene construct gave five fold higher transient β-glucuronidase activity than the uidA gene construct regulated by any of four other promoters initially chosen for high experession in monocotyledonous tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00231596
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  • 3
    Electronic Resource
    Electronic Resource
    Production of an extracellular polysaccharide byAgrobacterium sp DS3 NRRL B-14297 isolated from soil (1996)
    Springer
    Journal of industrial microbiology and biotechnology 16 (1996), S. 129-133 
    Details
    ISSN: 1476-5535
    Keywords: extracellular polysaccharide ; Agrobacterium ; viscous polysaccharide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A bacterium isolated from soil and identified asAgrobacterium sp produced a water-soluble extracellular polysaccharide capable of producing highly viscous solutions. Gas chromatographic analysis revealed a sugar composition of glucose, galactose and mannose in the molar ratio of 7.5∶2.4∶1, together with 3.7% (w/w) pyruvic acid. Methylation analyses showed the presence of (1→3)-, (1→4)- and (1→6)-linked glucose, (1→3)- and (1→4, 1→6)-linked galactose and a small portion of (1→3)-linked mannose residues. Succinic acid was not present. The molecular weight of the polysaccharide was estimated by light scattering to be 2×106 Da. The viscosity of solutions containing the polysaccharide remained constant from pH 3 to 11, and decreased by 50% when heated from 5 to 55°C. Maximum yield of the polysaccharide, 20 g L−1, was reached in 48 h at 30°C incubation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01570073
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  • 4
    Electronic Resource
    Electronic Resource
    Agrobacterium tumefaciens-mediated transformation of japonica and indica rice varieties (1996)
    Springer
    Planta 199 (1996), S. 612-617 
    Details
    ISSN: 1432-2048
    Keywords: Agrobacterium ; indica rice ; Inheritance ; Japonica rice ; Oryza ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic transformation of rice (Oryza sativa L.) mediated by Agrobacterium ttumefaciens has been confirmed for japonica varieties and extended to include the more recalcitrant indica varieties. Immature embryos were inoculated with either A. tumefaciens At656 (pCNL56) or LBA4404 (pTOK233). Experimental conditions were developed initially for immature embryos treated with strain At656, based upon both transient and stable β-glucuromdase (GUS) activities. However, plant regeneration following selection on G418 (pCNL56 contained the nptII gene) did not occur. Using the same basic protocol, but inoculating immature embryos of rice with LBA4404 (pTOK233), resulted in efficient (about 27%) production of transgenic plants of the japonica variety, Radon, and an acceptable efficiency (from 1–5%) for the indica varieties IR72 and TCS10. Transformation was based upon resistance to hygromycin (pTOK233 contains the hpt gene), the presence of GUS activity (from the gusA gene), Southern blots for detection of the integrated gusA gene, and transmission of GUS activity to progeny in a Mendelian 3:1 segregation ratio. Southern blots indicated two to three copies of the gene integrated in most transformants. Transgenic plants of both the japonica and indica varieties were self-fertile and comparable in this respect to seed-grown plants. Key factors facilitating the transformation of rice by Agrobacterium tumefaciens appeared to be the use of embryos as the expiant, the use of hygromycin as the selection agent (which does not interfere with rice regeneration), the presence of extra copies of certain vir genes on the binary vector of pTOK233, and maintaining high concentrations of acetosyringone for inducing the vir genes during co-cultivation of embryos with Agrobacterium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00195194
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  • 5
    Electronic Resource
    Electronic Resource
    Sequence and characterisation of a ribosomal RNA operon from Agrobacterium vitis (1996)
    Springer
    Molecular genetics and genomics 251 (1996), S. 99-107 
    Details
    ISSN: 1617-4623
    Keywords: Key words Ribosomal genes ; Agrobacterium ; Evolution ; 23S rRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  One of the four ribosomal RNA operons (rrnA) from the Agrobacterium vitis vitopine strain S4 was sequenced. rrnA is most closely related to the rrn operons of Bradyrhizobium japonicum and Rhodobacter sphaeroides and carries an fMet-tRNA gene downstream of its 5S gene, as in the case of R. sphaeroides. The 16S rRNA sequence of S4 differs from the A. vitis K309 type strain sequence by only one nucleotide, in spite of the fact that S4 and K309 have very different Ti plasmids. The predicted secondary structure of the S4 23S rRNA shows several features that are specific for the alpha proteobacteria, and an unusual branched structure in the universal B8 stem. The 3′ ends of the three other rrn copies of S4 were also cloned and sequenced. Sequence comparison delimits the 3′ ends of the four repeats and defines two groups: rrnA/rrnB and rrnC/rrnD.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02174350
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  • 6
    Electronic Resource
    Electronic Resource
    Transformation ofBrassica oleracea L.: a critical review (1996)
    Springer
    Molecular breeding 2 (1996), S. 185-210 
    Details
    ISSN: 1572-9788
    Keywords: Brassica oleracea ; Agrobacterium ; transformation ; direct gene transfer ; regeneration ; virulence ; flowering ; rol genes ; transgene expression ; transgene inactivation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Brassica oleracea is a highly polymorphic species encompassing a wide range of important vegetable and fodder crops. Gene transfer into cultivated forms of this species requires reproducible and efficient methods for genetic transformation and plant regeneration. In this review, we have collated the research experience on transformation ofB. oleracea to highlight the problems encountered. Most research effort has been directed at developingAgrobacterium-mediated transformation methods with relatively little emphasis to date on direct gene transfer techniques. Common procedures for the transformation ofB. oleracea have not emerged, due to the inherent variability between and amongst genotypes. Future progress would be facilitated by the use of genetically fixed material, such as double-haploid or inbred lines, to reduce variation of response within genotypes and would avoid the need for cultivar-specific transformation protocols if responsive lines amenable to crossing with cultivated forms could be identified. The principal difficulties relate to combining efficient plant regeneration with gene transfer. Methods that enhance bacterial virulence and increase the proportion of cells susceptible to transformation and competent for regeneration are discussed. Inefficient selection is a major cause of poor transformation frequencies inB. oleracea and has resulted in the regeneration of chimeric plants uponAgrobacterium tumefaciens-mediated transformation. Promising results have been obtained withAgrobacterium rhizogenes-mediated transformation but the impact of therol genes on flowering of primary transformants has not yet been fully assessed. Strategies to reduce the deleterious effects of therol genes on flowering are discussed. Few agronomically useful characters have been introduced, the majority of research having been confined to the introduction of marker and reporter genes; possible candidate genes are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00564197
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  • 7
    Electronic Resource
    Electronic Resource
    Sequence and characterisation of a ribosomal RNA operon fromAgrobacterium vitis (1996)
    Springer
    Molecular genetics and genomics 251 (1996), S. 99-107 
    Details
    ISSN: 1617-4623
    Keywords: Ribosomal genes ; Agrobacterium ; Evolution ; 23S rRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract One of the four ribosomal RNA operons (rrnA) from theAgrobacterium vitis vitopine strain S4 was sequenced.rrnA is most closely related to therrn operons ofBradyrhizobium japonicum andRhodobacter sphaeroides and carries an fMet-tRNA gene downstream of its 5S gene, as in the case ofR. sphaeroides. The 16S rRNA sequence of S4 differs from theA. vitis K309 type strain sequence by only one nucleotide, in spite of the fact that S4 and K309 have very different Ti plasmids. The predicted secondary structure of the S4 23S rRNA shows several features that are specific for the alpha proteobacteria, and an unusual branched structure in the universal B8 stem. The 3′ ends of the three otherrrn copies of S4 were also cloned and sequenced. Sequence comparison delimits the 3′ ends of the four repeats and defines two groups:rrnA/rrnB andrrnC/rrnD.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02174350
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  • 8
    Electronic Resource
    Electronic Resource
    Corresponding 16S rRNA gene segments in Rhizobiaceae and Aeromonas yield discordant phylogenies (1996)
    Springer
    Plant and soil 186 (1996), S. 69-74 
    Details
    ISSN: 1573-5036
    Keywords: Agrobacterium ; population genetics ; Rhizobium ; systematics ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Previous evidence has indicated that the 16S rRNA genes in certain species of Aeromonas may have a history of lateral transfer and recombination. A comparative analysis of patterns of 16S nucleotide sequence polymorphism among species of Rhizobium and Agrobacterium was conducted to determine if there is similar evidence for chimeric 16S genes in members of the Rhizobiaceae. Results from phylogenetic analyses and comparison of patterns of nucleotide sequence polymorphism in portions of rhizobial 16S genes revealed the same type of segment-dependent polymorphic site partitioning that was previously reported for Aeromonas. These results support the hypothesis that certain 16S segments in rhizobia may have a history of lateral transfer and recombination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00035057
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  • 9
    Electronic Resource
    Electronic Resource
    Identification of Agrobacterium and Rhizobium species based on cellular fatty acid composition (1996)
    Springer
    Plant and soil 184 (1996), S. 143-158 
    Details
    ISSN: 1573-5036
    Keywords: Agrobacterium ; FAME ; fatty acid analysis ; rapid identification ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The increasing number of phylogenetically defined species in the genera Agrobacterium, Rhizobium and Sinorhizobium suggests a need for a rapid identification method which will distinguish between these species. We have examined 65 strains of Agrobacterium representing: A. tumefacies, (34); A. rhizogenes, (16) A. vitis, (10) A. rubi (2) and some unclassified strains, and 150 strains of Rhizobium and Sinorhizobium representing: R. etli (21); R. galegae (20); R. huakuii (17); R. leguminosarum (20); R. loti (16); R. topici (18); S. fredii (19); and S. meliloti (20). Fatty acid methyl esters (FAME) were obtained from each strain, as previously described, and analysed by gas-chromatography using the MIDI Hewlett-Packard Microbial Identification System (MIS). Fatty acid profiles were recorded, characteristic fatty acids noted and the overall similarities between fatty acid profiles for each species calculated. Relationships between species were also derived from the fatty acid data by principal component analysis. This showed overlapping clusters for strains of R. leguminosarum and R. etli, R. topici and A. rhizogenes and S. fredii and S. meliloti within one supercluster. Strains of A. tumefaciens, A rubi, A. vitis and R. galegae formed a second supercluster while R. loti and R. huakuii strains formed a third cluster well separated from all the other strains. The fatty acid profiles were used to correctly identify at least 94% of the strains representing each species in the collection except R. etli. R. etli strains (23.8%) were misidentified as R. leguminosarum. This was attributed to the high similarity (44.7%) between R. etli and R. leguminosarum. It is concluded that whole cell fatty acid analysis should form part of the polyphasic description of new species of root nodule bacteria, with the proviso that growth conditions and analytical methods be carefully standardized. It is suggested that FAME-MIS system and the database we have compiled provide a basis for future development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029284
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  • 10
    Electronic Resource
    Electronic Resource
    Agrobacterium-mediated transformation of Javanica rice (1996)
    Springer
    Molecular breeding 2 (1996), S. 267-276 
    Details
    ISSN: 1572-9788
    Keywords: Agrobacterium ; Javanica rice ; regeneration ; rice ; TAIL-PCR ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Difficulties frequently encountered using direct DNA transfer methods for transformation of Javanica varieties of rice (Oryza sativa L.) have limited the application of biotechnology to these varieties. We now reportAgrobacterium-mediated transformation of Javanica cultivars Gulfmont and Jefferson that are, respectively, widely used or about to enter commercial cultivation in the southern USA. Vigorous, phenotypically normal, fertile plants expressing both the selectable marker and the gene of interest were obtained. Southern analysis showed that only one or two copies of the T-DNA insert were present. Sequence analysis of right border fragments of one line confirmed that insertion was into a coding region of rice nuclear DNA. This analysis also revealed the presence of relatively short regions of permuted T-DNA border sequences, similar to those found afterAgrobacterium-mediated transformation of dicots. Progeny analysis of lines bearing two copies showed co-segregation, indicating that they were located relatively closely on the same chromosome. The introduced genes were transmitted to the R1 and R2 generations in a Mendelian fashion, confirming the suitability of this approach for biotechnological improvement of elite rice cultivars.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00564204
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