Production of Transgenic Soybean Plants Using Agrobacterium-Mediated DNA Transfer

Hinchee, Maud A. W.; Connor-Ward, Dannette V.; Newell, C. A.; McDonnell, Raymond E.; Sato, Shirley; Gasser, Charles S.; Fischhoff, David A.; Re, Diane B.; Fraley, Robert T.; Horsch, Robert B.

doi:10.1038/nbt0888-915

Cited by 364 publications

(232 citation statements)

References 38 publications

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“…The same fragments were inserted into the plant transformation vector pMON850, creating the plasmid pMON601 (Figure 1). For construction of the /i-glucuronidase gene (GUS) fusion, the mutagenized heat shock promoter was fused to a 1.9-kb Hindlll fragment from pMON9749 (Hinchee et al, 1988) containing the entire GUS opening reading frame fused to the nopaline synthase 3' sequence.…”

Section: Vector Constructionmentioning

confidence: 99%

Alterations of Endogenous Cytokinins in Transgenic Plants Using a Chimeric Isopentenyl Transferase Gene.

et al. 1989

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Cytokinins, a class of phytohormones, appear to play an important role in the processes of plant development. We genetically engineered the Agrobacterium tumefaciens isopentenyl transferase gene, placing it under control of a heat-inducible promoter (maize hsp70). The chimeric hsp7O isopentenyl transferase gene was transferred to tobacco and Arabidopsis plants. Heat induction of transgenic plants caused the isopentenyl transferase mRNA to accumulate and increased the level of zeatin 52-fold, zeatin riboside 23-fold, and zeatin riboside 5'-monophosphate twofold. At the control temperature zeatin riboside and zeatin riboside 5'-monophosphate in transgenic plants accumulated to levels 3 and 7 times, respectively, over levels in wild-type plants. This uninduced cytokinin increase affected various aspects of development. In tobacco, these effects included release of axillary buds, reduced stem and leaf area, and an underdeveloped root system. In Arabidopsis, reduction of root growth was also found. However, neither tobacco nor Arabidopsis transgenic plants showed any differences relative to wild-type plants in time of flowering. Unexpectedly, heat induction of cytokinins in transgenic plants produced no changes beyond those seen in the uninduced state. The lack of effect from heat-induced increases could be a result of the transient increases in cytokinin levels, direct or indirect induction of negating factor(s), or lack of a corresponding level of competent cellular factors. Overall, the effects of the increased levels of endogenous cytokinins in non-heat-shocked transgenic plants seemed to be confined to aspects of growth rather than differentiation. Since no alterations in the programmed differentiation pattern were found with increased cytokinin levels, this process may be controlled by components other than absolute cytokinin levels.

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Section: Vector Constructionmentioning

confidence: 99%

Alterations of Endogenous Cytokinins in Transgenic Plants Using a Chimeric Isopentenyl Transferase Gene.

et al. 1989

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“…Principal targets have been increased resistance to disease and insect herbivory and increased tolerance to herbicide damage. Transformation and subsequent whole plant regeneration and characterization of field crop species including soybean, alfalfa, cotton, flax, oil seed rape, and tomato has now been demonstrated (6,13 and references therein). We are interested in modifying plants to reduce the content of undesirable heavy metals in crops.…”

Section: Methodsmentioning

confidence: 99%

Inheritance and Expression of the Mouse Metallothionein Gene in Tobacco

Maiti¹,

Wagner²,

Yeargan³

et al. 1989

Plant Physiol.

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Genetically engineered seedlings obtained from self-fertilized transgenic tobacco (Nkotlana tabacum) contained and expressed the mouse metallothionein and kanamycin resistance marker genes and were more tolerant to cadmium stress than untransformed controls. Cadmium accumulation in leaves of transgenic seedlings exposed to a low, field-like Cd concentration (0.02 micromolar) was about 20% lower than that in untransformed controls. (5, 8). Animal genes encoding MT are well characterized and are highly conserved (3,5,12). It is thought that MT functions in toxic heavy metal Plant Transformation, Regeneration, and Growth Transformation of Nicotiana tabacum cv Ky 14 was achieved using the binary vector pKYLX-7 (19) and an engineered Agrobacterium tumefaciens stain as described earlier (10). Primary transformants were regenerated from kanamycin resistant calli and propagated as described earlier (19). Segregation AnalysisSeeds from self-pollinated transgenic plants (RI and R2 progeny) were surface-sterilized with 70% ethanol for 2 min, then 0.5% sodium hypochlorite for 10 min, washed thoroughly with sterile water, then germinated on rooting medium (15) containing kanamycin (300 jAg/mL). Sensitivity to kanamycin was scored after 3 weeks and the x2 test was used to evaluate results.

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“…2). From a total of 2200 Ro plants tested, 16 showed NPT II enzyme activity in their protein extracts which comigrated with the bacterial derived NPT II enzyme; Figure 2 shows a reanalysis of NPT II enzyme activity in the protein extracts In contrast, the negative control plants, which resulted from sham inoculations of seeds with L-broth or with C58Z707 minus the binary plasmid (over 100 plants for each test), showed no comigrating NPT II enzyme activity in leaf extracts, even after prolonged autoradiographic exposures. In addition, no residual Agrobacteria could be recovered from leaf extracts obtained from the NPT II positive Ro plants.…”

Section: Identification Of Transformed Ro Soybean Plantsmentioning

confidence: 99%

“…The Agrobacterium strains which have been 1212 found to be the most effective for the infection of soybean cultivars are those known as the nopaline-type, which encode for the synthesis of the novel amino acid nopaline (8). Agrobacterium-mediated transformation ofsoybean tissue was first reported by Facciotti et aL (9); however, the first report to describe both gene transfer and regeneration of a whole soybean plant was more recent (16). The reason for the delay in obtaining a transformed soybean plant can be attributed to the difficulty of having the transformation and regeneration events occur in the same cell types (16).…”

mentioning

confidence: 99%

“…However, both procedures suffer because of several limitations. The Agrobacterium-mediated transformation described by Hinchee et al (16) is dependent on a large tissue culture effort, and the transformation and regeneration of transformed plants for many elite soybean lines may be very difficult. The microprojectile procedure described by McCabe et al (20) does require some tissue culture steps, but more importantly it requires a knowledge of the design of the acceleration instruments for those who wish to build their own, or at least access to an instrument which can control the acceleration of microprojectiles.…”

mentioning

confidence: 99%

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Transformation of Soybean (Glycine max) by Infecting Germinating Seeds with Agrobacterium tumefaciens

Chee¹,

Fober²,

Slightom³

1989

Plant Physiol.

133

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The transfer of genetic material into soybean tissue was accomplished by using an avirulent strain of Agrobacterlum tumefaciens which contained the binary vector pGA482. The method used for transformation requires no tissue culture steps as it involves the inoculation of the plumule, cotyledonary node, and adjacent cotyledon tissues of germinating seeds. The identification of neomycin phosphotransferase (NPT) II enzyme activity in the tissues of 16 (Ro) soybean plants indicated that the plant expressible Nos-NPT II gene, contained within the T-DNA region from pGA482, had been transferred at least into somatic tissues.Putative transformed Ro soybean plants were advanced to produce R1 plants which were also assayed for the presence of the transferred Nos-NPT II gene. The combined results of these assays indicated that about 0.7% of the surviving inoculated seeds yielded transformed tissues in the Ro plant, and that about 1/10 of these plants yielded transformed R1 plants. The presence of the Nos-NPT II gene in DNAs isolated from both Ro and R1 plant was demonstrated by using genomic blot hybridization and polymerase chain reaction methods. Integration of this gene into the soybean genome was demonstrated for three R1 soybean plants.

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Production of Transgenic Soybean Plants Using Agrobacterium-Mediated DNA Transfer

Cited by 364 publications

References 38 publications

Alterations of Endogenous Cytokinins in Transgenic Plants Using a Chimeric Isopentenyl Transferase Gene.

Alterations of Endogenous Cytokinins in Transgenic Plants Using a Chimeric Isopentenyl Transferase Gene.

Inheritance and Expression of the Mouse Metallothionein Gene in Tobacco

Transformation of Soybean (Glycine max) by Infecting Germinating Seeds with Agrobacterium tumefaciens

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