Expression of soybean nodulin 26 in transgenic tobacco. Targeting to the vacuolar membrane and effects on floral and seed development.
Nodulin 26 is an integral membrane protein of the symbiosome membrane of nitrogen-fixing soybean nodules. We expressed a nodulin 26 cDNA in transgenic tobacco (TN26 tobacco) under the control of the cauliflower mosaic virus 35S promoter to study subcellular targeting and the physiological effect(s) of its expression. Based on Northern and Western blots, the expression of nodulin 26 mRNA and protein in transgenic plants is high in apical shoot sections, flowers, and stems, low in mature leaves, and absent in roots. Western blot analysis revealed high levels of transgenic nodulin 26 protein in tonoplast membranes. In contrast, nodulin 26 protein was not found in isolated plasma membranes, the soluble fraction, nor in chloroplast and mitochondria-enriched membrane fractions. About 50-60% of the flowers and pods from TN26 tobacco plants abscised prematurely. Seed capsule size and seed fill per capsule from the remainder of surviving flowers were about 50% of that of control plants. Pollen viability was found to be normal, but flowers from TN26 tobacco plants showed shorter anther filaments compared with control plants. Normal seed production and capsule size was restored by manually crossing the stigmas from TN26 plants with isolated pollen from either transgenic or control plants. Thus, the aberrant filament growth could have resulted in the reproductive defects associated with the plants.
Zhang
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Department of Biochemistry, University of Tennessee, Knoxville 37996, USA.
Roberts
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DM
D001483:Base Sequence; D001692:Biological Transport; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D007425:Intracellular Membranes / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics; D009586:Nitrogen Fixation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012231:Rhizobium / Q000235:genetics; D012639:Seeds / Q000254:growth & development; D013025:Glycine max / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism*
D008565:Membrane Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C045625:nodulin