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  • 1
    Online Resource
    Online Resource
    Saint Louis :Elsevier Science & Technology,
    Keywords: Biological transport -- Congresses. ; Water in the body -- Congresses. ; Plant-water relationships -- Congresses. ; Plant translocation -- Congresses. ; Electronic books.
    Description / Table of Contents: Water Relations in Membrane Transport in Plants and Animals contains the presentations in a symposium dealing with Water Relations in Membranes in Plants and Animals, during the 27th Annual Fall Meeting of the American Physiological Society held at The University of Pennsylvania, 17-19 August 1976. The purpose of the symposium was to explore the common modes of water regulation in plants and animals. In these proceedings, the mechanisms employed to restrict water flow across plant and metazoan animal cells are described. Putative differences in mechanisms of water regulation retained by plant versus animal cells become inconsequential in the light of the numerous similarities: dependence upon bioelectric potentials maintained across cell membranes, energy dependence of uphill water movement, and solute coupling during water transport. The presentations can be organized into four. The first takes up specific mechanisms of water transport in plants. The second and third parts deal with specific mechanisms in invertebrates and vertebrates, respectively. The fourth part covers generalized mechanisms common to plants and animals.
    Type of Medium: Online Resource
    Pages: 1 online resource (408 pages)
    Edition: 1st ed.
    ISBN: 9781483273877
    DDC: 574.87/5
    Language: English
    Note: Front Cover -- Water Relations in Membrane Transport in Plants and Animals -- Copyright Page -- Table of Contents -- Contributors -- Preface -- Session I: Water and Solute Transport in Plant Cells -- CHAPTER 1. WATER AND SOLUTE TRANSPORT IN PLANT CELLS: CHAIRMAN'S INTRODUCTORY COMMENTS -- CHAPTER 2. ION TRANSPORT AND OSMOTIC REGULATIONIN GIANT ALGAL CELLS -- INTRODUCTION -- IDENTIFICATION OF THE EFFECTOR IN TURGOR REGULATION -- IDENTIFICATION OF THE PRIMARY SIGNAL FOR TURGOR REGULATION -- TUR'GOR-SENSITIVE K+ TRANSPORT IN Valonia -- THE TURGOR-PRESSURE TRANSDUCER -- SUMMARY AND CONCLUSIONS -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 3. TRANSCELLULAR ION MOVEMENTS AND GROWTH LOCALIZATIONIN FUCOID EGGS AND OTHER CELLS -- INTRODUCTION -- ULTRASTRUCTURE OF CELL WALL FORMATION -- ELECTRICAL CORRELATES OF DEVELOPMENT -- LOCAL CATION ENTRY AND ELECTROPHORESIS -- ELECTRICAL CURRENTS AND CALCIUM ACCUMULATIONS IN POLLEN -- SUMMARY -- REFERENCES -- CHAPTER 4. BIOPHYSICAL MODEL FOR PLANT CELL GROWTH: AUXIN EFFECTS -- INTRODUCTION -- IS WALL-YIELDING A COMPLEX PROCESS? -- IS THERE SELF-REGULATION IN HIGHER PLANTS? -- WHAT TYPES OF SELF-REGULATION COULD APPLY? -- PHYSIOLOGICAL PROCEDURES -- ANALYTICAL PROCEDURES -- NATURE OF SELF-STABILITY -- A MORE DETAILED MODEL -- DISCUSSION -- SUMMARY -- APPENDIX -- REFERENCES -- CHAPTER 5. THE OSMOTIC MOTOR OF STOMATAL MOVEMENT -- INTRUDUCTION: Function of Stomata -- CHANGES IN STOMATAL VOLUME, PRESSURE AND SOLUTE CONTENT -- TRANSPORT OF IONS -- METABOLISM OF ORGANIC ACIDS -- EFFECTS OF CO2 AND ABSCISIC ACID -- THE OSMOTIC MOTOR OF STOMATAL MOVEMENT: SUMMARY, HYPOTHESIS AND CONCLUSION -- Acknowledgements -- REFERENCES -- CHAPTER 6. CONTROL OF PROTON TRANSLOCATION IN THE CHLOROPLAST WATER OXIDATION SYSTEM -- INTRODUCTION -- CHEMICAL MODIFIERS AS PROBES FOR CONFORMATIONAL CHANGES. , A CHLOROPLAST MEMBRANE CONFORMATIONAL CHANGE -- PARTIAL ELECTRON TRANSPORT REACTIONS AND DABS BINDING -- H+ UPTAKE, PHOSPHORYLATI0N AND CONFORMATIONAL CHANGES -- RELATION TO MEMBRANE STRUCTURE -- SUMMARY -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 7. ENERGETICS AND CONTROL OF TRANSPORT IN NEUROSPORA -- INTRODUCTION -- THE CHEMIOSMOTIC MODEL -- CONTROL PHENOMENA -- SUMMARY AND CONCLUSIONS -- ACKNOWLEDGEMENTS -- REFERENCES -- Session II: Water and Solute Transport in Cells of Invertebrates -- CHAPTER 8. WATER AND SOLUTE REGULATION IN INVERTEBRATES: CHAIRMAN'S INTRODUCTORY REMARKS -- CHAPTER 9. COMPARATIVE ASPECTS OF INVERTEBRATE EPITHELIAL TRANSPORT -- INTRODUCTION -- EVALUATING OUABAIN INSENSITIVITY -- OUABAIN INSENSITIVITY AND HIGH HEMOLYMPH Na+ -- DISCUSSION -- SUMMARY -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 10. SOLUTE AND WATER MOVEMENT IN THE ROUNDWORM ASCARIS SUUM (NEMATODA) -- INTRODUCTION -- CYTOLOGY OF THE INTESTINE -- LIPIDS -- AMINO ACIDS -- SUGARS -- FLUID TRANSPORT -- MOVEMENT OF SOLUTES AND THE SHORT CIRCUIT CURRENT -- SUMMARY -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 11. SOLUTE AND WATER MOVEMENT IN FRESHWATER BIVALVE MOLLUSKS (Pelecypoda -- Unionidae -- Corbiculidae -- Margaritiferidae) -- INTRODUCTION -- ION LEVELS IN BIVALVES -- REGULATION OF IONS -- INULIN CLEARANCE -- KINETICS OF Na AND Cl TRANSPORT -- EFFECTS OF PHARMACOLOGICAL AGENTS -- CONCLUSION -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 12. ASPECTS OF FLUID MOVEMENT IN THE CRAYFISH ANTENNAL GLAND -- INTRODUCTION -- ROLE OF FORMED BODIES IN WATER METABOLISM -- UNIQUE FEATURES OF THE FORMED BODY MODEL -- PROCEDURES FOR QUANTITATING WATER MOVEMENT BY FORMED BODIES -- DISCUSSION -- REFERENCES -- CHAPTER 13. INTESTINAL WATER AND ION TRANSPORT IN FRESHWATER MALACOSTRACAN PRAWNS (CRUSTACEA) -- INTRODUCTION -- INCUBATION MEDIA -- NET WATER TRANSPORT. , TRANSMURAL FLUXES OF SODIUM AND CHLORIDE -- INFLUXES OF SODIUM AND CHLORIDE ACROSS THE APICAL CELL MEMBRANE -- A MODEL FOR WATER AND ION TRANSPORT -- CONCLUSIONS -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 14. MOLECULAR MECHANIS IS IN THE BRANCHIOPOD LARVAL SALT GLAND (CRUSTACEA) -- INTRODUCTION -- HYPOOSMOTIC REGULATION IN BRANCHIOPOD CRUSTACEANS (A. salina) -- ONTOGENY OF ELECTROLYTE REGULATION -- NATURE OF THE ELECTROLYTE ACTIVELY TRANSPORTED -- C-4 DICARBOXYLIC ACID PATHWAY AS A FACULTATIVE ANAEROBIC SHUNT -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 15. INSECT MIDGUT AS A MODEL EPITHELIUM -- INTRODUCTION -- THE ROUTE PROBLEM -- ELECTRICAL EVIDENCE FOR GOBLET CELL ROUTE -- LACK OF COUPLING BETWEEN CELLS -- KINETIC ANALYSIS OF POOL SIZES -- CONFIRMATION OF GOBLET ROUTE BY KINETIC ANALYSES -- MIDGUT MODEL AND PROSPECTS -- ACKNOWLEDGEMENT -- REFERENCES -- Session III: Epithelial Transport of Solutes and Water -- CHAPTER 16. SOLUTE AND WATER TRANSPORT ACROSS VERTEBRATE EPITHELIA: CHAIRMAN'S INTRODUCTORY REMARKS -- REFERENCES -- CHAPTER 17. THE SODIUM TRANSPORT POOL OF EPITHELIAL TISSUES -- INTRODUCTION -- LABELLING OF THE POOL WITH RADIOACTIVE SODIUM -- SUBCELLULAR IMMOBILIZATION -- SUBCELLULAR COMPARTMENTALIZATION -- SUMMARY -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 18. PASSIVE WATER TRANSPORT ACROSS EPITHELIA -- INTRODUCTION -- ORGANIZATION OF EPITHELIA AND ROUTES OF PERMEATION -- WATER IS JUST ANOTHER NON-ELECTROLYTE -- WATER PERMEABILITY OF EPITHELIA -- QUESTIONS ABOUT PERMEABILITY MEASUREMENTS -- CONCLUSION -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 19. IONIC ACTIVITIES AND SOLUTE TRANSFER IN EPITHELIAL CELLS OF THE SMALL INTESTINE -- INTRODUCTION -- THE MEASUREMENT OF IONIC ACTIVITIES IN EPITHELIAL CELLS IN THESMALL INTESTINE - SOME TECHNICAL CONSIDERATIONS -- K+ ACTIVITIES IN EPITHELIAL CELLS OF THE SMALL INTESTINE. , Na+ ACTIVITY IN THE SMALL INTESTINE -- Cl - ACTIVITY IN INTESTINAL EPITHELIA -- CONCLUSION -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 20. ANATOMICAL METHODS FOR STUDYING TRANSPORT ACROSS EPITHELIA -- INTRODUCTION -- I. TRADITIONAL ELECTRON MICROSCOPY -- II. AUTORADIOGRAPHIC LOCALIZATION OF ENZYMES -- III. ELECTRON MICROPROBE X-RAY ANALYSIS -- SUMMARY -- ACKNOWLEDGMENTS: -- REFERENCES -- CHAPTER 21. A MECHANISM FOR COUPLING ISOTONIC SALT AND WATER TRANSPORTIN THE MAMMALIAN PROXIMAL STRAIGHT TUBULE -- INTRODUCTION -- FLUID ABSORPTION AND TRANSEPITHELIAL VOLTAGE: SYMMETRICAL EXTERNAL SOLUTIONS -- ISOTOPIC Na+ FLUXES -- THE COUNTER-IONS FOR Na+ ABSORPTION -- FLUID ABSORPTION AND TRANSEPITHELIAL VOLTAGES: ASYMMETRICAL SOLUTIONS -- Jv AND Ve WITH ASYMMETRICAL SOLUTIONS -- Cl - BALANCE DURING FLUID ABSORPTION LINKED TO PASSIVE AND ACTIVEION FLUXES -- ANALYSIS OF SALT AND WATER ABSORPTIOH FROM ASYMMETRICAL SOLUTION -- MODEL -- THE DIFFUSION RESISTANCE FOR THE PARACELLULAR PATHWAY -- ASSUMPTIONS -- THE FLOW-DIFFUSION EQUATIONS -- THE CASE OF PASSIVE ION FLOWS -- THE CASE OF ACTIVE PLUS PASSIVE ION FLOWS -- EVALUATION IN TERMS OF FLOW-DIFFUSION EQUATIONS -- CONCLUDING REMARKS -- REFERENCES -- Session IV: General Session -- CHAPTER 22. INTRODUCTORY REMARKS OF THE GENERAL SESSION -- REFERENCES -- CHAPTER 23. ENERGY COUPLING IN ION AND WATER FLUXE SACROSS PLANT MEMBRANES -- INTRODUCTION -- ENERGY-LINKED MEMBRANE TRANSPORT -- SALT REGULATION BY PLANT CELLS -- SUMMARY -- REFERENCES -- CHAPTER 24. CELL VOLUME CONTROL -- Introduction -- VOLUME REGULATION IN DUCK ERYTHROCYTES -- CELL SHRINKAGE -- CELL ENLARGEMENT -- HORMONAL EFFECTS -- CONCLUSION -- SUMMARY -- REFERENCES -- CHAPTER 25. HORMONAL ACTION IN THE CONTROL OF FLUID AND SALT TRANSPORTING EPITHELIA -- INTRODUCTION -- INSECT SALIVARY GLAND -- ION-SELECTIVE MICROELECTRODES AND THE INSECT SALIVARY GLAND. , MAMMALIAN SALIVARY GLAND -- MALPIGHIAN TUBULES OF A BLOOD-SUCKING INSECT -- ELECTRON PROBE X-RAY MICROANALYSIS OF INSECT MALPIGHIAN TUBULE CELLS -- POSSIBLE INTRACELLULAR STORE OF CALCIUM IN MALPIGNIAN TUBULES -- CONCLUSIONS -- ACKNOWLEDGEMENTS -- REFERENCES -- CHAPTER 26. NON-EQUILIBRIUM THERMODYNAMICS OF WATER MOVEMENT -- INTRODUCTION -- NON-EQUILIBRIUM THERMODYNAMICS -- WATER TRANSPORT -- FACILITATED DIFFUSION -- ACTIVE TRANSPORT -- SOLVENT DRAG -- SUMMARY -- APPENDIX I -- APPENDIX II -- REFERENCES -- CHAPTER 27. PROBLEMS OF WATER TRANSPORT IN INSECTS -- INTRODUCTION -- GENERAL NATURE OF THE INSECT EXCRETORY PROCESS -- WATER ABSORPTION IN THE PAPILLATE RECTUM -- RECTAL ABSORPTION OF WATER VAPOUR -- THE SECRETING RECTUM OF SALINE WATER MOSQUITO LARVAE -- SUMMARY -- REFERENCES -- CHAPTER 28. THE ROLE OF HYDROSTATIC AND COLLOID-OSMOTIC PRESSURE ON THE PARACELLULAR PATHWAY AND ON SOLUTE AND WATER ABSORPTION -- INTRODUCTION -- THE PARACELLULAR PATH IS HIGHLY PERMEABLE -- EFFECTS OF HYDROSTATIC AND COLLOID OSMOTIC PRESSURE -- THE EFFECTS OF PRESSURE ON COUPLED SOLUTE-SOLVENT MOVEMENT -- REFERENCES -- AUTHOR INDEX -- SPECIES INDEX -- SUBJECT INDEX.
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  • 2
    ISSN: 1432-203X
    Keywords: Zea mays L. ; Transformation ; Protoplast ; Kanamycin ; β-glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The importance of cell culture conditions, including the use of feeder cells, on protoplast growth and transformation in maize (Zea mays L.) was investigated. Total GUS activity, measured two days after transformation, was five-fold higher in protoplasts cultured on feeder cells compared to those grown in the absence of feeder cells. Since the specific activity of GUS was only slightly higher in the transformed protoplasts plated over feeder cells, the stimulation in transient gene expression resulted mainly from the improved environment provided by the feeder system. For stable transformation, either PEG treatment or electroporation of protoplasts was used to introduce the neo gene. When PEG was used, over 85% of the putative transformants (resistant to kanamycin) contained the neo gene. The combination of PEG transformation and the optimized cell culture protocol using feeder cells enabled the selection of about 100 stably transformed lines per gFW of cells. Electroporation was less efficient.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Catharanthus ; Crown-gall ; Potassium uptake ; Tissue culture ; Vinca
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Potassium uptake in normal and crown-gall tissue culture cells of Vinca rosea L. (Cathalanthus roseus G. Don.) was studied. When grown on White's medium for 14–20 d, the tumor cells had a higher K+ content that did the normal cells. At this logarithmic stage of growth, the rate of K+ uptake was greater in normal cells than in tumor cells. K+ uptake in both normal and tumor cells was inhibited substantially by 2,4-dinitrophenol but only slightly by oligomycin, NaF, and N,N′-dicyclohexyl carbodiimide. When the internal K+ content of the cells was varied by growing the tissue in White's medium supplemented with varying concentrations of K+, the rate of K+ uptake in normal cells increased with decreasing internal K+ concentrations. In contrast both lines of tumor cells failed to show this normal increase in the rate of K+ uptake with decreasing internal K+ concentrations.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 9 (1990), S. 168-172 
    ISSN: 1432-203X
    Keywords: Oryza sativa L. ; indica rice, cotransformation ; protoplast ; kanamycin ; β ; glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of the indica rice (Oryza sativa L.) variety, IR54, were transiently transformed with the gusA gene and stably transformed with both the neo and gusA genes. We show that PEG-mediated co-transformation of protoplasts with two genes on separate plasmids coupled with selection on kanamycin is an effective way of transferring foreign gene(s) into the indica rice genome. The efficiency of co-transformation was generally 20–30%, i.e. the frequency of kanamycin-resistant calli having both the neo and gusA active genes. Southern blot analysis using a probe for gusA indicated integration of several copies of the gene, often as head to tail tandem repeats.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A system to regenerate fertile rice (Oryza sativa L.) plants (both indica and japonica varieties) from protoplasts isolated from anther-derived embryogenic haploid suspension cultures has been established. Green plants were regenerated from protoplast-derived cell clusters five months after suspension culture initiation. Protoplast yields and subsequent growth of the protoplast-derived microcalli were enhanced by transferring suspension cells into AA medium (Muller et al. 1978) three to four days prior to protoplast isolation. Protoplasts were cultured initially in Kao medium (Kao et al. 1977) and in association with nurse cells for four weeks. Protoplast-derived microcalli were transferred onto N6 (Chu et al. 1975) or MS (Murashige and Skoog 1962) media for callus proliferation. Callus growth was more rapid and the calli were more enbryogenic when grown on N6 medium. The 2,4-D concentration used to develop the suspension culture was important. Cell cultures grown in medium containing 0.5 mg/l 2,4-D released protoplasts whose plating efficiency was higher than for protoplasts obtained from suspension cultures grown in 2.0 mg/l 2,4-D. However, suspension cells grown in 2.0 mg/l 2,4-D were superior with regard to the ability of protoplast-derived calli to regenerate green plants. Amongst several hormone treatments evaluated, a combination of 0.5 mg/l NAA + 5.0 mg/l BAP resulted in the largest number of green plants regenerated. There were no significant differences between BAP or kinetin regarding total number of plants regenerated. More than 200 green plants have been produced form six independently initiated suspension cell lines. The number of regenerated plants per 106 protoplats plated anged from 0.4 to 20.0, and the average seed fertility of single panicles of these RO plants was about 40%.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-5028
    Keywords: Agrobacterium ; rice ; transformation ; Ti plasmid ; GUS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transient expression of GUS in rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens was characterized using binary vectors containing gusA genes that express minimal (pKIWI105 and pCNL1) or no (p35S-GUS-INT and pCNL56) GUS activity in bacteria. Four-day old seedlings obtained from seeds or immature embryos of rice were cut into shoot, root, and seed remnants and inoculated with various strains of A. tumefaciens. Transient GUS expression events were quantitated histochemically by determining the frequency of explants exhibiting blue spots indicative of GUS at four to six days after cocultivation with A. tumefaciens. A. tumefaciens strains that did not contain the gusA gene (At643) or a Ti-plasmid (At563 and At657) did not elicit any blue staining characteristic of GUS activity. Several parameters were important in obtaining efficient transient expression of GUS in rice mediated by A. tumefaciens. The growth regulator 2,4-D inhibited GUS expression if present during the seed germination period, but the presence of 6 mg/1 2,4-D during cocultivation of the explants with A. tumefaciens slightly enhanced GUS expression efficiency. All 21 rice cultivars tested expressed GUS after co-cultivation with A. tumefaciens. The GUS expression frequency was highest amongst the indica cultivars. The frequencies of GUS expression in japonica cultivars and in Oryza glaberrima cultivars (grown primarily in Africa) were generally one-half to one-third the level found for indica varieties. Leaf explants were more susceptible to A. tumefaciens-facilitated GUS expression than were roots or seed remnants. The vir genes of an agropine-type Ti-plasmid of A. tumefaciens were most effective in directing transient GUS expression in rice, whereas those of a nopaline-type and an octopine-type plasmid were less effective. We have also found that the frequency of transient expression of GUS was higher with pBIN19 as the precursor cloning vector than with pEND4K as the precursor cloning vector. Reasons for differences in effectiveness of these binary vectors are discussed. Using the conditions described here, A. tumefaciens-mediated frequencies of transient GUS expression in four-day old shoots of several rice cultivars were routinely in excess of 50%.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-5028
    Keywords: maize gene expression ; protoplasts ; transient assays ; transcriptional regulation ; zein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three DNA regions required for high levels of transcription were identified by transient gene expression analysis of the 5′ flanking region of a 19 kDa α-zein gene. For these analyses, the zein promoter region was fused to the β-glucuronidase (GUS) gene and assayed by transient expression in carrot protoplasts. A 107-bp sequence (-114/-8) containing the TATA box resulted in low levels of GUS activity. Addition of the proximal 75 bp (-189/-114) doubled the level of GUS expression, and a further increase in expression was obtained when additional upstream sequences (-483/-226) were placed 5′ of the zein promoters. Zein upstream sequences enhanced transcription independently of the-189/-114 region. Although the-189/-114 region was not essential for transcription, it was important to obtain maximum GUS activity. A 121 bp upstream sequence (-347/-226) that contains the conserved TGTAAAG sequence gave high levels of GUS activity when placed in either orientation 5′ of the zein promoter sequences. However, nucleotides-347 to-309, containing the TGTAAAG sequence, could be deleted from this fragment without a significant change in GUS activity. Zein upstream sequences did not promote transcription of the GUS gene in somatic maize protoplasts. The upstream activating sequence from the cauliflower mosaic virus (CaMV) 35S promoter placed 5′ of deletion mutants of the zein promoter also failed to produce GUS activity above background.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5028
    Keywords: bar ; herbicide resistance ; phosphinothricin acetyltransferase ; rice ; selectable marker ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have used the bar gene in combination with the herbicide Basta to select transformed rice (Oryza sativa L. cv. Radon) protoplasts for the production of herbicide-resistant rice plants. Protoplasts, obtained from regenerable suspension cultures established from immature embryo callus, were transformed using PEG-mediated DNA uptake. Transformed calli could be selected 2–4 weeks after placing the protoplast-derived calli on medium containing the selective agent, phosphinothricin (PPT), the active component of Basta. Calli resistant to PPT were capable of regenerating plants. Phosphinothricin acetyltransferase (PAT) assays confirmed the expression of the bar gene in plants obtained from PPT-resistant calli. The only exceptions were two plants obtained from the same callus that had multiple copies of the bar gene integrated into their genomes. The transgenic status of the plants was varified by Southern blot analysis. In our system, where the transformation was done via the protoplast method, there were very few escapes. The efficiency of co-transformation with a reporter gene gusA, was 30%. The To plants of Radon were self-fertile. Both the bar and gusA genes were transmitted to progeny as confirmed by Southern analysis. Both genes were expressed in T1 and T2 progenies. Enzyme analyses on T1 progeny plants also showed a gene dose response reflecting their homozygous and heterozygous status. The leaves of To plants and that of the progeny having the bar gene were resistant to application of Basta. Thus, the bar gene has proven to be a useful selectable and screenable marker for the transformation of rice plants and for the production of herbicide-resistant plants.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 13 (1989), S. 151-161 
    ISSN: 1573-5028
    Keywords: β-glucuronidase (gusA) gene ; maize ; protoplasts ; stable co-transformation ; transformation ; Zea mays L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient co-transformation protocol using polyethylene glycol was developed for Zea mays L. (cv. A188 × BMS) protoplasts isolated from suspension culture cells. Co-transformation was accomplished by using plasmid constructions containing β-glucuronidase (gusA) or neomycin phosphotransferase (neo) gene coding sequences; both were under control of the CaMV 35S promoter. Protoplast culture and transformation conditions were optimized to assure efficient recovery of transformed cells. The overall efficiency of transformation was 1 × 10−4 (calculated per viable protoplast plated). Among kanamycin-resistant lines, 50% showed a high level of GUS activity (above one unit). Southern blot hybridization confirmed the presence of numerous gusA and neo coding sequences in the maize genome. In two analyzed lines, integrated sequences appeared to be organized in tandem head-to-tail repeats. Results also indicated that the integrated sequences were partially methylated.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 27 (1995), S. 91-104 
    ISSN: 1573-5028
    Keywords: Oryza sativa L ; transgenic rice ; transgene inheritance ; gusA gene ; neo gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Inheritance of foreign genes neo and gusA in rice (Oryza sativa L. cv. IR54 and Radon) has been investigated in three different primary (T0) transformants and their progeny plants. T0 plants were obtained by co-transforming protoplasts from two different rice suspension cultures with the neomycin phosphotransferase II gene [neo or aph (3′) II] and the β-glucuronidase gene (uidA or gusA) residing on separate chimeric plasmid constructs. The suspension cultures were derived from callus of immature embryos of indica variety IR54 and japonica variety Radon. One transgenic line of Radon (AR2) contained neo driven by the CaMV 35S promoter and gusA driven by the rice actin promoter. A second Radon line (R3) contained neo driven by the CaMV 35S promoter and gusA driven by a promoter of the rice tungro bacilliform virus. The third transgenic line, IR54-1, contained neo driven by the CaMV 35S promoter and gusA driven by the CaMV 35S. Inheritance of the transgenes in progeny of the transgenic rice was investigated by Southern blot analysis and enzyme assays. Southern blot analysis of genomic DNA showed that, regardless of copy numbers of the transgenes in the plant genome and the fact that the two transgenes resided on two different plasmids before transformation, the introduced gusA and neo genes were stably transmitted from one generation to another and co-inherited together in transgenic rice progeny plants derived from self-pollination. Analysis of GUS and NPT II activities in T1 to T2 plants provided evidence that inheritance of the gusA and neo genes was in a Mendelian fashion in one plant line (AR2), and in an irregular fashion in the two other plant lines (R3 and IR54-1). Homozygous progeny plants expressing the gusA and neo genes were obtained in the T2 generation of AR2, but the homozygous state was not found in the other two lines of transgenic rice.
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