Plant Biotech / Plant Tissue Culture Time Line


MJ Schleiden hypothesized cell theory, suggesting totipotentiality of cells


Horse drawn harrows, seed drills, corn planters, horse hoes, 2-row cultivators, hay mowers, and rakes Industrially processed animal feed and inorganic fertilizer


Charles Darwin hypothesized that animal and plant populations adapt over time to best fit the environment


Gregor Mendel investigated how traits are passed from generation to generation - called them factors


Johann Meischer isolated DNA from the nuclei of white blood cells


J Sachs reported that plants synthesizing organ-forming substances that are polarly distributed


G Haberlandt made the first (but unsuccessful) attempt at tissue culture; Walter Suton coined the term "gene" and proposed that chromosomes carry genes (factors that Mendel said that could be passed from generation to generation) 


B Hanning attempted embryo culture (crucifers)


E Kuster achieved fusion of protoplasts although the products did not survive


Thomas H. Morgan proved that genes are carried on chromosomes; the term “biotechnology" coined


M Molliard cultured fragments of plant embryos


L Knudson germinated orchard seeds in vitro; WJ Robbins cultured root tips


Blumenthal and Meyer achieved callus formation on carrot root explants through the use of lactic acid


Laibach used embryo culture to rescue interspecific hybrids of Linum


FW Went demonstrated that there were growth substances in coleoptiles from Avena


Plant hybridization used widely in plant breeding


RJ Guatheret unsuccessfully cultured cambial tissues of trees and shrubs; PR White maintained long-term cultures of tomato roots; Kogl et al identified the first plant growth regulator, IAA


CR LaRue cultured embryos of conifers


Proteins and DNA studied by x-ray crystallography; Term “molecular biology" coined


Successful continuously growing cambial cultures of carrot and tobacco. Gautheret R. J., C. R. Acad. Sci. (Paris), 208: 118-120; Nobecourt P., C. R. Soc. Biol. (Paris), 130: 1270-1271; White P. R., Am. J. Bot., 26: 59-64


Gautheret cultured cambial tissues of Ulmus to study adventitious shoot formation


Van Overbeek used coconut milk to support growth and development of very young Datura embryos; Braun cultured crown gall tissues in vitro; George Beadle and Edward Tatum proposed the “one gene, one enzyme” hypothesis


Observation of secondary metabolites in callus cultures (Gautheret)


Braun elucidated the tumor-inducing principle of crown galls


Folke Skoog used tobacco cultures to study adventitious root formation


Transition from animal power to mechanical power on farms


E Ball produced the first whole plants from shoot tip cultures of Lupinus and Tropaeolum


Formation of adventitious shoots and roots in tobacco culture (Skoog and Tsui)


Culture of fruits in vitro (Nitch)


Ball regenerated organs from sequoia; Morel reported the first successful cultures of a monocot using coconut milk; Edwin Chargaff determined there is always a ratio of 1:1 adenine to thymine in DNA of many different organisms


Nitch cultured excised ovaries in vitro; Skoog demonstrated chemical control of growth and organ formation in vitro


First virus-free plants produced from meristem culture; first successful micrografts (Morel and Martin); Hershey and Chase used radioactive labeling to determine that DNA and not protein that carries the instructions for assembly of phages


James Watson and Frances Crick identify the helix structure of DNA; haploid callus was produced from pollen grains of ginko (Tulecke)


Muir et al regenerated calli from single cells using nurse cultures


Synthesis of kinetin (Miller et al)


Discovery of the regulation of organ formation by changing the ratio of auxin: cytokinin (Skoog and Miller); Culture of excised anthers of Allium (Vasil); Francis Crick and George Gamov explain how DNA functions to make protein


Regeneration of somatic embryos in vitro from the nucellus of Citrus ovules (Maheshwari and Rangaswamy); Proembryo formation in callus and cell suspensions of carrot (Reinert and Steward); Coenberg discovers DNA polymerase


First successful test tube fertilization in Papaver rhoeas (Kanta); Use of the microculture method for growing single cells in hanging drops in a conditioned medium (Jones et al.); Enzymatic degradation of cell walls to obtain large numbers of protoplasts (Cocking); Filtration of cell suspensions and isolation of single cells by plating (Bergmann); Isolation of mRNA


Development of Murashige and Skoog medium (Physiol Plant 15:473)


Production of first haploid plants from pollen grains of Datura (Guha and Maheshwari)


Regeneration of tobacco plants from single cells (Vasil and Hildebrant); Production of protocorms from orchids in vitro (Morel)


Marshall Nirenberg and Severo Ochoa determine that a sequence of 3 nucleotide bases determines each of the 20 amino acids


Haploid plants produced from pollen grains of tobacco (Bourgin and Nitch)


Enzymes involved in cleaving DNA termed restriction endonucleases (Meselson and Yuan)


Selection of biochemical mutants in vitro by the use of tissue culture derived

variation (Carlson); First achievement of protoplast fusion (Power et al.); Hybrid embryo culture and subsequent chromosome elimination for haploid production in barley (Kao and Kao); Discovery of first restriction endonuclease from Haemophillus influenzae Rd. It was later purified and named HindI (Smith)


Preparation of first restriction map using HindI to cut circular DNA of SV 40 into 11 specific fragments (Nathans); Regeneration of first plants from protoplasts (Takebe et al)


First report of interspecific hybridization through protoplast fusion in two species of Nicotiana (Carlson et al); Restriction fragments can be joined by DNA ligase regardless of their origin if they are cut with the same restriction enzyme (Mertz and Davis; Berg); Isolation of reverse transcriptase (Temin)?


Use of the Lobban and Kaiser technique to develop hybrid plasmid - insertion of EcoRl fragment of DNA molecule into circular plasmid DNA of bacteria using DNA ligase. Gene from African clawed toad inserted into plasmid DNA of bacteria (Herbert Boyer and Stanley Cohen) First recombinant DNAorganism - beginning of genetic engineering; Cytokinin found to be capable of breaking dormancy in Gerbera (Pierik et a Somatic hybridization of tomato and potato, resulting in pomato (Melchers et al.) Somatic hybridization of tomato and potato, resulting in pomato (Melchers et al.)l)


Regeneration of haploid petunia hybrids plants from protoplasts (Binding); Biotransformation in plant tissue cultures (Reinhard); Discovery that the Ti plasmid is the tumor inducing principle of Agrobacterium (Zaenen et al; Larebeke et al); Induction of branching by cytokinins in gerbera shoot tips


Positive in vitro selection of maize callus cultures resistant to T toxin of Helminthosporium maydis (Gengenbach and Green); Development of the high resolution two dimensional gel electrophoresis procedure, which led to the development of proteomics (O'Farrel); Moratorium on recombinant DNA techniques


Shoot initiation from cryopreserved shoot apices of carnation (Seibert); Octopine and nopaline synthesis and breakdown found to be genetically controlled by the Ti plasmid of A. tumefaciens (Bomhoff et al); National Institute of Health guidelines developed for study of recombinant DNA


Successful integration of the Ti plasmid DNA from A. tumefaciens in plants (Chilton et al); A method of DNA sequencing developed (Maxam, Gilbert); Discovery of split genes (Sharp Roberts); First practical application of genetic engineering – human growth hormone produced by bacterial cells


Genentech, Inc. uses genetic engineering techniques to produce human insulin in E. coli, became first biotech company on NY stock exchange; Somatic hybridization of tomato and potato, resulting in pomato (Melchers et al.)


Cocultivation procedure developed for the genetic transformation of plant protoplasts with Agrobacterium (Marton et al)


Use of immobilized whole cells for biotransformation of digitoxin into digoxin) (Alfermann et al); Commercial production of human insulin through genetic engineering in bacterial cells (Eli Lilly and Co.); Studies on the structure of T-DNA cloning the complete EcoRl digest of Ti, tobacco crown gall DNA into a phage vector, thus allowing the isolation a detailed study of T-DNA border sequence (Zambryski et al); US Supreme Court decides that manmade microbes can be patented


Introduction of the term somaclonal variation (Larkin and Scowcroft)


Naked DNA transformation of protoplasts (Krens et al); Electrofusion of protoplasts (Zimmerman)


Co-integrate type vectors designed for Agrobacterium transformation (Zambriski et al); The idea of polymerize chain reaction (PCR), a chemical DNA amplification process, is conceived (Kary Mullis)

Transgenic plants were first created in the early 1980s by four groups working independently at Washington University in St. Louis, Missouri, the Rijksuniversiteit in Ghent, Belgium, Monsanto Company in St. Louis, Missouri, and the University of Wisconsin. On the same day in January 1983, the first three groups announced at a conference in Miami, Florida, that they had inserted bacterial genes into plants. The fourth group announced at a conference in Los Angeles, California, in April 1983 that they had inserted a plant gene from one species into another species.

The Washington University group, headed by Mary-Dell Chilton, had produced cells of Nicotiana plumbaginifolia, a close relative of ordinary tobacco, that were resistant to the antibiotic kanamycin (Framond, A.J., M.W. Bevan, K.A. Barton, F. Flavell, and M.D. Chilton. 1983. Mini-Ti plasmid and a chimeric gene construct: new approaches to plant gene vector construction. Advances in Gene Technology: Molecular Genetics of Plants and Animals. Miami Winter Symposia Vol. 20:159-170).

Jeff Schell and Marc Van Montagu, working in Belgium, had produced tobacco plants that were resistant to kanamycin and to methotrexate, a drug used to treat cancer and rheumatoid arthritis (Schell, J., M. van Montagu, M. Holsters, P. Zambryski, H. Joos, D. Inze, L. Herrera-Estrella, A. Depicker, M. de Block, A. Caplan, P. Dhaese, E. Van Haute, J-P. Hernalsteens, H. de Greve, J. Leemans, R. Deblaere, L. Willmitzer, J. Schroder, and L. Otten. 1983. Ti plasmids as experimental gene vectors for plants. Advances in Gene Technology: Molecular Genetics of Plants and Animals. Miami Winter Symposia Vol. 20:191-209).

Robert Fraley, Stephen Rogers, and Robert Horsch at Monsanto had produced petunia plants that were resistant to kanamycin (Fraley, R.T., S.B. Rogers, and R.B. Horsch. 1983a. Use of a chimeric gene to confer antibiotic resistance to plant cells. Advances in Gene Technology: Molecular Genetics of Plants and Animals. Miami Winter Symposia Vol. 20:211-221.). The Wisconsin group, headed by John Kemp and Timothy Hall, had inserted a bean gene into a sunflower plant.

These discoveries were soon published in scientific journals. The Schell group's work appeared in Nature in May (Herrera-Estrella, L., A. Depicker, M. van Montagu, and J. Schell. 1983. Expression of chimaeric genes transfered into plant cells using a Ti-plasmid-derived vector. Nature 303:209-213) and the Chilton group's work followed in July (Bevan, M.W., R.B. Flavell, and M.D. Chilton. 1983. A chimaeric antibiotic resistance gene as a selectable marker for plant cell transformation. Nature 304:184-187). The Monsanto group's work appeared in August in Proceedings of the National Academy of Sciences (Fraley, R.T., S.G. Rogers, R.B. Horsch, P.R. Sanders, J.S. Flick, S.P. Adams, M.L. Bittner, L.A. Brand, C.L. Fink, J.S. Fry, G.R. Galluppi, S.B. Goldberg, N.L. Hoffmann, and S.C. Woo. 1983b. Expression of bacterial genes in plant cells. Proceedings of the National Academy of Sciences 80:4803-4807). The Hall group's work appeared in November in the journal Science (Murai, N., D.W. Sutton, M.G. Murray, J.L. Slightom, D.J. Merlo, N.A. Reichert, C. Sengupta-Gopalan, C.A. Stock, R.F. Barker, J.D. Kemp, and T.C. Hall. 1983. Phaseolin gene from bean is expressed after transfer to sunflower via tumor-inducing plasmid vectors. Science 222:476-482).


Infection and transformation of leaf discs with Agrobacterium tumefaciens and regeneration of transformed plants. Horsch RB et al., Science, 227:1229-1231; Development of disarmed Ti plasmid vector system for plant transformation. Fraley RT et al, Bio/Technol, 3:629-635; Development of binary vector system for plant transformation. An G. et al., EMBO J., 4:277-284; Gene transfer in protoplasts of dicot and monocot plants by electroporation. Fromm ME, PNAS (USA), 82:5824-5828; Plants can be patented


TMV virus-resistant tobacco and tomato ( transgenic plants developed using cDNA of coat protein gene of TMV (Powell-Abel et al); Transformation of tobacco protoplasts by direct DNA microinjection (Crossway); First field trials of transgenic plants


Development of biolistic gene transfer method for plant transformation (Sanford et al; Klein et al); Isolation of Bt gene from bacterium (Bacillus thuringiensis) (Barton et al); First monocot (Asparagus) transformation by Agrobacterium tumefaciens (Bytebier et al)


Recovery of stable transformants through particle bombardment (Klein et al); Automated mass propagation with organogenesis and embryogenesis( Levi R et al)


Formal launch of the Human Genome Program; Plant transformation by microinjection of intact plant cells (Neuhaus); Electroporation of intact plant tissues for direct DNA delivery (Dekeyser et al); Silicon carbide fiber-mediated DNA delivery into plant cells (Kaeppler et al)


Flavr Savr tomatoes sold to public


Sequencing of E coli genome (Blattner et al)


Sequencing of the genome of a multicellular organism (Caenorhabditis elegans)


Arabidopsis draft sequence completed


Sequencing of the human genome draft completed (Human Genome Project Consortium and Venter et al)

Toby Bradshaw’s lab is burned down; ELF claims responsibility


Of the soybeans grown in the US, 64% are transgenic; 34% of corn is. EU Union has had a 5 year ban on GMOs.