Austrian botanist. Russian Wikipedia. BnF authorities. Gottlieb Haberlandt.
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One of the stellar achievements of twentieth century plant biology was the genetic transformation of somatic cells enabling the regeneration of whole plants that were stably transformed and capable of transmitting the inserted genetic material to subsequent generations. This achievement grew out of three independent lines of research initiated early in the twentieth century: plant tissue culture, regeneration of plants from single somatic cells, and the study of crown gall disease.
The early discoveries made in these areas represent a combination of basic scientific research and technological innovations and led to the development of genetically transformed crop species expressing traits unobtainable by conventional breeding. Each of these fields can be traced back to a single research publication Haberlandt, ; Smith and Townsend, ; White, a that later came to be considered as the foundation of the field.
It is instructive to follow how these three fields were established, progressed, converged, and finally coalesced. Early workers, of course, could not know the ultimate way in which their discoveries would be applied to modern plant biotechnology Figure 1.
Instead, they posed questions that were of interest to them and to the scientific communities of which they were a part, and some of their discoveries led in directions that would later prove to be productive in the progression to biotechnology. This essay is intended to provide a review of the crucial discoveries that ultimately led to modern plant biotechnology and show how they contributed to this progression.
Some of the major steps and publications are listed. Further details, definitions, and additional citations are provided in the main text. Philip White worked at the Rockefeller Institute for Medical Research in Princeton, New Jersey in the s to develop an experimental system with which to study metabolism in a completely undifferentiated tissue where all cells are identical and hence exert similar influences on one another.
The most successful of the early attempts involved the culture of maize, pea, and cotton root tips Kotte, ; Robbins, These could be excised from the plant with minimal trauma and grown aseptically for a few weeks in nutrient media, but ultimately growth ceased. Tips were excised from seedling roots and subcultured at regular intervals. White a addressed this second question using a hybrid between Nicotiana glauca and N.
The hybrid plants produced tumor-like calluses and galls on the stem and leaves. Tissue removed aseptically from young stems was cultured on the same nutrient medium used for tomato roots.
Proliferated masses from the original explants were divided and subcultured at weekly intervals through 40 subcultures. The question remained as to whether the cells in the culture were undifferentiated. Histological examination revealed only mature parenchyma cells, regions of dividing meristematic cells, and occasional isolated xylem cells. Despite this level of cellular heterogeneity, White concluded that these tissues approximated an undifferentiated condition and grew for potentially unlimited periods and therefore were true tissue cultures.
Scientific results are greatly strengthened when other workers with appropriate expertise replicate the original findings. Remarkably, this occurred within 6 weeks of the publication of White's results. Neither cited White a , and presumably they were unaware of his results. Neither study was as detailed as White's.
White's objective, of studying metabolism in an undifferentiated tissue wherein all the cells are identical and presumably exert similar influences on one another, appears not to have been followed up by him or his contemporaries.
It was not until many years later that other scientists began using methods developed by White to study cellular metabolism. Notably, H. Street at Manchester University used cultured excised tomato roots in an extensive series of studies to examine the metabolism of inorganic ions, carbohydrates, amino acids, and hormones Street, A more direct approach to White's original intention was followed by F.
Steward and colleagues at Cornell University. They examined metabolic states in three contrasting carrot tap root cultures: secondary phloem cells as they existed in the intact root, excised tissue maintained on a minimal medium where cells grew predominantly by enlargement, and actively metabolizing cells stimulated by coconut milk in the medium to divide as rapidly as possible.
With White's original goal for plant tissue culture having been achieved and confirmed, attention turned in other directions. White b found that shoots were produced from Nicotiana tumor tissues submerged in a liquid medium, and Michael Levine Levine, reported the spontaneous formation of shoots and roots on cultured carrot tissue. These observations turned attention to the question of totipotency.
That is, did tissue cultured cells retain the full genetic competence of the zygote to form a complete plant? This question was addressed by Folke Skoog and his collaborators at the University of Wisconsin. Starting from White's discovery of shoot formation in submerged Nicotiana tumor cultures, they found that auxin IAA was a potent inhibitor of shoot formation in tobacco tissue cultures cv Wisconsin 38 , but high concentrations of auxin stimulated the formation of roots.
The nucleic acid base adenine supplied with low auxin concentrations stimulated shoot formation in these cultured tissues.
The amount of shoot and root formation depended on the proportions of auxin and adenine supplied in the culture medium Skoog and Tsui, In further work, they isolated from autoclaved DNA an adenine derivative, 6-furfurylaminopurine, that they named kinetin. Using tobacco callus cultures they found that by adjusting the relative concentrations of auxin and kinetin in the culture medium it was possible to induce the formation of shoots and roots or the growth of undifferentiated callus Skoog and Miller, Thus, they demonstrated that tobacco callus tissues retained the potentialities of the zygote to form both shoots and roots, but their research did not prove that a single cell had these potentialities.
Studies that approached this question were initiated in the laboratory of F. Steward at Cornell University. Previously this group had compared metabolism of carrot secondary phloem explants in basal medium or medium supplemented with coconut milk, a source of cell division factors.
Explants in the coconut milk medium produced new cells that became detached and proliferated freely in the culture medium, resulting in large numbers of single cells and cell aggregates.
Among these aggregates they noted some that formed roots and subsequently shoots to produce whole plants that could later be transferred to agar media and then to soil where they flowered and completed the life cycle Steward, ; Steward et al. The implication of this statement was that the origin of the regenerated plants was from single cells. If so, cellular totipotency would have been proven for carrot cells, at least.
However, other workers cited in Sussex, pointed out that there was no incontrovertible proof that single cells rather than cell aggregates that may have contained cells with different genetic potentialities were the source of the new plants that they obtained.
These tissue culture studies that demonstrated the potentially unlimited growth of undifferentiated cells and the production from them of roots, shoots, and entire plants did not contribute further to the questions that we are examining because of their failure to assure the single cell clonal origin of regenerated plants and thus the genetic totipotency of single cells.
However, the study of plant tissue and suspension cultures was continued in different directions, including the commercial production of secondary products Ramawat and Merillon, and commercial production of trees, crops, and horticultural plants, most notably species of orchids Arditti and Krikorian, Gottlieb Haberlandt, working in Graz, Austria, was the first to culture isolated somatic cells of higher plants in vitro.
He began these investigations in and published the results in Haberlandt, Recognizing the lack of knowledge of the nutrient requirements of higher plant cells, he used as a culture medium the seven inorganic elements that had been identified by Knop as sufficient for the water culture of higher plants, with additions of sucrose, dextrose, glycerine, asparagine, and peptone in various concentrations and combinations.
Haberlandt first attempted to culture green, photosynthetic cells from leaf bract mesophyll of Lamium purpureum. Bracts were teased apart in liquid until microscopy examination revealed numerous isolated palisade and spongy mesophyll parenchyma cells.
These were then transferred by finely drawn-out pipettes to hanging drops or dishes of culture medium. Microbial sterility was attempted by flaming instruments and glassware but usually failed to eliminate bacterial and fungal contamination completely. Cultures were maintained in lighted rooms at ambient temperature or in darkness.
Some cells remained alive for a month in lighted cultures but died soon in darkness. Haberlandt noted several changes in cell structure during the culture period. Cells expanded in length and girth. Plastids remained green in light, photosynthesized, and accumulated starch.
However, no cells were observed to divide. He then attempted to culture cells from other species: photosynthetic cells from Eichhorina crassipes , glandular hairs of Pulmonaria mollissima , stinging hairs of Urtica dioica , staminal filament hairs of Tradescantia virginica , and stomatal cells of Ornithogalum umbellatum, Erythronium dens-canis , and Fuschia magellanica with equal lack of success reviewed in Krikorian and Berquam, In retrospect, Haberlandt's failure to obtain dividing cells can be attributed to lack of microbial sterility, culture media that lacked hormones and growth factors that were unknown at that time, and his selection of highly differentiated mature cells.
However, he made immense contributions to plant and animal cell culture studies by his technical innovations, including the use of hanging drop culture methods and use of micropipettes to manipulate single cells. Similarly his prediction that cocultivation of vegetative cells with pollen tubes, that were then known to produce chemical stimuli that induced growth of orchid ovules, foreshadowed nurse culture technology, and his prediction that embryo sac fluids might be used as components of the culture medium to induce divisions in isolated vegetative cells foreshadowed the use of coconut milk.
Each of these predictions has led to advances in cell culture technology. Despite continued efforts by Haberlandt's collaborators and others, no significant progress on cultures derived from single cells was made for 56 years when W. They used tissue cultures of tobacco and carrot and crown gall cultures of grape, marigold, periwinkle, and sunflower. By testing the capacity for growth in liquid culture media, they identified several that produced large numbers of single cells.
Single cells were then transferred by micropipette to filter paper placed on nurse cultures of the same or other species growing on an agar medium. Those of tobacco, marigold, sunflower, and grape divided and produced macroscopic cultures, some of which were transferred through 25 or more agar subcultures without diminution of growth rate.
Marigold clones consistently produced roots, and hybrid tobacco clones produced shoots on media containing adenine and kinetin. Subsequently, Vimla Vasil and Hildebrandt Vasil and Hildebrandt, a , b transferred single tobacco hybrid cells to a drop of culture medium on a microscope slide that could be observed and photographed repeatedly under phase contrast microscopy.
Cells were observed to divide to form a filament and subsequently a microcallus mass that was transferred to an agar medium for further growth, where roots and leafy shoots were differentiated. Rooted shoots were transferred to soil where they produced buds and flowers. Thus, these studies demonstrated that plantlets derived from single cultured cells had the capacity to produce whole plants.
However, they did not prove that the whole plants were the direct product of a single cell, rather than the product of a tissue mass within which somaclonal or other genetic changes might have taken place during growth to produce a chimeric tissue mass.
They cultured single carrot cells from suspension cultures on microscope slides where they could be observed and photographed repeatedly. This early research in plant tissue culture demonstrated that tissues isolated from plants can be grown in culture for indefinite periods of time, they can produce shoots and roots, and finally, single isolated cells in culture can produce embryos. These studies provided the platform for genetic transformation of plants, as described below.
This was subsequently reclassified as Phytomonas tumefaciens and then as Agrobacterium tumefaciens Conn, Smith established that this bacterium was the cause of the disease by plating bacteria from galls onto an agar medium, inoculating uninfected plants with subcultures of the bacteria, reisolating bacteria from the galls produced, and inducing galls on new plants Koch's postulates.
Tumors were also produced on stems of tobacco, tomato, and potato and on roots of sugar beet and peach trees that were inoculated with B.
The latter galls closely resembled peach crown gall disease on which Smith had been working for several years without identifying the cause.
In a subsequent publication, Smith et al. Smith also observed secondary tumors that developed on stems or leaves of infected plants of some species at some distance from the primary gall. Based on histological examination, he concluded that secondary tumors developed from tumor strands that were root-like outgrowths from the primary gall Smith et al.
Smith believed that tumor strands might be comparable to certain types of metastases that occurred in malignant tumors of animals and humans Smith, In addition to establishing the cause of crown gall disease, Smith and Townsend suggested that their results might shed light on the origin of cancerous growths in animals.
Smith frequently alluded to the similarities between plant and animal tumors Smith, for which he received a Certificate of Honor from the American Medical Association in , and in , he was elected to the presidency of the American Association for Cancer Research. However, little progress was made on the nature of crown gall disease until Armin Braun at the Rockefeller Institute for Medical Research, began an investigation of crown gall disease in the s that lasted for 40 years and that laid the foundation for the molecular studies that were to come.
He began this investigation by examining the question of tumor strand connections between primary and secondary tumors in sunflower. By examining tissue sections cut between the primary and secondary tumors, he found no histological support for a tumor strand connection and concluded that the mechanism of formation of secondary tumors may not be identical to that concerned in the formation of the primary tumor Braun, Although crown gall tissues did not always yield cultures of the inducing bacterium, it had been assumed that the bacteria had been present at some stage in the development of the tumor.
Ungarisch-Altenburg, Hungary, 28 November ; d. Berlin, Germany, 30 January Despite great interest and talent in music, painting, and German literature , he studied botany. Julius Wiesner in Vienna became his first teacher and supervised the work on his Ph. Leitgeb in
Plant Cell and Tissue Cultures: The Role of Haberlandt
Gottlieb Haberlandt , born Nov. He returned to Austria in to teach botany at the Technical Academy in Graz. In Haberlandt succeeded Schwendener in the chair of plant physiology at the University of Berlin , where he established the Institute for Plant Study. Haberlandt decided that his students would profit from a system of classifying plants based on function. Although his system was not accepted by other botanists, the analysis of the relations between structure, function, and environment has been useful in the study of plant adaptations to different habitats. Gottlieb Haberlandt.
The Scientific Roots of Modern Plant Biotechnology
It seems that you're in Germany. We have a dedicated site for Germany. In the th anniversary of the publication on "Culturversuche mit isolierten Pflanzenzellen" by Gottlieb Haberlandt was celebrated. This book pays homage to a great Austrian scientist and the further development of his ideas. The first part of the book contains a facsimile of the original paper which is a true artistic masterpiece and its first translation into English from The fourth part of the book contains an overview of important topics of plant tissue culture with the most promising areas of application to date and an outlook into the future. Areas range from micropropagation, production of pharmaceutically interesting compounds, plant breeding, genetic engineering of crop plants, including trees, and cryopreservation of valuable germplasm.
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