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David Hopwood wants both to ‘capitalize on the power of microbial genetics combined with the facilities for plant research at the JII’ (Hopwood, 2007, p. 83), and diversify the work of the Genetics Department to satisfy the Agricultural Research Council which has reservations about the focus on Streptomyces genetics in his department. They do not suggest possible topics for research, so Hopwood decides to initiate three projects on the interface between microbiology and plant science: Rhizobium, Agrobacterium and plant mycoplasmas.
In 1970 John Beringer begins his PhD supervised by David Hopwood in the Genetics Department. Rhizobia are a group of bacteria that can form symbioses with members of the legume family. As we now know, the plants release chemical signals which attract the rhizobia, which in turn release chemicals that cause the plants to form small nodules on their roots. Rhizobia then live inside these nodules; the plants provide them with food (carbon) and the rhizobia ‘fix’ atmospheric nitrogen in a form which the plants can use. Rhizobium leguminosarum is potentially a particularly suitable species for use in genetic studies of symbiotic nitrogen fixation as it is the partner of the genetically well-known legume, the pea (Pisum sativum), a plant that is already well-established as a research organism in the Applied Genetics Department at JII. Beringer sets out to develop a workable system of genetic analysis in R. leguminosarum.
Initially his focus is to establish a genetic map for R. leguminosarum, which involves developing a method of plasmid-based (R-factor) transfer of chromosomal genes. The methods he develops are important for R. leguminosarum and his 1974 paper is one of the most highly cited papers ever published in this field. His early successes lead to the expansion of Rhizobium work and the appointment of Andrew Johnston in 1974 to begin work on plasmid-based gene transfer, and Nick Brewin (a biochemist) in 1975. The topic develops into a major theme of research at John Innes, continuing to the present day.
Agrobacterium tumefasciens is a plant pathogen that forms tumours (“crown galls”) on many dicotyledonous plants and there is already an indication that this is caused by the transfer of DNA from the bacterium into the plant, an unprecedented situation. Hopwood appoints David Cooper as a John Innes Fellow to work on this system, but the project is jeopardised by Cooper’s untimely death in 1975. It is continued by John Firmin, initially appointed on a Cancer Campaign grant that David Cooper had obtained, and later by Angus Hepburn. But the John Innes work is overshadowed by brilliant work in Ghent, Belgium, led by Jeff Schell, and later by Mike Bevan at the Plant Breeding Institute in Cambridge. Agrobacterium turns out to have been a far-sighted choice of topic when it becomes (in the late 1980s) the favoured method for the genetic engineering of the majority of plants, even if the project does not leave a lasting legacy at John Innes.
Mycoplasmas are wall-less bacteria that live inside the cells of multicellular animals and plants where they cause disease. Those of plants are so far unculturable outside the host and Hopwood reasons that if this could be shown to be due to the dependence of the bacterium on the host for aspects of macromolecular synthesis, rather than simply the provision of some specialised nutrient(s), this could represent a key step in the evolution of mitochondria, which are derived from bacteria that are no longer complete organisms but have become organelles within eukaryotic cells. Mike Daniels is appointed in the Genetics Department to work on this topic. He soon realises that the mycoplasmas are essentially intractable as a genetic system and switches his attention to another type of wall-less plant pathogens, the spiroplasmas, which are culturable. He makes progress in studying them, but later switches again to work on Xanthomonas campestris, a “normal” plant pathogenic bacterium, which turns out in his hands to be a tractable model for studying the molecular biology of plant-pathogen interactions. Daniels eventually obtains funding from the Gatsby Foundation and this leads to the setting up of The Sainsbury Laboratory in 1987 (formally opening in 1989) as a result of a major grant from the Foundation, with Daniels as its founding director.
J. E. Beringer, ‘R Factor transfer in Rhizobium leguminosarum’, Journal of General Microbiology 84 (1974): 188-198.
David A. Hopwood, Streptomyces in Nature and Medicine: The antibiotic makers, Oxford: Oxford University Press, 2007.
M. J. Daniels, P. G. Markham, B. M. Meddins, A. K. Plaskitt, R. Townsend and M. Bar-Joseph, ‘Axenic culture of a plant pathogenic Spiroplasma, Nature 244 (1973): 523-524.
In the 1970s two separate strands of work on tissue culture develop at the John Innes Institute: (1) Micropropagation mainly using axillary root propagation, and (2) Anther culture and the production of haploid plants.
In the late 1950s and early 1960s the development of tissue culture techniques in labs around the world enabled shoots to be regenerated from proliferating unorganised tissues known as callus cultures. Unfortunately such cultures were generally genetically very unstable and not suitable for cloning plants. Only a very few types of plant (such as orchids) could be successfully propagated using these methods. Micropropagation (a term that came into usage in the late 1960s to describe the use of tissue culture to propagate plants in vitro) was potentially useful for rapidly bulking up new genotypes or disease-free specimens of existing plants, and for storing valuable breeding material under disease-free conditions. What was needed to make this a workable technology was a method that did not share the main disadvantages of adventitious shoots regenerated from host tissue or most callus systems, namely the problem of genetic instablility.
In 1970 the first encouraging development at the John Innes Institute is a method devised by Roy Davies (Davies et al., 1971) of regenerating shoots of Freesia from callus which at first seem to be genetically stable. However, the method does not bring the hoped-for advance as the regenerating shoots eventually produce aberrant plants. A method pioneered in the United States in the 1970s and soon after in the UK, using proliferating axillary shoots with monocot and later woody species, becomes the method of choice. Techniques involve growing suitable pieces of each plant species in an appropriate culture medium and stimulating the precocious formation of multiple axillary shoots. Between 1970 and 1982 Graham Hussey and assistants Judith Hilton, Joan Turner, Carol Wyvill, Janet Hargreaves and N. J. Stacey in Applied Genetics establish micropropagation techniques for a wide range of monocot species including Narcissus, Allium, Iris, Alstroemeria, Gladiolus, Hippeastrum, Nerine, Cordyline and Amorphophallus. Many of these techniques pioneered at JII are adopted and developed by the horticultural industry (Hussey 1978, 1986).
(2) Anther culture
In parallel with but independent of the micropropagation work, Norman Sunderland, with co-workers Frances Wicks, James Dunwell, Mary Roberts and Linda Evans, researches improved methods of pollen and anther culture which are important techniques for genetic and mutation studies and have many potential applications in crop improvement. Sunderland, who joined the Institute in 1958 and started his career in the Department of Cell Biology, began tissue culture work initially ‘to see if cell cultures having large chromosomes, or small numbers, could be obtained that might be exploitable by the cytology department’. From callus cultures obtained from seeds Sunderland moved on to experiment with culturing anthers; his first success (with the tobacco plant Nicotiniana tabacum) was reported in Nature (Sunderland and Wicks 1969), and subsequently in The Sunday Times, The Guardian, and the New Scientist, and the flowering haploid plants were exhibited at the Chelsea Flower Show and at a conversazione of the Royal Society.
In the early 1970s visiting workers Sant S. Bhojwani (University of Delhi) and Glenn B. Collins (University of Kentucky, Lexington) collaborate with Sunderland’s team on aspects of embryogenic pollen, with Jim Dunwell (who joined JII in 1970) making the ultrastructural examinations. Sunderland early appreciates the potential of pollen for producing colonies of haploid cells and for the regeneration of haploid plants, useful in plant breeding; a notable success during this period is the first production of haploids from anther culture of potato, Solanum tuberosum (Sunderland and Dunwell, 1973). However, there is opposition to this technique from conventional plant breeders in that the plantlets derived from pollen calluses in some species (e.g. barley) are generally albinos, a problem which will be investigated by colleagues in the Department of Applied Genetics in the 1980s.
Anther culture spreads rapidly to laboratories worldwide, particularly to China where the objective is to apply the technique to major food crops (rice, wheat and maize) and where in 1978 Sunderland helps introduce plant scientists to culture procedures. As a result of this visit, Xu Zhihong, a research associate of Shanghai Institute of Plant Physiology, Chinese Academy of Sciences (CAS) and research student Huang Bin, from the Institute of Genetics, CAS, join JII in 1979 as visiting workers to work on float cultures of barley anthers (float cultures were pioneered at JII by Sunderland and Roberts in 1977). Chinese engagement with JIC is facilitated by an academic exchange agreement between the Royal Society and CAS.
Sunderland’s subsequent work on anther culture involves establishing the optimum stage for isolation and culture of the anther or pollen, the effects of temperature pre-treatments and the culture requirements of each species, and makes him an internationally recognised authority on the subject (see citation information in the link below).
Article on ‘Tissue culture’ by Norman Sunderland, with editorial insertions by Jim Dunwell, March 2010. A full bibliography of Jim Dunwell’s publications on anther culture is available from JIC archives on request.
D. R. Davies and P. Griffiths, ‘In vitro propagation of Freesia’, Sixty-Second Annual Report of the John Innes Institute (1971), p. 45.
G. Hussey, ‘The application of tissue culture to the vegetative propagation of plants’, Sci. Prog. Oxf., 65 (1978): 185-208.
G. Hussey, ‘Vegetative propagation of plants by tissue culture’, pp. 29-66 in M. M. Yeoman, Plant Cell Culture Technology. Botanical Monographs 23. Oxford: Blackwell Scientific Publications, 1986.
N. Sunderland and F. M. Wicks, ‘Cultivation of haploid plants from tobacco pollen’, Nature, 224 (1969): 1227-1229.
N. Sunderland and F. M. Wicks, ‘Embryoid formation in pollen grains of Nicotiniana tabacum, Journal of Experimental Botany, 22, no. 70 (1971): 213-226.
N. Sunderland and J. M. Dunwell, ‘Anther culture of Solanum tuberosum’, Euphytica, 22 (1973): 317-323.
N. Sunderland, ‘Pollen and anther culture’, pp. 205-239 in H. E. Street (ed.) Plant tissue and Cell culture, Oxford: Blackwell, 1973.
N. Sunderland and J. M. Dunwell, ‘Anther and pollen culture’, pp. 223-265 in H. E. Street (ed.) Plant tissue and Cell culture, 2nd edn., Oxford: Blackwell, 1977.
N. Sunderland and M. Roberts, ‘New approach to pollen culture’, Nature, 270, issue 5634 (1977): 236-238.
N. Sunderland, ‘The concept of morphogenetic competence with reference to anther and pollen culture’, pp. 125-140 in S. K. Sen and K. L Giles (eds.), Plant Cell Culture and Crop Improvement, New York: Plenum Press, 1983.
R. L. M. Pierik, In vitro cultivation of higher plants, Dordrecht, the Netherlands: Kluwer Academic Press, 1987; 4th revised printing 1997 has a timeline for the history of tissue culture on pp. 22-23.
For more background on the development of anther culture see:
The staff of the John Innes Institute’s new Virus Research Department begin the 1970s physically separated, with some based at JII and some still housed temporarily in the adjacent newly-established ARC Food Research Institute (FRI). The core staff of this department had moved from the ARC Virus Research Unit in Cambridge and was incorporated into Roy Markham’s Cell Biology Department at JII after 1967. By 1970 Cell Biology has been disbanded and its staff re-allocated to three departments according to their particular lines of work: Applied Genetics, Ultrastructural Studies and Virus Research. The FRI contingent of virologists joins the rest of their Department in new permanent laboratories at JII on Decimal Day, in February 1971
The Virus Research Department, under new Head Dr. John B. Bancroft from Purdue University, continues work on the structure and composition of plant viruses, increasingly using biochemical and molecular tools. Bancroft’s major interest is mutagenesis and protein studies with Cowpea chlorotic mottle virus (CCMV). Most members of the department contribute to this research which requires an ever increasing amount of amino acid analysis. A collaboration between viral protein chemist Maurice Rees and Ron Self of the FRI’s Mass Spectrometry Group leads to some very successful amino acid sequences using a method of derivatising peptides, so that they can be sequenced by mass spectrometry. One viral protein is sequenced mainly by mass spectrometry, a tour de force. CCMV work continues after Bancroft resigns and returns to Canada in 1973. With Rees as Acting Head, a transatlantic collaboration with Bancroft is sustained.
Overall the department’s work involves purifying and characterising a whole range of plant viruses including among others Papaya mosaic virus, Alfalfa mosaic virus, Pea enation mosaic virus, Beet yellows virus and Cauliflower mosaic virus (see section on Recombinant DNA technologies), and investigating the properties of virus particles using biophysical tools and electron microscopy. These methods include the analytical ultracentrifuge (Beckman Model E which Roy Markham helped to develop), optical diffraction techniques (using laser beams to reveal patterns that reflect virus particle structure), and new ideas of image reconstruction that are being pioneered in the Ultrastructural Studies Department and applied particularly to plant viruses. During the 1970s, Ultrastructural Studies and Virus Research work closely together and their achievements include new insights into the detailed structure of isometric virus particles (Horne et al. 1977) and bacilliform virus particles (Hull 1976). The interaction between the two departments also facilitates the successful infection of protoplasts with plant viruses, an area of research which is being spearheaded at JII (Motoyoshi et al. 1974 a, b; Motoyoshi et al. 1975). Not only is it hoped that this will be a route to getting foreign DNA into plant cells, but it is a valuable approach to understanding plant virus replication.
The work of the Virus Research Department attracts several prominent visiting scientists (including J. B. Bancroft (Canada), M. Delseny (France), R. M. Goodman (USA), Steve Howell (USA), Moshe Bar-Joseph (Israel), L. C. Lane (USA), F. Motoyoshi (Japan), M. Russo (Italy), J. G. Shaw (USA) and J. Tremaine (Canada)) who spend periods of one or two years in the department. With Jeff Davies’ appointment as Head in 1979 the emphasis of the Virus Research Department changes, and work begins on comoviruses (the Cowpea mosaic virus group) and geminiviruses.
S. Howell and R. Hull, ‘Replication of cauliflower mosaic virus and transcription of its genome in turnip leaf protoplasts’, Virology, 86 (1978): 468-481.
R. W. Horne, J. M. Harnden, R. Hull, ‘The in vitro crystalline formations of turnip rosette virus. 1. Electron Microscopy of two- and three-dimensional arrays, Virology, 82 (1977): 150-162.
R. W. Horne, Structure and Function of Viruses. London: Edward Arnold, 1978.
R. Hull, ‘The structure of tubular viruses’, Advances in Virus Research, 20 (1976): 1-32.
F. Motoyoshi and R. Hull, ‘The infection of tobacco protoplasts with pea enation mosaic virus’, Journal of General Virology, 24 (1974a): 89-99.
F. Motoyoshi, J. W. Watts and J. B. Bancroft, ‘Factors influencing the infection of tobacco protoplasts by cowpea chlorotic mottle virus’, Journal of General Virology, 24 (1974b): 245-256.
F. Motoyoshi and R. Hull, ‘The infection of tobacco mesophyll protoplasts by alfalfa mosaic virus’, Journal of General Virology, 27 (1975): 263-266.
I. Takebe, ‘Protoplasts in the study of plant virus replication’, pp. 237-283 in H. Fraenkel-Conrat and R. R. Wagner (eds.), Comprehensive Virology, Volume XI. New York: Plenum Press, 1977.
R. L. M. Synge, ‘Maurice William Rees, 1915-1978’, Sixty-Ninth Report of the John Innes Institute (1978): 17-20.
M. N. Short, ‘Origins, 1944-1984’, Memoir covering Margaret Short’s years at VRU, Cambridge and in the Virus Research Department, JII (Unpublished, c. 1989, John Innes Archives).
R. Hull, ‘The Phenotypic Expression of a Genotype: Bringing Muddy Boots and Micropipettes together’, Annual Review of Phytopathology, 46 (2008): 1-11.
One of the important resources available to JII’s virologists is a large collection of plant virus cultures, started in the 1930s by Kenneth Smith in Cambridge and considered to be the best in the country. An inventory of the Plant Virus Collection compiled in 1970 lists around 54 different viruses, for many of these JII holds several interesting strains. It includes, for example, the Alfalfa mosaic virus and Cauliflower mosaic virus collections which prove invaluable to Roger Hull’s investigations. By 1979 the JII collection has grown to around 150 separate virus isolates and there are an increasing number of requests for virus material from researchers around the world.
The large size of the collection (which is augmented over the years by the collection of new virus cultures) means that only a small proportion of the viruses can be maintained in stock plants. To conserve glasshouse space and reduce the risks of contamination most of the sap-transmissible viruses are stored in dried leaf tissue. The collection is managed by Margaret Short, who had been Smith’s assistant at the Virus Research Unit (VRU) in Cambridge. Here she received diseased plants from all over the country, and occasionally overseas. Many of the early samples were sent by the National Agricultural Advisory Service (NAAS) in Trumpington, Cambridge, whose plant pathologist channelled material from growers to VRU. Short would test the samples by inoculation into a variety of seedlings to detect possible ‘new’ viruses. She also researched the method for storing viruses in use at JII (after H. H. McKinney 1953) back in 1957. Roger Hull takes over the collection Margaret Short has managed for nearly 40 years on her retirement in 1984.
M. N. Short, ‘Origins, 1944-1984’, Memoir covering Margaret Short’s years at VRU, Cambridge and in the Virus Research Department, JII (Unpublished, c. 1989, John Innes Archives).
The John Innes Institute’s new Ultrastructural Studies Department (inaugurated in 1970 with R.W. (Bob) Horne as Head), begins work from a temporary building on the Colney site. Too cold in the winter and too hot in the summer, working conditions are made worse because the windows have to be taped up to keep out the dust. Nevertheless the Department, which comprises staff from the Electron Microscopy section reinforced by workers from the old Cell Biology Department, makes significant advances on the structure of nuclear pores and provides some of the best images available at the time (Lacour and Wells 1974- the first Electron Microscopy paper published from work at Colney). The change in organisation signals a move to a technology based department (significantly Horne is a physicist by training), rather than a biological subject-oriented department. Horne, who was co-founder of the Electron Microscopy Section of the Royal Microscopical Society in 1965, cultivates an ethos of working collaboratively with scientists, rather than running the Department as a service organisation as happens in many universities and other institutes. This far-sighted policy helps the staff gain a feel for what is likely to be real and what is likely to be artefact in the images they produce. Their ultramicroscopical expertise is applied to the understanding of the structure and assembly of cell walls, the mechanisms of cell and nuclear division, and the recognition of viruses and mycoplasmas infecting plant tissues, among other topics.
Until the permanent buildings become fully operational at the end of November 1970, the electron microscopes are physically separated with a Siemens I and an English EM, the AE1 6B, located at the adjacent Food Research Institute, and another Siemens at JII (these models are among the first commercial EMs available in Britain). The new permanent building houses all the EMs together and is designed with special foundations to safeguard against vibrations affecting the EM equipment, and with materials that will not introduce undesirable magnetic fields, important features given the crude mechanics and imperfect magnetic screening of these early machines. There is also ample space for teaching in the Department, which is responsible for most of the structural work at JII, from cytology studied by optical microscopy to high resolution EM work (in 1970 ‘a true resolution by electron microscopy of crystal structure at 0.14nm has been realised’ which is close to the theoretical limits of the EM at the time - about 0.08nm). Ultrastructural Studies at JII (which includes Brian Wells, Len Lacour, Graham Hills, Kitty Plaskitt, John Watts and John McCleish) is designated as an Agricultural Research Council centre of excellence, and supplies training in EM techniques to other institutes through a series of annual ARC (later AFRC) EM users’ meetings; these are ‘hands-on-technology’ based rather than formal scientific meetings. Summer schools and ARC EM Conferences are also held at JII. In addition the John Innes Trustees finance a ‘Royal Microscopical Society John Innes Lecture’ series; the first is given by Bob Horne in 1971 on ‘The application of Electron Microscopy to Virology’.
Horne brings to JII an established reputation in the investigation of viral particles at the ultrastructural level, a field which opened up after he and Sydney Brenner published a technique which became known as the ‘negative staining’ method (negative contrast electron microscopy) in 1959. This dramatically improved the electron microscopy of viruses and its usefulness as a diagnostic technique in medical virology. In Horne’s previous post, the EM Laboratory at the Institute of Animal Physiology at Babraham, Cambridge, his expertise was applied to the structure of viral particles and bacterial cells in animal tissues. On joining JII he deploys negative staining techniques in the investigation of Tobacco mosaic virus and other plant viruses, producing high-resolution micrographs that reveal protein structure. With Roy Markham he collaborates on the application of optical diffraction and image reconstruction techniques to electron micrographs. Another important Ultrastructural technique that becomes identified with JII in the 1970s is serial sectioning (including three-dimensional reconstructions based on serial sectioning of small objects), developed by Brian Wells.
Audio clip from Keith Roberts’ interview with Brian Wells, 12 January 2000. The full interview is available in the John Innes Archives
Sydney Brenner on Bob Horne and the development of negative staining (filmed April-May 1994)
A.W. Agar, ‘The story of European electron microscopes’, pp. 415–584 in T. Mulvey (ed.), Advances in Imaging and Electron Physics (1st ed.). Academic Press: London, 1996.
B. E. P. Beeston, R. W. Horne, R. Markham, ‘Electron diffraction and optical diffraction techniques’, in (ed.) Audrey Glauert, Electron Diffraction and Optical Diffraction Techniques: Practical Methods in Electron Microscopy. Amsterdam.
S. Brenner and R.W. Horne, ‘A negative staining method for high resolution electron microscopy of viruses’, Biochem. Biophys. Acta, 34 (1959): 103–110.
R. W. Horne, ‘Completion of the building for the Department of Ultrastructural Studies’, Sixty-First Annual Report of the John Innes Institute (1970), pp. 12-17.
R. W. Horne, ‘Early developments in the negative staining technique for electron microscopy’, Micron et Microscopica Acta, 22, 4 (1991): 321-326.
R. W. Horne and I. Pasquali-Ronchetti (1974). ‘A negative stain-carbon film technique for studying viruses in the electron microscope. I. Preparative procedures for examining icosahedral and filamentous viruses’, Journal of Ultrastructure Research, 47 (1974): 361-383.
R. W. Horne, I. Pasquali-Ronchetti and J. M. Hobart, ‘A negative stain-carbon film technique for studying viruses in the electron microscope. II. Application to adenovirus type 5’, Journal of Ultrastructure Research, 51 (1975): 233-252.
R. W. Horne and P. Wildy, ‘An historical account of the development and applications of negative staining technique to the electron microscopy of viruses’, Journal of Microscopy, 117 (1979): 103-122.
L. F. Lacour and B. Wells, ‘Nuclear pores at prophase of meiosis in plants’, Philosophical Transactions of the Royal Society of London ‘B’, 268 (1974): 95-100.
B. Wells, ‘A convenient technique for the collection of ultra-thin serial sections’, Micron, 5 (1974): 79-81.
I. A. McQuade and B. Wells, ‘The synaptinemal complex in Rhoeo spathacea, Journal of Cell Science, 17 (1975): 349-69.
Although the Agricultural Research Council has since the early 1960s had the objective of extending the range of glasshouse flowers as an alternative crop for the glasshouse industry, which was then mainly dependent on tomatoes, only recently has there been any publicly funded work on the breeding and genetics of ornamental plants for the British horticultural industry, a market which in 1972-3 is worth £72.5 million (more than double the value of the glasshouse vegetable sector at this date).
Several flower breeding programmes are introduced at the John Innes Institute in the 1970s. These include genetic studies of Freesias to solve difficulties in propagating by seed, to increase their colour range, and to investigate the inheritance of doubleness, a very desirable character in freesias; of Carnations, to produce plants resistant to Fusarium wilt, a fungal disease (the germ plasm from this project is sold off in the early 80s for commercial development to Brooke Bond who have a side-line in carnation breeding and growing in Kenya and Uganda); of Antirrhinums (Snapdragons), to produce acceptable forms for forcing as cut flowers, and of Anenome and Streptocarpus (Cape Primrose).
Eddie Arthur leads the work on breeding improved forms of Anenome coronaria, the florists’ anemone. His aim is to analyse the genetic components of productivity, including quality, uniformity and yield, and to increase the hardiness of the plants. Hardiness is important to Cornish growers who use corms raised from seed in Holland; the Dutch varieties are not well-adapted to English conditions and suffer from winter damage. At the close of the programme (1979) Arthur’s team deliver inbred parents of 15 selected F1 hybrids to the National Seed Development Organisation (which acts as a seed merchant for research institutes) for further commercial development.
Gavin Brown leads the Streptocarpus breeding programme. Streptocarpus was first brought to England from Southern Africa by plant collectors in 1818 - a large blue-flowered plant S. rexii. After more than a hundred years of moderate use as an indoor plant a new wave of interest in Streptocarpus began in 1947 with a John Innes variety ‘Constant Nymph’, bred by William Lawrence while he was working on colour inheritance. Produced by crossing a hybrid streptocarpus called ‘Merton Blue’ (S. hybridus) with a species called Streptocarpus johannis (introduced just before World War II), this plant flowers continuously from April to November and with additional light could be kept in bloom throughout the year. Soon it was common to see ‘Constant Nymph’ on display in florists’ windows in Paris, Amsterdam and other major European cities. The Dutch popularised and made a good market in Europe and the US with this original blue variety and a number of blue mutations, and also a natural white sport (‘Maasen’s White’); however, there were no other colours commercially available.
Gavin Brown’s project, which starts in 1969, is to increase the colour range of Streptocarpus. The cross that made ‘Constant Nymph’ is remade several times, but a pink or red form of S. hybridus is substituted for one of the parents each time. All the first-generation seedlings are blue, but when these are cross-bred with each other in the second generation, white, pink, red, magenta, purple, and pale blue flowers are produced and all retain the constant flowering habit. Ten of these hybrids are selected for naming and further testing. In 1972 JII releases its first batch of seven new varieties of Streptocarpus named Diana, Fiona, Karen, Louise, Marie, Paula and Tina through the National Seed Development Organisation (NSDO). Later Brown develops further varieties, both by traditional breeding methods and by X-ray- and chemical-induced mutation. The new JII varieties receive considerable attention from plant breeders in Holland (where they are awarded a gold medal), but the British market for these new houseplants is slow to develop. Brown’s retirement in 1975 ends the programme and this, together with JII’s overall policy of closing down the breeding work, removes the incentive for JII to pursue the marketing of the new varieties through NSDO. Nevertheless Louise, Paula, and Tina were all awarded the Royal Horticultural Society’s Award of Merit and in 2010 several John Innes varieties are still available from specialists.
Anon. ‘JI hybrids take top Dutch award’, Commercial Grower, 5 July 1974, p. 10 [Gold medal at Aalsmeer Experimental Station show].
Gavin Brown’s breeding material was eventually passed to the specialist nursery Dibleys, who are the main breeder of new varieties in the UK and holders of the UK National Collection of Streptocarpus
G. W. Cooke (ed.), Agricultural Research 1931-1981, London, ARC, 1981.
The John Innes Institute programme for breeding apple varieties with resistance to scab (Venturia inaequalis) concludes with several John Innes selections sent to National Fruit Trials at Faversham in Kent. Gavin Brown collects a number of accolades for this research, including an MBE and the Royal Horticultural Society’s Lindley Medal in 1971 for an exhibition illustrating the successful breeding of disease-resistant dessert apples at JII. Gavin’s retirement in 1975 effectively ends the apple- and pear-breeding at the John Innes Institute, with the work being transferred to the top-fruit East Malling Research Station in Kent. No longer needed for fruit work, John Innes Institute’s Field Station and Farm, Stanfield, near East Dereham is sold in 1976.
Fruit research at JII in Colney is now focussed on cherries and strawberries. Peter Matthews and Paddy Dow are working to breed cherry varieties resistant to bacterial canker. Matthews began this work in 1958 when he transferred from the Department of Genetics to Applied Genetics, taking over the then unused cherry seedling- and variety-collections resulting from the earlier incompatibility and breeding work of Morley Benjamin Crane and Dan Lewis. Alongside breeding he introduces laboratory studies of the causative agents of bacterial canker, Pseudomonas morsprunorum and Ps. syringae, work on dwarfing rootstocks, and incorporates the self-fertile mutation produced by Dan Lewis in the 1950s. For his work on cherry breeding Peter Matthews is awarded the Royal Horticultural Society’s Jones-Bateman cup in 1973. Matthews achieves his primary objective of producing a range of cherry varieties with resistance to bacterial canker, the ‘second generation’ varieties including ‘Merchant’, ‘Hertford’ and ‘Colney’, which are released in the early 1980s. Matthews’ subsidiary programmes also have delivered high quality, self-fertile selections and dwarf or small tree scion varieties. However the potential of these developments will be exploited elsewhere since in 1982 the JII’s cherry germplasm collection and breeding programme is transferred to East Malling Research Station.
Hedley Williams leads the strawberry breeding which has been ongoing at John Innes (initially at Bayfordbury) since its transfer from the Horticultural Research Station at Cambridge University in 1951. Strawberry breeding had first begun at Cambridge in 1928, and the UK crop then dominated by just two varieties was increasingly succumbing to what was subsequently recognised as a virus. The aim of the Cambridge programme (on which Hedley Williams worked as an assistant to Daniel Boyes) was to widen the choice of varieties, while at the same time producing a range of similar seedlings that could replace initial selections lost to disease. Hedley Williams’ JII breeding programme has the additional aim of selecting for disease resistance and uses micro-propagation techniques to produce disease-free stocks. ‘Merton Dawn’, one of the six strawberry varieties bred at the Institute, will be one of the last of the Institute’s fruit to be named with the prefix ‘Merton’ due to recent EEC regulations. Hedley Williams’ tragic death in a cycling accident in 1979 brings the strawberry breeding programme to an end, and the breeding material is sent to Long Ashton Research Station, Bristol.
P. Matthews, ‘Recent advances in breeding sweet cherries at the John Innes Institute’, Fruit Present and Future, volume 2, (1973), pp. 100-113. London: Royal Horticultural Society.
H. Williams, ‘The ever changing strawberry’, ibid., pp. 140-144.
P. Matthews, ‘Progress in breeding cherries for resistance to bacterial canker’, Proceedings of the Eucarpia Fruit Section Symposium, 1979 (1980), pp. 157-174. Angers: Eucarpia.
P. Matthews, ‘X-ray induced small tree mutants in sweet cherry selections resistant to bacterial canker’, ibid., pp. 173-179.
The foundation of the work undertaken on the cytology and genetics of peas over the previous decades at the John Innes Institute is the JI Pisum collection which by the end of the 1970s numbers about 1,400 distinct lines from around the world. The collection includes material from wild, cultivated and semi-cultivated sources, some dating back to the nineteenth century, many of which have arisen as spontaneous mutants. The parents of the new generation of ‘semi-leafless’ and ‘leafless’ pea varieties come from this large collection. As the pea collection has grown, it has become essential to catalogue phenotypes and genotypes properly, using a variety of methods which in 1970 leads to the first computer-based information retrieval system for a pea gene bank. This is developed by Brian Snoad in Applied Genetics who devises a standardised method for recording the location, origin, pedigree and genotype of the pea accessions. Designed exclusively for peas, but compatible with Food and Agriculture Organisation (FAO) and International Atomic Energy Authority (IAEA) systems for wheat and rice, the database is set up on IBM cards to permit, with invaluable help from Mick Johnson in Virology, computer storage, processing and retrieval, usable on an international basis. This work coincides with the recognition that the Pisum collection is a public resource and has to be responsive to external requests.
The growth of the John Innes Pisum collection, and the increasing number of contacts being made, lead to the decision to participate in founding an international group, the Pea Genetics Association, which from 1969 arranges meetings, exchanges material and ideas, and begins publishing its own newsletter. The Association comes to play an important role in international research on peas, in some cases making possible collaborative trials of new lines in other countries.
During the 1970s Pisum sativum continues to be developed as the major crop plant for genetical and physiological research in Applied Genetics at JII and it is the range of genetic variability being revealed by the collection that underpins co-operative research projects within and outside the Institute, with investigations on pea root and leaf growth, interaction with the environment, pathogen resistance, and storage proteins underway. Snoad’s co-authors and co-operators include Cliff Hedley and Mike Ambrose (Physiology); Eddie Arthur (Genetics and Biometrics); Peter Matthews (Pathology and Genetics), and Derek Harvey (Physiology) at JII, and visiting workers, S T Ali-Khan, Canada (Genetics); G P Gent, PGRO (Agronomy); Luigi Monti, Italy (Genetics); Luigi Fursciante, Italy (Genetics); D J Rogers, USA (Genebanks) and L. Seidewitz, Germany (Genebanks). In addition there are significant international collaborations associated with germplasm and genetic stocks with Stig Blixt (Weibullsholm Collection, Sweden) and Gerry Marx (Cornell, USA). The maintenance of the collection and servicing requests are undertaken by Peter Matthews who acts as unofficial Curator, a duty he manages alongside his primary research.
B. Snoad, ‘Taking the leaf out of peas’, Supplement to GROWER (March 19th 1981).
M. W. Johnson and B. Snoad, ‘Pisum germ plasm records’, Sixtieth Annual Report of the John Innes Institute (1969): 17.
M. W. Johnson, ‘Computing’, Sixty-First Annual Report of the John Innes Institute (1970): 20.
The final chapter of William Hayes’ influential textbook The Genetics of Bacteria and their Viruses (2nd ed. 1968, p. 746) begins: ‘One of the most interesting, and certainly the strangest, of the discoveries to issue from the genetic study of bacteria is the existence of a new kind of genetic element, the sex factor, which determines the ability of E. coli K-12 bacteria to conjugate and to transfer genetic material to recipient cells’. Hayes refers to the discovery in the 1950s that sex or ‘fertility’ plasmids promote conjugation (mating) in the bacterium Escherichia coli (Hayes, 1953; Jacob and Wollman 1958), an observation that turns out to be a special case of a very general phenomenon among bacteria. How the mechanism of conjugation and genetic transfer works remains a controversial and unresolved question in 1970.
One of the main priorities of the John Innes Institute’s new Streptomyces group is to contribute to this debate by looking for the involvement of plasmids in gene exchange in Streptomyces coelicolor. Alan Vivian, following up David Hopwood’s data on the fertility of various recombinant strains, finds that differences in mating behaviour are transmitted between strains at a much higher frequency than chromosomal genes. In other words, he identifies an infectious conversion of one fertility type to another, behaviour which implicates the involvement of a plasmid (a DNA molecule separate from the main bacterial chromosome) which he names SCP1 (Vivian and Hopwood 1970). Soon Hopwood’s group have examples of strains with and without the plasmid, as well as strains with SCP1 integrated into the chromosome. Crosses between two strains lacking the plasmid are the least fertile, and crosses between bacteria with and without SCP1 are more fertile. Most fertile of all are the crosses between a strain with SCP1 in the chromosome and one lacking the plasmid. The discovery of a conjugation system, at least in part plasmid controlled, in a complex, Gram positive bacterium, S. coelicolor, is of great interest because it indicates the widespread importance of plasmids as genetic determinants in bacteria. While Hopwood’s team has provided genetic evidence for SCP1, finding physical or biochemical evidence for the plasmid proves impossible until new techniques for separating large linear fragments of DNA become available in the 1980s – SCP1 then turns out, unprecedentedly, to be a linear DNA molecule.
A key step in securing the future of Streptomyces research at John Innes had been the appointment of Keith Chater to the Genetics Department in 1969, the first staff appointment made by David Hopwood on coming to John Innes. Keith brings his skills as an experienced microbiologist, having graduated in bacteriology at Birmingham University and studied the genetics of Salmonella typhimurium there for his PhD. Hopwood had initiated a study of morphogenesis while still in Glasgow and handed over this topic, along with his collection of morphological mutants (the whi mutants) to Chater, who quickly begins, in the 1970s, to make his mark on their study (Chater 1972). This is the start of a career-long and definitive project which will expand to occupy many PhD students and post-docs, as well as to catalyse the work of other project leaders worldwide, including, later, Mark Buttner at John Innes. Chater had also acquired an interest in bacteriophages while still in Birmingham and this will eventually lead to his development of Streptomyces phages as cloning vectors.
William Hayes, The Genetics of Bacteria and their Viruses: Studies in basic genetics and molecular biology, Oxford and Edinburgh: Blackwell Scientific Publications, 2nd edition, 1968.
D. A. Hopwood, K. F. Chater, J. E. Dowding and A. Vivian, ‘Advances in Streptomyces coelicolor Genetics’, Bacteriological Reviews 37, 3 (1973): 371-405.
K. F. Chater, ‘A morphological and genetic mapping study of white colony mutants of Streptomyces coelicolor’, Journal of General Microbiology, 72 (1972): 9-28.
David A. Hopwood, Streptomyces in Nature and Medicine: The antibiotic makers, Oxford: Oxford University Press, 2007, pp.84-85.
A joint application for a computing system to the Agricultural Research Council by R. Self of the Food Research Institute and Mick Johnson of the John Innes Institute is successful and an IBM 1130 computer is purchased and installed at FRI for the use of both Institutes. The main purpose of the system is to process data from the mass spectrometer at FRI and data from amino acid analyses and gas chromatographs, but it is also used for general computing. Two years after installation the computer has become heavily used and many of the routine programs have to be run overnight. The Applied Genetics Department (‘three regular users’) are making increasing use of the statistical programs available and the JII/FRI Computer Group is gradually ‘converting’ other members of staff to using the computing facilities, with staff of both institutes being offered lectures in FORTRAN programming. New JII computing projects include a pea gene data retrieval program for Applied Genetics and the development of programs to handle data from Roy Markham’s digital recording microdensitometer (the first computer-aided cell image analysis at JII).
Roy Markham had been investigating problems of ultrastructure and had designed a simple and effective diffractometer for electron microscope image analysis. He uses the computer as a means of minimizing the ‘random noise’ in these images and programmes are developed to analyse and then enhance electron micrographs. This involves the conversion of optical information into digital form and Markham works to improve on the complex and expensive equipment required for the transformation of optical data. He designs and builds a simple ‘densitometer’ which scans electron micrographs and converts the densities across the scan into digital data. His first model is so successful that it provides the foundation for an application for a larger and more powerful computer to develop JII’s image processing projects. The ARC agree to purchase a new and more appropriate computer for JII in December 1978, and the Trustees of the John Innes Charity agree to fund the provision of a ‘computer suite’ as part of an extension to the Ultrastructural Studies laboratory. A large multi-user ‘mini-computer’ is installed in the new suite at JII and a smaller one at FRI as a ‘satellite’ in 1979; the joint computer group staff move across to the new suite at JII in April 1980. By this date there are about 60 computer users at JII and 100 at FRI.
Annual Reports of the John Innes Institute, 1970-1979.
The first John Innes Symposium is held on 6th July 1972. Taking the theme of the ‘Generation of Subcellular Structures’, it covers the assembly of organelles and viruses and includes as topics cell walls, spores, mitochondria, chloroplasts, ribosomes, flagella, bacteriophages and plant viruses. This inaugurates a biannual tradition at the John Innes Institute and the result is an almost unbroken series of biannual Symposia until the 2000s.The second John Innes Symposium, on the ‘Modification of the Information Content of Plant Cells’ (1974), is chosen for its current interest and ‘controversial nature’, and is attended by over 150, including visitors from 19 different countries. This is the first time that the new Conference Hall in the Recreation Centre has been used for a meeting.
A couple of years prior to this event it had been announced in the US that novel hybrid plants had been successfully produced from the fusion of protoplasts of Nicotiana glauca and N. langsdorfii (Carlson et al. 1972). Additional examples presented by visiting speakers at this Symposium confirm that the breakthrough has taken place and somatic hybrids can be obtained by artificial treatment between related and unrelated species. However, despite some reports suggesting the contrary at this meeting (Ledoux et al. 1974), achieving plant transformation by application of heterologous DNA to plant species is not yet possible. Work on possible plant transformation systems (bacterial and viral) is in progress at the John Innes Institute.
R. Markham, D. R. Davies, D. A. Hopwood, and R. W. Horne (eds.), Modification of the Information Content of Plant Cells. Proceedings of the Second John Innes Symposium held in Norwich, July 1974. Amsterdam and Oxford: North Holland Publishing Co., 1974.
P. S. Carlson, H. H. Smith, and R. D. Dearing, ‘Parasexual interspecific plant hybridization’, Proceedings of the National Academy of Sciences, USA, 69 (1972): 2292-2294.
L. Ledoux, R. Huart, M. Mergeay and P. Charles, ‘DNA mediated genetic correction of thiamineless Arabidopsis thaliana’, in Markham et al. (eds.) 1974, pp. 67-89.
Although phages that attack actinomycetes (‘actinophages’) have been known for some considerable time, and were first isolated in 1936, none have previously been isolated for S. coelicolor A3(2), the only genetically well-known streptomycete. As recently as 1970 a report by microbiologist C. A. Collard (in Maurice Welsch’s lab at the University of Liège, Belgium) on a variety of methods to isolate such phages concluded unsuccessfully; this inability has severely hampered opportunities to study interactions between host and phage genomes. Phenomena such as host-controlled modification and restriction that were first recognised in other bacteria by the use of phages are as yet beyond the reach of the Streptomyces group.
This is the problem that John Dowding, PhD student at John Innes, has been studying since 1969. He first has to overcome the difficulties commonly encountered when plating phages isolated from soil samples; by carefully adjusting the growing medium he is able to improve plating efficiencies. By introducing and where necessary adapting classical phage methods to the study of S. coelicolor A3(2) (namely introducing a specific enrichment technique), Dowding successfully isolates a group of 28 phages, including both temperate and virulent examples. He characterises one of the virulent phages, VP11, in a series of experiments that explode the common belief that ‘actinophages’ are somehow different from ‘bacteriophages’; all his experiments indicate VP11 to be a typical bacteriophage.
The demonstration of a workable phage system in S. coelicolor A3 (2) has the potential to make available many new technical approaches to the study of this organism that are in common use in other bacteria, including the ability to transfer DNA between different strains. In particular Hopwood and Dowding hope that further studies of the temperate phages isolated in the lab will eventually yield unambiguous evidence of transduction in a streptomycete, the process in which a bacteriophage picks up bacterial genes from its host and transfers them to a new host, where they are incorporated into its genome by crossing-over. Transduction was first observed and named in Salmonella typhimurium in 1952, but it has eluded Streptomyces researchers so far.
By the end of the 1970s research in other labs will add generalized transduction to the other two modes of gene transfer known in Streptomyces (plasmid-mediated conjugation and artificial transformation) but in the species S. venezulae and S. fradiae, not in S. coelicolor where a workable system of natural transduction remains elusive. However, in the twenty-first century the ease of transduction in S. venezuelae will be an important stimulus for its exploitation as an alternative model streptomycete, along with several other experimentally convenient attributes that are lacking in the classical S. coelicolor.
C. A. Collard, ‘Étude comparative des méthods d’isolement d’actinophages à partir de leurs habitats naturels’, Comptes rendues des séances de la Société de Biologie 164 (1970): 465-468.
J. E Dowding, Studies on bacteriophages in Streptomyces coelicolor. PhD thesis, University of East Anglia, Norwich, 1972.
D. A. Hopwood, K. F. Chater, J. E. Dowding and A. Vivian, ‘Advances in Streptomyces coelicolor Genetics’, Bacteriological Reviews 37, 3 (1973): 371-405.
D. A. Hopwood, ‘Forty years of genetics with Streptomyces: from in vivo through in vitro to in silico’, Microbiology, 145 (1999): 2183-2202.
The John Innes Institute’s staff has grown from 17 in 1967 (Elsden 1979, p. 21) to 154, not including the increasing number of visiting workers (the low initial figure reflects the Institute’s recent move from Bayfordbury, Hertfordshire, with many of the former staff choosing to leave JII rather than re-locate with it. Staff recruitment for Norwich could not proceed until the new Departmental Heads had been appointed). To house staff, a building programme has increased the original office and laboratory accommodation of 16,000 square feet to 91,000 square feet. The new Virus Research wing is built and, despite strikes by electricity and building workers, the new laboratories for Genetics and Applied Genetics are finally completed in 1973. The Recreation Centre and Swimming Pool open in June 1973 having also been delayed by strike action in the autumn of 1972. The swimming pool is an innovation which director Roy Markham has been planning since 1970; he is reputed to have negotiated this with the John Innes Charity on the basis that John Innes was a keen swimmer!
Another innovation to the Institute’s permanent facilities is added by Head of Genetics, David Hopwood: a centralised ‘Media Kitchen’ for the cleaning and sterilisation of glassware and the preparation of agar and other media for growing bacteria. Hopwood had access to a similar facility at Glasgow University before coming to JII and the JII facility is planned by Hopwood and his assistant from Glasgow, Helen Ferguson (later Wright, then Kieser). Though common in major hospitals, it is rare for small groups of microbiologists to have a media kitchen provided, especially one purpose-built according to the designs such as those recommended in Methods in Microbiology (1969). JII’s Media Kitchen is an efficient means of relieving the scientists from routine preparatory tasks and provides an off-the-shelf service, an important contribution to the success of Hopwood’s department and later all the microbiological and plant tissue culture work at JII.
Despite these promising new developments at JII there is considerable uncertainty about the future funding of science, with anticipated financial and fuel crises affecting the UK economy. At the end of October 1974 the ARC announce a ‘stand still’ on new appointments so that posts left vacant through resignations and retirement cannot be filled.
E. C. Elliott and D. L. Georgala, ‘Sources, handling and storage of media and equipment’, pp. 2-20 in J. R. Norris and D. W. Ribbons (eds), Methods in Microbiology, Volume 1, London and New York: Academic Press, 1969.
S. R. Elsden, ‘Professor Roy Markham, F.R.S’, Seventieth Annual Report of the John Innes Institute (1979): 15-23.
A revolution in the technology available to geneticists begins with the experiments of Paul Berg, Herb Boyer, Stanley Cohen and Annie Chang and their colleagues at Stanford University and the University of California at San Francisco. Their research shows how new combinations of genes can be made in the test tube by breaking open the cells of an organism, purifying the DNA and using enzymes to manipulate the DNA. Restriction enzymes recognize a precisely defined sequence and can cut the two strands at offset positions producing ends that are ‘sticky’ and can be readily joined in new combinations, so allowing fragments of the DNA of a donor organism to be inserted into a plasmid as a recombinant DNA (rDNA) molecule that is then introduced into a new bacterial host; the bacterium then replicates the foreign DNA as part of its genetic endowment. In 1980 Paul Berg is awarded the Nobel Prize in Chemistry (shared with Walter Gilbert and Frederick Sanger for their work on DNA sequencing) for his ‘fundamental studies of the biochemistry of nucleic acids, with particular regard to recombinant DNA’. It comes as a surprise to many scientists that Cohen and Boyer are not honoured along with Berg.
Cohen influences the course of Streptomyces genetics at the John Innes Institute when he spends a sabbatical period with David Hopwood learning about Streptomyces (in 1975, see below) and contributes to the work, mainly of Mervyn Bibb, leading to the invention of transformation of Streptomyces protoplasts, which will underpin the proliferation of recombinant DNA techniques in these bacteria.
Within four years after the discovery of rDNA, genetically engineered bacteria are making insulin, somatostatin and growth hormone. An explosion in investment activity creates a new biotechnology industry dedicated to innovation through genetic approaches (Demain 1988).
On Paul Berg’s Nobel prize: http://nobelprize.org/nobel_prizes/chemistry/laureates/1980/
Oral histories for Paul Berg, Herbert Boyer and Stanley Cohen: http://bancroft.berkeley.edu/ROHO/projects/biosci/oh_list.html
A. L. Demain, ‘Contribution of genetics to the production and discovery of microbial pharmaceuticals’, Pure and Applied Chemistry, 60, 6 (1988): 833-836.
Roger Hull, virologist at the John Innes Institute, has recently returned from a sabbatical year at the University of California, Davis where he has been studying Cauliflower mosaic virus (CaMV) in co-operation with R. J. Shepherd in the Department of Plant Pathology. CaMV is of interest because it is one of the very few plant DNA viruses known, a double-stranded DNA molecule (most are RNA viruses). On his return Hull begins to set up the new recombinant DNA technologies, essentially introducing them to John Innes.
‘In the middle of 1974 I heard rumours about some strange new enzymes called restriction endonucleases that could be used to characterize DNA by cutting it at specific sites. I took some CaMV DNA to Herb Boyer’s lab in San Francisco and was introduced to the strange new world of EcoRI, SalI, and BamHI (the three enzymes then available), flat-bed agarose gels, and ethidium bromide. We did the first cuts on CaMV DNA, which in retrospect, were probably the first cuts on any plant-related DNA’.
Hull and his colleagues at John Innes have to build their own equipment and extract their own enzymes before they can begin to employ rDNA technology. The CaMV work initially focuses on the physical structure of the genome and transcription from it using the new gel techniques, experiments which are first reported in 1977. By 1978 CaMV is being promoted both as one of the most promising candidates as a vector DNA in the genetic manipulation of plants, and as an alternate model system to be compared with the by now well-known animal viruses whose genomes are of the same size, such as Simian virus 40 (SV40). Ultimately CaMV is superseded as a system for getting the genes into plants (vector DNA) but a region of the CaMV sequence comes to play a key role as a promoter - that is, helping to get transferred genes expressed in GM plants (a ‘switch’ to turn the genes on) - that is used in nearly all GM crops. Simon Covey, George Lomonossoff, and Roger Hull at JII play a prominent role in the discovery of the ‘35S promoter’ in the early 1980s.
Roger Hull, ‘The Phenotypic Expression of a Genotype: Bringing Muddy Boots and Micropipettes together’, Annual Review of Phytopathology, 46 (2008): 1-11.
R. Hull, S. H. Howell and D. Aldous, ‘Mapping the genome of cauliflower mosaic virus’, Sixty-Eighth Annual Report of the John Innes Institute, (1977).
R. Hull and R. J. Shepherd, ‘The structure of cauliflower mosaic virus genome’, Virology, 79 (1977): 216-230.
R. Hull, ‘The possible use of plant viral DNA’s in genetic manipulation of plants’, Trends in Biochemical Sciences, 3, 4 (1978): 254-56.
S. N. Covey, G. P. Lomonossoff, and R. Hull, ‘Characterisation of cauliflower mosaic virus DNA sequences which encode major polyadenylated transcripts’, Nucleic Acids Research, 9 (1981): 6735-6746.
The UK is leading the European scientific community in the early phase of recombinant DNA regulation, being the first EU country to introduce a temporary moratorium on certain kinds of rDNA experiments in 1974 after the publication in the US of letters to Science in 1973-4 calling for such a move. As in the US, consensus is initially built around the adoption of flexible, voluntary guidelines (Dunlop, 2000). The conduct of GM research is considered by two different working groups in government: the Ashby Working Party (1975) and the Williams Working Party (1976), of which David Hopwood was a member. Both groups recommend the setting up of a central advisory service on GM to provide guidance to laboratories (Vickers, 1978). Acceptance of the recommendations leads to the setting up of a central advisory service in 1975-6, the Genetic Manipulation Advisory Group (GMAG), by the Medical Research Council to look at work involving the construction of genetically modified organisms. GMAG establishes rules for ranking the level of risk and assigning GM research and products to different levels of containment. Initially advice is given to laboratories on a voluntary basis, but Health and Safety regulations which come into force in 1978, make it a legal requirement to set up local safety committees and for scientists to notify GMAG of proposed rDNA experiments.
GMAG was shaped by a conference on 11-13 July 1975 in Oxford jointly organised by the Medical Research Council, Agricultural Research Council and the Science Research Council on Recombinant DNAs. The purpose of the meeting was to bring together workers broadly representative of research in DNA recombination in the UK, to discuss the present state of the art and plans for the future. Another aim was to identify specific problems, such as the need for programmes of training in the safe handling of pathogens, and for ensuring the ready availability of enzymes and safe vectors. Over 120 participants (including representatives from the John Innes Institute) were invited across medical and agricultural research institutes and University bioscience departments in the UK. Their discussions were to provide guidance to the major funding bodies on how this area could be effectively supported and encouraged within constraints considered to be appropriate to reduce and contain potential hazards.
Also in 1975 the European Molecular Biology Organisation or EMBO (founded in 1964), forms a Standing Advisory Committee on Recombinant DNA (de Chadarevian, 2002; Strasser 2003). Prompted by a letter from 12 European virologists concerned about the possibility of restrictions being placed on rDNA technology, the EMBO Council sends delegates to the Asilomar Conference (see below) and then sets up its advisory committee to discuss the implications of the guidelines laid down by National Institutes of Health in the US for rDNA work in Europe. The outcome, a joint EMBO-NIH workshop to ‘assess the risks of recombinant DNA experiments involving the genomes of animal, plant and insect viruses’, has been described as a ‘turning point’ in the history of rDNA regulation (de Chadarevian, 2002, p. 334) . Roger Hull, John Innes virologist, attended the British workshop meeting, held in a hotel in Ascot near London in January 1978. The EMB0-NIH workshop report forms the basis for an international regulatory structure for GM crops.
The Health and Safety (Genetic Manipulation) Regulations 1978 [Statutory Instrument 1978 No. 752] Section 5; this SI came into force in August 1978.
T. Vickers, ‘Flexible DNA regulation: the British model’, Bulletin of the Atomic Scientists, 34, 1 (January 1978): 4-5.
‘Genetic manipulation: new guidelines for UK’, Nature, 276 (9 November 1978): 104-108 http://www.nature.com/nature/journal/v276/n5684/abs/276104a0.html
Sir G. Wolstenholme, ‘Public confidence in scientific research: the British response to genetic engineering’, Technology in Society, 6, 1 (1984): 9-16. (Wolstenholme was the first Chairman of GMAG).
National Institutes of Health (US), US-EMBO Workshop to assess risks for recombinant DNA experiments involving the genomes of animal, plant, and insect viruses, [Report] Washington, US: US Government Print Office.
S. de Chadarevian, Designs for Life: Molecular Biology after World War II, Cambridge: Cambridge University Press, 2002.
B. Strasser, ‘The transformation of the biological sciences in post-war Europe’, EMBO report, 4, 6 (June 2003): 540-543.
Roger Hull, ‘The Phenotypic Expression of a Genotype: Bringing Muddy Boots and Micropipettes together’, Annual Review of Phytopathology, 46 (2008): 1-11.
Claire Dunlop, 'GMOs and Regulatory Styles', Environmental Politics, 9, 2, 2000, pp. 149-155.
David Hopwood had been in touch with Stanley Cohen of Stanford University ever since he had heard Cohen speak about his revolutionary studies on recombinant DNA at a meeting in New Orleans in January 1974 and Cohen had been excited by the differences in the behaviour of Streptomyces plasmids compared with the paradigm provided by the plasmids of Gram-negative bacteria such as Escherichia coli. Cohen spends a sabbatical period in the Genetics Department from May to November 1975. Although attempts to introduce DNA into Streptomycetes, to open the way to genetic engineering in them, had not yet succeeded, Cohen’s visit did much to introduce the department – and the John Innes Institute more generally – to the technical and ethical issues surrounding the new science. In one successful experiment the SCP2 plasmid was found to have one cut-site for the restriction enzyme EcoRI and two for HindIII. This augured well for the later development of the plasmid into a cloning vector for Streptomyces because an ideal vector has single or very small numbers of cut-sites for a given enzyme, allowing the vector to be converted easily to a full-length linear form with sticky ends into which foreign DNA can be inserted.
Cohen later appoints Mervyn Bibb from JII to establish Streptomyces genetics and cloning in his lab at Stanford, and goes on to maintain a significant contribution to research on Streptomyces for more than thirty years.
Stanley N. Cohen, M.D., “Science, Biotechnology, and Recombinant
DNA: A Personal History,” an oral history conducted by Sally Smith
Hughes in 1995, Regional Oral History Office, The Bancroft
Library, University of California, Berkeley, 2009
The safety and ethics of the new tools of molecular biology that allow the cutting and splicing of DNA together from different species is a subject causing concern among US scientists. The National Academy of Sciences responds by asking Paul Berg, a leading biochemist in the field of recombinant DNA technology at Stanford University, to chair a committee in April 1974 to consider the potential hazards. An international conference is subsequently convened in February 1975, one of the Berg committee’s key recommendations.
On the conference organizing committee with Berg are David Baltimore, Sydney Brenner, Richard O. Roblin III, and Maxine F. Singer. Around 140 professionals (mostly biologists, but also a few physicians, lawyers and journalists) gather at the Asilomar Conference Center in Pacific Grove, California, USA to discuss voluntary guidelines to secure safe practice in laboratories manipulating DNA from different species. This landmark ‘International Conference on Recombinant DNA Molecules’ becomes known simply as ‘Asilomar’.
Despite its acknowledged potential for research in molecular biology, recombinant DNA research has been put on hold in the US while safety concerns over the potential risks are addressed. This followed letters to the journal Science in 1973-4 calling for a temporary moratorium on rDNA research, a call that was endorsed by the Berg committee and the National Institutes of Health (the main funding agency for rDNA research in the US). The outcome of Asilomar is the resumption of recombinant research but under very cautious safeguards which are laid out in the first set of National Institutes of Health ‘Guidelines for Research Involving Recombinant DNA Molecules’ (1976).The main principles adopted include making containment of modified organisms an essential consideration in experimental design.
Hailed by many as a landmark of self-regulation by scientists, and the first example of what came to be known as the ‘precautionary principle’, the legacy of Asilomar remains controversial three decades later. It establishes mechanisms for decision-making about rDNA technologies in the US, and heavily influences guidelines adopted by other countries, but some critics argue that the guidelines adopted were too cautious and perhaps alarmist, while others critique Asilomar for marginalising social, political and ethical questions.
P. Berg, D. Baltimore, S. Brenner, R. O Roblin III, and M. F. Singer, ‘Summary Statement of the Asilomar Conference on Recombinant DNA Molecules’, Proc. National Academy of Sciences, 72, 6 (1975): 1981-1984.
Judith A. Johnson, ‘The NIH recombinant DNA Guidelines’, Library of Congress, Congressional Research Service (1982).
Susan Wright, Molecular Politics: Developing American and British Regulatory Policy for Genetic Engineering, 1972-1982. Chicago: University of Chicago Press, 1994.
Claire Dunlop, 'GMOs and Regulatory Styles', Environmental Politics, 9, 2, 2000, pp. 149-155.
Paul Berg, ‘Asilomar and Recombinant DNA’ (2004)
Adam Briggle, ‘Asilomar Conference’, in Encyclopedia of Science, Technology and Ethics, ed. Carl Mitcham, 4 vols. Farmington Hills, MI: Macmillan Reference (2005): Vol. 1, pp. 118-121
For photographs, archives, and oral histories:
The only evidence for the existence of the plasmid SCP1, first published in 1971, is still genetic despite all the effort expended in David Hopwood’s lab trying to find physical evidence of it and to estimate its molecular weight. Although so far without successful conclusion, the search has begun to reveal the existence of a second sex factor.
The first clue is the identification by Hildgund Shrempf et al. (1975) of covalently closed circular (ccc) DNA in strains of Streptomyces coelicolor of known fertility types; their experiments also show that this biochemically isolated plasmid is not SCP1. More evidence is provided by Mervyn Bibb, a UEA graduate and now PhD student at JII. Using Hopwood’s plate-crossing technique, Bibb’s genetic experiments show that there is a second fertility determinant in S. coelicolor that exists as an autonomous plasmid. Further experiments confirm that the ccc DNA is the second plasmid (SCP2). Unlike SCP1, this second plasmid is a conventional circle of DNA that can be isolated and visualized in the electron microscope by standard methods.
Although extra-chromosomal DNA elements, which are either plasmids or defective phage genomes, have been isolated from a wide variety of bacterial species, this is the first supercoiled DNA to be identified in streptomycetes. Its discovery and physical characterisation is an important step in understanding sex in Streptomyces and is a prerequisite for developing gene cloning. The discovery of SCP2 was an essential stepping stone towards introducing plasmid DNA into Streptomyces hosts by transformation, thereby opening the way for gene cloning in these organisms.
In another breakthrough in 1975, marking the start of JII’s involvement in the genetics of antibiotic synthesis, PhD students Ralph Kirby and L. F. (Fred) Wright show in their experiments with SCP1 that both methylenomycin antibiotic production and resistance in S. coelicolor are determined by genes borne on the plasmid. The methylenomycin gene cluster continues to be a source of novelty right up to the present day, being key to the development of the mutational cloning method (see Chater and Bruton, 1983), as well as proving to be regulated by a novel kind of furan autoinducer molecule (a kind of signalling molecule).
H. Schrempf, H. Bujard, D. A. Hopwood, and W.Goebel, ‘Isolation of covalently closed circular deoxyribonucleic acid from Streptomyces coelicolor
A3(2)’, Journal of Bacteriology, 121(1975):416-421.
M. J. Bibb, R. F. Freeman, and D. A. Hopwood, ‘Physical and genetical characterisation of a second sex factor, SCP2, for Streptomyces coelicolor A3 (2)’, Molecular and General Genetics, 154, 2 (1977): 155-166.
K. F. Chater and C. J. Bruton, ‘Mutational cloning in Streptomyces and the isolation of antibiotic production genes’, Gene, 26 (1983): 67-78.
R. Kirby, L. F. Wright and D. A. Hopwood, ‘Plasmid-determined antibiotic synthesis and resistance in Streptomyces coelicolor’, Nature, 254 (20 March 1975): 265-267.
Newfound Farm, a 56-hectare site close to the John Innes Institute on Colney Lane, is acquired to provide the land necessary for pea trials and the associated crop rotation. Farm buildings designed for the pea programme are built and equipped for vining and dried pea trials.
One of the new lines of research in Applied Genetics at JII is on mutant peas carrying the gene af, first reported in Finland in 1953, which converts pea leaflets into tendrils and produces what have become known in agriculture as ‘semi-leafless’ forms. Following observations of these mutant forms in small plots, Brian Snoad begins to investigate the commercial potential of this new type of plant which is so unlike those normally grown. A survey he had carried out in 1969 among pea growers, other farmers, canners and freezers had revealed that harvesting difficulties caused by the lodging of crops are the key problems that the pea industry wants solving. The improved standing ability of the af mutants suggests to Snoad that modifying the architecture of pea plants might be a way to aid the drying and therefore the machine-harvesting of commercial crops.
The usefulness of a semi-leafless pea is not immediately apparent to producers, and there is some scepticism about their commercial value: one author in Grower quips that ‘when someone finds a market for tinned tendrils we may be able to understand all the ooh-aah over the John Innes Genotype peas’. Undaunted, a ‘pre-breeding’ programme is started at John Innes, in which the af gene begins to be incorporated into pea varieties. This is confined to pre-breeding because at this time it is Agricultural Research Council policy to leave breeding itself (which requires scaled-up field trials) to commercial companies. However, Snoad’s pre-breeding work requires land other than that available at John Innes’s Stanfield Farm and here he is helped by the Processors and Growers Research Organisation near Peterborough, a research and advisory body founded by pea producers and buyers. Not only do they provide land and facilities but, even more importantly, they encourage the development of mutant peas and give more guidance and instruction about the practical aspects of farming than is available at JII. Their resources also enable JII peas to be professionally trialled at a variety of locations in the UK from 1974-76 (to supplement the trials at Newfound Farm).
Snoad and his team of assistants (Margaret Cobb, Judith Harnden, Ann Payne, Sharon Negus, Denise Caston, Peter Watkins, Paddy Dow and Kim Graves) at JII develop a wide range of entirely novel forms of pea plant, including the most extreme type, with all its leaflets converted into tendrils (af) and its stipules reduced in size and altered in shape (st), becoming known in the UK as the ‘leafless pea’. The unexpected discovery that some of these mutant pea crops are capable of giving yields of dry seed at the same level as many of the existing crops of leafed peas but with the advantage of being easier to harvest with a combine harvester leads, in 1975, to the decision to market a small number of purely experimental lines. It is hoped that this move will improve awareness and activity among UK commercial breeders and encourage them to increase the use of lines such as these as parents.
Between 1976 and 1977 six John Innes lines are launched through the National Seed Development Organisation: a leafless type (Filby), and five semi-leafless types. Filby and the other lines are granted Plant Breeders’ Rights in 1977 and are accepted for the National List of vegetable varieties in 1978. In the late 1970s the ARC makes a radical change of approach and finances a full pea breeding programme using the JII stocks. It is decided that the JII breeding will focus on dried peas (suitable for canning, packeting, milling and for animal feed) to redress their relative neglect compared with the more widely-researched vining peas, and to fit in with the strategic need to develop home-produced protein to reduce UK dependency on imported American soya. In order to speed up the development of lines, co-operation is established with the Department of Scientific and Industrial Research at Lincoln in New Zealand, giving two seed multiplications each year.
Subsequent research work around the world will prove these new pea plant models, especially the semi-leafless form, to be particularly advantageous (for standing ability, machine-harvesting, water-efficiency and yield), and in many countries, as the result of extensive breeding, they come to represent the majority of new varieties being registered and grown. In the UK, for example, semi-leafless accounts for 100% of UK dried pea varieties in 2009.
V. Kujala, ‘Felderbse, bei welcher die ganze Blattspreite in Ranken umgewandelt ist’, Arch. Soc. Zool. Bot. Fen., 8 (1953): 44-45.
Anon., ‘Extension of PGRO’, Grower (Vegetable Section commentary), 12th August [1974?]. Press cutting in John Innes Centre archives.
Anon., ‘Agriculture: Pea-breeding progress’, The Times, 21st July 1975.
G. P. Gent, ‘Peas for the 80s’, Field Vegetable Quarterly, June 1978.
B. Snoad, ‘The origin, performance and breeding of leafless peas’, ADAS Quarterly Review, 37 (1980): 69-86.
B. Snoad, ‘Taking the leaf out of peas’, Supplement to GROWER (March 19th 1981).
In July, David Hopwood, Helen Wright, and Mervyn Bibb at JII, together with Stanley N. Cohen who was on sabbatical from the Department of Medicine, Stanford University School of Medicine, California in 1975, publish a letter in the premier science journal Nature announcing that they have a simple and generally applicable procedure to recombine actinomycete strains at high frequency. This is important news because it promises to facilitate the routine use of recombination in strain improvement, which until this development had only been used to a very limited extent. Because members of the Streptomyces genus already yield over 60% of known antibiotics, the new procedure has important implications for industrial microbiology and medicine.
The method devised in Hopwood’s lab depends on polyethylene glycol (PEG)-induced protoplast fusion and regeneration. A protoplast, in this case, is a bacterial cell that has had its cell wall removed by enzymic treatment, and protoplast fusion involves the formation of hybrid cells that can grow into a mature hybrid bacterium. Protoplast fusion as a new method of recombining bacterial genes (using Bacillus cells) was first reported in 1976 by Katalin Fodor and Lajos Alföldi at the Hungarian Academy of Sciences Institute of Biological Sciences in Szeged, and Pierre Schaeffer, Brigitte Cami, and Rollin Hotchkiss at the University of Paris in Orsay. Hopwood’s team are also able to build on the experience of Masanori Okanishi and his colleagues at the National Institute of Health in Tokyo who discovered how to prepare protoplasts in several Streptomyces species a few years before. Especially important is Okanishi’s group’s study of the components required in a special medium to allow protoplasts to re-synthesize the cell wall and become mycelium again, methods that prove successful for S. coelicolor.
Helen Wright, in Hopwood’s lab, is the first to take the important next step of trying to fuse Streptomyces protoplasts. Although it would have been natural to begin by applying the protocol devised for Bacillus, Helen instead uses a technique that Guido Pontecorvo found to work for animal cells at the Imperial Cancer Research Fund laboratories in London, borrowed from botanical literature where he had read about the use of PEG to fuse plant protoplasts. Pontecorvo, now on the governing council of JII, passed on the method he and Anne Hales had perfected, before it was published, on a short visit to Hopwood’s lab in October 1976. Helen successfully uses it for S. coelicolor and finds that protoplast fusion yields an amazing number of recombinants. Hopwood’s team hope that the high recombination frequencies achieved by protoplast fusion will make it easy to create new combinations of genes affecting antibiotic production, as a tool for breeding superior strains. However, although the technique certainly comes to be used extensively for strain improvement, the commercial interests of pharmaceutical companies will prevent them discovering just how widely their work on protoplast fusion is applied.
R. H Baltz, ‘Genetic recombination in Streptomyces fradiae by protoplast fusion and regeneration’, Journal of General Microbiology, 107 (1978): 93-102. (In parallel with JII work Baltz independently developed protoplast fusion at the US pharmaceutical company Eli Lilly).
D. A. Hopwood, H.M. Wright, M. J. Bibb and S. N. Cohen, ‘Genetic recombination through protoplast fusion in Streptomyces’, Nature 268 (14 July 1977): 171-174.
David A. Hopwood, Streptomyces in Nature and Medicine: The antibiotic makers, Oxford: Oxford University Press, 2007, pp.86-90.
Genetic manipulation is identified as a high priority area by the Agricultural Research Council’s Priority Working Party in 1977. Additional funds obtained from the Department of Education and Science enable the programme to be formally initiated in 1978 as the ‘ARS programme on genetic manipulation’, catalysed by nine new appointments, spread over three years. (The Agricultural Research Service (ARS) was made up of the institutes and units grant-aided by the ARC and the, then, Department of Agriculture for Scotland (DAFS), plus those wholly owned by the ARC). The main emphasis of the programme is ‘to establish the technology necessary for the genetic manipulation of plants, which requires the production of vector systems for the transformation of plant cells’. However, the programme is pursued on a broad front, from the production of somatic hybrids of plants (hybridisation by protoplast fusion) to the genetic manipulation of bacteria, so that GM technology may be exploited wherever relevant to the agricultural and food industries.
The programme is built on existing expertise within the ARS with groups at the Plant Breeding Institute, Cambridge, the John Innes Institute, Rothamsted Experimental Station, Harpenden, the Welsh Plant Breeding Station, Aberystwyth, and also at the University of Nottingham; parts of the work at the Unit of Nitrogen Fixation, University of Sussex, are also included. Altogether about 50 staff are involved in the programme, which is planned as a whole and co-ordinated centrally. JII’s staff expands under the scheme.
JII is responsible for the major bacterial interests of the ARS programme, in Streptomyces and Rhizobium, together with the possible plant transformation systems Agrobacterium tumefaciens, DNA viruses, and endogenous flax sequences. To achieve these aims molecular cloning methods are being developed to transfer genes between unrelated streptomycetes. John Innes research on the possible vehicles for the transformation of plant cells is focussing on the tumour-inducing (Ti) plasmid of Agrobacterium tumefaciens and Cauliflower Mosaic Virus (CaMV) DNA.
As noted (see 1970-74, above) Agrobacterium work at John Innes makes little progress in the face of a strong lead at the Plant Breeding Institute in Cambridge, but genetic engineering in Streptomyces develops very successfully. The objectiveis to develop a cloning system to enable rDNA to facilitate biological studies and develop industrial strains with increased antibiotic yield or capable of yielding new antibiotics. Work is already in progress with a grant from the National Research Development Corporation (‘NRDC’- a body established by the Government in 1948 to transfer technology from the public sector to the private sector) which funds Mervyn Bibb to develop a system for introducing DNA into Streptomyces. When Bibb leaves in November 1978 to take up a post-doctoral position with Stanley Cohen at Stanford, United States he is replaced by Charles Thomson from the University of Wisconsin, Madison. Thompson works in parallel (in competition even) with Bibb to be the first to successfully clone genes in Streptomyces using plasmids.
The entry of JII into this wide field now known as ‘genetic engineering’ means that containment facilities have to be provided on site.
R. Markham, ‘Director’s Report’, Sixty-Eighth Annual Report of the John Innes Institute (1977): 15.
Report of the Agricultural Research Council, 1979-80, London: HMSO, 1980, pp. 7-13.
M. J. Bibb, J. L. Schottel and S. N. Cohen, ‘A DNA cloning system for interspecies gene transfer in antibiotic-producing Streptomyces’, Nature, 284 (1980): 526–531.
C. J. Thompson, Ward, J. M. and D. A. Hopwood, ‘DNA cloning in Streptomyces: resistance genes from antibiotic-producing species’, Nature, 286 (1980): 525-527.
Gavin Brown, former Curator of the Gardens at JII, is awarded the Veitch Memorial Gold Medal of the Royal Horticultural Society for his services to horticulture, particularly for his work on breeding fruit and flowering plants. The Veitch Medal is awarded to "persons of any nationality who have made an outstanding contribution to the advancement and improvement of the science and practice of horticulture." The award follows Brown’s earlier notable honours: the Royal Horticultural Society’s Jones-Bateman Cup in 1960, and an MBE in 1971.
(Andrew) Gavin Brown originally came to the John Innes Horticultural Institution as a student gardener in 1930-32. He returned in 1935, on a Fruiterers’ Company Bursary, and for many years was a technical assistant in the Pomology Department. His main research was from 1936-1947 the time and period of blossoming in apples and pears; and from 1948-51 incompatibility, breeding and selection in various fruits, haricot beans and tomatoes (with Morley Benjamin Crane). After Crane’s retirement in 1953 Gavin led the top fruit breeding work, especially the breeding of apples for mildew and scab resistance. However, during his time at Colney (1967-75) his main interest was in flower breeding. He represented JII on the National Fruit Trials Apple and Pear sub-committee, and other fruit-related and horticultural bodies. In 1967 he became Acting Head of Applied Genetics and Curator of the Gardens; he retired in 1975. Gavin was one of the few people to work at all three sites of the John Innes: Merton, Bayfordbury and Norwich.
Professor Roy Markham dies after a long illness on November 16th. Professor D. Roy Davies is appointed Acting Director. Markham’s key legacies include the re-building and expansion of the John Innes Institute on the Colney site (effectively retaining its independence as a research institute), the creation of a new Department of Virus Studies with staff transferred from Cambridge University, and (from staff re-organisation and new recruitment) the founding of an Ultrastructural Studies Department equipped as an ARC centre of excellence in electron microscopy. Markham’s vision was of an institute whose future research would be interdepartmental and interdisciplinary, and this was reflected in the arrangement of the JII Annual Reports which abandoned the old departmental reporting structure. He aimed to create an institute with a relatively small permanent staff but state-of-the-art facilities, which would be a haven for visiting scientists to carry out collaborative research.
Markham had left his own distinguished career in virus research to concentrate on administration, his aim to create an environment at JII where plant and microbial science could develop and flourish, but his interest in technology (he listed himself under the ‘Electronics Section’ of the Annual Reports) led him also to make significant contributions to ultrastructural studies, including the first computer-aided cell image analysis at JII. This work included a significant contribution to virus structure using optical diffraction and image reconstruction with crystalline arrays of viruses.
S. R. Elsden, ‘Professor Roy Markham, F.R.S’, Seventieth Annual Report of the John Innes Institute (1979): 15-23.
R. E. F. Matthews, ‘Roy Markham: pioneer in plant pathology’, Annual Review of Phytopathology, 27 (1989): 13-22.
Mervyn Bibb, Judith Ward and David Hopwood report the development of a plasmid transformation system for Streptomyces which they anticipate will allow the cloning of any DNA sequence into these organisms and which, potentially, provides a means of directly manipulating the pathways of antibiotic production. The system involves the uptake of covalently closed circular (ccc) DNA by protoplasts in the presence of polyethylene glycol (PEG) and the visual detection of transformants, at high resolution, after regeneration of the protoplasts. This deploys Bibb’s powerful method of finding plasmid-carrying cells by looking for ‘pock’ formation when the spores are added to a confluent mass of plasmid-free cells. The introduction of DNA into Streptomyces coelicolor by plasmid transformation (using SCP2) in Hopwood’s lab is a crucial step in opening up Streptomyces to genetic manipulation and ensures that S. coelicolor retains its leading role as a model organism, even as the terrain shifts from formal genetics to genetic engineering. In parallel, it proves possible to introduce bacteriophage DNA into protoplasts by PEG-assisted transfection (Suarez and Chater, 1980), which will allow the collaborative investigation of bacteriophage phiC31 genetics in Norwich and Moscow (Lomovskaya et al., 1980), resulting in the development of valuable phage-based cloning systems over the next few years.
Thus, the 1970s see the genetics of industrial microorganisms beginning to shift from its original focus, the production of chromosome maps and biosynthetic pathways of secondary metabolites, to a science of plasmid and virus vectors for exporting and re-importing genes from various sources. The GM techniques that Hopwood’s lab pioneers provide the impetus for antibiotics companies to begin working on the fundamental science of Streptomyces. The discovery by Brian Rudd and David Hopwood that genes for antibiotic production are clustered also greatly enhances the prospects of their successful manipulation (making it much easier to isolate and manipulate the gene set). A major outcome is that Hopwood’s lab becomes hugely attractive to scientists in the industry who are both keen to spend time on sabbatical in Norwich and to translate JII’s research into improved antibiotic production. Hitherto industrial practice has been to use mutagenesis (applied at random) to come up with higher producing strains of Streptomyces. Hopwood’s lab offers another way through genetic manipulation.
Important in facilitating JII’s interactions with industry is the Streptomyces Club founded by Hopwood in 1968: ‘many companies contributed financial support, at various levels and for varying periods of time, as Club members. There was no exclusivity, and no rights were ever transferred to a company. They were all entitled to receive “available” strains, plasmids and phages (in practice almost every one of the group’s stocks with an occasional exception for IP reasons), and to use them for their own commercial purposes. They could also ask for reasonable amounts of advice from members of the JII group and sometimes they made occasional visits to JII to help solve specific problems’ (Hopwood 2009). The Streptomyces Club proves of enormous benefit to JII as for many years all the PhD students (and many post-docs) in the Streptomyces group are funded from this source, leaving studentships from the Agricultural Research Council and the John Innes Trustees for the other two major groups in the Genetics Department working on Rhizobium and Antirrhinum. Perhaps one of the most far-reaching appointments, in 1979, using Club funds is that of Tobias Kieser who had obtained his PhD at the Swiss Federal Institute of Technology (ETH) in Zürich. He had devised imaginative methods for studying Streptomyces plasmids and his expertise is to be invaluable for the future development of these and other studies at John Innes. Kieser stays until leaving to become a school teacher in 2005.
C. Ball (ed.), Genetics and Breeding of Industrial Microorganisms. Florida, USA: CRC Press, 1984.
M. J. Bibb, J. M. Ward, and D. A. Hopwood, ‘Transformation of plasmid DNA into Streptomyces at high frequency’, Nature, 274 (1979): 398-400.
D. A. Hopwood, ‘Industrial interactions between the JIC Streptomyces group and commercial companies, 1968-1998’, 2009. Typescript available in JIC Archives.
David A. Hopwood, Streptomyces in Nature and Medicine: The antibiotic makers, Oxford: Oxford University Press, 2007, pp.90-95.
N. D. Lomovskaya, K. F. Chater, N. M. Mkrtumian. ‘Genetics and molecular biology of Streptomyces bacteriophages’, Microbiological Reviews, 44 (1980): 206-229.
B. A. M. Rudd and D. A. Hopwood, ‘Genetics of actinorhodin biosynthesis by Streptomyces coelicolor A3 (2)’, Journal of General Microbiology, 114 (1979): 35-43.
J. E. Suarez, K. F. Chater, DNA cloning in Streptomyces: a bifunctional replicon comprising pBR322 inserted into a Streptomyces phage, Nature, 286 (1980): 527-529.
Professor David Hopwood is elected a Fellow of the Royal Society in recognition of his pioneering genetic studies of Streptomycetes, the soil bacteria that produce the great majority of medically- and agriculturally-important antibiotics. When Hopwood started his work in 1954, Streptomycetes were usually regarded as a group of microbes ‘intermediate’ between bacteria and fungi. Hopwood established their true phylogenetic relationships - that Streptomycetes are bacteria (a bacterium or ‘prokaryote’ lacks a nuclear membrane) that have evolved a filamentous life-style independently of the fungi - and went on to pioneer the mapping of Streptomyces coelicolor genes. Interestingly, all of the achievements acknowledged in Hopwood’s FRS election certificate (see below) are based on in vivo genetics and precede the later advances in Hopwood’s group towards the recombinant DNA revolution.
The certificate reads: ‘…he discovered genetic recombination of Streptomyces and developed original systems of genetic mapping which led him to the demonstration of a circular linkage group. This mapping work was important both in strengthening the generalization that the prokaryotes in general have circular chromosomes and in showing a tendency towards symmetry in the map suggestive of evolution by genome doubling. His electron microscope studies (with Audrey Glauert) showed beyond doubt the prokaryotic affinities of Streptomyces and demonstrated for the first time the existence of membranous "organelles" in continuity with the plasma membrane. He has shown that the fertility system of Streptomyces coelicolor involves a sex factor associated with a plasmid. In the course of these studies he has discovered the first clear example of a plasmid-encoded antibiotic synthesis. Hopwood and his group have also extended the genetic analysis to other species of Streptomyces and Nocardia and demonstrated efficient DNA-mediated transformation of Thermoactinomyces. Current studies are directed towards the genetic analysis of development in S. coelicolor. While Giuseppe Sermonti made some of the basic observations on recombination independently, Hopwood has been the prime mover in most of the advances. He now has an established international reputation as the leading pioneer and authority in what has become a very important aspect of microbial genetics’.
David A. Hopwood, ‘Forty years of genetics with Streptomyces: from in vivo through in vitro to in silico’, Microbiology, 145 (1999): 2183-2202.
David A. Hopwood, Streptomyces in Nature and Medicine: The antibiotic makers, Oxford: Oxford University Press, 2007.