Nature of the resource

Although culture collections of filamentous fungi date back to the late nineteenth century and one of the largest of them, the Centraalbureau voor Schimmelcultures (CBS) in The Netherlands was established in 1904, rather little interest had been shown in their funding, and proposals to set up such collections often received only token support. With the advent of biotechnology the search for microorganisms which have properties suitable for commercial exploitation has led to a renewed interest in culture collections because of the appreciation of the valuable resource they represent.

The term ‘filamentous fungi’ is used for species producing filamentlike hyphae. It includes almost the entire fungal kingdom and is used in contradistinction to ‘yeasts’, which are essentially unicellular fungi with vegetative cells capable of repeated budding. The yeast fungi are not considered further here as they are treated in a companion volume in this series. While some fungus culture collections maintain both filamentous fungi and yeasts, most specialise in one growth form or the other. This is a consequence of the different uses made of them (and so of their relevance to particular industries), the diverse growth conditions, and the dissimilar ways in which they are currently characterised (physiologically and biochemically in the yeasts, but largely morphologically in all other fungi).

Precisely how many strains of filamentous fungi are maintained in the living state in culture collections throughout the world is unknown. However, the number certainly exceeds 170 000 scattered through over 200 collections, and it has been estimated that around 7000 different species are represented (Hawksworth, 1985a).

Introduction

This chapter is intended to give the reader who is unfamiliar with patents an introduction to the patent system as it applies to biotechnology, and a general guide to the procedures and pitfalls involved in obtaining patent protection for biotechnological inventions. For a detailed discussion of the whole subject of patents in biotechnology and a review of the variety of national patent systems the reader is referred to the excellent texts by Crespi (1982), Beier, Crespi & Straus (1985) and Straus (1985). It is not possible here to provide a step-by-step guide to getting a patent in every country in the world, for, despite an overall similarity, variations between different national patent laws are manifold, and professional help is necessary to guide even the experienced inventor through their complexities. The present account does no more than skim the surface of what is a complex and often fascinating subject; for this reason a short list of selected publications which illustrate in more detail many of the points raised here is given in Section 6.6, Further reading.

Basis of the patent system

Principles

The principle (if not the practice) of the patent system is straightforward: the inventor of a new product or process publicly discloses the details of his invention and in return he is granted for a limited period a legally enforceable right to exclude others from exploiting it. In this way the inventor's ingenuity is acknowledged and rewarded, while at the same time further technical progress is encouraged by the public dissemination of information about the invention.

Supply of cultures

Terms and conditions

Service collections supply cultures from their collections with no terms or conditions of use attached. The purchaser is therefore free to use the culture in any way he wishes. These ‘open’ collections comprise cultures listed in catalogues that are available for sale to the public at stated prices.

However, not all cultures held in collections are available without restriction. In addition to patent strains, where special conditions apply (Chapter 6), or safe-deposit strains held on a confidential basis for a depositor (Chapter 7), strains isolated by collection staff or on which research has been carried out may also be held in ‘reserve’. Such ‘reserve’ strains, which may be of potential industrial importance, are not included in catalogues nor made generally available to the public, but may be supplied on special terms on a case-by-case basis. They are usually supplied exclusively to a single industrial purchaser, together with data on their attributes. In addition to an initial payment for the exclusive rights, involvement in the further development of the strain and access to data derived, a small royalty on any resultant income may be stipulated. Some culture collections that have used their own resources to discover new biochemical or microbial activities of strains, feel it is appropriate that they receive some return should these strains prove to be commercially rewarding.

Introduction

It is essential that the most appropriate growth conditions and preservation techniques are used to ensure the viability, purity and stability of maintained microorganisms. Ideally, the methods used should retain all characteristics throughout storage.

Many filamentous fungi grow on culture media and can be kept viable by periodic transfer. However, the properties of fungi in culture may be unstable through loss of plasmids, spontaneous mutations or genetic recombination (due to the presence of heterokaryons, the parasexual cycle or normal sexual events). These phenomena can result in modification of a strain's characteristics, and conditions of preservation and storage should be selected to minimise the risk of such changes.

Growth of cultures

The major factors affecting growth are medium, temperature, light, aeration, pH and water activity.

Media

The growth requirements for fungi may vary from strain to strain, although cultures of the same species and genera usually grow best on similar media. The source of isolates can give an indication of suitable growth conditions, thus isolates from jam can be expected to grow well on high-sugar media, species from leaves may sporulate best in light, those from marine situations may require salt and those from deserts and the tropics, high growth temperatures.

Cultures are usually best grown on agar slopes in test-tubes or culture bottles. A list of recommended media and growth temperatures for common species is given in Smith & Onions (1983). Details of the most frequently used media are given in the Appendix at the end of this book.

The filamentous fungi represent the group of microorganisms with the largest number of species, showing an immense variety not only in morphology but also in physiological and biochemical attributes. About 63 700 species (excluding yeasts) are currently known, but around 1500 are described as new to science each year, and the number in nature may well exceed 250 000. Over 3000 secondary metabolites have already been characterised, but actual numbers are far in excess of this and the biological activities of most remain to be determined. The biotechnological importance of the filamentous fungi is, therefore, of considerable significance, and the potential of this vast resource is only now starting to be appreciated by biotechnologists. Fortunately they have at their disposal over 170 000 strains maintained in culture collections throughout the world.

This book provides an introduction to these resources and describes how information can be obtained on what is available, how filamentous fungi can be preserved and identified, how collections operate, and the additional support services available from them.

In preparing this volume, I have been fortunate in securing the assistance of colleagues from different parts of the world who are experienced with the work of culture collections, as curators or users, as well as from the individual collections themselves. Without their help the international overview this text aims to provide could not have been presented.

Introduction

Microbiologists are faced with consideration of exponential growth in their laboratories on a daily basis. As users of a chapter on information resources for biotechnology they are exposed to a double dose of exponential growth. First, the explosion of information technology itself is due to the massive amounts of computing power available at ever diminishing cost. In turn, a population of computeraware and computer-literate microbiologists present a growing demand for more sophisticated access to modern information technology. The community of information technologists in concert with microbiologists are responding to this demand with a multiplicity of initiatives using various strategies.

The resulting activity induces feelings of inadequacy in the authors of such chapters as this, since at the moment of delivery to the editors the information is out of date. Resources previously known only by rumour are tested. Simple facilities being tested as pilot projects are quickly made available to the community. Local data banks open their doors to regional and even world-wide participation. Databases on databases spring up because of the need to discover available resources. Occasionally, resources fall by the wayside. The net result is an ever increasing base of information resources for biotechnologists.

While the information about information presented in this chapter is out of date as soon as it is written, the resources described are most likely to be improved and be more useful than the descriptions indicate. For information on new developments the listed resources should be contacted.

Introduction

In response to the needs of users, many culture collections provide a range of services to the scientific, technological and commercial world. This chapter provides an introduction to the types of services available from culture collections, but it is beyond its scope to give a comprehensive list of such services. As the range of work that can be undertaken is increasing at many of the collections, the reader should contact individual collections to find out whether they can offer particular services.

Types of services

Directly associated and customer services

The two major services which are intrinsically part of culture collection work are those concerning the identification and preservation of organisms. Collections of necessity need expertise in these fields to be able to function, and many provide comprehensive services in these areas. Aspects of culture identification methods (Chapter 5), sales of cultures (Chapter 3), preservation techniques (Chapter 4) and patent deposits (Chapter 6) are covered elsewhere in this volume.

Safe-deposits. Many collections hold organisms which are not listed in their catalogues. These cultures are held for a variety of reasons: the organisms may not be fully identified, their taxonomic status may be unclear, their stability in preservation may be suspect or they may be held at the request of the depositor who wishes to have back-up material and yet retain ownership and confidentiality, not releasing the strain to other parties. Many collections have introduced safe-deposit services as a back-up to the depositor's working collection, providing a service intermediate between an open collection deposit and a deposit for patent purposes.

The rapid advances taking place in biotechnology have introduced large numbers of scientists and engineers to the need for handling microorganisms, often for the first time. Questions are frequently raised concerning sources of cultures, location of strains with particular properties, requirements for handling the cultures, preservation and identification methods, regulations for shipping, or for the deposit of strains for patent purposes. For those in industry, research institutes or universities with little experience in these areas, resolving such difficulties may seem overwhelming. The purpose of the World Federation for Culture Collections’ (WFCC) series, Living Resources for Biotechnology, is to provide answers to these questions.

Living Resources for Biotechnology is a series of practical books that provide primary data and guides to sources for further information on matters relating to the location and use of different kinds of biological material of interest to biotechnologists. A deliberate decision was taken to produce separate volumes for each group of microorganism rather than a combined compendium, since our enquiries suggested that inexpensive specialised books would be of more general value than a larger volume containing information irrelevant to workers with interests in one particular type of organism. As a result each volume contains specialised information together with material on general matters (information centres, patents, consumer services, the international coordination of culture collection activities) that is common to each.

A large family well represented in West Africa, at least half of the genera containing species which are widespread throughout West Africa, many of these, in addition, in Gambia.

Introduced garden species give a useful introduction to the general appearance of members of this family. Allamanda spp., oleander (Nerium oleander), Madagascar periwinkle (Catharanthus roseus), frangipani (Plumeria spp.) and herald's trumpet (Beaumontia grandiflora) are all widely available.

Members of the family may be recognised by their opposite, simple, entire, generally exstipulate leaves (though often with interpetiolar ridges), possession of latex, and cymes of fragrant, gamopetalous 5-part flowers with contorted corolla lobes. The fruit is a berry of two carpels, two berries (of one carpel each), or a pair of follicles (or one by abortion) with plumed seeds. Most of the widespread species are climbers (some with branch tendrils) or under-storey trees in forest, a few only being open woodland savanna species.

Four genera diverge sufficiently from the ‘family pattern’ as to be recognisable on sight. The only species with spirally arranged leaves (and prominent leaf scars) is a fleshy stemmed deciduous shrub, Adenium obesum, found (and often planted) in the northern guinea and sudan zones of Nigeria.

Carissa edulis, in the same zone (Ghana–Nigeria) has branch spines, the only species in the family in West Africa to do so.

A tropical and temperate family, represented in West Africa by large woody or fibrous perennial herbs up to 3 m high, less often by shrubs and rarely by trees. Most species are deep rooting, withstand drought, and are prominent in drier savannas.

The decorative introductions include several Hibiscus spp.: H. mutabilis (blushing Hibiscus) and H. rosa-sinensis (rose of China, shoe flower) from China, H. schizopetalus (frilled Hibiscus) from East Africa.

Members of the family may be recognised by their alternate, stipulate simple leaves with digitate venation (pinnate in Malvastrum and Sida) with stellate hairs on young growth (scales in Thespesia). Ptyxis is mostly conduplicate, conduplicate-plicate in Abutilon and Pavonia. On the back of the leaf, glands occur in patches, depressions or pits (concealed in small swellings) (absent in Pavonia). The flowers are axillary, five-part and often large, lasting a day only, their colour often changing during that time. The petals are veined or basally blotched, and the column of anthers in the centre of the flower is prominent.

Flowers ⊕ (rarely dioecious) 5-part. The flower is usually surrounded by 3(–13) united bracteoles, adnate to the calyx and forming an epicalyx in Abelmoschus, Pavonia and Urena etc., free from the calyx and forming an involucel in most genera and often associated with extrafloral nectaries (Cienfuegosia); bracteoles free in Gossypium and Thespesia; in Malachra, there is an involucre round each head of flowers, while bracteoles are lacking in Abutilon etc.

A family of woody plants in all the warmer parts of the world, represented in West Africa by trees, some shrubs and shrubby epiphytes, and a few stranglers.

Members of the family may be recognised by their latex and alternate stipulate leaves. The leaves may be pinnately veined, relatively small, and conduplicate in the bud, when the stipules are also small (but see Ficus carica and Artocarpus heterophyllus) or the stipules may be intrapetiolar, large and hoodlike, leaving circular scars on the stem. The leaves are then rolled (supervolute) in the bud (Ficus etc.) or pleated (plicate) in the bud and later large and digitately divided or compound (Artocarpus communis (now A. altilis), Musanga). The leaves are usually leathery or rough. The flowers are small, ♂ or ♀, apetalous and often four-part, closely packed in cymose inflorescences, the inflorescence axis (the receptacle, cf. Euphorbia) often enlarged. It may be shaped like a spike, a disc or a bag. The fruit is false, sometimes a syncarp or a syconium.

The boundary between the Moraceae and the Urticaceae rests on rather few characters, and Musanga and Myrianthus are now generally considered to be part of the latter family as a consequence of their possession of clear (not milky) sap, which turns black, basal ovules, small achenes and mineral bodies (cystoliths) in the epidermal cells. In conformity with the second edition of the Flora of West tropical Africa, however, the two genera are here retained in the Moraceae.

A large woody tropical family represented in West Africa by c. 60 genera, mainly trees. In this family, and the two that follow, the Mimosaceae and the Papilionaceae, the pod (legume) is regarded as the characteristic fruit. It is formed from the single superior carpel, and becomes dry, opening by splitting down both margins. Both pods and the seeds they contain are commonly referred to as ‘beans’. In some classifications, the three families are united under the name Leguminosae (Fabaceae) (with subfamilies), thus forming the second largest family of flowering plants.

Members of the Caesalpiniaceae may be recognised by their alternate, stipulate, compound pinnate leaves without stipels (except Amphimas and Pellegrinodendron). The conduplicate leaflets may be alternate or paired, and there may (imparipinnate) or may not (paripinnate) be an odd terminal leaflet. A pulvinus occurs at the base of the petiole, a pulvinule at the base of the petiolule. Both have a wrinkled surface. Compound racemose inflorescences of zygomorphic five-part flowers each with a single superior carpel are usual. The fruit is generally a pod.

Unifoliate leaves are uncommon, most often being seen in three whiteflowered savanna species: Bauhinia (now Adenolobus) rufescens (Schmitz, 1973) (with flowers) in riparian woodland, and, with ♂ and ♀ flowers on separate plants, Piliostigma reticulatum in sudan savanna and P. thonningii in forestsavanna mosaic, both also in northern guinea savanna. The leaves are twolobed in all cases. Half a dozen other genera have species with entire unifoliate leaves.

Except for some species of the genus Garcinia, these are upper-storey, evergreen forest trees. Most are glabrous, the slash exuding yellow or orange resinous sap or latex. The family is a pantropical one, of about 600 species. No purely decorative species have been introduced. The Hypericaceae may be included as a subfamily.

Members of the family may be recognised by their pairs of exstipulate, simple entire leaves, which are streaked and spotted with resin glands, and carried on branches which appear to be whorled. The flowers are axillary, solitary or in cymes, red or white and conspicuous, with numerous stamens often in bundles. The fruits are large and berry like, and of economic importance.

Flowers ⊕, ♂, ♀ and, often 5-part, though 4-part in Calophyllum etc., when the sepals are orthogonal and the petals diagonal. Anthers mostly united in bundles each opposite a petal and alternating with the stigmata, but free in Garcinia sp., united only basally in Calophyllum and Mammea, and united into a staminal tube in Symphonia, cf. Chapter 18, Meliaceae; present as staminodes in ♀ flowers. An extrastaminal disc is usually present. G(2)–(5)–(8) with the same number of cells and axile placentation, the terminal style sometimes very short; 1-celled with 1 basal ovule in Calophyllum and 5-celled with parietal placentae in Allanblackia.

Pollination The disc in Allanblackia, Garcinia and Symphonia, thought to be staminodal in origin, and the numerous stamens suggest insect pollination, the shape of the flower providing ‘open access’ to short-tongued insects.