Joan W. Bennett

Fungal secondary metabolism — from biochemistry to genomics

Much of natural product chemistry concerns a group of compounds known as secondary metabolites. These low-molecular-weight metabolites often have potent physiological activities. Digitalis, morphine and quinine are plant secondary metabolites, whereas penicillin, cephalosporin, ergotrate and the statins are equally well known fungal secondary metabolites. Although chemically diverse, all secondary metabolites are produced by a few common biosynthetic pathways, often in conjunction with morphological development. Recent advances in molecular biology, bioinformatics and comparative genomics have revealed that the genes encoding specific fungal secondary metabolites are clustered and often located near telomeres. In this review, we address some important questions, including which evolutionary pressures led to gene clustering, why closely related species produce different profiles of secondary metabolites, and whether fungal genomics will accelerate the discovery of new pharmacologically active natural products.

Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus.

Aspergillus fumigatus is exceptional among microorganisms in being both a primary and opportunistic pathogen as well as a major allergen. Its conidia production is prolific, and so human respiratory tract exposure is almost constant. A. fumigatus is isolated from human habitats and vegetable compost heaps. In immunocompromised individuals, the incidence of invasive infection can be as high as 50% and the mortality rate is often about 50% (ref. 2). The interaction of A. fumigatus and other airborne fungi with the immune system is increasingly linked to severe asthma and sinusitis. Although the burden of invasive disease caused by A. fumigatus is substantial, the basic biology of the organism is mostly obscure. Here we show the complete 29.4-megabase genome sequence of the clinical isolate Af293, which consists of eight chromosomes containing 9,926 predicted genes. Microarray analysis revealed temperature-dependent expression of distinct sets of genes, as well as 700 A. fumigatus genes not present or significantly diverged in the closely related sexual species Neosartorya fischeri, many of which may have roles in the pathogenicity phenotype. The Af293 genome sequence provides an unparalleled resource for the future understanding of this remarkable fungus.

Genome sequencing and analysis of Aspergillus oryzae

The genome of Aspergillus oryzae, a fungus important for the production of traditional fermented foods and beverages in Japan, has been sequenced. The ability to secrete large amounts of proteins and the development of a transformation system have facilitated the use of A. oryzae in modern biotechnology. Although both A. oryzae and Aspergillus flavus belong to the section Flavi of the subgenus Circumdati of Aspergillus, A. oryzae, unlike A. flavus, does not produce aflatoxin, and its long history of use in the food industry has proved its safety. Here we show that the 37-megabase (Mb) genome of A. oryzae contains 12,074 genes and is expanded by 7–9 Mb in comparison with the genomes of Aspergillus nidulans and Aspergillus fumigatus. Comparison of the three aspergilli species revealed the presence of syntenic blocks and A. oryzae-specific blocks (lacking synteny with A. nidulans and A. fumigatus) in a mosaic manner throughout the genome of A. oryzae. The blocks of A. oryzae-specific sequence are enriched for genes involved in metabolism, particularly those for the synthesis of secondary metabolites. Specific expansion of genes for secretory hydrolytic enzymes, amino acid metabolism and amino acid/sugar uptake transporters supports the idea that A. oryzae is an ideal microorganism for fermentation.

Clustered Pathway Genes in Aflatoxin Biosynthesis

Aflatoxins, a group of polyketide-derived furanocoumarins (Fig. [1][1]), are the most toxic and carcinogenic compounds among the known mycotoxins. Among the at least 16 structurally related aflatoxins characterized, however, there are only four major aflatoxins, B1, B2, G1, and G2 (AFB1, AFG1, AFB2

Seeing red : The story of prodigiosin

Publisher Summary S. marcescens has played an important role in the history of bacterial taxonomy, in research on the transmission of bacterial aerosols, in the study of emerging nosocomial infections, and in the understanding of secondary metabolite biosynthesis. The prodigiosin pigments have intrigued organic chemists and pharmacologists, and play roles in the treatment of infectious diseases such as malaria, and perhaps as immunosuppressant agents. Undecylprodiginine played an important role in the first cloning of a gene, playing a defined role in the biosynthesis of an antibiotic. An O-methyltransferase gene was isolated by complementation and the color of undecylprodiginine was used as the selectable phenotype. The regulation of prodigiosin biosynthesis is complex, being influenced by increased glucose levels and decreased by increased phosphate level. The antibiotic resistance of many strains of S. marcescens is a serious problem with rapid horizontal transfer of drug resistance by plasmids.

Microbial stimulation of plant growth and protection from disease

Certain microbes stimulate plant growth. The mechanisms used by such microbes to achieve this are: biofertilization (i.e. increasing the supply of mineral nutrients to the plant); biocontrol (i.e. elimination of the plants enemies including microbial pathogens, insects and weeds); and direct plant growth promotion (e.g. by delivering plant growth hormones to the plant). In all of these mechanisms, plant root colonization is crucial as the delivery system.

New Perspectives on Aflatoxin Biosynthesis

Publisher Summary Aflatoxins are a family of toxic secondary metabolites produced by certain strains of the common molds Aspergillus flavus and A. parasiticus. The general biosynthetic pathway of aflatoxin appears to be as follows: acetate/malonate norsolorinic acid → averantin → averufin → versiconal hemiacetal acetate → versicolorin A → sterigmatocystin → aflatoxins. Within the context of this general pathway, some mechanistic details concerning formation and subsequent conversion of these individual metabolites have been provided by NMR studies-in particular, 13C-NMR. The chapter focuses on the studies that have provided information concerning bonds broken and formed during the biosynthetic process. The preliminary data on toxicity and mutagenicity indicate that the evolution of the aflatoxin pathway may be a response to selective pressures from mycophagous species in nature. Because of the abundant biosynthetic data and the availability of blocked mutants, this system may be uniquely suited among micrcbial secondary pathways for testing this ecological hypothesis.

Aspergillus flavus genomics: gateway to human and animal health, food safety, and crop resistance to diseases

Aspergillus flavus is an imperfect filamentous fungus that is an opportunistic pathogen causing invasive and non-invasive aspergillosis in humans, animals, and insects. It also causes allergic reactions in humans. A. flavus infects agricultural crops and stored grains and produces the most toxic and potent carcinogic metabolites such as aflatoxins and other mycotoxins. Breakthroughs in A. flavus genomics may lead to improvement in human health, food safety, and agricultural economy. The availability of A. flavus genomic data marks a new era in research for fungal biology, medical mycology, agricultural ecology, pathogenicity, mycotoxin biosynthesis, and evolution. The availability of whole genome microarrays has equipped scientists with a new powerful tool for studying gene expression under specific conditions. They can be used to identify genes responsible for mycotoxin biosynthesis and for fungal infection in humans, animals and plants. A. flavus genomics is expected to advance the development of therapeutic drugs and to provide information for devising strategies in controlling diseases of humans and other animals. Further, it will provide vital clues for engineering commercial crops resistant to fungal infection by incorporating antifungal genes that may prevent aflatoxin contamination of agricultural harvest.

Mycotoxins, mycotoxicoses, mycotoxicology andMycopathologia

SummaryMycotoxins are fungal poisons. This definition does not stipulate whether fungi are the targets of poisoning or are the producers of the poisons. The following is suggested as a useful working definition: Mycotoxins are natural products produced by fungi that evoke a toxic response when introduced in low concentration to higher vertebrates and other animals by a natural route. Some mycotoxins have multiple effects, and may cause phytotoxic and antimicrobial syndromes in addition to animal toxicity. By convention, mushroom and yeast poisons are usually excluded from discussions of mycotoxins.The eclectic nature of the discipline and the international scope of the problem has attracted scientists from many different backgrounds. The publishers and editors ofMycopathologia intend for this journal to become a major forum for mycotoxin research.

Association of aflatoxin biosynthesis and sclerotial development in Aspergillus parasiticus

Secondary metabolism in fungi is frequently associated with asexual and sexual development. Aspergillus parasiticus produces aflatoxins known to contaminate a variety of agricultural commodities. This strictly mitotic fungus, besides producing conidia asexually, produces sclerotia, structures resistant to harsh conditions and for propagation. Sclerotia are considered to be derived from the sexual structure, cleistothecia, and may represent a vestige of ascospore production. Introduction of the aflatoxin pathway-specific regulatory gene, aflR, and aflJ, which encoded a putative co-activator, into an O-methylsterigmatocystin (OMST)-accumulating strain,A. parasiticus SRRC 2043, resulted in elevated levels of accumulation of major aflatoxin precursors, including norsolorinic acid (NOR), averantin (AVN), versicolorin A (VERA) and OMST. The total amount of these aflatoxin precursors, NOR, VERA, AVN and OMST, produced by the aflR plus aflJ transformants was two to three-fold that produced by the aflR transformants. This increase indicated a synergisticeffect of aflR and aflJ on the synthesis of aflatoxin precursors. Increased production of the aflatoxin precursors was associated with progressive decrease in sclerotial size, alteration in sclerotial shape and weakening in the sclerotial structure of the transformants. The results showed that sclerotial development and aflatoxin biosynthesis are closely related. We proposed that competition for a common substrate, such as acetate, by the aflatoxin biosynthetic pathway could adversely affect sclerotial development in A. parasiticus.