Isolation and nucleotide sequence of a sesquiterpene cyclase gene from the trichothecene-producing fungus Fusarium sporotrichioides

Höhn, Thomas; Beremand, Phillip D.

doi:10.1016/0378-1119(89)90098-x

Cited by 218 publications

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“…Trichodiene synthase is the enzyme involved in trichodiene biosynthesis. A gene encoding trichodiene synthase has been isolated from Fusarium sporotrichioides (8) and expressed in Escherichia coli. Expression of trichodiene synthase in E. coli resulted in the production of active enzyme and in the accumulation of trichodiene (10).…”

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confidence: 99%

Expression of a Fungal Sesquiterpene Cyclase Gene in Transgenic Tobacco

Höhn¹,

Ohlrogge²

1991

Plant Physiol.

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The complete coding sequence for the trichodiene synthase gene from Fusarium sporotrichioides was introduced into tobacco (Nicotiana tabacum) under the regulation of the cauliflower mosiac virus 35S promoter. Expression of trichodiene synthase was demonstrated in the leaves of transformed plants. Leaf homogenates incubated with [3H]famesyl pyrophosphate produced trichodiene as a major product. Trichodiene was detected in the leaves of a transformed plant at a level of 5 to 10 nanograms per gram fresh weight. The introduction of a fungal sesquiterpene cyclase gene into tobacco has resulted in the expression of an active enzyme and the accumulation of low levels of its sesquiterpenoid product.Sesquiterpene cyclases participate in the biosynthesis of most sesquiterpenoids. This group of enzymes converts the isoprenoid pathway intermediate, FPP,2 into an estimated 100 to 200 different cyclic sesquiterpenoids (2). Information concerning the properties of sesquiterpene cyclases has been reported for isolated or partially purified enzyme preparations from plants (4, 13), fungi (9, 11), and a streptomycete (1). Comparisons among these enzymes indicate that they are mechanistically similar but differ with respect to properties such as molecular weight and subunit composition.Plants, fungi, insects, and marine invertebrates all produce large numbers of structurally distinct sesquiterpenoids. In many cases, distantly related organisms accumulate the same sesquiterpene cyclase product or produce sesquiterpenoids derived from the same parent compound. This means that genes coding for any particular sesquiterpene cyclase might be available from several sources.The modification of plant sesquiterpenoid biosynthesis could result in altered resistance to disease and insect pests. Potential targets of such efforts include the production of sesquiterpenoid phytoalexins within the Solanaceae (16) (12). Trichodiene synthase is the enzyme involved in trichodiene biosynthesis. A gene encoding trichodiene synthase has been isolated from Fusarium sporotrichioides (8) and expressed in Escherichia coli. Expression of trichodiene synthase in E. coli resulted in the production of active enzyme and in the accumulation of trichodiene (10).The expression of foreign sesquiterpene cyclase genes in plants represents one possible approach for altering sesquiterpenoid biosynthesis. To determine the feasibility of this approach, we introduced the trichodiene synthase gene into tobacco. Expression in tobacco may provide insights into both the regulation of sesquiterpenoid biosynthesis and the role of these compounds in disease resistance. MATERIALS AND METHODS Transformation of TobaccoThe BamHI fragment from pTS37-4A(470-529), containing the complete coding sequence for the trichodiene synthase gene of Fusarium sporotrichioides (10), was inserted into the plasmid, pBI221 (Clontech3), from which the ,B-glucuronidase-coding sequence had been deleted (SmaI-SstI fragment EXPRESSION OF TRICHODIENE SYNTHASE IN TOBACCOmosaic virus 35S promoter, a u...

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confidence: 99%

Expression of a Fungal Sesquiterpene Cyclase Gene in Transgenic Tobacco

Höhn¹,

Ohlrogge²

1991

Plant Physiol.

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“…68,69 He then identified the structural genes for each of these two enzymes, becoming the first to clone any terpene synthase. 70,71 Zhen Wu, a graduate student in my laboratory, now with Roche Biosciences in Shanghai, then overexpressed both the trichodiene and aristolochene synthases in Escherichia coli. 72,73 Jae-Kyung Sohng, another graduate student who is now Professor at Sun Moon University in Korea, purified pentalenene synthase from Streptomyces exfoliatus, which allowed us within two years to clone and overexpress the corresponding pentalenene synthase structural gene.…”

Section: Enzymology and Molecular Genetics Of Natural Product Biosyntmentioning

confidence: 99%

Nature as organic chemist

Cane

2016

J Antibiot

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Figure 15 Stereospecific reduction of (2R)-2-methyl-3-ketopentanoyl-ACP by recombinant ketoreductase (KR) domains: EryKR6, from module 6 of the 6dEB synthase (DEBS); TylKR1, from module 1 of the tylactone synthase; EryKR1, from DEBS module 1; RifKR7, from module 7 of the rifamycin PKS. The (2R)-2-methyl-3-ketopentanoyl-ACP substrate is generated in situ by a reconstituted mixture of dissected PKS domains, Ery[KS6][AT6] (ketosynthase-acyl transferase didomain) and EryACP6 are both from DEBS module 6. Editorial

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“…Hohn and coworkers pioneered the research by purifying the branch-point enzyme, trichodiene synthase (Hohn and Vanmiddlesworth, 1986), and isolating the encoding gene Tri5 (Hohn and Beremand, 1989). Although Tri5 was not linked to Tri1 (i.e., a genetic locus related to the C-8 hydroxylation step) required for T-2 toxin biosynthesis (Beremand and Desjardins, 1988;Desjardins et al, 1993), the Tri5-containing cosmids did complement the Tri3 Ϫ and Tri4 Ϫ mutations of F.…”

Section: Trichothecene Biosynthetic and Regulatory Genes From F Spormentioning

confidence: 99%

Microbial toxins in plant-pathogen interactions: Biosynthesis, resistance mechanisms, and significance.

Kimura

Anzai

Yamaguchi

2001

J. Gen. Appl. Microbiol.

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In the history of phytopathology, microbial toxins have been the objects of extensive studies as possible pathogenicity or virulence factors for the producer pathogens. The recent development of molecular genetic techniques provided an experimental basis to thoroughly test the role of these secondary metabolites in pathogenesis. Some of them did prove to be highly associated with disease initiation or enhanced virulence in certain plant-pathogen interactions. In this review, we describe recent progresses in the field of plant-pathogen interactions focusing on two toxins; i.e., tabtoxin from Pseudomonas syringae and trichothecenes from Fusarium and other fungi. These microbial toxins have convincingly been shown to play causal roles in plant disease development. Studies on the biosynthesis and resistance mechanisms of these producers are outlined, and the significance of this knowledge is discussed in relation to practical applications in agriculture.Key Words--disease-resistant plants; gene cluster evolution; Fusarium mycotoxins; horizontal gene transfer; Pseudomonas syringae; self-protection mechanism; tabtoxin; trichothecene biosynthesis J. Gen. Appl. Microbiol., 47, 149-160 (2001) Invited Review * Address reprint requests to: Dr. Isamu Yamaguchi, Environmental Plant Research Group, Plant Science Center, RIKEN, 2-1 Hirosawa, Wako, 351-0198, Japan. E-mail: isyama@postman.riken.go.jp pathogen of the ability to produce the toxic substances in question or to endow the host with the ability to inactivate them, leaving all other properties of these organisms exactly the same in either case. Molecular genetic techniques served as powerful tools to prove that some of the toxins are indeed required for pathogenicity or virulence. Perhaps the most well-known examples for this are the host-specific toxins produced by three Cochliobolus species; i.e., HCtoxin of C. carbonum, T-toxin of C. heterostrophus, and victorin of C. victoriae, all of which proved to be the causal agents of severe blight diseases of economically important crops. The remarkable progresses in understanding the biological systems of these toxin producers have been summarized by the researchers involved (Cheng and Walton, 2000;Kodama et al., 1999;Walton, 1996;Walton and Panaccione, 1993).In this review, we deal with the recent progress of research on toxin biosynthesis and self-resistance mechanisms of plant pathogenic microorganisms. The following two sections summarize studies on two toxins, tabtoxin produced by Pseudomonas syringae and trichothecenes produced by a wide variety of fungal genera comprising mainly Fusarium. Tabtoxin Characteristics of tabtoxinTobacco leaves diseased by infection with wildfire bacteria result in a small necrotic spot containing bacterial cells, surrounded by a chlorotic zone that is free from bacteria. The wildfire bacterium P. syringae pv. tabaci produces a pathogenic toxin called tabtoxin. When a tobacco leaf was treated with tabtoxin, this toxin caused chlorotic spots similar to the halos on leaves infected with th...

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Isolation and nucleotide sequence of a sesquiterpene cyclase gene from the trichothecene-producing fungus Fusarium sporotrichioides

Cited by 218 publications

References 20 publications

Expression of a Fungal Sesquiterpene Cyclase Gene in Transgenic Tobacco

Expression of a Fungal Sesquiterpene Cyclase Gene in Transgenic Tobacco

Nature as organic chemist

Microbial toxins in plant-pathogen interactions: Biosynthesis, resistance mechanisms, and significance.

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