Chemical Diversity and Defence Metabolism: How Plants Cope with Pathogens and Ozone Pollution

Chemical defences represent a main trait of the plant innate immune system. Besides regulating the relationship between plants and their ecosystems, phytochemicals are involved both in resistance against pathogens and in tolerance towards abiotic stresses, such as atmospheric pollution. Plant defence metabolites arise from the main secondary metabolic routes, the phenylpropanoid, the isoprenoid and the alkaloid pathways. In plants, antibiotic compounds can be both preformed (phytoanticipins) and inducible (phytoalexins), the former including saponins, cyanogenic glycosides and glucosinolates. Chronic exposure to tropospheric ozone (O₃) stimulates the carbon fluxes from the primary to the secondary metabolic pathways to a great extent, inducing a shift of the available resources in favour of the synthesis of secondary products. In some cases, the plant defence responses against pathogens and environmental pollutants may overlap, leading to the unspecific synthesis of similar molecules, such as phenylpropanoids. Exposure to ozone can also modify the pattern of biogenic volatile organic compounds (BVOC), emitted from plant in response to herbivore feeding, thus altering the tritrophic interaction among plant, phytophagy and their natural enemies. Finally, the synthesis of ethylene and polyamines can be regulated by ozone at level of S-adenosylmethionine (SAM), the biosynthetic precursor of both classes of hormones, which can, therefore, mutually inhibit their own biosynthesis with consequence on plant phenotype.     

The Physiological Importance of Glucosinolates on Plant Response to Abiotic Stress in Brassica

Glucosinolates, a class of secondary metabolites, mainly found in Brassicaceae, are affected by the changing environment. This review is focusing on the physiological significance of glucosinolates and their hydrolysis products in the plant response to different abiotic stresses. Special attention is paid to the crosstalk between some of the physiological processes involved in stress response and glucosinolate metabolism, with the resulting connection between both pathways in which signaling mechanisms glucosinolate may act as signals themselves. The function of glucosinolates, further than in defense switching, is discussed in terms of alleviating pathogen attack under abiotic stress. The fact that the exogenous addition of glucosinolate hydrolysis products may alleviate certain stress conditions through its effect on specific proteins is described in light of the recent reports, but the molecular mechanisms involved in this response merit further research. Finally, the transient allocation and re-distribution of glucosinolates as a response to environmental changes is summarized.     

Phenolic Phytoalexins in Rice: Biological Functions and Biosynthesis

Phytoalexins are inducible secondary metabolites possessing antimicrobial activity against phytopathogens. Rice produces a wide array of phytoalexins in response to pathogen attacks and environmental stresses. With few exceptions, most phytoalexins identified in rice are diterpenoid compounds. Until very recently, flavonoid sakuranetin was the only known phenolic phytoalexin in rice. However, recent studies have shown that phenylamides are involved in defense against pathogen attacks in rice. Phenylamides are amine-conjugated phenolic acids that are induced by pathogen infections and abiotic stresses including ultra violet (UV) radiation in rice. Stress-induced phenylamides, such as N-trans-cinnamoyltryptamine, N-p-coumaroylserotonin and N-cinnamoyltyramine, have been reported to possess antimicrobial activities against rice bacterial and fungal pathogens, an indication of their direct inhibitory roles against invading pathogens. This finding suggests that phenylamides act as phytoalexins in rice and belong to phenolic phytoalexins along with sakuranetin. Phenylamides also have been implicated in cell wall reinforcement for disease resistance and allelopathy of rice. Synthesis of phenolic phytoalexins is stimulated by phytopathogen attacks and abiotic challenges including UV radiation. Accumulating evidence has demonstrated that biosynthetic pathways including the shikimate, phenylpropanoid and arylmonoamine pathways are coordinately activated for phenolic phytoalexin synthesis, and related genes are induced by biotic and abiotic stresses in rice.     

The Desert Locust, <Emphasis Type="Italic">Schistocerca gregaria</Emphasis>, Detoxifies the Glucosinolates of <Emphasis Type="Italic">Schouwia purpurea</Emphasis> by Desulfation

Desert locusts (Schistocerca gregaria) occasionally feed on Schouwia purpurea, a plant that contains tenfold higher levels of glucosinolates than most other Brassicaceae. Whereas this unusually high level of glucosinolates is expected to be toxic and/or deterrent to most insects, locusts feed on the plant with no apparent ill effects. In this paper, we demonstrate that the desert locust, like larvae of the diamondback moth (Plutella xylostella), possesses a glucosinolate sulfatase in the gut that hydrolyzes glucosinolates to their corresponding desulfonated forms. These are no longer susceptible to cleavage by myrosinase, thus eliminating the formation of toxic glucosinolate hydrolysis products. Sulfatase is found throughout the desert locust gut and can catalyze the hydrolysis of all of the glucosinolates present in S. purpurea. The enzyme was detected in all larval stages of locusts as well as in both male and female adults feeding on this plant species. Glucosinolate sulfatase activity is induced tenfold when locusts are fed S. purpurea after being maintained on a glucosinolate-free diet, and activity declines when glucosinolates are removed from the locust diet. A detoxification system that is sensitive to the dietary levels of a plant toxin may minimize the physiological costs of toxin processing, especially for a generalist insect herbivore that encounters large variations in plant defense metabolites while feeding on different species.     

Secondary Metabolites of the Rice Blast Fungus Pyricularia oryzae: Biosynthesis and Biological Function

Plant pathogenic fungi produce a wide variety of secondary metabolites with unique and complex structures. However, most fungal secondary metabolism genes are poorly expressed under laboratory conditions. Moreover, the relationship between pathogenicity and secondary metabolites remains unclear. To activate silent gene clusters in fungi, successful approaches such as epigenetic control, promoter exchange, and heterologous expression have been reported. Pyricularia oryzae, a well-characterized plant pathogenic fungus, is the causal pathogen of rice blast disease. P. oryzae is also rich in secondary metabolism genes. However, biosynthetic genes for only four groups of secondary metabolites have been well characterized in this fungus. Biosynthetic genes for two of the four groups of secondary metabolites have been identified by activating secondary metabolism. This review focuses on the biosynthesis and roles of the four groups of secondary metabolites produced by P. oryzae. These secondary metabolites include melanin, a polyketide compound required for rice infection; pyriculols, phytotoxic polyketide compounds; nectriapyrones, antibacterial polyketide compounds produced mainly by symbiotic fungi including endophytes and plant pathogens; and tenuazonic acid, a well-known mycotoxin produced by various plant pathogenic fungi and biosynthesized by a unique NRPS-PKS enzyme.     

Strategies to Modulate Specialized Metabolism in Mediterranean Crops: From Molecular Aspects to Field

Plant specialized metabolites (SMs) play an important role in the interaction with the environment and are part of the plant defense response. These natural products are volatile, semi-volatile and non-volatile compounds produced from common building blocks deriving from primary metabolic pathways and rapidly evolved to allow a better adaptation of plants to environmental cues. Specialized metabolites include terpenes, flavonoids, alkaloids, glucosinolates, tannins, resins, etc. that can be used as phytochemicals, food additives, flavoring agents and pharmaceutical compounds. This review will be focused on Mediterranean crop plants as a source of SMs, with a special attention on the strategies that can be used to modulate their production, including abiotic stresses, interaction with beneficial soil microorganisms and novel genetic approaches.     

Rhizopus and Fusarium are Selected as Dominant Fungal Genera in Rhizospheres of Brassicaceae

We isolated several strains of Rhizopus and Fusarium spp. as dominant fungi in the rhizospheres of Brassicaceae plants. The Fusarium isolates showed a higher tolerance of the antifungal constituents of "mustard oil," which originates from the glucosinolates that are characteristic secondary metabolites of the Brassicaceae, than other Fusarium isolates from non-Brassicaceae plants. In contrast, the Rhizopus isolates showed a high tolerance regardless of their source. Myrosinase activity was found in Bn-R-1-1 (Rhizopus sp.) isolated from the rhizoplane of Brassica napus and Ls-F-in-4-1 (Fusarium sp.) isolated from a surface-disinfected root of Lepidium sativum (Brassicaceae). Ls-F-in-4-1 was the Fusarium most tolerant of the Brassicaceae antifungal constituents. These results suggest that fungi in the rhizospheres of Brassicaceae plants may be selected because of secondary metabolites exuded from the roots of host plants.     

An Unusual Thioesterase Promotes Isochromanone Ring Formation in Ajudazol Biosynthesis

The ajudazols are antifungal secondary metabolites produced by a hybrid polyketide synthase (PKS)‐nonribosomal peptide synthetase (NRPS) multienzyme “assembly line” in the myxobacterium Chondromyces crocatus Cm c5. The most striking structural feature of these compounds is an isochromanone ring system; such an aromatic moiety is only known from two other complex polyketides, the electron transport inhibitor stigmatellin and the polyether lasalocid. The cyclization and aromatization reactions in the stigmatellin pathway are presumed to be catalyzed by a cyclase domain located at the end of the PKS, while the origin of the lasalocid benzenoid ring remains obscure. Notably, the ajudazol biosynthetic machinery does not incorporate a terminal cyclase, but instead a variant thioesterase (TE) domain. Here we present detailed phylogenetic and sequence analysis, coupled with experiments both in vitro and in vivo, that suggest that this TE promotes formation of the isochromanone ring, a novel reaction for this type of domain. As the ajudazol TE has homologues in several other secondary‐metabolite pathways, these results are likely to be generalizable.     

Definition of the common and divergent steps in carbapenem β-lactam antibiotic biosynthesis

Approximately 50 naturally occurring carbapenem β-lactam antibiotics are known. All but one of these have been isolated from Streptomyces species and are disubstituted structural variants of a simple core that is synthesized by Pectobacterium carotovorum (Erwinia carotovora), a phylogenetically distant plant pathogen. While the biosynthesis of the simple carbapenem, (5R)-carbapen-2-em-3-carboxylic acid, is impressively efficient requiring only three enzymes, CarA, CarB and CarC, the formation of thienamycin, one of the former group of metabolites from Streptomyces, is markedly more complex. Despite their phylogenetic separation, bioinformatic analysis of the encoding gene clusters suggests that the two pathways could be related. Here we demonstrate with gene swapping, stereochemical and kinetics experiments that CarB and CarA and their S. cattleya orthologues, ThnE and ThnM, respectively, are functionally and stereochemically equivalent, although their catalytic efficiencies differ. The biosynthetic pathways, therefore, to thienamycin, and likely to the other disubstituted carbapenems, and to the simplest carbapenem, (5R)-carbapen-2-em-3-carboxylic acid, are initiated in the same manner, but share only two common steps before diverging.     

Desulfation followed by sulfation: metabolism of benzylglucosinolate in Athalia rosae (Hymenoptera: Tenthredinidae)

The sawfly species Athalia rosae (L.) (Hymenoptera: Tenthredinidae) is phytophagous on plants of the family Brassicaceae and thus needs to cope with the plant defence, the glucosinolate-myrosinase system. The larvae sequester glucosinolates in their haemolymph. We investigated how these compounds are metabolized by this specialist. When larvae were fed with ([(14) C]-labelled) benzylglucosinolate, one major degradation metabolite, with the same sum formula as benzylglucosinolate, was defecated. This metabolite was also found in the haemolymph along with desulfobenzylglucosinolate, which continuously increased in concentration. NMR spectroscopy in conjunction with LC-TOF-MS measurements revealed the major degradation metabolite to be desulfobenzylglucosinolate-3-sulfate, probably converted from desulfobenzylglucosinolate after sulfation at the sugar moiety. The enzymes responsible must be located in the haemolymph. Additionally, a putative sulfotransferase forms benzylglucosinolate sulfate in the gut from intact, non-sequestered glucosinolate. The corresponding desulfoglucosinolate sulfates were also detected in faeces after feeding experiments with phenylethylglucosinolate and prop-2-enylglucosinolate, which indicates a similar degradation mechanism for various glucosinolates in the larvae. This is the first report on glucosinolate metabolism of a glucosinolate-sequestering insect species.     

Light and singlet oxygen in plant defense against pathogens: phototoxic phenalenone phytoalexins

Plants defend themselves from pathogen infections or mechanical injury by a number of mechanisms, including the induced biosynthesis of antimicrobial secondary metabolites. These compounds, termed phytoalexins, represent a very economical way to counteract hazard, because the carbon and energy resources are diverted to phytoalexin synthesis only at the early period of attack and only at its site. The occurrence of phenalenone chromophores in phytoalexins of plants originally nonphototoxic suggests that these plants respond to pathogen attacks by biosynthesizing singlet oxygen photosensitizers able to use solar energy for defense. This concept may have implications for the development of novel crop protection strategies.     

Differential Effects of Indole and Aliphatic Glucosinolates on Lepidopteran Herbivores

Glucosinolates are a diverse group of defensive secondary metabolites that is characteristic of the Brassicales. Arabidopsis thaliana (L.) Heynh. (Brassicaceae) lines with mutations that greatly reduce abundance of indole glucosinolates (cyp79B2 cyp79B3), aliphatic glucosinolates (myb28 myb29), or both (cyp79B2 cyp79B3 myb28 myb29) make it possible to test the in vivo defensive function of these two major glucosinolate classes. In experiments with Lepidoptera that are not crucifer-feeding specialists, aliphatic and indole glucosinolates had an additive effect on Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) larval growth, whereas Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) and Manduca sexta (L.) (Lepidoptera: Sphingidae) were affected only by the absence of aliphatic glucosinolates. In the case of two crucifer-feeding specialists, Pieris rapae (L.) (Lepidoptera: Pieridae) and Plutella xylostella (L.) (Lepidoptera: Plutellidae), there were no major changes in larval performance due to decreased aliphatic and/or indole glucosinolate content. Nevertheless, choice tests show that aliphatic and indole glucosinolates act in an additive manner to promote larval feeding of both species and P. rapae oviposition. Together, these results support the hypothesis that a diversity of glucosinolates is required to limit the growth of multiple insect herbivores.     

植物次级代谢产物研究

近年来，人们对植物的次级代谢过程以及代谢产物进行了研究，并取得了较大的进展，研究表明，可以用基因编码的生物合成酶和基因编码的诱变蛋白来改变不同的合成路径，而且还使用抗敏抑制竞争性路径，依此来提高目标次级代谢产物的产率^[1]。     

Phytotoxin Production and Phytoalexin Elicitation by the Phytopathogenic Fungus Sclerotinia sclerotiorum

The fungus Sclerotinia sclerotiorum (Lib.) de Bary causes rot disease in a vast range of plant families, including Cruciferae (Brassicaceae). We investigated the production of phytotoxins by S. sclerotiorum by using a bioassay-guided isolation, as well as the phytoalexins produced by the resistant wild crucifer Erucastrum gallicum under elicitation by S. sclerotiorum and other agents. We established for the first time that S. sclerotiorum produces a somewhat selective phytotoxin, sclerin, which is phytotoxic to three cruciferous species (Brassica napus, B. juncea, and Sinapis alba) susceptible to Sclerotinia stem rot disease, causing severe necrosis and chlorosis, but not to a resistant species (Erucastrum gallicum). In addition, we have shown that oleic acid, the major fatty acid isolated from sclerotia of S. sclerotiorum is responsible for the toxic activity of extracts of sclerotia to brine shrimp larvae (Artemia salina). Phytoalexin elicitation in leaves of E. gallicum led to the isolation of three known phytoalexins: indole-3-acetonitrile, arvelexin, and 1-methoxyspirobrassinin. Considering that resistance of E. gallicum to S. sclerotiorum is potentially transferable to B. rapa, a susceptible canola species, and that arvelexin, and 1-methoxyspirobrassinin are not produced by B. rapa, these phytoalexins may become useful markers for resistance against S. sclerotiorum.     

毛状根生产次生代谢产物的研究进展

近30年来利用毛状根生产植物次生代谢产物的研究不断增加,但毛状根商业化生产还受到限制,其主要原因是对毛状根合成次生代谢产物的影响机制了解得还不细致。本文讨论了物理、化学因素对毛状根生长和次生代谢产物合成的影响机制,提高毛状根合成代谢物产量的调控技术,以及生物反应器中关键影响因素的研究进展。虽然借助于代谢工程将使得毛状根大量表达次生代谢产物成为可能,但由于毛状根的形态和生理特性,深入研究相应的反应器并提高传质效率将为毛状根法工业化提供可靠依据。     

Plant Antimicrobial Agents and Their Effects on Plant and Human Pathogens

To protect themselves, plants accumulate an armoury of antimicrobial secondary metabolites. Some metabolites represent constitutive chemical barriers to microbial attack (phytoanticipins) and others inducible antimicrobials (phytoalexins). They are extensively studied as promising plant and human disease-controlling agents. This review discusses the bioactivity of several phytoalexins and phytoanticipins defending plants against fungal and bacterial aggressors and those with antibacterial activities against pathogens affecting humans such as Pseudomonas aeruginosa and Staphylococcus aureus involved in respiratory infections of cystic fibrosis patients. The utility of plant products as “antibiotic potentiators” and “virulence attenuators” is also described as well as some biotechnological applications in phytoprotection.     

Antinutritive Compounds in Different Oilseeds

Different oilseeds, usable as renewable resources, are analyzed for their content of condensed tannins, glucosinolates, phytic acid and its degradation products as well as for sinapine, and the results are presented. Seeds of sunflower, flax, coriander, Euphorbia lagascae, iberic. dragon head, safflor, Camelina sativa, soy, mustard, hemp and rapeseeds are used. Apart from hemp, coriander and Euphorbia lagascae different cultivars or genotypes of each species are investigated. The variation of the results of each cultivar or genotype is shown. Tannins and phytic acid are found in different amounts in all oilseeds, whereas sinapine and glucosinolates are limited to seeds from species belonging to the plant family Brassicaceae.     

Effects of Allelochemicals from First (Brassicaceae) and Second (Myzus persicae and Brevicoryne brassicae) Trophic Levels on Adalia bipunctata

Three Brassicaceae species, Brassica napus (low glucosinolate content), Brassica nigra (including sinigrin), and Sinapis alba (including sinalbin) were used as host plants for two aphid species: the generalist Myzus persicae and the specialist Brevicoryne brassicae. Each combination of aphid species and prey host plant was used to feed the polyphagous ladybird beetle, Adalia bipunctata. Experiments with Brassicaceae species including different amounts and kinds of glucosinolates (GLS) showed increased ladybird larval mortality at higher GLS concentrations. When reared on plants with higher GLS concentrations, the specialist aphid, B. brassicae, was found to be more toxic than M. persicae. Identification of GLS and related degradation products, mainly isothiocyanates (ITC), was investigated in the first two trophic levels, plant and aphid species, by high-performance liquid chromatography and gas chromatography–mass spectrometry, respectively. While only GLS were detected in M. persicae on each Brassicaceae species, high amounts of ITC were identified in B. brassicae samples (allyl-ITC and benzyl-ITC from B. nigra and S. alba, respectively) from all host plants. Biological effects of allelochemicals from plants on predators through aphid prey are discussed in relation to aphid species to emphasize the role of the crop plant in integrated pest management in terms of biological control efficacy.