Phylloplane microfungal metabolite influences activity of RuBisCO

The phylloplane microbes are known to affect systemic acquired resistance in plants against a broad spectrum of pathogens. The influence of phylloplane microbes and their metabolites on physiology and biochemistry of plants is well-documented but their effect on the physiology and functioning of chloroplasts and mitochondria is not well understood. The present study was conducted to understand the impact of metabolite of a dominant phylloplane microfungi (Fusarium oxysporum) isolated from the phylloplane of Lycopersicon esculentum on activity of RuBisCO. Activity of RuBisCO (EC 4.1.1.39) in chloroplasts of fungal metabolite treated and untreated eight-week old tomato plants were studied. Significant increase in the activity of RuBisCO (P???0.01), isolated from plants inoculated with the fungal metabolite was observed. The fungal metabolite could elicit the enzyme activity by about two folds (P???0.05) within 72–96?h of treatment of tomato plants. The result demonstrates that probably phylloplane microfungi and/or their metabolites influence the photosynthetic efficiency of plants.     

The effect of rhizosphere microbes outweighs host plant genetics in reducing insect herbivory

Rhizosphere microbes affect plant performance, including plant resistance against insect herbivores; yet, a direct comparison of the relative influence of rhizosphere microbes versus plant genetics on herbivory levels and on metabolites related to defence is lacking. In the crucifer Boechera stricta, we tested the effects of rhizosphere microbes and plant population on herbivore resistance, the primary metabolome, and select secondary metabolites. Plant populations differed significantly in the concentrations of six glucosinolates (GLS), secondary metabolites known to provide herbivore resistance in the Brassicaceae. The population with lower GLS levels experienced ~60% higher levels of aphid (Myzus persicae) attack; no association was observed between GLS and damage by a second herbivore, flea beetles (Phyllotreta cruciferae). Rhizosphere microbiome (disrupted vs. intact native microbiome) had no effect on plant GLS concentrations. However, aphid number and flea beetle damage were respectively about three‐ and seven‐fold higher among plants grown in the disrupted versus intact native microbiome treatment. These differences may be attributable to shifts in primary metabolic pathways previously implicated in host defence against herbivores, including increases in pentose and glucoronate interconversion among plants grown with an intact microbiome. Furthermore, native microbiomes with distinct community composition (as estimated from 16s rRNA amplicon sequencing) differed two‐fold in their effect on host plant susceptibility to aphids. The findings suggest that rhizosphere microbes, including distinct native microbiomes, can play a greater role than population in defence against insect herbivores, and act through metabolic mechanisms independent of population.     

In silico,in vitro and in vivo approach in understanding the functional relationship between ergosterol and Rubisco

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) is one of the key enzymes involved in assimilation of CO₂ in chloroplasts. Phylloplane microfungi and their metabolites have been reported to affect the physiology of host plants, particularly, their photosynthesis. However, information is lacking on the effect of these microflora on the physiology of chloroplasts. The current study emphasized the impact of two dominant phylloplane fungi, Aspergillus niger and Fusarium oxysporum, on activity of Rubisco in tomato chloroplasts. Ergosterol, which is a component of only fungal cell membranes and is not synthesized by plants, have been demonstrated to elicit activity of Rubisco. In the present study, it was demonstrated through in silico, in vitro, and in vivo approaches. Results demonstrated that the fungal metabolites, which contained ergosterol, could double Rubisco activity. Maximum carboxylation rate of Rubisco increased also in ergosterol-treated plants. Michaelis-Menten constant of Rubisco was also slightly affected. Ergosterol was found also to influence and enhance the binding of CO₂ and ribulose-1,5-bisphosphate to Rubisco. Therefore we can postulate that the physiology of the chloroplast is probably influenced by phylloplane microfungi.     

Ecochemical Studies of Interrelationships between Epiphytic Bacteria and Host Plants via Secondary Metabolites

The plant surface, which is representative of the phylloplane and rhizoplane, is a characteristic habitat for microorganisms. In this review, the ecological roles of phytoepiphytic bacteria will be described. The phylloplane and rhizoplane, which are adjacent to the atmosphere and soil sphere respectively, accumulate topically and/or selectively release secondary metabolites that are specific to the plant genera and species which reside within these regions. Some epiphytes have abilities to decarboxylate xenobiotic phenolic acids that have accumulated in the plant tissues and surfaces as a majority of such secondary metabolites. In physicochemically stressed soil, rhizosphere microflora often remedy such microenvironments within the rhizosphere in order to assist in the survival of the host, and some of the microfloral compositions behave as if they were symbionts. Specifically, some Sphingomonas spp., which are frequently isolated from the rhizosphere of acidic soil-tolerant plants in tropical zones, make possible the development of a rhizo-biocomplex. In this review, the possibility of rhizosphere regulation utilizing such a rhizo-biocomplex is discussed.     

Monitoring of the chitinolytic microbial complex of the phylloplane

A comprehensive study of chitinolytic microbial complexes of the phylloplane from cultured and forest plants has been conducted. An increase of the number and biomass of metabolically active cells of the representatives of the domain Bacteria and a decrease in fungal biomass in the experimental microcosms have been shown to occur after the introduction of chitin. The characteristic features of the taxonomic structure of metabolically active chitinolytic complexes of the phylloplane of the plants studied have been elucidated. Representatives of the phyla Proteobacteria, Bacteroidetes, and Verrucomicrobia have been shown to play important roles in the chitinolytic complexes of green leaf samples, while mycelial actinobacteria of the phylum Actinobacyteria played a similar role in needles of coniferous trees. A collection of chitinolytic microorganism cultures isolated from the phylloplane of different plant species has been created.     

Importance of plant‐ and microbe‐driven metabolic pathways for plant defence

Expression of plant phenotypes can depend on both plant genomes and interactions between plants and the microbes living in, on and near their roots. We understand a growing number of the mechanistic links between plant genotypes and phenotypes, such as defence against herbivory (see brief review in Hubbard et al., 2019), yet the links between root microbiomes and the comprehensive swathe of plant phenotypes they affect (Friesen et al., 2011) remain less clear. In this issue of Molecular Ecology, Hubbard et al. (2019) follow microbe‐ and plant‐driven changes in plant defence against hervibory from molecular underpinnings to ecological consequences, contrasting both the metabolites affected and the magnitude of defensive impact. Naively, we might expect plant genomes to drive more variation in phenotype than the root microbiome, but Hubbard et al. (2019) find the opposite, implying profound consequences for plant trait evolution and ecological interactions.     

Systematic analysis of rice (Oryza sativa) metabolic responses to herbivory

Plants defend against attack from herbivores by direct and indirect defence mechanisms mediated by the accumulation of phytoalexins and release of volatile signals, respectively. While the defensive arsenals of some plants, such as tobacco and Arabidopsis are well known, most of rice's (Oryza sativa) defence metabolites and their effectiveness against herbivores remain uncharacterized. Here, we used a non‐biassed metabolomics approach to identify many novel herbivory‐regulated metabolic signatures in rice. Most were up‐regulated by herbivore attack while only a few were suppressed. Two of the most prominent up‐regulated signatures were characterized as phenolamides (PAs), p‐coumaroylputrescine and feruloylputrescine. PAs accumulated in response to attack by both chewing insects, i.e. feeding of the lawn armyworm (Spodoptera mauritia) and the rice skipper (Parnara guttata) larvae, and the attack of the sucking insect, the brown planthopper (Nilaparvata lugens, BPH). In bioassays, BPH insects feeding on 15% sugar solution containing p‐coumaroylputrescine or feruloylputrescine, at concentrations similar to those elicited by heavy BPH attack in rice, had a higher mortality compared to those feeding on sugar diet alone. Our results highlight PAs as a rapidly expanding new group of plant defence metabolites that are elicited by herbivore attack, and deter herbivores in rice and other plants.     

UVB radiation and 17‐hydroxygeranyllinalool diterpene glycosides provide durable resistance against mirid (Tupiocoris notatus) attack in field‐grown Nicotiana attenuata plants

Depending on geographical location, plants are exposed to variable amounts of UVB radiation and herbivore attack. Because the role(s) of UVB in the priming and/or accumulation of plant defence metabolites against herbivores are not well understood, we used field‐grown Nicotiana attenuata plants to explore the effects of UVB on herbivore performance. Consistent with previous reports, UVB‐exposed plants accumulated higher levels of ultraviolet (UV)‐absorbing compounds (rutin, chlorogenic acid, crypto‐chlorogenic acid and dicaffeoylspermidine). Furthermore, UVB increased the accumulation of jasmonic acid, jasmonoyl‐L‐isoleucine and abscisic acid, all phytohormones which regulate plant defence against biotic and abiotic stress. In herbivore bioassays, N. attenuata plants experimentally protected from UVB were more infested by mirids in three consecutive field seasons. Among defence metabolites measured, 17‐hydroxygeranyllinalool diterpene glycosides (HGL‐DTGs) showed strongly altered accumulation patterns. While constitutive HGL‐DTGs levels were higher under UVB, N. attenuata plants exposed to mirid bugs (Tupiocoris notatus) had still more HGL‐DTGs under UVB, and mirids preferred to feed on HGL‐DTGs‐silenced plants when other UVB protecting factors were eliminated by UVB filters. We conclude that UVB exposure not only stimulates UV protective screens but also affects plant defence mechanisms, such as HGL‐DTGs accumulation, and modulates ecological interactions of N. attenuata with its herbivores in nature.     

Prevention of Bacterial Wilt of Grasses by Phylloplane Bacteria

Various chromogenic bacteria were isolated from the mowing site in natural and sown meadows and in pastures. Their ability to protect young ryegrass plants from wilt by simultaneous inoculation with Xanthomonas campestris pv. graminis was tested under low concentrations of the parasite inoculum. From about 200 isolates tested 35% had a very high protective effect, 45% a moderate one, and only 20% had no significant effect. Mixtures of different species of phylloplane bacteria usually gave very good protection. Most of these bacteria live between the leaf-sheaths. In vitro antagonism of the phylloplane bacteria was not related to their protective effect. Avirulent mutant strains of X. c. pv. graminis and pathovars, of other grass species, e.g. pv. poae, pv. phlei, and pv. arrhenatheri, have the same protecting effect. Heat killed bacteria were no longer effective. These observations suggest an unspecific reaction of the plant, such as the induction of a general defense mechanism which results in resistance also to the parasite.     

Bacterial diversity in the rhizosphere of maize and the surrounding carbonate‐rich bulk soil

Maize represents one of the main cultivar for food and energy and crop yields are influenced by soil physicochemical and climatic conditions. To study how maize plants influence soil microbes we have examined microbial communities that colonize maize plants grown in carbonate‐rich soil (pH 8.5) using culture‐independent, PCR‐based methods. We observed a low proportion of unclassified bacteria in this soil whether it was planted or unplanted. Our results indicate that a higher complexity of the bacterial community is present in bulk soil with microbes from nine phyla, while in the rhizosphere microbes from only six phyla were found. The predominant microbes in bulk soil were bacteria of the phyla Acidobacteria, Bacteroidetes and Proteobacteria, while Gammaproteobacteria of the genera Pseudomonas and Lysobacter were the predominant in the rhizosphere. As Gammaproteobacteria respond chemotactically to exudates and are efficient in the utilization of plants exudate products, microbial communities associated to the rhizosphere seem to be plant‐driven. It should be noted that Gammaproteobacteria made available inorganic nutrients to the plants favouring plant growth and then the benefit of the interaction is common.     

Sekundärstoffe – die Geheimwaffen der Pflanzen

Secondary metabolites Already 400 million years ago when land plants evolved, they probably produced secondary metabolites as means of defence against herbivores, microbes and competing plants. Secondary metabolites usually are bioactive agents, which can interfere with molecular targets in animals and microbes. Therefore, many plants and substances isolated from them can serve as valuable drugs in medicine and pharmacy. Some secondary metabolites also serve as signal compounds to attract pollinating animals and seed‐dispersing animals, but also for UV protection, as antioxidants or mobile nitrogen stores. Biology and evolution but also physiological and genetic bases of secondary metabolism are discussed in this overview.     

Tryptophan‐derived secondary metabolites in Arabidopsis thaliana confer non‐host resistance to necrotrophic Plectosphaerella cucumerina fungi

A defence pathway contributing to non‐host resistance to biotrophic fungi in Arabidopsis involves the synthesis and targeted delivery of the tryptophan (trp)‐derived metabolites indol glucosinolates (IGs) and camalexin at pathogen contact sites. We have examined whether these metabolites are also rate‐limiting for colonization by necrotrophic fungi. Inoculation of Arabidopsis with adapted or non‐adapted isolates of the ascomycete Plectosphaerella cucumerina triggers the accumulation of trp‐derived metabolites. We found that their depletion in cyp79B2 cyp79B3 mutants renders Arabidopsis fully susceptible to each of three tested non‐adapted P. cucumerina isolates, and super‐susceptible to an adapted P. cucumerina isolate. This assigns a key role to trp‐derived secondary metabolites in limiting the growth of both non‐adapted and adapted necrotrophic fungi. However, 4‐methoxy‐indol‐3‐ylmethylglucosinolate, which is generated by the P450 monooxygenase CYP81F2, and hydrolyzed by PEN2 myrosinase, together with the antimicrobial camalexin play a minor role in restricting the growth of the non‐adapted necrotrophs. This contrasts with a major role of these two trp‐derived phytochemicals in limiting invasive growth of non‐adapted biotrophic powdery mildew fungi, thereby implying the existence of other unknown trp‐derived metabolites in resistance responses to non‐adapted necrotrophic P. cucumerina. Impaired defence to non‐adapted P. cucumerina, but not to the non‐adapted biotrophic fungus Erysiphe pisi, on cyp79B2 cyp79B3 plants is largely restored in the irx1 background, which shows a constitutive accumulation of antimicrobial peptides. Our findings imply differential contributions of antimicrobials in non‐host resistance to necrotrophic and biotrophic pathogens.     

中国北部湾光裸方格星虫可培养微生物的分离鉴定及其活性代谢物产生菌筛选

【目的】本研究从北部湾海域光裸方格星虫(Sipunculus nudus)肠道中分离鉴定可培养微生物,并对筛选菌株的代谢物活性进行研究,为后续开发和利用光裸方格星虫肠道微生物代谢产物提供理论支持。【方法】通过微生物培养、菌株分离纯化和16S rRNA基因序列分析,分析鉴定湛江、北海、防城港三地光裸方格星虫肠道可培养微生物;采用透明圈法、可见分光光度法、平板打孔法等对产胞外活性代谢物的菌株进行筛选和活性分析。【结果】中国北部湾不同海域光裸方格星虫肠道可培养微生物包括弧菌属(Vibrio)、希瓦氏菌属(Shewanella)、假交替单胞菌属(Pseudoalteromonas)、发光杆菌属(Photobacterium)和芽孢杆菌属(Bacillus)等12个细菌属。弧菌属(Vibrio)是3个地区样本共有的优势菌群。具有产胞外水解蛋白酶、壳聚糖酶、多糖以及抑菌活性等能力的菌株主要来自假交替单胞菌属(Pseudoalteromonas)、发光杆菌属(Photobacterium)和芽孢杆菌属(Bacillus)。【结论】中国北部湾不同海域光裸方格星虫肠道可培养微生物在属的种类上存在显著性差异,且光裸方格星虫肠道菌株具有产生多种胞外活性代谢物的能力,是一种良好的海洋活性代谢物来源。     

Glucosinolates in oilseed rape: secondary metabolites that influence interactions with herbivores and their natural enemies

Glucosinolates are sulphur‐containing secondary metabolites characteristic of Brassicaceous plants. Glucosinolate breakdown products, which include isothiocyanates, are released following tissue damage when hydrolytic enzymes act on them. The isothiocyanates have toxic effects on generalist herbivores when they attempt to feed on oilseed rape, Brassica napus, and also function as repellents. However, specialist herbivores such as Brevicoryne brassicae aphids, flea beetles, Psylliodes chrysocephala and the Lepidopteran pest, Pieris rapae, are adapted to the presence of glucosinolates and thrive on plants containing them. They may do this by avoiding tissue damage to prevent the formation of isothiocyanates or by metabolising or tolerating glucosinolates. For many specialist herbivores, the isothiocyanates function as attractants and glucosinolates can even be sequestered for defence against predatory insects. Thus, these herbivores have evolved resistance to host‐plant secondary metabolites and this type of evolutionary history may have given some insects an enhanced ability to adapt to xenobiotics. In an agricultural context, this may make pests better able to evolve resistance to artificially applied pesticides. The effect of increased glucosinolate content in making oilseed rape cultivars more susceptible to specialist pests was highlighted in a seminal article in the Annals of Applied Biology in 1995. This review of the literature considers developments in this area since then.     

Rhizosphere microbes facilitate redox homeostasis in Cicer arietinum against biotic stress

The aim of this study was to evaluate the potentiality of three compatible rhizosphere microbes, viz. fluorescent Pseudomonas aeruginosa (PHU094), Trichoderma harzianum (THU0816) and Mesorhizobium sp. (RL091), in community to mobilise antioxidant mechanisms in chickpea under the challenge of Sclerotium rolfsii. The microbes were applied as seed treatment in different combinations in two sets and the pathogen was inoculated in one of the sets after 3 weeks of sowing. A comparative study was conducted on the effect of the microbial combinations on host antioxidant mechanisms between the two sets. In pathogen challenged plants host defence responses included higher accumulation of hydrogen peroxide (H₂O₂) at petiolar and interveinal regions of leaf and high activities of catalase (CAT), glutathione reductase (GR) and guaiacol peroxidase (GPx) compared to unchallenged plants. The antioxidant enzyme activities increased 1.8‐3.3 and 1.9‐3.1 folds at 48 and 72 h, respectively, in the triple microbe treated challenged plants compared to unchallenged ones. Although, ascorbate peroxidase (APX) activity was significantly low, ascorbic acid (AA) and chitinase accumulation was high in the pathogen challenged plants. Antioxidant flavonols associated with host defence namely myricetin, quercetin and kaempferol also accumulated in high amounts in pathogen challenged plants. Among the microbial treatments, the triple microbe combination induced the highest response in all parameters as compared to dual or single application of the same microbes. The triple microbe consortium modulated the chickpea antioxidant mechanisms more efficiently and modulation of oxidative stress was directly correlated with lower plant mortality, thus demonstrating the synergistic behaviour of the microbes in protecting chickpea from the pathogen.     

Dual herbivore attack and herbivore density affect metabolic profiles of Brassica nigra leaves

Plant responses to dual herbivore attack are increasingly studied, but effects on the metabolome have largely been restricted to volatile metabolites and defence‐related non‐volatile metabolites. However, plants subjected to stress, such as herbivory, undergo major changes in both primary and secondary metabolism. Using a naturally occurring system, we investigated metabolome‐wide effects of single or dual herbivory on Brassica nigra plants by Brevicoryne brassicae aphids and Pieris brassicae caterpillars, while also considering the effect of aphid density. Metabolomic analysis of leaf material showed that single and dual herbivory had strong effects on the plant metabolome, with caterpillar feeding having the strongest influence. Additionally, aphid‐density‐dependent effects were found in both the single and dual infestation scenarios. Multivariate analysis revealed treatment‐specific metabolomic profiles, and effects were largely driven by alterations in the glucosinolate and sugar pools. Our work shows that analysing the plant metabolome as a single entity rather than as individual metabolites provides new insights into the subcellular processes underlying plant defence against multiple herbivore attackers. These processes appear to be importantly influenced by insect density.     

Microbes in Helicoverpa armigera oral secretions contribute to increased senescence around plant wounds

1. Plants have long been exposed to insect herbivore attack. Crucial to the plant's ability to defend itself is its ability to identify specific signals associated with attacking insects. Distinctive chemical cues, such as those associated with chewing insect oral secretions (OS), activate targeted defence responses to chewing insect herbivores. 2. Herbivore-associated cues can be complicated by the fact that many herbivores form associations with microbes that produce their own specific signals, which may induce alternative defence processes. 3. Here we report that OS of the global pest, the cotton bollworm (Helicoverpa armigera), induce senescence around wounds in Brachypodium distachyon leaves. Crude OS activate greater levels of senescence than OS with reduced microbial abundance or mechanical wounding alone. Nonetheless, plants closed mechanical wounds more rapidly when treated with H. armigera OS regardless of the microbial component. 4. This study concludes that H. armigera OS can activate senescence and wound closure in plant tissues and that microbes within OS have an important role in shaping plant-herbivore interactions through additional increases in senescence.     

Chitinolytic microbes confer Meloidogyne incognita resistance and augment secondary metabolites in Bacopa monnieri (L.) Pennell

Meloidogyne incognita (Kofoid and White) Chitwood a calamitous plant pathogen affects almost all the commercial crops, especially medicinal plants viz. Bacopa monnieri L. In this study, the efficacy of indigenous chitinolytic microbes against M. incognita in B. monnieri was evaluated under sterilised and natural soil conditions. The plant growth was observed for various parameters viz. nutrient uptake, phytochemical activities and secondary metabolites (bacoside). In the natural soil under green house conditions, the isolates significantly (P ≤ 0.05) reduced M. incognita infestation (1.38–2.78-folds) with significant increase in secondary metabolites (1.28–1.88-fold) over the untreated control. The potential strains were identified through 16S-rRNA as Streptomyces sp., Microbacterium arabinogalactanolyticum, Cellulosimicrobium cellulans, Actinomycetales bacterium, Chitiniphilus sp. and Flavobacterium johnsoniae. These findings thus highlight the potential of indigenous chitinolytic microbes that can be utilised for overall fitness of the plant along with the reduced root-knot nematode infestation in B. monnieri.