The cross-pathway control system regulates production of the secondary metabolite toxin, sirodesmin PL, in the ascomycete, Leptosphaeria maculans

<p>Abstract</p> <p>Background</p> <p>Sirodesmin PL is a secondary metabolite toxin made by the ascomycetous plant pathogen, <it>Leptosphaeria maculans</it>. The sirodesmin biosynthetic genes are clustered in the genome. The key genes are a non-ribosomal peptide synthetase, <it>sirP</it>, and a pathway-specific transcription factor, <it>sirZ</it>. Little is known about regulation of sirodesmin production.</p> <p>Results</p> <p>Genes involved in regulation of sirodesmin PL in <it>L. maculans </it>have been identified. Two hundred random insertional T-DNA mutants were screened with an antibacterial assay for ones producing low levels of sirodesmin PL. Three such mutants were isolated and each transcribed <it>sirZ </it>at very low levels. One of the affected genes had high sequence similarity to <it>Aspergillus fumigatus cpcA</it>, which regulates the cross-pathway control system in response to amino acid availability. This gene was silenced in <it>L. maculans </it>and the resultant mutant characterised. When amino acid starvation was artificially-induced by addition of 3-aminotriazole for 5 h, transcript levels of <it>sirP </it>and <it>sirZ </it>did not change in the wild type. In contrast, levels of <it>sirP </it>and <it>sirZ </it>transcripts increased in the silenced <it>cpcA </it>mutant. After prolonged amino acid starvation the silenced <it>cpcA </it>mutant produced much higher amounts of sirodesmin PL than the wild type.</p> <p>Conclusions</p> <p>Production of sirodesmin PL in <it>L. maculans </it>is regulated by the cross pathway control gene, <it>cpcA</it>, either directly or indirectly via the pathway-specific transcription factor, <it>sirZ</it>.</p

Digitalna procjena lisne površine krošnje stijenke vinove loze (Vitis vinifera cv. Sauvignon) korištenjem LIDAR mjerne tehnologije

A dosage rate reduction of plant protection products mixed with water, i.e. spray mixture, in a prescribed concentration in the vineyard will only be possible in the future, if the natural characteristics of vine canopy structures (leaf wall area) and canopy management are taken into account. In a practical experiment in the vineyard we evaluated the leaf wall area of the vine cv. Sauvignon on different segments on the left and right side of the vine canopy. We compared the results of manual measurements and laser measuring technology (LIDAR) with the corresponding algorithm, with which we enabled the digital reconstruction of the leaf wall area of the vine. The manual measurement of the leaf wall area was carried out using an automated image analyser. The digital system for measuring the leaf wall area on different segments consisted of a LIDAR sensor and a Differential Global Positioning System (hereinafter DGPS). To determine the exact DGPS position of the LIDAR sensor during the measurement, we set up a DGPS base station. Using the Excel software (CORREL function), we estimated the relationship between the dependent variable (digital number of points in the cloud) and an independent variable (leaf wall area, manually measured). An analysis of six randomly selected vines in the vineyard revealed the maximum value of the correlation coefficient r = 0.80 for the left side and r = 0.90 for the right side of the leaf wall area of the vine, respectively. In the near future the virtual three-dimensional space will provide more even control of spray mixture over the entire structure of the leaf wall area in the vineyard based on autonomous decision-making models.Smanjenje količine utroška sredstava za zaštitu bilja i same smjese za prskanje u budućnosti će biti moguće samo ako se uzmu u obzir prirodne karakteristike krošnje vinove loze tj. lisne površine krošnje trsa. U praktičnom pokusu u vinogradu procijenjena je lisna površinu krošnje vinove loze cv. Sauvignon na različitim segmentima s lijeve i desne strane krošnje uz pomoć ručnih mjerenja i laserske mjerne tehnologije (LIDAR). Dobiveni rezultati uspoređeni su s pripadajućim algoritmom čime je dobivena digitalna rekonstrukcija lisne površine vinove loze. Ručno mjerenje površine listova provedeno je u laboratoriju pomoću digitalnog lisnog skenera nakon što je lišće ručno pobrano s trsova i dopremljeno u sam laboratorij. Digitalni sustav za mjerenje lisne površine na različitim segmentima krošnje sastojao se od LIDAR senzora i DGPS navigacijskog sustava. Da bi se odredio točan DGPS položaj LIDAR senzora tijekom mjerenja, postavljena je DGPS bazna stanica. Pomoću regresijske metode utvrđen je odnos između zavisne varijable (digitalni broj točaka u oblaku) i nezavisne varijable (površina listova izmjerena skenerom). Rezultati analize imeđu dvije uspoređivane metode na šest slučajno odabranih trsova vinove loze otkrivaju vrijednost koeficijenta korelacije r = 0,80 za lijevu i r = 0,90 za desnu stranu krošnje. U bliskoj budućnosti virtualni trodimenzionalni prostor pružit će ravnomjerniju kontrolu smjese raspršivača preko cijele strukture područja stijenke lišča u vinogradu na temelju autonomnih modela odlučivanja

Origin and distribution of epipolythiodioxopiperazine (ETP) gene clusters in filamentous ascomycetes

<p>Abstract</p> <p>Background</p> <p>Genes responsible for biosynthesis of fungal secondary metabolites are usually tightly clustered in the genome and co-regulated with metabolite production. Epipolythiodioxopiperazines (ETPs) are a class of secondary metabolite toxins produced by disparate ascomycete fungi and implicated in several animal and plant diseases. Gene clusters responsible for their production have previously been defined in only two fungi. Fungal genome sequence data have been surveyed for the presence of putative ETP clusters and cluster data have been generated from several fungal taxa where genome sequences are not available. Phylogenetic analysis of cluster genes has been used to investigate the assembly and heredity of these gene clusters.</p> <p>Results</p> <p>Putative ETP gene clusters are present in 14 ascomycete taxa, but absent in numerous other ascomycetes examined. These clusters are discontinuously distributed in ascomycete lineages. Gene content is not absolutely fixed, however, common genes are identified and phylogenies of six of these are separately inferred. In each phylogeny almost all cluster genes form monophyletic clades with non-cluster fungal paralogues being the nearest outgroups. This relatedness of cluster genes suggests that a progenitor ETP gene cluster assembled within an ancestral taxon. Within each of the cluster clades, the cluster genes group together in consistent subclades, however, these relationships do not always reflect the phylogeny of ascomycetes. Micro-synteny of several of the genes within the clusters provides further support for these subclades.</p> <p>Conclusion</p> <p>ETP gene clusters appear to have a single origin and have been inherited relatively intact rather than assembling independently in the different ascomycete lineages. This progenitor cluster has given rise to a small number of distinct phylogenetic classes of clusters that are represented in a discontinuous pattern throughout ascomycetes. The disjunct heredity of these clusters is discussed with consideration to multiple instances of independent cluster loss and lateral transfer of gene clusters between lineages.</p

Potential effects of oilseed rape expressing oryzacystatin-1 (OC-1) and of purified insecticidal proteins on larvae of the solitary bee Osmia bicornis

Despite their importance as pollinators in crops and wild plants, solitary bees have not previously been included in non-target testing of insect-resistant transgenic crop plants. Larvae of many solitary bees feed almost exclusively on pollen and thus could be highly exposed to transgene products expressed in the pollen. The potential effects of pollen from oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) were investigated on larvae of the solitary bee Osmia bicornis (= O. rufa). Furthermore, recombinant OC-1 (rOC-1), the Bt toxin Cry1Ab and the snowdrop lectin Galanthus nivalis agglutinin (GNA) were evaluated for effects on the life history parameters of this important pollinator. Pollen provisions from transgenic OC-1 oilseed rape did not affect overall development. Similarly, high doses of rOC-1 and Cry1Ab as well as a low dose of GNA failed to cause any significant effects. However, a high dose of GNA (0.1%) in the larval diet resulted in significantly increased development time and reduced efficiency in conversion of pollen food into larval body weight. Our results suggest that OC-1 and Cry1Ab expressing transgenic crops would pose a negligible risk for O. bicornis larvae, whereas GNA expressing plants could cause detrimental effects, but only if bees were exposed to high levels of the protein. The described bioassay with bee brood is not only suitable for early tier non-target tests of transgenic plants, but also has broader applicability to other crop protection products

Genome analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea

Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38–39 Mb genomes include 11,860–14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared t

A global-scale expert assessment of drivers and risks associated with pollinator decline.

Pollinator decline has attracted global attention and substantial efforts are underway to respond through national pollinator strategies and action plans. These policy responses require clarity on what is driving pollinator decline and what risks it generates for society in different parts of the world. Using a formal expert elicitation process, we evaluated the relative regional and global importance of eight drivers of pollinator decline and ten consequent risks to human well-being. Our results indicate that global policy responses should focus on reducing pressure from changes in land cover and configuration, land management and pesticides, as these were considered very important drivers in most regions. We quantify how the importance of drivers and risks from pollinator decline, differ among regions. For example, losing access to managed pollinators was considered a serious risk only for people in North America, whereas yield instability in pollinator-dependent crops was classed as a serious or high risk in four regions but only a moderate risk in Europe and North America. Overall, perceived risks were substantially higher in the Global South. Despite extensive research on pollinator decline, our analysis reveals considerable scientific uncertainty about what this means for human society.University of Reading’s Building Outstanding Impact Support Programm

Distinct Roles of Jasmonates and Aldehydes in Plant-Defense Responses

BACKGROUND: Many inducible plant-defense responses are activated by jasmonates (JAs), C(6)-aldehydes, and their corresponding derivatives, produced by the two main competing branches of the oxylipin pathway, the allene oxide synthase (AOS) and hydroperoxide lyase (HPL) branches, respectively. In addition to competition for substrates, these branch-pathway-derived metabolites have substantial overlap in regulation of gene expression. Past experiments to define the role of C(6)-aldehydes in plant defense responses were biased towards the exogenous application of the synthetic metabolites or the use of genetic manipulation of HPL expression levels in plant genotypes with intact ability to produce the competing AOS-derived metabolites. To uncouple the roles of the C(6)-aldehydes and jasmonates in mediating direct and indirect plant-defense responses, we generated Arabidopsis genotypes lacking either one or both of these metabolites. These genotypes were subsequently challenged with a phloem-feeding insect (aphids: Myzus persicae), an insect herbivore (leafminers: Liriomyza trifolii), and two different necrotrophic fungal pathogens (Botrytis cinerea and Alternaria brassicicola). We also characterized the volatiles emitted by these plants upon aphid infestation or mechanical wounding and identified hexenyl acetate as the predominant compound in these volatile blends. Subsequently, we examined the signaling role of this compound in attracting the parasitoid wasp (Aphidius colemani), a natural enemy of aphids. PRINCIPAL FINDINGS: This study conclusively establishes that jasmonates and C(6)-aldehydes play distinct roles in plant defense responses. The jasmonates are indispensable metabolites in mediating the activation of direct plant-defense responses, whereas the C(6)-aldehyes are not. On the other hand, hexenyl acetate, an acetylated C(6)-aldehyde, is the predominant wound-inducible volatile signal that mediates indirect defense responses by directing tritrophic (plant-herbivore-natural enemy) interactions. SIGNIFICANCE: The data suggest that jasmonates and hexenyl acetate play distinct roles in mediating direct and indirect plant-defense responses. The potential advantage of this "division of labor" is to ensure the most effective defense strategy that minimizes incurred damages at a reduced metabolic cost

Insertion of an Esterase Gene into a Specific Locust Pathogen (Metarhizium acridum) Enables It to Infect Caterpillars

An enduring theme in pathogenic microbiology is poor understanding of the mechanisms of host specificity. Metarhizium is a cosmopolitan genus of invertebrate pathogens that contains generalist species with broad host ranges such as M. robertsii (formerly known as M. anisopliae var. anisopliae) as well as specialists such as the acridid-specific grasshopper pathogen M. acridum. During growth on caterpillar (Manduca sexta) cuticle, M. robertsii up-regulates a gene (Mest1) that is absent in M. acridum and most other fungi. Disrupting M. robertsii Mest1 reduced virulence and overexpression increased virulence to caterpillars (Galleria mellonella and M. sexta), while virulence to grasshoppers (Melanoplus femurrubrum) was unaffected. When Mest1 was transferred to M. acridum under control of its native M. robertsii promoter, the transformants killed and colonized caterpillars in a similar fashion to M. robertsii. MEST1 localized exclusively to lipid droplets in M. robertsii conidia and infection structures was up-regulated during nutrient deprivation and had esterase activity against lipids with short chain fatty acids. The mobilization of stored lipids was delayed in the Mest1 disruptant mutant. Overall, our results suggest that expression of Mest1 allows rapid hydrolysis of stored lipids, and promotes germination and infection structure formation by M. robertsii during nutrient deprivation and invasion, while Mest1 expression in M. acridum broadens its host range by bypassing the regulatory signals found on natural hosts that trigger the mobilization of endogenous nutrient reserves. This study suggests that speciation in an insect pathogen could potentially be driven by host shifts resulting from changes in a single gene

Transcriptional Responses of Arabidopsis thaliana during Wilt Disease Caused by the Soil-Borne Phytopathogenic Bacterium, Ralstonia solanacearum

Bacterial wilt is a common disease that causes severe yield and quality losses in many plants. In the present study, we used the model Ralstonia solanacearum-Arabidopsis thaliana pathosystem to study transcriptional changes associated with wilt disease development. Susceptible Col-5 plants and RRS1-R-containing resistant Nd-1 plants were root-inoculated with R. solanacearum strains harbouring or lacking the matching PopP2 avirulence gene. Gene expression was marginally affected in leaves during the early stages of infection. Major changes in transcript levels took place between 4 and 5 days after pathogen inoculation, at the onset of appearance of wilt symptoms. Up-regulated genes in diseased plants included ABA-, senescence- and basal resistance-associated genes. The influence of the plant genetic background on disease-associated gene expression is weak although some genes appeared to be specifically up-regulated in Nd-1 plants. Inactivation of some disease-associated genes led to alterations in the plant responses to a virulent strain of the pathogen. In contrast to other pathosystems, very little overlap in gene expression was detected between the early phases of the resistance response and the late stages of disease development. This observation may be explained by the fact that above-ground tissues were sampled for profiling whereas the bacteria were applied to root tissues