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Encasements formed around haustoria and biotrophic hyphae as well as hypersensitive reaction (HR) cell death are essential plant immune responses to filamentous pathogens. In this study we examine the components that may contribute to the absence of these responses in susceptible barley attacked by the powdery mildew fungus. We find that the effector CSEP0162 from this pathogen targets plant MONENSIN SENSITIVITY1 (MON1), which is important for the fusion of multivesicular bodies to their target membranes. Overexpression of CSEP0162 and silencing of barley MON1 both inhibit encasement formation. We find that the Arabidopsis ecotype No-0 has resistance to powdery mildew, and that this is partially dependent on MON1. Surprisingly, we find the MON1-dependent resistance in No-0 not only includes an encasement response, but also an effective HR. Similarly, silencing of MON1 in barley also blocks Mla3-mediated HR-based powdery mildew resistance. Our results indicate that MON1 is a vital plant immunity component, and we speculate that the barley powdery mildew fungus introduces the effector CSEP0162 to target MON1 and hence reduce encasement formation and HR.

Self-forming double-crosslinked (DC) hydrogels were designed by incorporating enzyme-mimicking metal coordination polymer crosslinks and catechol chemistry. A macromolecular tris-histidine copper complex acted both as part of the hydrogel network and as a catalyst of catechol oxidation to induce a second hydrogel network by covalent crosslinking of catechol functionalized polymers thus giving catalytic control over hydrogel crosslinking.

Excess cellular sterol is harmful in mammals and plants, but the mechanisms why are awaiting clarification. Here we find a strong autoimmune response to be associated to excess endoplasmic reticulum (ER) sterols. This was obtained by studying a plant peroxisome lipase, SSD5, required for the lesion phenotype of the Arabidopsis syntaxin mutant, pen1 syp122. SSD5 is a lipase with a catalytic triad including a GxSxG motif localized to a subdomain of the peroxisome periphery. Lipidomics revealed reduced steryl ester levels in pen1 syp122 when SSD5 is mutated. This involvement in sterol homeostasis was confirmed by a requirement of SSD5 for the lesions of hise1 psat1 that suffers from excess ER sterol. These data suggest SSD5 is contributing to a peroxisome-located segment of the sterol biosynthesis pathway. SSD5’s contribution to the pen1 syp122 autoimmunity is not associated with nine highly diverse down-stream immune components, and SSD5 does not influence general plant disease levels and immunity. Therefore, our data indicated SSD5 as well as ER sterol functions up-stream of immune activation. This in turn suggests plant excess ER sterol to activate one or more immune receptors.

Saffron quality is determined by the content of three apocarotenoids, including crocin, picrocrocin, and safranal, giving saffron culinary, industrial, and medical importance. The level of these secondary metabolites is also affected by environmental factors such as light, which play a pivotal role in regulating plants' signaling pathways. In this study, we investigated the effects of different ratios of blue (B) to red (R) and white (W) light on physiological, biochemical, and molecular responses of saffron in the flowering stage. Flowers' morphological properties were improved in plants grown under monochromatic B light. The highest content of total carotenoids, anthocyanins, and flavonoids was detected in plants grown under a high proportion of B light. The highest crocin content, especially the trans-crocetin ester isomer, was recorded in monochromatic B light-grown plants. However, the highest picrocrocin content was in both monochromatic R- and B light-grown plants. The highest safranal content was detected in plants grown under a high proportion of R light. Transcriptome analysis of secondary metabolism pathways showed that the transcript level of the genes was highly correlated with the content of the target metabolites. Monochromatic B light upregulated the expression of genes involved in crocin production (CsCCD2, CsALDH31l, and CsUGT2). However, the expression of CsUGT709G1, which is involved in picrocrocin and safranal pathways, was upregulated in plants grown under a high ratio of R light and W lights. In conclusion, monochromatic B light enhances the flowering rate, crocin (trans-crocetin ester), and picrocrocin content, which increases the quantity and quality of saffron products under controlled conditions.

Encasements formed around haustoria and biotrophic hyphae as well as hypersensitive reaction (HR) cell death are essential plant immune responses to filamentous pathogens. Here we study a possible reason why these responses are absent in susceptible barley attacked by the powdery mildew fungus. We find that the effector CSEP0162 from this pathogen targets plant MON1, important for fusion of multivesicular bodies to their target membranes. Over-expression of CSEP0162 and silencing of barley MON1 both inhibit encasement formation. We find that the Arabidopsis ecotype No-0 has partial resistance to powdery mildew, and that this is dependent on MON1. Surprisingly, we find the MON1-dependent resistance in No-0 not only include an effective encasement response, but also HR. Similarly, silencing of MON1 in barley also blocked Mla3-mediated HR-based powdery mildew resistance. These data indicate that MON1 is a vital plant immunity component, and we speculate that the barley powdery mildew fungus introduces the effector CSEP0162 to target MON1 and reduce encasement formation and HR.HighlightMON1 is essential for MVB fusion to plasma membrane. We find that MON1 also is important for immunity, and that it is targeted by the barley powdery mildew effector CSEP0162.

Plant disease resistance often occurs upon direct or indirect recognition of pathogen effectors by host nucleotide-binding leucine-rich-repeat (NLR) receptors. The barley powdery mildew fungus,Blumeria graminisf. sp.hordei(Bgh), secretes hundreds of candidate secreted effector proteins (CSEPs) to facilitate pathogen infection and colonization. One of these, CSEP0008, is directly recognized by the barley NLR, MLA1, and therefore designated AVRA1. Here we show that AVRA1and the sequence-unrelatedBgheffector BEC1016 (CSEP0491) suppress immunity in barley cells. We then used yeast 2-hybrid next-generation-interaction screens (Y2H-NGIS), followed by binary Y2H and bimolecular fluorescence complementation, to identify a common barley target of AVRA1and BEC1016, the endoplasmic reticulum (ER)-localized J-domain protein,HvERdj3B. This is an ER protein quality control (ERQC) protein and silencing it increased theBghpenetration rate in barley.HvERdj3B is localized to the ER lumen, and AVRA1and BEC106 translocation into the ER was confirmed using a split GFP system. Together, these results suggest that the barley innate immunity, preventingBghentry into epidermal cells, is dependent on ERQC, which in turn is regulated by J-domain proteinHvERdj3B and the two effectors. Previous work has shown that AVRA1is directly recognized in the cytosol by the immune receptor, MLA1. We speculate whether the AVRA1J-domain target being inside the ER, where it is inapproachable by NLRs, has forced the plant to evolve this challenging direct recognition.

From One Cold War to Another Klaus Carsten Pedersen (Det Udenrigspolitiske Selskab,) reviews Från ett kallt krig till ett annat (From One Cold War to Another) written by Thomas Bertelman.

This article explores the politics behind the promise of ‘blue growth’. Reframing it as a ‘blue fix’, we argue that the blue growth discourse facilitates new opportunities for capital accumulation, while claiming that this accumulation is compatible with social and ecological aims as well. The blue fix is made up of three underlying sub-fixes. First of all, the conservation fix quenches the social thirst for action in the face of climate change. Here we see how protecting marine areas can be an important part of mitigating climate change, but in practice, gains at the national level are overshadowed by the ongoing expansion of offshore drilling for oil and gas. Second, the protein fix satisfies the growing global demand for healthy food and nutrition through the expansion of capital-intensive large-scale aquaculture, while ignoring the negative socio-ecological impacts, which effectively squeeze small-scale capture fishing out, while industrial capture fishing remains well positioned to expand into as well as supply industrial aquaculture with fish feed from pelagic fish. And third, an energy fix offers a burst of wind energy and a splash of new deep-sea minerals without disturbing the familiar and persistent foundation of oil and gas. This dimension of the blue fix emphasizes the transition to wind and solar energy, but meanwhile the deep sea mining for minerals required by these new technologies launches us into unknown ecological territories with little understood consequences. The synergy of these three elements brought together in a reframing of ocean politics manifests as a balancing act to frame blue growth as ‘sustainable’ and in everyone’s interest, which we critically analyze and discuss in this article.

BACKGROUND: Studies typically highlight area level variation in the incidence of non-affective but not affective psychoses. We compared neighbourhood-level variation for both types of disorder, and the specific effects of neighbourhood urbanicity and ethnic density, using Danish national registry data.METHODS: Population based cohort (2,224,464 people) followed from 1980 to 2013 with neighbourhood exposure measured at age 15 and incidence modelled using multilevel Poisson regression.RESULTS: Neighbourhood variation was similar for both disorders with an adjusted median risk ratio of 1.37 (95% CI 1.34-1.39) for non-affective psychosis and 1.43 (1.38-1.49) for affective psychosis. Associations with neighbourhood urbanicity differed: living in the most compared to the least urban quintile at age 15 was associated with a minimal increase in subsequent affective psychosis, IRR 1.13 (1.01-1.27) but a substantial increase in rates of non-affective psychosis, IRR 1.66 (1.57-1.75). Mixed results were found for neighbourhood ethnic density: for Middle Eastern migrants there was an increased average incidence of both affective, IRR 1.54 (1.19-1.99), and non-affective psychoses, 1.13 (1.04-1.23) associated with each decrease in ethnic density quintile, with a similar pattern for African migrants, but for European migrants ethnic density appeared to be associated with non-affective psychosis only.CONCLUSIONS: While overall variation and the effect of neighbourhood ethnic density were similar for both types of disorder, associations with urbanicity were largely confined to non-affective psychosis. This may reflect differences in aetiological pathways although the mechanism behind these differences remains unknown.

Cytosolic GS1 (Gln synthetase) is central for ammonium assimilation in plants. High ammonium treatment enhanced the expression of the GS1 isogene Gln-1;2 encoding a low-affinity high-capacity GS1 protein in Arabidopsis (Arabidopsis thaliana) shoots. Under the same conditions, the expression of the high-affinity low-capacity isoform Gln-1;1 was reduced. The expression of Gln-1;3 did not respond to ammonium treatment while Gln-1;4 and Gln-1;5 isogenes in all cases were expressed at a very low level. Gln-2 was highly expressed in shoots but only at a very low level in roots. To investigate the specific functions of the two isogenes Gln-1;1 and Gln-1;2 in shoots for ammonium detoxification, single and double knock-out mutants were grown under standard N supply or with high ammonium provision. Phenotypes of the single mutant gln1;1 were similar to the wild type, while growth of the gln1;2 single mutant and the gln1;1:gln1;2 double mutant was significantly impaired irrespective of N regime. GS1 activity was significantly reduced in both gln1;2 and gln1;1:gln1;2. Along with this, the ammonium content increased while that of Gln decreased, showing that Gln-1;2 was essential for ammonium assimilation and amino acid synthesis. We conclude that Gln-1;2 is the main isozyme contributing to shoot GS1 activity in vegetative growth stages and can be up-regulated to relieve ammonium toxicity. This reveals, to our knowledge, a novel shoot function of Gln-1;2 in Arabidopsis shoots.

In the fall of 2016, an unusual phenological event occurred in Quercus garryana Douglas ex Hook. in Victoria, British Columbia. After normal autumn leaf drop, some trees burst bud and leafed out prematurely in late October. This allowed a comparison of the cold hardiness of the prematurely flushed and non-flushed trees over the following year. Cold hardiness of five tree pairs (premature fall flush and non-flush) in three locations in Victoria was assessed bi-weekly over the dehardening period in January–March 2017 and again over the hardening period in September–December 2017. Cold hardiness of 10 non-flushed trees from the most northerly population of Q. garryana was also assessed twice in spring 2017. Between January and March, all trees dehardened, but cold hardiness was greatest in non-flushed trees on the first sampling date, and thereafter, the non-flushed trees dehardened more rapidly than the prematurely flushed trees. Index of injury was consistently 10% greater in Victoria than in northern trees. In fall 2017, trees that had flushed prematurely in fall 2016 had the same cold hardiness as non-flushed trees. Hardiness of all trees decreased from mid-September to the end of October, followed by rapid hardening in November and December of 2017.

This chapter looks at what may be deduced about the possible military missions of the armed forces of the Kaliningrad Special Defence Area (KOOR) simply on the basis of their size and composition. It explores how relevant and realistic these missions may be under various conditions. The chapter aims to place Kaliningrad and the KOOR forces in the general context of Russian security policy and foreign relations. KOOR was formed in 1994 as a unique formation in the Russian defence structure. The academic analyses of alternative sovereignty arrangements for Kaliningrad therefore seem redundant - and potentially almost as provocative as the territorial claims by some political extremists. From a military point of view the location of Kaliningrad offers opportunities and presents problems - different sets of opportunities and problems depending on who is looking at them and depending on the situation.

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The barley powdery mildew fungus, Blumeria graminis f. sp. hordei (Bgh), expresses 491 candidate for secreted effector proteins (CSEPs), identified based on presence of a predicted signal peptide and relative sequence uniqueness. Virtually nothing is known about the mechanism of action of CSEPs. The present study used host-induced gene silencing (HIGS) to identify genes involved in early fungal aggressiveness. Screening of 22 selected CSEP-encoding genes suggested that CSEP0007, CSEP0025, CSEP0128, CSEP0211, CSEP0247, CSEP0345, CSEP0420 and CSEP0422 are individually required for normal levels of aggressiveness of Bgh. Expression data showed that these genes were up-regulated at initial stages of infection, suggesting their involvement in early fungal aggressiveness. Altogether, the findings expand the current knowledge of genes contributing to Bgh pathogenesis and provide the basis for future functional characterization.

Pathogens secrete effector proteins to establish a successful interaction with their host. Here, we describe two barley (Hordeum vulgare) powdery mildew candidate secreted effector proteins, CSEP0105 and CSEP0162, which contribute to pathogen success and appear to be required during or after haustorial formation. Silencing of either CSEP using host-induced gene silencing significantly reduced the fungal haustorial formation rate. Interestingly, both CSEPs interact with the barley small heat shock proteins, Hsp16.9 and Hsp17.5, in a yeast two-hybrid assay. Small heat shock proteins are known to stabilize several intracellular proteins, including defense-related signaling components, through their chaperone activity. CSEP0105 and CSEP0162 localized to the cytosol and the nucleus of barley epidermal cells, whereas Hsp16.9 and Hsp17.5 are cytosolic. Intriguingly, only those specific CSEPs changed localization and became restricted to the cytosol when coexpressed with Hsp16.9 and Hsp17.5, confirming the CSEP-small heat shock protein interaction. As predicted, Hsp16.9 showed chaperone activity, as it could prevent the aggregation of Escherichia coli proteins during thermal stress. Remarkably, CSEP0105 compromised this activity. These data suggest that CSEP0105 promotes virulence by interfering with the chaperone activity of a barley small heat shock protein essential for defense and stress responses.

Plant “nucleotide-binding leucine-rich repeat” receptor proteins (NLRs) detect alterations in host targets of pathogen effectors and trigger immune responses. The Arabidopsis thaliana mutant pen1 syp122 displays autoimmunity, and a mutant screen identified the deubiquitinase “associated molecule with the SH3 domain of STAM3” (AMSH3) to be required for this phenotype. AMSH3 has previously been implicated in ESCRT-mediated vacuolar targeting. Pathology experiments show that AMSH3 activity is required for immunity mediated by the CC-NLRs, RPS2 and RPM1. Co-expressing the autoactive RPM1 D505V and the catalytically inactive ESCRT-III protein SKD1 E232Q in Nicotiana benthamiana supports the requirement of ESCRT-associated functions for this CC-NLR-activated immunity. Meanwhile, loss of ESCRT function in A. thaliana is lethal, and we find that AMSH3 knockout-triggered seedling lethality is “enhanced disease susceptibility 1” (EDS1) dependent. Future studies may reveal whether AMSH3 is monitored by a TIR-NLR immunity receptor. NLR receptors are important for plant immunity as they monitor plant processes that are targeted by pathogens. Schultz-Larsen et al. find that the AMSH3 deubiquitinase enzyme is important for immunity activated by specific NLRs. Furthermore, their results suggest that AMSH3 itself is monitored by other NLRs.

Members of the rural communities living around the edges of the Kosi Lake system in northern KwaZulu Natal have been subsistence users of the resources of the lakes, estuary and surrounding swamp forest for many generations.Women and children dig up a variety of crabs in the mangrove ecosystem, and in the late 1980s collected as many as 638 000 crabs a year. The red mangrove crab Neosarmatium meinerti de Man, 1884 (Crustacea: Decapoda: Grapsidae: Sesarminae) constituted the major proportion of these catches, while Scylla serrata and Cardisoma carnifex made up the rest. However, subsistence fishing effort and catches have been reduced steadily, declining from about 5000 outings p.a. and between 2 and4×105 crabs p.a. in the 1980s, to about 1000 outings and 1.3 × 105 crabs in the late 1990s. CPUE has increased from 66 to 145 crabs/person/outing during this period. Crab densities ranged between 0.35/m2 on the high shore and 11.4/m2 on a densely vegetated island, but it was not clear what factors most affected crab density. Densities inside and outside a ‘sanctuary area’ were not significantly different, although there were significant differences in densities among sites within the ‘sanctuary area’. Stock size within the entire mangrove area of Kosi was estimated as 728 514 N. meinerti, but this is almost certainly an underestimate of the total stock size. Sex ratios in the N. meinerti population were not significantly different from 1:1 but above 38 mm carapace width (CW) male crabs considerably outnumbered females. Females matured at 24.9 mm CW and mean sizes of crabs captured by subsistence harvesters were 33.4mmand 31.3mmCWfor males and females, respectively. Instantaneous natural mortality was estimated at M = 2.27 and fishing mortality F = 0.83. The N. meinerti population in the Kosi mangrove system appears to be sustainably exploited.Key words: subsistence fishery, sex ratios, crab density, Brachyura, Crustacea.

Series Crocus comprises ten autumn-flowering species, including the cultivated Crocus sativus, Saffron-Crocus. Interspecific genetic variation was examined in all species of the series, except for C. naqabensis. Intraspecific genetic and morphological variation was considered in the three Greek endemics, C. cartwrightianus, C. hadriaticus and C. oreocreticus. Genetic variation was evaluated based on amplified fragment length polymorphism and simple sequence repeats analyses, including 94 and 233 specimens, respectively. Analysis of molecular variance demonstrated significant genetic variation within populations, compared with low genetic variation between populations suggesting substantial gene flow between populations. In Neighbour-Net analysis, C. hadriaticus samples from mainland Greece were separated from Peloponnesian samples; C. cartwrightianus, C. hadriaticus and C. oreocreticus generally were grouped with C. sativus samples. Pollination and maintenance of genetic variation are discussed. The large intraspecific variation found within the three specifically studied species reflects dynamic population structures with potential to meet future ecological fluctuations. It emphasises a large gene pool which should be considered by including a larger number of accessions in genetic diversity studies.

Salicylic acid (SA) is an important signaling hormone in plant immunity. It can be synthesized by either the phenylpropanoid pathway or the isochorismate pathway, but mutant studies of this have been scarce in other species than Arabidopsis. Here we identified a mutation that introduced a stop-codon early in the barley gene for isochorismate synthase (ICS). We found that homozygous ics plants wilted if not sprayed with 1,4-dihydroxy-2-naphthoic acid, a precursor of phylloquinone, also synthesized via the isochorismate pathway. Interestingly, ics had unchanged SA, suggesting that the basal level of SA is synthesized via the phenylpropanoid pathway. Previous studies have failed seeing increased SA levels in barley after attack by the powdery mildew fungus, Blumeria graminis f.sp. hordei (Bgh), and indeed, we saw no changes in the interaction of ics with this fungus. Overall, we hope this mutant will be useful for other studies of SA in barley.

Characterization of apple germplasm is important for conservation management and breeding strategies. A set of 448 Malus domestica accessions, primarily of local Danish origin, were genotyped using 15 microsatellite markers. Ploidy levels were determined by flow cytometry. Special emphasis was given to pedigree reconstruction, cultivar fingerprinting and genetic clustering. A reference set of cultivars, mostly from other European countries, together with a private nursery collection and a small set of Malus sieversii, Malus sylvestris and small-fruited, ornamental Malus cultivars, was also included. The microsatellite markers amplified 17–30 alleles per loci with an average degree of heterozygosity at 0.78. We identified 104 (23%) duplicate genotypes including colour sports. We could infer first-degree relationships for many cultivars with previously unknown parentages. STRUCTURE analysis provided no evidence for a genetic structure but allowed us to present a putative genetic assembly that was consistent with both PCA analysis and parental affiliation. The Danish cultivar collection contains 10% duplicate genotypes including colour sports and 22% triploids. Many unique accessions and considerable genetic diversity make the collection a valuable resource within the European apple germplasm. The findings presented shed new light on the origin of Danish apple cultivars. The fingerprints can be used for cultivar identification and future management of apple genetic resources. In addition, future genome-wide association studies and breeding programmes may benefit from the findings concerning genetic clustering and diversity of cultivars.

Powdery mildew fungi are parasites that cause disease on a wide range of important crops. Plant resistance (R) genes, which induce host defences against powdery mildews, encode proteins that recognise avirulence (AVR) molecules from the parasite in a gene-for-gene manner. To gain insight into how virulence evolves in Blumeria graminis f.sp. hordei, associations between segregating AVR genes were established. As a prerequisite to the isolation of AVR genes, two loci were selected for further analysis. AVR(a22) is located in a tightly linked cluster comprising AVR(a10) and AVR(k1) as well as up to five other AVR genes. The ratio between physical and genetic distance in the cluster ranged between 0.7 and 35 kB/cM. The AVR(a22) locus was delimited by the previously isolated gene AVR(a10) and two cleaved amplified polymorphic sequence (CAPS) markers, 19H12R and 74E9L. By contrast, AVR(a12) was not linked to other AVR genes in two crosses. Bulk segregant analysis of over 100,000 AFLP fragments yielded two markers, ETAMTG-285 and PAAMACT-473, mapping 10 and 2cM from AVR(a12), respectively, thus delimiting AVR(a12) on one side. All markers obtained for AVR(a12) mapped proximal to it, indicating that the gene is located at the end of a chromosome. Three more AVR(a10) paralogues were identified at the locus interspersed among genes for metabolic enzymes and abundant repetitive elements, especially those homologous to the CgT1 class of retrotransposons. The flanking and close markers obtained will facilitate the isolation of AVR(a22) and AVR(a12) and provide useful tools for studies of the evolution of powdery mildew fungi in agriculture and nature.

Summary We identified a wheat stripe rust (Puccinia striiformis) effector candidate (PEC6) with pattern-triggered immunity (PTI) suppression function and its corresponding host target. PEC6 compromised PTI host species-independently. In Nicotiana benthamiana, it hampers reactive oxygen species (ROS) accumulation and callose deposition induced by Pseudomonas fluorescens. In Arabidopsis, plants expressing PEC6 were more susceptible to Pseudomonas syringae pv. tomato (Pto) DC3000 ΔAvrPto/ΔAvrPtoB. In wheat, PEC6-suppression of P. fluorescens-elicited PTI was revealed by the fact that it allowed activation of effector-triggered immunity by Pto DC3000. Knocking down of PEC6 expression by virus-mediated host-induced gene silencing decreased the number of rust pustules, uncovering PEC6 as an important pathogenicity factor. PEC6, overexpressed in plant cells without its signal peptide, was localized to the nucleus and cytoplasm. A yeast two-hybrid assay showed that PEC6 interacts with both wheat and Arabidopsis adenosine kinases (ADKs). Knocking down wheat ADK expression by virus-induced gene silencing reduced leaf growth and enhanced the number of rust pustules, indicating that ADK is important in plant development and defence. ADK plays essential roles in regulating metabolism, cytokinin interconversion and methyl transfer reactions, and our data propose a model where PEC6 may affect one of these processes by targeting ADK to favour fungal growth.

Summary A large number of effector candidates have been identified recently in powdery mildew fungi. However, their roles and how they perform their functions remain unresolved. In this study, we made use of host-induced gene silencing and confirmed that the secreted barley powdery mildew effector candidate, CSEP0055, contributes to the aggressiveness of the fungus. This result suggests that CSEP0055 is involved in the suppression of plant defence. A yeast two-hybrid screen indicated that CSEP0055 interacts with members of the barley pathogenesis-related protein families, PR1 and PR17. Interaction with PR17c was confirmed by bimolecular fluorescence complementation analyses. Down-regulation and over-expression of PR17c in epidermal cells of barley confirmed that this protein is important for penetration resistance against the powdery mildew fungus. In line with this, PR17c was found to be apoplastic, localizing to the papillae formed in response to this fungus. The CSEP0055 transcript did not start to accumulate until 24 h after inoculation. This suggests that this gene is expressed too late to influence primary penetration events, but rather sustains the fungus at sites of secondary penetration, where PR17c appears to be able to accumulate.

We compare HER2 receptor amplification analysis by immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and real-time polymerase chain reaction (real-time PCR) DNA copy-number assay following laser capture microdissection (LCM) in formalin-fixed paraffin embedded tissue from 40 women with verified ovarian cancer. We speculate that LCM should result in a more accurate assessment of HER2 amplification in our real-time PCR assay compared with IHC and FISH. HER2 overexpression measured by IHC, FISH, or real-time PCR was found in 5.0%, 5.0%, and 22.5%, respectively. HER2 negative results measured by IHC, FISH, or real-time PCR were found in 95%, 92.5%, and 60.0%, respectively. Analysis failed for IHC, FISH, or real-time PCR in 0%, 2.5%, or 17.5% of cases. Concordance between IHC and FISH, IHC and real-time PCR, or FISH and real-time PCR were 89.7%, 72.7%, or 78.1%, respectively. Only few ovarian cancer patients were HER2 overexpressed measured by IHC or FISH and thus could be eligible for antibody-based therapy with trastuzumab (Herceptin). Interestingly, we find an increased number of HER2 positive patients by real-time PCR analysis on microdissected cancer cells, suggesting a number of HER2 positive patients not detected by current methods. Thus, the concept of quantitative measurement of HER2 on microdissected cancer cells should be explored further.

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The lesion-mimic Arabidopsis mutant, syp121 syp122, constitutively expresses the salicylic acid (SA) signaling pathway and has low penetration resistance to powdery mildew fungi. Genetic analyses of the lesion-mimic phenotype have expanded our understanding of programmed cell death (PCD) in plants. Inactivation of SA signaling genes in syp121 syp122 only partially rescues the lesion-mimic phenotype, indicating that additional defenses contribute to the PCD. Whole genome transcriptome analysis confirmed that SA-induced transcripts, as well as numerous other known pathogen-response transcripts, are up-regulated after inactivation of the syntaxin genes. A suppressor mutant analysis of syp121 syp122 revealed that FMO1, ALD1, and PAD4 are important for lesion development. Mutant alleles of EDS1, NDR1, RAR1, and SGT1b also partially rescued the lesion-mimic phenotype, suggesting that mutating syntaxin genes stimulates TIR-NB-LRR and CC-NB-LRR-type resistances. The syntaxin double knockout potentiated a powdery mildew-induced HR-like response. This required functional PAD4 but not functional SA signaling. However, SA signaling potentiated the PAD4-dependent HR-like response. Analyses of quadruple mutants suggest that EDS5 and SID2 confer separate SA-independent signaling functions, and that FMO1 and ALD1 mediate SA-independent signals that are NPR1-dependent. These studies highlight the contribution of multiple pathways to defense and point to the complexity of their interactions.

Mange serbere er tilbøjelige til at tro, at deres vanskeligheder skyldes en ond omverden og ikke deres egne valg. De mener, at de har retten til Kosovo, skønt de ikke aner, hvad de i givet fald skulle stille op med to millioner albanere

EU-landene bør blive bedre til at beskytte deres fælles interesser og fremme deres fælles værdier, og verden har brug for, at EU træder tydeligere og mere enigt frem og påtager sig et ansvar

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Penetration resistance is often the first line of defence against fungal pathogens. Subsequently induced defences are mediated by the programmed cell death (PCD) reaction pathway and the salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) signalling pathways. We previously demonstrated that full penetration resistance in Arabidopsis against the non-host barley powdery mildew fungus (Blumeria graminis f.sp. hordei) requires the syntaxin SYP121 (PEN1). Here we report that SYP121, together with SYP122, functions as a negative regulator of subsequently induced defence pathways. The SA level in the syntaxin double mutant syp121-1 syp122-1 is dramatically elevated, resulting in necrosis and dwarfism. This phenotype is partially rescued by introducing the SA-signalling mutations eds1-2, eds5-3, sid2-1 and npr1-1 as well as the NahG transgene. These partially rescued triple mutants have an unknown defence to Pseudomonas syringae pv. tomato, and have increased HR-like responses to non-host and host powdery mildew fungi. The HR-like responses cause efficient resistance to the latter. These defence pathways are SA-independent. Furthermore, the JA/ET signalling marker, PDF1.2, is highly upregulated in the triple mutants. Thus SYP121 and SYP122 are negative regulators of PCD, SA, JA and ET pathways through a molecular function distinct from that of SYP121 in penetration resistance. Our data suggest that individual cells preferentially express either penetration resistance or the subsequently induced defences.

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