Genetic Disease Resistance Research Articles

B cell signatures of BCWD-resistant and susceptible lines of rainbow trout: A shift towards more EBF-expressing progenitors and fewer mature B cells in resistant animals

Bacterial cold water disease (BCWD) is a chronic disease of rainbow trout, and is caused by the Gram-negative bacterium Flavobacterium psychrophilum (Fp), a common aquaculture pathogen. The National Center for Cool and Cold Water Aquaculture has bred two genetic lines of rainbow trout: a line of Fp-resistant trout (ARS-Fp-R or R-line trout) and a line of susceptible trout (ARS-Fp-S, or S-line). Little is known about how phenotypic selection alters immune response parameters or how such changes relate to genetic disease resistance. Herein, we quantify interindividual variation in the distribution and abundance of B cell populations (B cell signatures) and examine differences between genetic lines of naive animals. There are limited trout-specific cell surface markers currently available to resolve B cell subpopulations and thus we developed an alternative approach based on detection of differentially expressed transcription factors and intracellular cytokines. B cell signatures were compared between R-line and S-line trout by flow cytometry using antibodies against transcription factors early B cell factor-1 (EBF1) and paired domain box protein Pax5, the pro-inflammatory cytokine IL-1β, and the immunoglobulin heavy chain mu. R-line trout had higher percentages of EBF+ B myeloid/ progenitor and pre-B cells in PBL, anterior and posterior kidney tissues compared to S-line trout. The opposite pattern was detected in more mature B cell populations: R-line trout had lower percentages of both IgM+ mature B cells and IgM-secreting cells in anterior kidney and PBL compared to S-line trout. In vitro LPS-activation studies of PBL and spleen cell cultures revealed no significant induction differences between R-line and S-line trout. Together, our findings suggest that selective resistance to BCWD may be associated with shifts in naive animal developmental lineage commitment that result in decreased B lymphopoiesis and increased myelopoiesis in BCWD resistant trout relative to susceptible trout.

Genetic diversity, population structure, and resistance to Phytophthora capsici of a worldwide collection of eggplant germplasm.

Eggplant (Solanum melongena L.) is an important solanaceous crop with high phenotypic diversity and moderate genotypic diversity. Ninety-nine genotypes of eggplant germplasm (species (S. melongena, S. incanum, S. linnaeanum and S. gilo), landraces and heirloom cultivars) from 32 countries and five continents were evaluated for genetic diversity, population structure, fruit shape, and disease resistance to Phytophthora fruit rot. Fruits from each line were measured for fruit shape and evaluated for resistance to two Phytophthora capsici isolates seven days post inoculation. Only one accession (PI 413784) was completely resistant to both isolates evaluated. Partial resistance to Phytophthora fruit rot was found in accessions from all four eggplant species evaluated in this study. Genetic diversity and population structure were assessed using 22 polymorphic simple sequence repeats (SSRs). The polymorphism information content (PIC) for the population was moderate (0.49) in the population. Genetic analyses using the program STRUCTURE indicated the existence of four genetic clusters within the eggplant collection. Population structure was detected when eggplant lines were grouped by species, continent of origin, country of origin, fruit shape and disease resistance.

Absence of detectable yield penalty associated with insensitivity to Pleosporales necrotrophic effectors in wheat grown in the West Australian wheat belt

Genetic disease resistance is widely assumed, and occasionally proven, to cause host yield or fitness penalties due to inappropriate activation of defence response mechanisms or diversion of resources to surplus preformed defences. The study of resistance gene trade‐offs has so far been restricted to biotrophic pathogens. In some Pleosporales necrotrophic interactions, quantitative resistance is positively associated with insensitivity to effectors. Host lines that differ in sensitivity can easily be identified amongst current cultivars and advanced breeding lines. Large wheat cultivar trials were used to test whether lines sensitive or insensitive to three necrotrophic effectors from Pyrenophora tritici‐repentis and Parastagonospora nodorum differed in yield when subjected to natural disease and stress pressures in the West Australian wheat belt. There was no significant yield penalty associated with insensitivity to the fungal effectors ToxA, SnTox1 and SnTox3. Some yield gains were associated with insensitivity and some of these gains could be attributed to increased disease resistance. It is concluded that insensitivity to these effectors does not render such plants more vulnerable to any relevant biotic or abiotic stress present in these trials. These results suggest that the elimination of sensitivity alleles for necrotrophic effectors is a safe and facile strategy for improving disease resistance whilst maintaining or improving other desirable traits.

Pathotype variation of barley powdery mildew in Western Australia

Barley powdery mildew caused by the fungus Blumeria graminis f. sp. hordei (Bgh) has emerged as the most damaging disease of barley in Western Australia (WA). Many of the available cultivars display high levels of disease in the field when climatic conditions are conducive. As a result, fungicides have become the main method of disease control in the last 10 years. Different types and sources of genetic disease resistance are available but to optimise their deployment it is necessary to evaluate the spectrum of pathotypes present in the pathogen population. Sixty isolates of Bgh were collected in the 2009 season from 9 locations, single spored and characterised by infection on reference barley lines and cultivars. Eighteen unique pathotypes were resolved. Virulence against many of the R-genes in the reference lines was present in at least one pathotype. Isolates were virulent against 16 out of a total of 23 resistance gene combinations. Undefeated resistance genes included the major R-genes Mla-6, Mla-9, Ml-ra and the combinations of Mla-1 plus Mla-A12 and Mla-6 plus Mla-14 and Mla-13 plus Ml-Ru3 together with the recessive resistance gene mlo-5. There was significant pathotype spatial differentiation suggesting limited gene flow between different regions with WA or localised selection pressures and proliferation. On the basis of the results we recommend a number of strategies to manage powdery mildew disease levels within WA.

Registration of the Restorer Oilseed Sunflower Germplasm RHA 464 Possessing Genes for Resistance to Downy Mildew and Sunflower Rust

Sunflower rust (caused by Puccinia helianthi Schwein.) has become a more significant threat to U.S. sunflower (Helianthus annuus L.) production as a result of the multiplicity of races, increasing incidence, and lack of commercial hybrids with resistance. Thus, there is a need for new sources of effective rust resistance in the sunflower breeding community. At the same time, resistance to downy mildew [caused by Plasmopara halstedii (Farl.) Berl. & De Toni] is also critical to parental line development because of the major impact of this disease over years. Our objective was to produce a restorer sunflower germplasm with resistance to the most virulent race of sunflower rust, race 777, and combine with it the best currently available downy mildew resistance in a high‐yielding genetic background. We achieved this objective by using the pedigree breeding method with early generation selection for rust and downy mildew resistance and late‐generation testcross evaluation for yield. The resulting restorer oilseed sunflower (Helianthus annuus L.) germplasm, ‘RHA 464’ (Reg. No. GP‐325; PI 655015; experimental ‘05 187’), which has resistance to the two pathogens of interest, was released by the USDA‐ARS and the North Dakota Agricultural Experiment Station, Fargo, ND, to fill the urgent need in the sunflower industry for breeding lines with elite genetic background and disease resistance.

Multiple gene loci affecting genetic background-controlled disease resistance conferred by R gene Xa3/Xa26 in rice

The function of bacterial-blight resistance gene Xa3/Xa26 in rice is influenced by genetic background; the Oryza sativa L. ssp. japonica background can increase Xa3/Xa26 expression, resulting in an enhanced resistance. To identify whether Xa3/Xa26 transcript level is the only factor contributing to genetic background-controlled resistance, we screened an F(2) population that was developed from a cross between Oryza sativa L. ssp. indica and japonica rice lines and was segregating for Xa3/Xa26, and compared the expression profiles of a pair of indica and japonica rice lines that both carried Xa3/Xa26. Eight quantitative trait loci (QTLs), in addition to Xa3/Xa26, were identified as contributing to the bacterial resistance of this population. Four of the eight QTLs were contributed to the japonica line. The resistance of this population was also affected by epistatic effects. Some F(2) individuals showed significantly increased Xa3/Xa26 transcripts, but the increased transcripts did not completely correlate with the reduced disease in this population. The analysis of the expression profile of Xa3/Xa26-mediated resistance using a microarray containing approximate 7,990 rice genes identified 44 differentially expressed genes. Thirty-five genes were rapidly activated in the japonica background, but not in the indica background, during disease resistance. These results suggest that multiple factors, including the one resulting in increased Xa3/Xa26 expression, may contribute to the enhanced resistance in the japonica background. These factors can cause a variation in gene expression profile that differs from that in the indica background during disease resistance.

FITNESS TRADE-OFFS MEDIATED BY IMMUNOSUPPRESSION COSTS IN A SMALL MAMMAL

Trade-offs are widespread between life-history traits, such as reproduction and survival. However, their underlying physiological and behavioral mechanisms are less clear. One proposed physiological factor involves the trade-off between investment in male reproductive effort and immunity. Based on this hypothesis, we investigated differences in fitness between artificially selected immune response bank vole groups, Myodes glareolus. Significant heritability of immune response was found and a correlated response in testosterone levels to selection on immune function. Male reproductive effort, reproductive success, and survival of first generation offspring were assessed and we demonstrate a relationship between laboratory measured immune parameters and fitness parameters in field enclosures. We identify a trade-off between reproductive effort and survival with immune response and parasites as mediators. However, this trade-off results in equal male fitness in natural conditions, potentially demonstrating different male signaling strategies for either reproductive effort or survival. Females gain indirect genetic benefits for either genetic disease resistance or male reproductive effort, but not both. Immune response is genetically variable, genetically linked to testosterone and may indirectly maintain genetic variation for sexually selected traits. Evidence for both a genetic and a field trade-off between reproductive effort and survival indicates an evolutionary constraint on fitness traits.

Investigations on genetic disease resistance in swine—A contribution to the reduction of pain, suffering and damage in farm animals

This review deals with genetic disease resistance in pigs as a prospective opportunity to reduce pain, suffering and damage in swine production. Even under favourable terms of housing and management, infectious diseases are wide-spread, and have to be ranked among the major sources for suffering in international pig production. There is abundant variability in resistance/susceptibility of pig populations and breeds to a number of pathogens, including several viruses, bacteria, and parasites, even at similar performance levels. Practical applications of disease resistance in farm animals and established knowledge on disease resistance in swine are presented. The difficulty lies in the identification of carriers of favourable gene variants. With tools and technologies developed in the context of genome projects, new opportunities to dissect the genetic control of host response to infection are presented with QTL analysis of Sarcocystis miescheriana resistance given as an exemplary parasite model: this model demonstrates the existence of outstanding gene loci and their role for polygenic trait variability. It further provides evidence for common mechanisms involved in resistance to different pathogens. Although a long way from the model to markers for disease resistance, pain, suffering, and damage might be reduced by the applications of genetic disease resistance in pig production in the future.

Association of Avenanthramide Concentration in Oat (Avena sativa L.) Grain with Crown Rust Incidence and Genetic Resistance

ABSTRACTAvenanthramides (avn) are antioxidant compounds found in oat tissues, including the grain, that are of interest from a nutritional standpoint. In this study, we have measured avenanthramide concentration in the grain of 18 oat genotypes grown in six environments. These genotypes varied widely in crown rust (Puccinia coronata) resistance. Crown rust infected two of the six environments studied. The grain avenanthramide concentrations in the crown rust environments were significantly higher than those in the uninfected environments. Avenanthramide concentrations in the crown rust infected environments was also significantly correlated with their genetic crown rust resistance, as evaluated in the field. These results suggest that avenanthramide accumulation in the grain is associated with crown rust infection and that, in most of the cultivars evaluated, the extent of their accumulation also correlated with their genetic disease resistance; avenanthramide accumulation in the grain was, with noted exceptions, highest in those cultivars showing the greatest genetic resistant to crown rust.

Genetic Diversity and Disease Resistance of Wild Malus orientalis from Turkey and Southern Russia

Genetic diversity and disease resistance are described for 496 seedlings from wild populations of Malus orientalis Uglitzh. collected in southern Russia and Turkey in 1998 and 1999. Eighty-five half-sib families were genotyped using seven microsatellite markers, and disease resistance was determined for apple scab (Venturia inaequalis Cooke), cedar apple rust (Gymnosporangium juniperi-virginianae Schwein), and fire blight (Erwinia amylovora Burrill). Individuals from the two Russian Caucasus collection locations were homogeneous compared with populations from the four Turkish collection locations. Within three of the Turkish collection locations, some half-sib families were highly diverse and several of these families had unusually high levels of disease resistance. In all, twenty individuals exhibited resistance to all three diseases. Bayesian analyses of the population structure revealed six distinct clusters. Most of the individuals segregated into two clusters, one containing individuals primarily from southern Russia and the other containing individuals from both Russia and northern Turkey. Individuals in the four small clusters were specific to Turkish collection locations. These data suggest wild populations of M. orientalis from regions around the Black Sea are genetically distinguishable and show high levels of diversity.

Comparing Explanations of Polygyny

Polygyny is common in the ethnographic record. The vast majority of cultures known to anthropology allowed at least some men to have more than one wife simultaneously. This article compares various explanations of nonsororal polygyny, by far the most common type of polygyny. Multiple regression analyses of data for the societies in the Standard Cross-Cultural Sample indicate that the two main independent cross-cultural predictors of appreciable (i.e., more than occasional) nonsororal polygyny are high male mortality in war (resulting in an excess of females) and high pathogen stress, which seems to favor nonsororal polygyny to maximize genetic variation and disease resistance in progeny. High male mortality in war predicts particularly in nonstate societies. High pathogen stress predicts particularly in more densely populated state societies.

Passerine MHC: genetic variation and disease resistance in the wild

The passerine major histocompatibility complex (MHC) class I and IIB genes are different from those of the avian model species the chicken because passerines have (1) a larger number of MHC genes, (2) MHC genes with longer introns, and (3) MHC genes that are pseudo-genes. Most passerine MHC genes are transcribed (coding), extremely variable and subject to balancing selection. The high genetic diversity of the MHC genes of passerines is most likely maintained by selection from a large number of different pathogens. Association between MHC alleles and resistance to avian malaria infections have been reported in House Sparrows and Great Reed Warblers. Passerines are infected by a large number of different avian malaria infections. Therefore passerines and avian malaria is a study system that is well-suited to investigations of balancing selection and associations between MHC genes and disease resistance.

Enhanced resistance to Phoma tracheiphila and Botrytis cinerea in transgenic lemon plants expressing a Trichoderma harzianum chitinase gene

AbstractAntifungal genes from micro‐organisms were inserted into the genome of a number of plant species, representing a promising strategy for conferring genetic disease resistance against a broad range of plant pathogenic fungi. In the present study, the chit42 gene from Trichoderma harzianum (codifying the antifungal protein endochitinase) was introduced into the ‘Femminello siracusano’ lemon by Agrobacterium tumefaciens, in order to regenerate transgenic plants resistant to fungal disease. Three polymerase chain reaction (PCR)‐positive clones were obtained. Southern blot confirmed the integration of the transgene in the lemon genome and revealed that one or two copies had been inserted. Reverse transcriptase‐PCR, Northern blot and Western analysis were performed and the results confirmed the expression of the inserted gene. The transgenic clones were tested in vitro and in vivo for disease resistance. Conidia germination and fungal growth of Phoma tracheiphila were strongly inhibited in vitro by the transgenic foliar proteins, while no effects were observed with the controls. Disease resistance assays were performed in vivo with Botrytis cinerea, the causal agent of grey mould in fruit. Transgenic lemon plants, inoculated with lemon petals infected by a single‐conidial isolate of B. cinerea, showed significantly less lesion development than the controls. On the whole, the results indicate that the transformation with the antifungal endochitinase gene may represent a strategy for disease control in citrus crops.

Genetic differences in natural antibody levels in common carp ( Cyprinus carpio L.)

In mammals, natural antibodies (Nabs) are mostly of the IgM isotype and can bind to a particular antigen or pathogen even if the host has never been exposed. Despite their early detection and abundance, the exact role and genetic control of Nabs remain unclear. We have used an indirect ELISA with three different antigens (keyhole limpet haemocyanin, chicken ovalbumin and bovine serum albumin) to demonstrate the ubiquitous presence of Nabs in common carp. Serum levels of Nabs increased with age, i.e. 10-month-old fish showed higher levels than 4-month-old fish. Also, fish grown in earth ponds showed higher levels of Nabs than fish grown in a clean environment of UV-treated water. Furthermore, we show that Nabs are present in different levels in the serum of carp lines with a different genetic background, suggestive of a genetic control. These genetic differences were independent of antigen, age and environment. Genetic differences in levels of Nabs could not unequivocally be related to differences in survival under farmed conditions. The possibilities for using levels of Nabs as marker criterion for selection for genetic disease resistance are discussed.

Acquired Immunity and Stochasticity in Epidemic Intervals Impede the Evolution of Host Disease Resistance

Disease can generate intense selection pressure on host populations, but here we show that acquired immunity in a population subject to repeated disease outbreaks can impede the evolution of genetic disease resistance by maintaining susceptible genotypes in the population. Interference between the life-history schedule of a species and periodicity of the disease has unintuitive effects on selection intensity, and stochasticity in outbreak period further reduces the rate of spread of disease-resistance alleles. A general age-structured population genetic model was developed and parameterized using empirical data for phocine distemper virus (PDV) epizootics in harbor seals. Scenarios with acquired immunity had lower levels of epizootic mortality compared with scenarios without acquired immunity for the first PDV outbreaks, but this pattern was reversed after about five disease cycles. Without acquired immunity, evolution of disease resistance was more rapid, and long-term population size variation is efficiently dampened. Acquired immunity has the potential to significantly influence rapid evolutionary dynamics of a host population in response to age-structured disease selection and to alter predicted selection intensities compared with epidemiological models that do not consider such feedback. This may have important implications for evolutionary population dynamics in a range of human, agricultural, and wildlife disease settings.

Functional analysis of a single nucleotide polymorphism in a potential binding site for GATA transcription factors in the ovine interleukin 2 gene

The transcription factor GATA-3 is one regulator of Th1/Th2 differentiation. In sheep, we recently discovered a putative GATA-binding site (WGATAR) in the second intron of the Th1-cytokine gene interleukin 2 ( IL2), showing a single nucleotide polymorphism (G/C). As genetic variations in cytokine genes are thought to regulate cytokine production, we studied the significance of this polymorphism for IL2 transcription. Sheep with different IL2 genotypes were identified by single-strand conformation polymorphism (SSCP)-analysis and IL2 transcription levels in peripheral blood mononuclear cells (PBMC) isolated from these animals were compared. For this purpose, transcription of IL2 mRNA was quantified by real-time polymerase chain reaction in unstimulated PBMC and in PBMC incubated for 4 h in the presence of concanavalin A (ConA) or phorbol 12-myristate 13-acetate plus ionomycin (PMA/I). Compared to unstimulated cells, stimulation with ConA and PMA/I increased the IL2 mRNA transcription in average by 300- and 20-fold, respectively. Nevertheless, no significant differences in IL2 transcription between the genotypes could be detected. These findings were confirmed by band shift studies using different oligonucleotides containing variations of the potential binding motif, which showed no differences in the gel mobility after incubation with nuclear extract containing GATA-3. The obtained results argue against an impact of this polymorphism on the IL2 transcription and the genetic disease resistance in sheep.

Mate Choice, Male Condition-Dependent Ornamentation and MHC in the Pheasant

The Hamilton and Zuk model predicts that genes for resistance to various pathogens may be continuously heritable due to selection disequilibria caused by coadaptional cycles of hosts and pathogens. The model further suggests that the expression of male secondary ornaments is condition-dependent and that only individuals with superior genetic disease resistance and vigour can fully express exaggerated secondary ornaments. Female choice is therefore expected to discriminate among males on the basis of secondary sexual characters in order to pass on genes for disease resistance that improve fitness in the offspring. In wild ring-necked pheasants, Phasianus colchicus, of the Revinge area in southern Sweden, females prefer to mate with long-spurred males and data on reproductive success indicate that they may improve their chicks' survival rate by doing so. Male spur length is positively correlated with age, body size and viability. MHC genotyping for both class I and class II B of pheasant males trapped in the study area identified eight MHC haplotypes and a total of 17 different MHC genotypes. Multivariate analyses revealed that MHC genotype is significantly associated with variation in both male spur length and male viability. These data indicate that polymorphic genes with a central role in immune recognition can be associated with viability and the expression of a condition-dependent intersexually selected male trait, thus supporting essential parts of the Hamilton and Zuk model. (Less)

The role and regulation of programmed cell death in plant-pathogen interactions.

It is commonly known that animal pathogens often target and suppress programmed cell death (pcd) pathway components to manipulate their hosts. In contrast, plant pathogens often trigger pcd. In cases in which plant pcd accompanies disease resistance, an event called the hypersensitive response, the plant surveillance system has learned to detect pathogen-secreted molecules in order to mount a defence response. In plants without genetic disease resistance, these secreted molecules serve as virulence factors that act through largely unknown mechanisms. Recent studies suggest that plant bacterial pathogens also secrete antiapoptotic proteins to promote their virulence. In contrast, a number of fungal pathogens secrete pcd-promoting molecules that are critical virulence factors. Here, we review recent progress in determining the role and regulation of plant pcd responses that accompany both resistance and susceptible interactions. We also review progress in discerning the mechanisms by which plant pcd occurs during these different interactions.

Population dynamics of epiphytic mycoparasites of the genera Clonostachys and Fusarium for the biocontrol of black pod (Phytophthora palmivora) and moniliasis ( Moniliophthora roreri) on cocoa ( Theobroma cacao)

Mycoparasites collected from aerial parts of the cocoa plant (Theobroma cacao) have shown great promise in the control of black pod, caused by Phytophthora palmivora, and moniliasis, caused by Moniliophthora roreri. However, the ecology of epiphytic mycoparasites is still poorly understood although it has a direct bearing on applied biocontrol practices, ranging from the identification and isolation of promising biocontrol candidates to formulation needs and required application frequency. One objective of this study was to determine the natural abundance of mycoparasites on cocoa flowers and pods in relation to crop development stage and cultivar. For this purpose, native mycoparasites were detected on cocoa flowers and pods using the precolonised plate baiting technique. Furthermore, the survival of an applied Clonostachys rosea isolate on cocoa pods on shaded and non-shaded trees was compared as well as the recolonisation patterns of surface-sterilised pods by native mycoparasites under these conditions. Clonostachys spp. were the most commonly isolated native mycoparasites, followed by Fusarium spp. No differences in the occurrence of native, epiphytic mycoparasites were observed between the three main cocoa cultivars, 'Criollo', 'Forastero' and 'Trinitario', nor between clones within these groups. Thus, a single biocontrol inoculum can be suitable for application to cultivar mixtures of cocoa commonly grown together in a field. Different susceptibility classes of segregating F1 populations of hybrids with resistance against M. roreri and P. palmivora supported similar population levels and taxonomic assemblages of mycoparasites. Therefore, we reject the hypothesis that these antagonists mediate resistance. Mycoparasite abundance and genetic disease resistance to black pod and moniliasis are independent phenomena and should lead to additive effects if employed simultaneously in an integrated disease management programme. The survival of applied C. rosea was not affected by the shading regime or any other meteorological parameter measured. On the other hand, recolonisation of surface-sterilised cocoa pods by most native mycoparasites was faster in the shade. Only Trichoderma spp. colonised pods exposed to direct sunlight faster than shaded ones. The implications for the design of biocontrol inocula and formulation technology are discussed.

Genetic diversity and disease resistance: some considerations for research, breeding, and deployment

Fungal pathogens present a complex challenge for genetic management of forest trees. The need is for disease resistance that withstands mutations and genetic shifts in pathogens. Also desirable are defences against new and dangerous pathogens. An understanding of how hosts and pathogens can continue to coexist should help. Experience from agriculture has allowed modelling of pathosystems, the genetic variations within hosts and pathogens that permit coexistence. While it is impractical to construct a comprehensive model, two phenomena seem generally conducive to stability: a cost of "virulence" in pathogen fitness and a multiplicity of host resistance mechanisms. However, other factors, notably indirect costs of resistance, are very difficult to model. Overall, the diversity of behaviour of models, of the nature of resistance and virulence genes, and of biology of both hosts and pathogens precludes any unique formula for stability. For current crops, genetic diversity offers risk spread for susceptibility to a new and serious pathogen or pathotype. For longer-term breeding, relatively rare resistance may be useful, but pedigreed breeding populations typically entail very finite population sizes. Providing for selecting within improved production populations may therefore be needed. This would give breeders technical challenges, and give forest managers opportunity costs and major logistical challenges.