Fine mapping of root lesion nematode (Pratylenchus thornei) resistance loci on chromosomes 6D and 2B of wheat.

KEY MESSAGE: Resistance QTL to root lesion nematode (Pratylenchus thornei) in wheat (Triticum aestivum), QRlnt.sk-6D and QRlnt.sk-2B, were mapped to intervals of 3.5 cM/1.77 Mbp on chromosome 6D and 1.4 cM/2.19 Mbp on chromosome 2B, respectively. Candidate resistance genes were identified in the QTL regions and molecular markers developed for marker-assisted breeding. Two previously known resistance QTL for root lesion nematode (Pratylenchus thornei) in bread wheat (Triticum aestivum), QRlnt.sk-6D and QRlnt.sk-2B, were fine-mapped using a Sokoll (moderately resistant) by Krichauff (susceptible) doubled haploid (DH) population and six newly developed recombinant inbred line populations. Bulked segregation analysis with the 90K wheat SNP array identified linked SNPs which were subsequently converted to KASP assays for mapping in the DH and RIL populations. On chromosome 6D, 60 KASP and five SSR markers spanned a total genetic distance of 23.7 cM. QRlnt.sk-6D was delimited to a 3.5 cM interval, representing 1.77 Mbp in the bread wheat cv. Chinese Spring reference genome sequence and 2.29 Mbp in the Aegilops tauschii genome sequence. These intervals contained 42 and 43 gene models in the respective annotated genome sequences. On chromosome 2B, 41 KASP and 5 SSR markers produced a map spanning 19.9 cM. QRlnt.sk-2B was delimited to 1.4 cM, corresponding 3.14 Mbp in the durum wheat cv. Svevo reference sequence and 2.19 Mbp in Chinese Spring. The interval in Chinese Spring contained 56 high-confidence gene models. Intervals for both QTL contained genes with similarity to those previously reported to be involved in disease resistance, namely genes for phenylpropanoid biosynthetic pathway-related enzymes, NBS-LRR proteins and protein kinases. The potential roles of these candidate genes in P. thornei resistance are discussed. The KASP markers reported in this study could potentially be used for marker-assisted breeding of P. thornei-resistant wheat cultivars.

A QTL on the short arm of wheat (Triticum aestivum L.) chromosome 3B affects the stability of grain weight in plants exposed to a brief heat shock early in grain filling.

BACKGROUND: Molecular markers and knowledge of traits associated with heat tolerance are likely to provide breeders with a more efficient means of selecting wheat varieties able to maintain grain size after heat waves during early grain filling. RESULTS: A population of 144 doubled haploids derived from a cross between the Australian wheat varieties Drysdale and Waagan was mapped using the wheat Illumina iSelect 9,000 feature single nucleotide polymorphism marker array and used to detect quantitative trait loci for heat tolerance of final single grain weight and related traits. Plants were subjected to a 3 d heat treatment (37 °C/27 °C day/night) in a growth chamber at 10 d after anthesis and trait responses calculated by comparison to untreated control plants. A locus for single grain weight stability was detected on the short arm of chromosome 3B in both winter- and autumn-sown experiments, determining up to 2.5 mg difference in heat-induced single grain weight loss. In one of the experiments, a locus with a weaker effect on grain weight stability was detected on chromosome 6B. Among the traits measured, the rate of flag leaf chlorophyll loss over the course of the heat treatment and reduction in shoot weight due to heat were indicators of loci with significant grain weight tolerance effects, with alleles for grain weight stability also conferring stability of chlorophyll ('stay-green') and shoot weight. Chlorophyll loss during the treatment, requiring only two non-destructive readings to be taken, directly before and after a heat event, may prove convenient for identifying heat tolerant germplasm. These results were consistent with grain filling being limited by assimilate supply from the heat-damaged photosynthetic apparatus, or alternatively, accelerated maturation in the grains that was correlated with leaf senescence responses merely due to common genetic control of senescence responses in the two organs. There was no evidence for a role of mobilized stem reserves (water soluble carbohydrates) in determining grain weight responses. CONCLUSIONS: Molecular markers for the 3B or 6B loci, or the facile measurement of chlorophyll loss over the heat treatment, could be used to assist identification of heat tolerant genotypes for breeding.

Modelling and genetic dissection of staygreen under heat stress.

KEY MESSAGE: Staygreen traits are associated with heat tolerance in bread wheat. QTL for staygreen and related traits were identified across the genome co-located with agronomic and physiological traits associated to plant performance under heat stress. Plant chlorophyll retention-staygreen-is considered a valuable trait under heat stress. Five experiments with the Seri/Babax wheat mapping population were sown in Mexico under hot-irrigated environments. Normalized difference vegetation index (NDVI) during plant growth was measured regularly and modelled to capture the dynamics of plant greenness decay, including staygreen (Stg) at physiological maturity which was estimated by regression of NDVI during grainfilling. The rate of senescence, the percentage of plant greenness decay, and the area under the curve were also estimated based on NDVI measurements. While Stg and the best fitted curve were highly environment dependent, both traits showed strong (positive for Stg) correlations with yield, grainfilling rates, and extended grainfilling periods, while associations with kernel number and kernel weight were weak. Stg expression was largely dependent on rate of senescence which was related to the pattern of the greenness decay curve and the initial NDVI. QTL analyses revealed a total of 44 loci across environments linked to Stg and related traits, distributed across the genome, with the strongest and most repeatable effects detected on chromosomes 1B, 2A, 2B, 4A, 4B and 7D. Of these, some were common with regions controlling phenology but independent regions were also identified. The co-location of QTL for Stg and performance traits in this study confirms that the staygreen phenotype is a useful trait for productivity enhancement in hot-irrigated environments.

Variation in the interaction between alleles of HvAPETALA2 and microRNA172 determines the density of grains on the barley inflorescence.

Within the cereal grasses, variation in inflorescence architecture results in a conspicuous morphological diversity that in crop species influences the yield of cereal grains. Although significant progress has been made in identifying some of the genes underlying this variation in maize and rice, in the temperate cereals, a group that includes wheat, barley, and rye, only the dosage-dependent and highly pleiotropic Q locus in hexaploid wheat has been molecularly characterized. Here we show that the characteristic variation in the density of grains along the inflorescence, or spike, of modern cultivated barley (Hordeum vulgare) is largely the consequence of a perturbed interaction between microRNA172 and its corresponding binding site in the mRNA of an APELATA2 (AP2)-like transcription factor, HvAP2. We used genome-wide association and biparental mapping to identify HvAP2. By comparing inflorescence development and HvAP2 transcript abundance in an extreme dense-spike mutant and its nearly isogenic WT line, we show that HvAP2 turnover driven by microRNA 172 regulates the length of a critical developmental window that is required for elongation of the inflorescence internodes. Our data indicate that this heterochronic change, an altered timing of developmental events caused by specific temporal variation in the efficiency of HvAP2 turnover, leads to the striking differences in the size and shape of the barley spike.

HVP10 encoding V-PPase is a prime candidate for the barley HvNax3 sodium exclusion gene: evidence from fine mapping and expression analysis.

In cereals, a common salinity tolerance mechanism is to limit accumulation of Na(+) in the shoot. In a cross between the barley variety Barque-73 (Hordeum vulgare ssp. vulgare) and the accession CPI-71284 of wild barley (H. vulgare ssp. spontaneum), the HvNax3 locus on chromosome 7H was found to determine a ~10-25 % difference in leaf Na(+) accumulation in seedlings grown in saline hydroponics, with the beneficial exclusion trait originating from the wild parent. The Na(+) exclusion allele was also associated with a 13-21 % increase in shoot fresh weight. The HvNax3 locus was delimited to a 0.4 cM genetic interval, where it cosegregated with the HVP10 gene for vacuolar H(+)-pyrophosphatase (V-PPase). Sequencing revealed that the mapping parents encoded identical HVP10 proteins, but salinity-induced mRNA expression of HVP10 was higher in CPI-71284 than in Barque-73, in both roots and shoots. By contrast, the expression of several other genes predicted by comparative mapping to be located in the HvNax3 interval was similar in the two parent lines. Previous work demonstrated roles for V-PPase in ion transport and salinity tolerance. We therefore considered transcription levels of HVP10 to be a possible basis for variation in shoot Na(+) accumulation and biomass production controlled by the HvNax3 locus under saline conditions. Potential mechanisms linking HVP10 expression patterns to the observed phenotypes are discussed.

Fine mapping and chromosome walking towards the Ror1 locus in barley (Hordeum vulgare L.).

KEY MESSAGE: The Ror1 gene was fine-mapped to the pericentric region of barley chromosome 1HL. Recessively inherited loss-of-function alleles of the barley (Hordeum vulgare) Mildew resistance locus o (Mlo) gene confer durable broad-spectrum disease resistance against the obligate biotrophic fungal powdery mildew pathogen Blumeria graminis f.sp. hordei. Previous genetic analyses revealed two barley genes, Ror1 and Ror2, that are Required for mlo-specified resistance and basal defence. While Ror2 was cloned and shown to encode a t-SNARE protein (syntaxin), the molecular nature or Ror1 remained elusive. Ror1 was previously mapped to the centromeric region of the long arm of barley chromosome 1H. Here, we narrowed the barley Ror1 interval to 0.18 cM and initiated a chromosome walk using barley yeast artificial chromosome (YAC) clones, next-generation DNA sequencing and fluorescence in situ hybridization. Two non-overlapping YAC contigs containing Ror1 flanking genes were identified. Despite a high degree of synteny observed between barley and the sequenced genomes of the grasses rice (Oryza sativa), Brachypodium distachyon and Sorghum bicolor across the wider chromosomal area, the genes in the YAC contigs showed extensive interspecific rearrangements in orientation and order. Consequently, the position of a Ror1 homolog in these species could not be precisely predicted, nor was a barley gene co-segregating with Ror1 identified. These factors have prevented the molecular identification of the Ror1 gene for the time being.

The WtmsDW Locus on Wheat Chromosome 2B Controls Major Natural Variation for Floret Sterility Responses to Heat Stress at Booting Stage.

Heat stress at booting stage causes significant losses to floret fertility (grain set) and hence yield in wheat (Triticum aestivum L.); however, there is a lack of well-characterized sources of tolerance to this type of stress. Here, we describe the genetic analysis of booting stage heat tolerance in a cross between the Australian cultivars Drysdale (intolerant) and Waagan (tolerant), leading to the definition of a major-effect tolerance locus on the short arm of chromosome 2B, Wheat thermosensitive male sterile Drysdale/Waagan (WtmsDW). WtmsDW offsets between 44 and 65% of the losses in grain set due to heat, suggesting that it offers significant value for marker-assisted tolerance breeding. In lines lacking the WtmsDW tolerance allele, peaks in sensitivity were defined with reference to auricle distance, for various floret positions along the spike. Other (relatively minor) floret fertility response effects, including at the Rht-D1 dwarfing locus, were considered likely escape artifacts, due to their association with height and flowering time effects that might interfere with correct staging of stems for heat treatment. Heat stress increased grain set at distal floret positions in spikelets located at the top of the spike and increased the size of spikelets at the base of the spike, but these effects were offset by greater reductions in grain set at other floret positions. Potentially orthologous loci on chromosomes 1A and 1B were identified for heat response of flowering time. The potential significance of these findings for tolerance breeding and further tolerance screening is discussed.

HvNax3--a locus controlling shoot sodium exclusion derived from wild barley (Hordeum vulgare ssp. spontaneum).

Previous work identified the wild barley (Hordeum vulgare ssp. spontaneum) accession CPI-71284-48 as being capable of limiting sodium (Na(+)) accumulation in the shoots under saline hydroponic growth conditions. Quantitative trait locus (QTL) analysis using a cross between CPI-71284-48 and a selection of the cultivated barley (H. vulgare ssp. vulgare) cultivar Barque (Barque-73, a moderate Na(+) excluder) attributed the control of the Na(+) exclusion trait from CPI-71284-48 to a single locus on the short arm of chromosome 7H, which was named HvNax3. The locus reduced shoot Na(+) accumulation by 10-25% in plants grown in 150 mM NaCl. Markers generated using colinearity with rice and Brachypodium, together with the analysis of introgression lines and F(2) and F(3) families, enabled HvNax3 to be mapped to a 1.3-cM interval. Genes from the corresponding rice and Brachypodium intervals encode 16 different classes of proteins and include several plausible candidates for HvNax3. The potential of HvNax3 to provide a useful trait contributing to salinity tolerance in cultivated barley is discussed.

SNARE-protein-mediated disease resistance at the plant cell wall.

Failure of pathogenic fungi to breach the plant cell wall constitutes a major component of immunity of non-host plant species--species outside the pathogen host range--and accounts for a proportion of aborted infection attempts on 'susceptible' host plants (basal resistance). Neither form of penetration resistance is understood at the molecular level. We developed a screen for penetration (pen) mutants of Arabidopsis, which are disabled in non-host penetration resistance against barley powdery mildew, Blumeria graminis f. sp. hordei, and we isolated the PEN1 gene. We also isolated barley ROR2 (ref. 2), which is required for basal penetration resistance against B. g. hordei. The genes encode functionally homologous syntaxins, demonstrating a mechanistic link between non-host resistance and basal penetration resistance in monocotyledons and dicotyledons. We show that resistance in barley requires a SNAP-25 (synaptosome-associated protein, molecular mass 25 kDa) homologue capable of forming a binary SNAP receptor (SNARE) complex with ROR2. Genetic control of vesicle behaviour at penetration sites, and plasma membrane location of PEN1/ROR2, is consistent with a proposed involvement of SNARE-complex-mediated exocytosis and/or homotypic vesicle fusion events in resistance. Functions associated with SNARE-dependent penetration resistance are dispensable for immunity mediated by race-specific resistance (R) genes, highlighting fundamental differences between these two resistance forms.

Flt-2L, a locus in barley controlling flowering time, spike density, and plant height.

Flowering time represents an important adaptive trait for temperate cereal crops and may also impact on frost damage in cereal reproductive tissues by enabling escape or by influencing accumulation of genuine tolerance. The Flowering time-2L (Flt-2L) quantitative trait locus (QTL) on the distal end of barley chromosome arm 2HL overlaps with QTL for rachis internode length and reproductive frost damage. Flt-2L was also found to be associated with plant height. By combining marker analysis with phenotyping in progeny families of selected Amagi Nijo x WI2585 F(6) recombinants, we were able to map quantitative flowering time, rachis internode length, and plant height effects on 2HL as discrete Mendelian traits. The three developmental characters showed codominant modes of expression and perfectly cosegregated with one another in a 1.3-cM marker interval, indicating control by the same gene or closely linked genes. Twelve genes were identified in the related intervals in the rice and Brachypodium distachyon genomes. The HvAP2 gene cosegregated with Flt-2L and represents a plausible candidate for Flt-2L, since it is highly similar to the wheat domestication gene Q which has similar developmental effects. These data will contribute to isolation of the Flt-2L gene(s) and help establish the basis of the frost damage QTL.

Structure-function analysis of the barley genome: the gene-rich region of chromosome 2HL.

A major gene-rich region on the end of the long arm of Triticeae group 2 chromosomes exhibits high recombination frequencies, making it an attractive region for positional cloning. Traits known to be controlled by this region include chasmogamy/cleistogamy, frost tolerance at flowering, grain yield, head architecture, and resistance to Fusarium head blight and rusts. To assist these cloning efforts, we constructed detailed genetic maps of barley chromosome 2H, including 61 polymerase chain reaction markers. Colinearity with rice occurred in eight distinct blocks, including five blocks in the terminal gene-rich region. Alignment of rice sequences from the junctions of colinear chromosome segments provided no evidence for the involvement of long (>2.5 kb) inverted repeats in generating inversions. However, reuse of some junction sequences in two or three separate evolutionary breakage/fusion events was implicated, suggesting the presence of fragile sites. Sequencing across 91 gene fragments totaling 107 kb from four barley genotypes revealed the highest single nucleotide substitution and insertion-deletion polymorphism levels in the terminal regions of the chromosome arms. The maps will assist in the isolation of genes from the chromosome 2L gene-rich region in barley and wheat by providing markers and accelerating the identification of the corresponding points in the rice genome sequence.

Genes and traits associated with chromosome 2H and 5H regions controlling sensitivity of reproductive tissues to frost in barley.

Frost at flowering can cause significant damage to cereal crops. QTL for low temperature tolerance in reproductive tissues (LTR tolerance) were previously described on barley 2HL and 5HL chromosome arms. With the aim of identifying potential LTR tolerance mechanisms, barley Amagi Nijo x WI2585 and Haruna Nijo x Galleon populations were examined for flowering time and spike morphology traits associated with the LTR tolerance loci. In spring-type progeny of both crosses, winter alleles at the Vrn-H1 vernalization response locus on 5H were linked in coupling with LTR tolerance and were unexpectedly associated with earlier flowering. In contrast, tolerance on 2HL was coupled with late flowering alleles at a locus we named Flt-2L. Both chromosome regions influenced chasmogamy/cleistogamy (open/closed florets), although tolerance was associated with cleistogamy at the 2HL locus and chasmogamy at the 5HL locus. LTR tolerance controlled by both loci was accompanied by shorter spikes, which were due to fewer florets per spike on 5HL, but shorter rachis internodes on 2HL. The Eps-2S locus also segregated in both crosses and influenced spike length and flowering time but not LTR tolerance. Thus, none of the traits was consistently correlated with LTR tolerance, suggesting that the tolerance may be due to some other visible trait or an intrinsic (biochemical) property. Winter alleles at the Vrn-H1 locus and short rachis internodes may be of potential use in barley breeding, as markers for selection of LTR tolerance at 5HL and 2HL loci, respectively.

Varietal and chromosome 2H locus-specific frost tolerance in reproductive tissues of barley (Hordeum vulgare L.) detected using a frost simulation chamber.

Exposure of flowering cereal crops to frost can cause sterility and grain damage, resulting in significant losses. However, efforts to breed for improved low temperature tolerance in reproductive tissues (LTR tolerance) has been hampered by the variable nature of natural frost events and the confounding effects of heading time on frost-induced damage in these tissues. Here, we establish conditions for detection of LTR tolerance in barley under reproducible simulated frost conditions in a custom-built frost chamber. An ice nucleator spray was used to minimize potential effects arising from variation in naturally occurring extrinsic nucleation factors. Barley genotypes differing in their field tolerance could be distinguished. Additionally, an LTR tolerance quantitative trait locus (QTL) on the long arm of barley chromosome 2H could be detected in segregating families. In a recombinant family, the QTL was shown to be separable from the effects of the nearby flowering time locus Flt-2L. At a minimum temperature of -3.5 degrees C for 2 h, detection of the LTR tolerance locus was dependent on the presence of the nucleator spray, suggesting that the tolerance relates to freezing rather than chilling, and that it is not the result of plant-encoded variation in ice-nucleating properties of the tiller surface.

Lazy males? Bioenergetic differences in energy acquisition and metabolism help to explain sexual size dimorphism in percids.

1. Differences in energy use between genders is a probable mechanism underlying sexual size dimorphism (SSD), but testing this hypothesis in the field has proven difficult. We evaluated this mechanism as an explanation for SSD in two North American percid species--walleye Sander vitreus and yellow perch Perca flavescens. 2. Data from 47 walleye and 67 yellow perch populations indicated that SSD is associated with the onset of maturation: typically, males of both species matured smaller and earlier and attained a smaller asymptotic size than females. Males also demonstrated equal (perch) or longer (walleye) reproductive life spans compared with females. 3. To examine whether reduced post-maturation growth in males was due to lower energy acquisition or higher reproductive costs we applied a contaminant mass-balance model combined with a bioenergetics model to estimate metabolic costs and food consumption of each sex. Mature males exhibited lower food consumption, metabolic costs and food conversion efficiencies compared with females. 4. We propose that slower growth in males at the onset of maturity is a result of decreased feeding activity to reduce predation risk. Our finding that SSD in percids is associated with the onset of maturity is supported by laboratory-based observations reported elsewhere, showing that changes in growth rate, consumption and food conversion efficiency were elicited by oestrogen (positive effects) or androgen (negative effects) exposure in P. flavescens and P. fluviatilis. 5. Researchers applying bioenergetic models for comparative studies across populations should use caution in applying bioenergetic models in the absence of information on population sex ratio and potential differences between the sexes in energetic parameters.

Diverging temperature responses of CO2 assimilation and plant development explain the overall effect of temperature on biomass accumulation in wheat leaves and grains.

There is a growing consensus in the literature that rising temperatures influence the rate of biomass accumulation by shortening the development of plant organs and the whole plant and by altering rates of respiration and photosynthesis. A model describing the net effects of these processes on biomass would be useful, but would need to reconcile reported differences in the effects of night and day temperature on plant productivity. In this study, the working hypothesis was that the temperature responses of CO2 assimilation and plant development rates were divergent, and that their net effects could explain observed differences in biomass accumulation. In wheat (Triticum aestivum) plants, we followed the temperature responses of photosynthesis, respiration and leaf elongation, and confirmed that their responses diverged. We measured the amount of carbon assimilated per "unit of plant development" in each scenario and compared it to the biomass that accumulated in growing leaves and grains. Our results suggested that, up to a temperature optimum, the rate of any developmental process increased with temperature more rapidly than that of CO2 assimilation and that this discrepancy, summarised by the CO2 assimilation rate per unit of plant development, could explain the observed reductions in biomass accumulation in plant organs under high temperatures. The model described the effects of night and day temperature equally well, and offers a simple framework for describing the effects of temperature on plant growth.

Truncation of grain filling in wheat (Triticum aestivum) triggered by brief heat stress during early grain filling: association with senescence responses and reductions in stem reserves.

Short heat waves during grain filling can reduce grain size and consequently yield in wheat (Triticum aestivum L.). Grain weight responses to heat represent the net outcome of reduced photosynthesis, increased mobilisation of stem reserves (water-soluble carbohydrates, WSC) and accelerated senescence in the grain. To compare their relative roles in grain weight responses under heat, these characteristics were monitored in nine wheat genotypes subjected to a brief heat stress at early grain filling (37°C maximum for 3 days at 10 days after anthesis). Compared with the five tolerant varieties, the four susceptible varieties showed greater heat-triggered reductions in final grain weight, grain filling duration, flag leaf chla and chlb content, stem WSC and PSII functionality (Fv/Fm). Despite the potential for reductions in sugar supply to the developing grains, there was little effect of heat on grain filling rate, suggesting that grain size effects of heat may have instead been driven by premature senescence in the grain. Extreme senescence responses potentially masked stem WSC contributions to grain weight stability. Based on these findings, limiting heat-triggered senescence in the grain may provide an appropriate focus for improving heat tolerance in wheat.

RAR1, ROR1, and the actin cytoskeleton contribute to basal resistance to Magnaporthe grisea in barley.

The fungus Magnaporthe grisea, the causal agent of rice blast disease, is a major pathogen of rice and is capable of producing epidemics on other cultivated cereals, including barley (Hordeum vulgare). We explored the requirements for basal resistance of barley against a compatible M. grisea isolate using both genetic and chemical approaches. Mutants of the RAR1 gene required for the function of major resistance gene-mediated resistance and mutants of the ROR1 and ROR2 genes required for full expression of cell-wall-penetration resistance against powdery mildew pathogens were examined for macroscopic and microscopic alterations in M. grisea growth and symptoms. RAR1 contributed to resistance in epidermis and mesophyll at different stages of fungal infection dependent on the MLO/mlo-5 status. Whereas no ROR2 effect was detected, ROR1 was found to contribute to cell-wall-penetration resistance, at least in the epidermis. Application of the actin agonist cytochalasin E promoted cell wall penetration by M. grisea in a dose-dependent manner, demonstrating an involvement of the actin cytoskeleton in penetration resistance.

Genetic and molecular characterization of the maize rp3 rust resistance locus.

In maize, the Rp3 gene confers resistance to common rust caused by Puccinia sorghi. Flanking marker analysis of rust-susceptible rp3 variants suggested that most of them arose via unequal crossing over, indicating that rp3 is a complex locus like rp1. The PIC13 probe identifies a nucleotide binding site-leucine-rich repeat (NBS-LRR) gene family that maps to the complex. Rp3 variants show losses of PIC13 family members relative to the resistant parents when probed with PIC13, indicating that the Rp3 gene is a member of this family. Gel blots and sequence analysis suggest that at least 9 family members are at the locus in most Rp3-carrying lines and that at least 5 of these are transcribed in the Rp3-A haplotype. The coding regions of 14 family members, isolated from three different Rp3-carrying haplotypes, had DNA sequence identities from 93 to 99%. Partial sequencing of clones of a BAC contig spanning the rp3 locus in the maize inbred line B73 identified five different PIC13 paralogues in a region of approximately 140 kb.

Balancing risks? Responses and non-responses of mayfly larvae to fish and stonefly predators.

In a series of laboratory experiments we examined the hypothesis that larvae of stream mayflies would respond to the presence of two different types of predators in such a way as to minimize their risk of being consumed by each. Positioning of larvae (whether they frequent the top, sides, or bottom of stones) of Baetis tricaudatus and Ephemerella subvaria was altered by the presence of predaceous stoneflies (Agnetina capitata) with a larger proportion of the population occurring on the upper surfaces, where the probability of encountering the predator was lowest. The presence of a benthivorous fish (Cottus bairdi) had no significant effects on positioning of the mayfly larvae. Lack of fish effects may reflect an inability of the mayflies to detect or respond to sculpins, or alternately may indicate that sculpins do not normally present a important predation risk for these mayflies. Failure of mayfly prey to account for fish predators when responding to the presence of stoneflies appcars to explain facilitation previously observed between stoneflies and sculpins.

A mechanism for interference between stream predators: responses of the stonefly Agnetina capitata to the presence of sculpins.

Mottled sculpins (Cottus bairdi) have a strong negative effect on the ability of the stonefly Agnetina capitata to capture some types of mayfly prey. To determine the mechanism for this interference effect, behavior of Agnetina in the presence and absence of sculpins was observed over 24 h periods (12 h light, 12 h dark), using an infra-red sensitive camera and a time-lapse video recorder. Agnetina larvae reacted to the presence of sculpins by significantly reducing the time they spent off the bottom of the substrate, and by significantly decreasing the amount of time spent moving on the substrate. These experiments suggest that in the presence of fish, stonefly diets may contain a smaller proportion of prey that tend to frequent tops and sides of stones. This behavioral flexibility may be important in streams in that it allows stoneflies to advantageously shift their diets when fish population densities are low.