Cherry picking by pseudomonads: After a century of research on canker,genomics provides insights into the evolution of pathogenicity towards stone fruits

Bacterial canker disease is a major limiting factor in the growing of cherry and other Prunus species worldwide. At least five distinct clades within the bacterial species complex Pseudomonas syringae are known to be causal agents of the disease. The different pathogens commonly coexist in the field. Reducing canker is a challenging prospect as the efficacy of chemical controls and host resistance may vary against each of the diverse clades involved. Genomic analysis has revealed that the pathogens use a variable repertoire of virulence factors to cause the disease. Significantly, strains of P. syringae pv. syringae possess more genes for toxin biosynthesis and fewer encoding type III effector proteins. There is also a shared pool of key effector genes present on mobile elements such as plasmids and prophages that may have roles in virulence. By contrast, there is evidence that absence or truncation of certain effector genes, such as hopAB, is characteristic of cherry pathogens. Here we highlight how recent research, underpinned by the earlier epidemiological studies, is allowing significant progress in our understanding of the canker pathogens. This fundamental knowledge, combined with emerging insights into host genetics, provides the groundwork for development of precise control measures and informed approaches to breed for disease resistance.     

Soil carbon distribution and quality in a montane rangeland-forest mosaic in northern Utah

Relatively little is known about soil organic carbon (SOC) dynamics in montane ecosystems of the semi-arid western U.S. or the stability of current SOC pools under future climate change scenarios. We measured the distribution and quality of SOC in a mosaic of rangeland-forest vegetation types that occurs under similar climatic conditions on non-calcareous soils at Utah State University's T.W. Daniel Experimental Forest in northern Utah: the forest types were aspen [Populus tremuloides] and conifer (mixture of fir [Abies lasiocarpa] and spruce [Picea engelmannii]); the rangeland types were sagebrush steppe [Artemisia tridentata], grass-forb meadow, and a meadow-conifer ecotone. Total SOC was calculated from OC concentrations, estimates of bulk density by texture and rock-free soil volume in five pedons. The SOC quality was expressed in terms of leaching potential and decomposability. Amount and aromaticity of water-soluble organic carbon (DOC) was determined by water extraction and specific ultra violet absorbance at 254 nm (SUVA) of leached DOC. Decomposability of SOC and DOC was derived from laboratory incubation of soil samples and water extracts, respectively.

Although there was little difference in total SOC between soils sampled under different vegetation types, vertical distribution, and quality of SOC appeared to be influenced by vegetation. Forest soils had a distinct O horizon and higher SOC concentration in near-surface mineral horizons that declined sharply with depth. Rangeland soils lacked O horizons and SOC concentration declined more gradually. Quality of SOC under rangelands was more uniform with depth and SOC was less soluble and less decomposable (i.e., more stable) than under forests. However, DOC in grass-forb meadow soils was less aromatic and more bioavailable, likely promoting C retention through cycling. The SOC in forest soils was notably more leachable and decomposable, especially near the soil surface, with stability increasing with soil depth. Across the entire dataset, there was a weak inverse relationship between the decomposability and the aromaticity of DOC. Our data indicate that despite similar SOC pools, vegetation type may affect SOC retention capacity under future climate projections by influencing potential SOC losses via leaching and decomposition.     

Potential errors in measurement of nonuniform sap flow using heat dissipation probes

The empirical calibration of Granier-type heat dissipation sap flow probes that relate temperature difference (DeltaT) to sap velocity (v) was reevaluated in stems of three tropical tree species. The original calibration was confirmed when the entire heated probe was in contact with conducting xylem, but mean v was underestimated when part of the probe was in contact with nonconducting xylem or bark. Analysis of the effects of nonuniform sap velocity profiles on heat dissipation estimates showed that errors increased as v and the proportion of the probe in nonconducting wood increased. If half of a 20-mm probe is in sapwood with a v of 0.15 mm s(-1) and the other half is in nonconducting wood, then mean v for the whole probe can be underestimated by as much as 50%. A correction was developed that can be used if the proportion of the probe in nonconducting wood is known. Even with the entire heated probe in contact with conducting xylem, v would be underestimated when radial velocity gradients are present. In this case, the error would be smaller except when velocity gradients are very steep, as can occur in species with ring-porous wood anatomy. Errors occur because the relationship between DeltaT and v is nonlinear. Mean DeltaT along the probe is therefore not a measure of mean v, and users of heat dissipation probes should not assume that v is integrated along the length of the probe. The same type of error can occur when DeltaT is averaged through time while v is changing, but the error is small unless there are sudden, step changes between zero and high sap velocity. It is recommended that relatively short probes (20 mm or less) be used and that probes longer than the depth of conducting sapwood be avoided. Multiple probes inserted to a range of depths should be used in situations where steep gradients in v are expected. If these conditions are met, heat dissipation probes remain useful and widely applicable for measuring sap flow in woody stems.     

Establishment techniques affect productivity,nutritive value and atmospheric N2 fixation of two sunn hemp cultivars

Nitrogen fertilization is a common practice for sustaining forage production in forage systems in southeastern United States. Warm-season annual legumes may be an alternative forage to warm-season perennial grasses that do not require N fertilization. Sunn hemp (Crotalaria juncea L.) is a fast-growing, warm-season annual legume native to India and Pakistan. The objective of this 2-year study was to assess the herbage accumulation (HA), atmospheric N₂ fixation (ANF) and nutritive value of sunn hemp. Treatments were the factorial arrangement of two sunn hemp cultivars (“Crescent Sun” and “Blue Leaf”), three seeding rates (17, 28 and 39 kg seed/ha) and seed inoculation (inoculated or non-inoculated seeds), distributed in a randomized complete block design with four replicates. Crescent sun had greater HA (3,218 vs. 1764 kg DM/ha) and ANF (41 vs. 25 kg N/ha). Blue leaf had greater crude protein (CP) (188 vs. 176 g/kg) and in vitro digestible organic matter (IVDOM) concentrations (564 vs. 531 g/kg) than crescent sun. Non-inoculated seed had greater CP than inoculated seed, 188 and 177 g/kg, respectively, and inoculation did not affect HA. Intermediate seeding rate (28 kg/ha) decreased HA (2002 kg DM/ha), while HA from high and low seeding rates (17 and 39 kg/ha, respectively) did not differ (2,863 and 2,615 kg DM/ha respectively). Planting non-inoculated crescent sun at 17 kg/ha seeding rate is a feasible management practice to produce sunn hemp in subtropical regions; however, inoculation should always be recommended for proper establishment.     

Glycine max and Glycine soja are capable of cold acclimation

Soybean has been considered a cold intolerant species; based largely upon seed germination and soil emergent evaluations. This study reports a distinct acquisition of cold tolerance, in seedlings, following short acclimation periods. Diversity in cold responses was assessed in eight cultivars of Glycine max and six accessions of G. soja. All varieties of soybean significantly increased in freezing tolerance following acclimation. This study indicates soybean seedlings are indeed capable of sensing cold and acquiring cold tolerance. Germination rates after cold imbibition were negatively correlated with maturity group, but positively correlated with cold acclimation potential in G. soja. Seed fatty acid composition was varied between the species, with Glycine soja accessions containing about 2‐times more linolenic acid (18:3) than G. max. Furthermore, high levels of linoleic acid (18:2) in seeds were positively correlated with germination rates following cold imbibition in G. soja only. We suggest that domestication has not impacted the overall ability of soybean to cold acclimate at the seedling stage and that there is little variation within the domesticated species for ability to cold acclimate. Thus, this brief comparative study reduces the enthusiasm for the “wild” species as an additional source of genetic diversity for cold tolerance.     

Comparison of the α-amylase inhibitor-1 from common bean (Phaseolus vulgaris) varieties and transgenic expression in other legumes--post-translational modifications and immunogenicity

The seeds of peas (Pisum sativum) and chickpeas (Cicer arietinum) expressing a gene for α-amylase inhibitor-1 (αAI) from the common bean (Phaseolus vulgaris) are protected from damage by old world bruchids (pea and cowpea weevils). Here, we used electrospray ionization time-of-flight mass spectrometry to compare the post-translational modifications of αAI from transgenic sources with the processed forms of the protein from several bean varieties. All sources showed microheterogeneity with differences in the relative abundance of particular variants due to differences in the frequency of addition of glycans, variable processing of glycans, and differences of C-terminal exopeptidase activity. The structural variation among the transgenics was generally within the range of the bean varieties. Previously, mice showed allergic reactions following ingestion of transgenic pea αAI but not bean αAI. Here, only minor differences were observed following intraperitoneal sensitization. Both of the transgenic pea and bean forms of αAI elicited Th1 and Th2 antibody isotype responses, suggesting that both proteins are immunogenic and could potentially be allergenic.     

Validity and reliability of the Block98 food-frequency questionnaire in a sample of Canadian women

OBJECTIVE: To assess the validity and reliability of the most recent adaptation of Block's full-diet food-frequency questionnaire (FFQ) among a sample of Canadian women. DESIGN: Participants completed a self-administered FFQ (FFQ1), two unannounced 24-hour recalls (weekday and weekend) and a second FFQ (FFQ2) between October 2003 and February 2004. FFQs and recalls were analysed for 32 nutrients using Block Dietary Data Systems and the University of Minnesota's Nutrient Data System. Mean and median intakes were computed, along with crude and deattenuated Pearson correlation coefficients between FFQ1 and the average of two recalls (validity) and between FFQ1 and FFQ2 (reliability). SETTING: Ontario, Canada. SUBJECTS: A random population-based sample (n = 166) of women aged 25 to 74 years. RESULTS: One hundred and fifteen (69%) women completed FFQ1, 96 completed FFQ1 and both recalls, and 93 completed both FFQs, about 56 days apart. Mean intakes were similar for most nutrients. FFQ reliability was high, with Pearson correlation coefficients having a median of 0.75, ranging from 0.57 to 0.90 (macronutrients) and from 0.65 to 0.88 (micronutrients from supplements and food). FFQ validity was moderate to high, with deattenuated Pearson correlation coefficients having a median of 0.59, ranging from 0.11 to 0.73 (macronutrients) and from 0.50 to 0.76 (micronutrients from supplements and food). Our micronutrient correlations were similar to or higher than those of other studies that included supplements. Two correlations <0.40 were associated with fats. CONCLUSIONS: The validity and reliability of this full-diet version of the Block FFQ were moderate to high, supporting its use in future studies among Canadian women.     

Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria x ananassa)

An octaploid (Fragaria x ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria x ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways.     

Detecting interaction(s) and assessing the impact of component subsets in a chemical mixture using fixed-ratio mixture ray designs

An important environmental and regulatory issue is the protection of human health from potential adverse effects of cumulative exposure to multiple chemicals. Earlier literature suggested restricting inference to specific fixed-ratio rays of interest. Based on appropriate definitions of additivity, single chemical data are used to predict the relationship among the chemicals under the zero-interaction case. Parametric comparisons between the additivity model and the model fit along the fixed-ratio ray(s) are used to detect departure from additivity. Collection of data along reduced fixed-ratio rays, where subsets of chemicals of interest are removed from the mixture and the remaining compounds are at the same relative ratios as considered in the full ray, allow researchers to make inference about the effect of the removed chemicals. Methods for fitting simultaneous confidence bands about the difference between the best fitting model and the model predicted under additivity are developed to identify regions along the rays where significant interactions occur. This general approach is termed the “single chemicals required” (SCR) method of analysis. A second approach, termed “single chemicals not required” (SCNR) method of analysis, is based on underlying assumptions about the parameterization of the response surface. Under general assumptions, polynomial terms for models fit along fixed-ratio rays are associated with interaction terms. Consideration is given to the case where only data along the mixture rays are available. Tests of hypotheses, which consider interactions due to subsets of chemicals, are also developed.