Jennifer Johnson-Cicalese

Tracing the history of plant traits under domestication in cranberries: potential consequences on anti-herbivore defences

The process of selecting certain desirable traits for plant breeding may compromise other potentially important traits, such as defences against pests; however, specific phenotypic changes occurring over the course of domestication are unknown for most domesticated plants. Cranberry (Vaccinium macrocarpon) offers a unique opportunity to study such changes: its domestication occurred recently, and we have access to the wild ancestors and intermediate varieties used in past crosses. In order to investigate whether breeding for increased yield and fruit quality traits may indirectly affect anti-herbivore defences, the chemical defences have been examined of five related cranberry varieties that span the history of domestication against a common folivore, the gypsy moth (Lymantria dispar). Direct defences were assessed by measuring the performance of gypsy moth caterpillars and levels of phenolic compounds in leaves, and indirect defences by assaying induced leaf volatile emissions. Our results suggest that breeding in cranberry has compromised plant defences: caterpillars performed best on the derived NJS98-23 (the highest-yielding variety) and its parent Ben Lear. Moreover, NJS98-23 showed reduced induction of volatile sesquiterpenes, and had lower concentrations of the defence-related hormone cis-jasmonic acid (JA) than ancestral varieties. However, induced direct defences were not obviously affected by breeding, as exogenous JA applications reduced caterpillar growth and increased the amounts of phenolics independent of variety. Our results suggest that compromised chemical defences in high-yielding cranberry varieties may lead to greater herbivore damage which, in turn, may require more intensive pesticide control measures. This finding should inform the direction of future breeding programmes.

Endophytic and pathogenic fungi of developing cranberry ovaries from flower to mature fruit: diversity and succession

Culturable fungal population diversity and succession was investigated in developing cranberry ovaries of fruit rot-resistant and rot-susceptible cranberry selections, from flower through mature fruit. Fungi were recovered in culture from 1185 of 1338 ovary tissues collected from June to September, yielding 2255 isolates that represented 42 morphotaxa. During the season, species richness varied from 2 to 17 and 2 to 18 in rot-resistant and rot-susceptible selections, respectively, increasing from wk1 to wk10 and then gradually declining to wk14. Shannon-Wiener diversity index varied from 0.27 to 2.32 in rot-resistant and 0.18 to 2.38 in rot-susceptible, and Pielou’s evenness index varied from 0.11 to 0.63 and 0.06 to 0.64 in rot-resistant and rot-susceptible selections, respectively, confirming that diversity of fungi in developing ovaries was similar among rot-resistant and rot-susceptible selections, but varied among sampling time points. Principal component analysis grouped samples collected at the same sampling time point together regardless of rot susceptibility of the selections, and detected the predominant fungal species associated with each stage of development. Successional changes were observed in populations of endophytic, pathogenic and saprophytic fungi throughout the season as ovaries matured.

Characterization and quantification of flavonoids and organic acids over fruit development in American cranberry (Vaccinium macrocarpon) cultivars using HPLC and APCI-MS/MS

Cranberry flavonoids, including anthocyanins, flavonol glycosides and proanthocyanidins, and organic acids were characterized and quantified by HPLC and LC-MS/MS during fruit development and ripening in eight cranberry cultivars. Anthocyanin biosynthesis initiated at early fruit development and reached highest level in mature fruit, with significant differences between cultivars. Major flavonol glycosides, including the most abundant quercetin-3-galactoside and myricetin-3-galactoside, showed consistent concentrations during the season with moderate fluctuation, and were at similar levels in mature fruits of the eight cultivars. Proanthocyanidins declined during fruit development and then increased slightly in later maturation stages. Levels of various proanthocyanidin oligomers/polymers with different degree-of-polymerization were highly correlated within a cultivar during fruit development. Cultivars with coancestry exhibited similar levels (high/low) of anthocyanins or proanthocyanidins, indicating genetic effects on biosynthesis of such flavonoids. All cultivars showed similar levels of malic and citric acids, and declining levels of quinic acid during fruit development. Benzoic acid was extremely low early in the season and increased sharply during fruit ripening. Levels of quinic and citric acids were significantly different among cultivars in the mature fruit. Concentrations of proanthocyanidins, anthocyanins, quinic acid and benzoic acid have a strong developmental association in developing ovaries.

Identification and mapping of fruit rot resistance QTL in American cranberry using GBS

Sustainability of the cranberry industry is threatened by widespread and increasing losses due to fruit rot in the field as well as increasing restrictions on fungicide inputs. Breeding for resistance offers a partial solution but is challenging because fruit rot is caused by a complex of pathogenic fungi that can vary by location and from year to year. We identified four genetically diverse germplasm accessions that exhibit broad-spectrum fruit rot resistance under field conditions. Three of these accessions were used in biparental crosses to develop four populations segregating for resistance. Genotyping by sequencing was used to generate single-nucleotide polymorphism (SNP) markers for development of high-density genetic maps and quantitative trait locus (QTL) analyses. Nineteen QTL associated with fruit rot resistance, distributed on nine linkage groups, were discovered in our populations. Three of these QTL matched previously reported fruit rot resistance QTL. Four newly reported QTL found on linkage group 8 (Vm8), which explain between 21 and 33% of the phenotypic variance for fruit rot, are of particular interest to our breeding program. The populations described herein were also phenotyped for other horticulturally important traits, and QTL associated with yield and berry weight were identified. These QTL provide markers for candidate gene discovery and for future breeding efforts to enhance and pyramid disease resistance and other traits into elite horticultural backgrounds.

Interactions between cranberries and fungi: the proposed function of organic acids in virulence suppression of fruit rot fungi.

Cranberry fruit are a rich source of bioactive compounds that may function as constitutive or inducible barriers against rot-inducing fungi. The content and composition of these compounds change as the season progresses. Several necrotrophic fungi cause cranberry fruit rot disease complex. These fungi remain mostly asymptomatic until the fruit begins to mature in late August. Temporal fluctuations and quantitative differences in selected organic acid profiles between fruit of six cranberry genotypes during the growing season were observed. The concentration of benzoic acid in fruit increased while quinic acid decreased throughout fruit development. In general, more rot-resistant genotypes (RR) showed higher levels of benzoic acid early in fruit development and more gradual decline in quinic acid levels than that observed in the more rot-susceptible genotypes. We evaluated antifungal activities of selected cranberry constituents and found that most bioactive compounds either had no effects or stimulated growth or reactive oxygen species (ROS) secretion of four tested cranberry fruit rot fungi, while benzoic acid and quinic acid reduced growth and suppressed secretion of ROS by these fungi. We propose that variation in the levels of ROS suppressive compounds, such as benzoic and quinic acids, may influence virulence by the fruit rot fungi. Selection for crops that maintain high levels of virulence suppressive compounds could yield new disease resistant varieties. This could represent a new strategy for control of disease caused by necrotrophic pathogens that exhibit a latent or endophytic phase.

Massive phenotyping of multiple cranberry populations reveals novel QTLs for fruit anthocyanin content and other important chemical traits

Because of its known phytochemical activity and benefits for human health, American cranberry (Vaccinium macrocarpon L.) production and commercialization around the world has gained importance in recent years. Flavonoid compounds as well as the balance of sugars and acids are key quality characteristics of fresh and processed cranberry products. In this study, we identified novel QTL that influence total anthocyanin content (TAcy), titratable acidity (TA), proanthocyanidin content (PAC), Brix, and mean fruit weight (MFW) in cranberry fruits. Using repeated measurements over the fruit ripening period, different QTLs were identified at specific time points that coincide with known chemical changes during fruit development and maturation. Some genetic regions appear to be regulating more than one trait. In addition, we demonstrate the utility of digital imaging as a reliable, inexpensive and high-throughput strategy for the quantification of anthocyanin content in cranberry fruits. Using this imaging approach, we identified a set of QTLs across three different breeding populations which collocated with anthocyanin QTL identified using wet-lab approaches. We demonstrate the use of a high-throughput, reliable and highly accessible imaging strategy for predicting anthocyanin content based on cranberry fruit color, which could have a large impact for both industry and cranberry research.

Multivariate GBLUP Improves Accuracy of Genomic Selection for Yield and Fruit Weight in Biparental Populations of Vaccinium macrocarpon Ait

The development of high-throughput genotyping has made genome-wide association (GWAS) and genomic selection (GS) applications possible for both model and non-model species. The exploitation of genome-assisted approaches could greatly benefit breeding efforts in American cranberry (Vaccinium macrocarpon) and other minor crops. Using biparental populations with different degrees of relatedness, we evaluated multiple GS methods for total yield (TY) and mean fruit weight (MFW). Specifically, we compared predictive ability (PA) differences between univariate and multivariate genomic best linear unbiased predictors (GBLUP and MGBLUP, respectively). We found that MGBLUP provided higher predictive ability (PA) than GBLUP, in scenarios with medium genetic correlation (8–17% increase with corg~0.6) and high genetic correlations (25–156% with corg~0.9), but found no increase when genetic correlation was low. In addition, we found that only a few hundred single nucleotide polymorphism (SNP) markers are needed to reach a plateau in PA for both traits in the biparental populations studied (in full linkage disequilibrium). We observed that higher resemblance among individuals in the training (TP) and validation (VP) populations provided greater PA. Although multivariate GS methods are available, genetic correlations and other factors need to be carefully considered when applying these methods for genetic improvement.