Project description:Terpene volatiles play an important role in the interactions among specialized pathogens and fruits. Citrus Black Spot (CBS), caused by the fungus Phyllosticta citricarpa, is associated with losses in different citrus-growing areas worldwide. The pathogen may infect the fruit for 20-24 weeks after petal fall but the typical hard spot symptoms appear when the fruit become almost mature, as consequence of fungal colonization and induction of cell lyses around essential oil cavities. D-limonene represents about 95% of total oil gland content in mature orange fruit. We have used here d-limonene synthase downregulated orange fruit generated via an antisense (AS) approach to investigate whether d-limonene decrease content in peel oil glands may affect fruit interaction with P. citricarpa compared to that of empty-vector (EV) controls. AS fruit showed enhanced resistance to the fungus compared to EV ones. Because of d-limonene decreased content, an over-accumulation of linalool and other monoterpene alcohols was found in AS compared to EV fruit. A global gene expression analysis at 2h and 8d after inoculation with P. citricarpa revealed activation of defence responses in AS fruit, via the upregulation of different PR genes, likely due to the enhanced constitutive accumulation of linalool and other alcohols. When assayed in vitro and in vivo, the main altered terpene volatile compounds including linalool at concentrations emulating those present in AS fruit, showed strong antifungal activity. We show here that terpene engineering in fruit peels could be a promising method for developing new strategies to obtain resistance to fruit diseases.
Project description:Devastating citrus disease Huanglongbing (HLB) is without existing cures. Herein, we present results demonstrating the possible mechanisms (hypoxia stress) behind HLB-triggered shoot dieback by comparing the transcriptomes, hormone profiles, and key enzyme activities in buds of severely and mildly symptomatic ‘Hamlin’ sweet orange (Citrus sinensis). Within six months (October – May) in field conditions, severe trees had 23% bud dieback, greater than mild trees (11%), with a concomitant reduction in canopy density. In February, differentially expressed genes (DEGs) associated with responses to osmotic stress, low oxygen levels, and cell death were upregulated, with those for photosynthesis and cell cycle downregulated in severe versus mild trees. For severe trees, not only were the key markers for hypoxia, including anaerobic fermentation, reactive oxygen species (ROS) production, and lipid oxidation, transcriptionally upregulated, but also alcohol dehydrogenase activity was significantly greater compared to mild trees, indicating a link between bud dieback and hypoxia. Tricarboxylic acid cycle revival, given the upregulation of glutamate dehydrogenase and alanine aminotransferase DEGs, suggests that ROS may also be generated during hypoxiareoxygenation. Greater (hormonal) ratios of abscisic acid to cytokinins and jasmonates and upregulated DEGs encoding NADPH oxidases in severe versus mild trees indicate additional ROS production under limited oxygen availability due to stomata closure. Altogether, our results provided evidence that as HLB progresses, excessive ROS produced in response to hypoxia and during hypoxia-reoxygenation likely intensify the oxidative stress in buds leading to cell death, contributing to marked bud and shoot dieback and decline of the severely symptomatic sweet orange trees.
Project description:To identify genes associated with citrus peel development and manifestation of peel disorders, we analyzed flavedo, albedo and juice sac tissues from navel orange displaying, and not displaying, the puff disorder. Symptomatic and healthy M-bM-^@M-^\NavelM-bM-^@M-^] orange fruits were harvested from an orchard located in in Pauma Valley, San Diego County, California, USA. Sampling for all analysis (healthy or disordered Navel orange) was performed at the same time, from trees grown under the same agronomic, soil, and environmental conditions. Healthy and disordered fruits were analyzed at the mature stage. All transcriptome analysis was performed on mature fruit. For each type of fruit, three tissues (flavedo, albedo, and juice sacs) from three different trees (biological replicates) were separately analyzed. Four symptomatic fruits comprised one biological replicate each. Two healthy fruits comprised two biological replicates of control samples. A 1 cm-thick equatorial disc and four sections (N, S, E, and W) were cut per fruit. Each section of flavedo, albedo, and juice sac tissue was dissected. gene expression variation underlying quality trait, different genotypes
Project description:Navel orange trees were grafted with budwood infected with the citrus greening bacterium, 'Candidatus Liberibacter asiaticus', or control budwood, and leaf samples were collected every two weeks post graft for LC/MS analysis.
Project description:We evaluated the effect on citrus trees of two newly-identified molecules, benzbromarone and tolfenamic acid, used as antimicrobials in commercial groves of sweet orange (Citrus sinensis). We delivered the molecules by trunk injection and evaluated safety and efficacy parameters by performing RNAseq of the citrus host responses.
Project description:With the aid of a biochip, carrying representative sequences from approximately 2200 sequences from the genome of isolate 9a5c from X. fastidiosa (Xf), microarray-based comparisons have been performed with 6 different Xf isolates obtained from citrus plants (Table 1). Four of these isolates (56a, 9.12c, 187b, and 36f) were obtained from CVC-affected trees and are representatives of the most prevalent Xf haplotypes found in sweet orange orchards across the state of São Paulo, while isolate CV21 was obtained from a non-symptomatic tree from the same region. Isolate Fb7, on the other hand, was obtained from a sweet orange tree that displayed symptoms of “Pecosita”, a disease similar to CVC that occurs in some citrus-growing regions of Argentina. Keywords: Comparative Genomic Hybridization