Linking mechanistic and behavioral responses to sublethal esfenvalerate exposure in the endangered delta smelt; Hypomesus transpacificus (Fam. Osmeridae)

<p>Abstract</p> <p>Background</p> <p>The delta smelt (<it>Hypomesus transpacificus</it>) is a pelagic fish species listed as endangered under both the USA Federal and Californian State Endangered Species Acts and considered an indicator of ecosystem health in its habitat range, which is limited to the Sacramento-San Joaquin estuary in California, USA. Anthropogenic contaminants are one of multiple stressors affecting this system, and among them, current-use insecticides are of major concern. Interrogative tools are required to successfully monitor effects of contaminants on the delta smelt, and to research potential causes of population decline in this species. We have created a microarray to investigate genome-wide effects of potentially causative stressors, and applied this tool to assess effects of the pyrethroid insecticide esfenvalerate on larval delta smelt. Selected genes were further investigated as molecular biomarkers using quantitative PCR analyses.</p> <p>Results</p> <p>Exposure to esfenvalerate affected swimming behavior of larval delta smelt at concentrations as low as 0.0625 μg.L^-1, and significant differences in expression were measured in genes involved in neuromuscular activity. Alterations in the expression of genes associated with immune responses, along with apoptosis, redox, osmotic stress, detoxification, and growth and development appear to have been invoked by esfenvalerate exposure. Swimming impairment correlated significantly with expression of aspartoacylase (ASPA), an enzyme involved in brain cell function and associated with numerous human diseases. Selected genes were investigated for their use as molecular biomarkers, and strong links were determined between measured downregulation in ASPA and observed behavioral responses in fish exposed to environmentally relevant pyrethroid concentrations.</p> <p>Conclusions</p> <p>The results of this study show that microarray technology is a useful approach in screening for, and generation of molecular biomarkers in endangered, non-model organisms, identifying specific genes that can be directly linked with sublethal toxicological endpoints; such as changes in expression levels of neuromuscular genes resulting in measurable swimming impairments. The developed microarrays were successfully applied on larval fish exposed to esfenvalerate, a known contaminant of the Sacramento-San Joaquin estuary, and has permitted the identification of specific biomarkers which could provide insight into the factors contributing to delta smelt population decline.</p

A two-genome microarray for the rice pathogens Xanthomonas oryzae pv. oryzae and X. oryzae pv. oryzicola and its use in the discovery of a difference in their regulation of hrp genes

<p>Abstract</p> <p>Background</p> <p><it>Xanthomonas oryzae </it>pv. <it>oryzae </it>(<it>Xoo</it>) and <it>X. oryzae </it>pv. <it>oryzicola </it>(<it>Xoc</it>) are bacterial pathogens of the worldwide staple and grass model, rice. <it>Xoo </it>and <it>Xoc </it>are closely related but <it>Xoo </it>invades rice vascular tissue to cause bacterial leaf blight, a serious disease of rice in many parts of the world, and <it>Xoc </it>colonizes the mesophyll parenchyma to cause bacterial leaf streak, a disease of emerging importance. Both pathogens depend on <it>hrp </it>genes for type III secretion to infect their host. We constructed a 50–70 mer oligonucleotide microarray based on available genome data for <it>Xoo </it>and <it>Xoc </it>and compared gene expression in <it>Xoo </it>strains PXO99^Aand <it>Xoc </it>strain BLS256 grown in the rich medium PSB vs. XOM2, a minimal medium previously reported to induce <it>hrp </it>genes in <it>Xoo </it>strain T7174.</p> <p>Results</p> <p>Three biological replicates of the microarray experiment to compare global gene expression in representative strains of <it>Xoo </it>and <it>Xoc </it>grown in PSB vs. XOM2 were carried out. The non-specific error rate and the correlation coefficients across biological replicates and among duplicate spots revealed that the microarray data were robust. 247 genes of <it>Xoo </it>and 39 genes of <it>Xoc </it>were differentially expressed in the two media with a false discovery rate of 5% and with a minimum fold-change of 1.75. Semi-quantitative-RT-PCR assays confirmed differential expression of each of 16 genes each for <it>Xoo </it>and <it>Xoc </it>selected for validation. The differentially expressed genes represent 17 functional categories.</p> <p>Conclusion</p> <p>We describe here the construction and validation of a two-genome microarray for the two pathovars of <it>X. oryzae</it>. Microarray analysis revealed that using representative strains, a greater number of <it>Xoo </it>genes than <it>Xoc </it>genes are differentially expressed in XOM2 relative to PSB, and that these include <it>hrp </it>genes and other genes important in interactions with rice. An exception was the <it>rax </it>genes, which are required for production of the host resistance elicitor AvrXa21, and which were expressed constitutively in both pathovars.</p

A two-genome microarray for the rice pathogens Xanthomonas oryzae pv. oryzae and X. oryzae pv. oryzicola and its use in the discovery of a difference in their regulation of hrp genes.

Xanthomonas oryzae pv. oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc) are bacterial pathogens of the worldwide staple and grass model, rice. Xoo and Xoc are closely related but Xoo invades rice vascular tissue to cause bacterial leaf blight, a serious disease of rice in many parts of the world, and Xoc colonizes the mesophyll parenchyma to cause bacterial leaf streak, a disease of emerging importance. Both pathogens depend on hrp genes for type III secretion to infect their host. We constructed a 50-70 mer oligonucleotide microarray based on available genome data for Xoo and Xoc and compared gene expression in Xoo strains PXO99A and Xoc strain BLS256 grown in the rich medium PSB vs. XOM2, a minimal medium previously reported to induce hrp genes in Xoo strain T7174.Three biological replicates of the microarray experiment to compare global gene expression in representative strains of Xoo and Xoc grown in PSB vs. XOM2 were carried out. The non-specific error rate and the correlation coefficients across biological replicates and among duplicate spots revealed that the microarray data were robust. 247 genes of Xoo and 39 genes of Xoc were differentially expressed in the two media with a false discovery rate of 5% and with a minimum fold-change of 1.75. Semi-quantitative-RT-PCR assays confirmed differential expression of each of 16 genes each for Xoo and Xoc selected for validation. The differentially expressed genes represent 17 functional categories.We describe here the construction and validation of a two-genome microarray for the two pathovars of X. oryzae. Microarray analysis revealed that using representative strains, a greater number of Xoo genes than Xoc genes are differentially expressed in XOM2 relative to PSB, and that these include hrp genes and other genes important in interactions with rice. An exception was the rax genes, which are required for production of the host resistance elicitor AvrXa21, and which were expressed constitutively in both pathovars