Research Accomplishment Reports 2007

Advanced Genetic Technologies, KY

C.L. Schardl, E.M. D'Angelo, S.C. Artiushin, J.F. Timoney, A. Downie
Department of Plant Pathology

 

Project Description

The facility was instrumental in expanding the studies at the University of Kentucky on soil metagenomes, molecular phylogenetics of wasps, the molecular basis for pathogenicity of the horse pathogen, Streptococcus equi, virus structures and functions, plant defenses against pathogens and insects, and the mechanisms of variation in secondary metabolism biosynthesis by plants and their symbionts.

Biosolid wastes from animal production facilities and wastewater treatment plants are widely disposed to soils in Kentucky and other states in the nation. One of the main concerns with this practice is that pathogens in the wastes will acquire antibiotic resistance by gene transfer from soil bacteria and other processes in soil environments, thereby affecting the utility of the drugs to combat disease. A comprehensive survey of soil samples for such antibiotic resistance genes was conducted. Genetic variations were investigated in Streptococcus zooepidemicus and S. canis isolated from outbreaks of fatal respiratory disease of dogs and cats respectively. Sequence data acquire following shotgun sequence of S. zooepidemicus W60 genomic library have also been used in projects aimed at development of new generation of assays for diagnosis of strangles of horses.

Impact

Agricultural productivity depends on the genetic potential of the crops and livestock, and their interactions with the agro-ecosystem. The results of this project have enhanced techniques of genetic analysis, and through such techniques, increased understanding of plant genes, fungal symbionts of plants, bacterial pathogens of livestock, and bacterial consortia in the soil environment. Out of 15 resistance genes analyzed in agricultural soils, one gene (vatD) was identified that codes for an acetyltransferase that confers resistance to streptogramin antibiotics, one of the last resort antibiotics for treating multiple antibiotic resistant bacteria in hospital settings. Several different soil bacteria that possess the vat D gene were isolated and characterized. Studies are underway to determine the soil conditions that affect vat D gene transfer to a common pathogen, Enterococcus faecium. Results from the study have important implications on the health of humans and animals that live near areas where biosolid wastes are generated and applied to soils.

Publications

Allen, G.P. 2006. Antemortem detection of horses latently infected with neuropathogenic strains of equine herpesvirus-1. American Journal of Veterinary Research 67: 1401-1405.

Bai, H., Parthasarathy, R., and Palli, S.R. 2007. Identification and characterization of juvenile hormone esterase gene from the yellow fever mosquito, Aedes aegypti. Insect Biochemistry and Molecular Biology 37(8): 829-837.

Caston, J.R., Luque, D., Trus, B. l., Rivas, G., Alfonso, C., Roca, R., Gonzalez, J. M, Carrascosa, J. L., Annamalai, P. and Ghabrial, S. A. 2006. Three-dimensional structure and stoichiometry of Helminthosporium victoriae 190S totivirus. Virology 347: 323-332.

Chandra-Shekara, A.C, Gupte, M., Navarre D.A., Raina, R., Klessig D.F., Kachroo, P. 2006. Light-dependent hypersensitive response and resistance signaling against the Turnip Crinkle Virus in Arabidopsis. The Plant Journal 45: 320-335.

Chandra-Shekara, A.C, Venugopal, S.C., Barman, S.R., Kachroo, A., Kachroo, P. 2007. Plastidial fatty acid levels regulate resistance gene-dependent defense signaling in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America 104: 7277-7282.

Cheng, C.P., Jaag, H.M., Jonczyk, M., Serviene, E., and Nagy, P.D. 2007. Expression of the Arabidopsis Xrn4p 5 '-3 ' exoribonuclease facilitates degradation of tombusvirus RNA and promotes rapid emergence of viral variants in plants. Virology 368(2): 238-248.

Greenhagen B.T., O'Maille, P.E., Noel, J.P., and Chappell, J. 2006. Identifying and manipulating structural determinates linking catalytic specificities in terpene synthases. Proceedings of the National Academy of Sciences of the United States of America 103: 9826-9831.

Gu, H., Zhang, C. and Ghabrial, S.A. 2007. Novel naturally occurring Bean pod mottle virus reassortants with mixed heterologous RNA1 genomes. Phytopathology 97: 79-86.

Hildebrand, D. 2007. Biotechnology and Crop Improvement in Agriculture. In: . Weselake (ed.), Teaching Innovations in Lipid Science. Taylor & Francis, N.Y.

Jaag, H.M., Stork, J., and Nagy, P.D. 2007. Host transcription factor Rpb11p affects tombusvirus replication and recombination via regulating the accumulation of viral replication proteins. Virology 368(2): 388-404.

Jiang, Y., Serviene, E., Gal, J., Panavas, T., and Nagy, P.D. 2006. Identification of essential host factors affecting tombusvirus RNA replication based on the yeast Tet promoters Hughes collection. Journal of Virology 80(15): 7394-7404.

Jonczyk, M., Pathak, K.B., Sharma, M., and Nagy, P.D. 2007. Exploiting alternative subcellular location for replication: Tombusvirus replication switches to the endoplasmic reticulum in the absence of peroxisomes. Virology 362(2): 320-330.

Kachroo, A., Shanklin, J., Lapchyk, L., Whittle, E., Hildebrand, D., Kachroo, P. 2007. The Arabidopsis stearoyl-acyl carrier protein-desaturase family and the contribution of leaf isoforms to oleic acid synthesis. Plant Molecular Biology 63: 257-271.

Kutil, B.L., Greenwald, C., Liu, G., Spiering, M.J., Schardl, C.L., and Wilkinson H.H. 2007. Comparison of loline alkaloid gene clusters across fungal endophytes: Predicting the co-regulatory sequence motifs and the evolutionary history. Fungal Genetics and Biology 44: 1002-1010.

Li, Y., Robinson, G.E., and Palli, S.R. 2007. Identification and Characterization of DmJHRE. Journal of Biological Chemistry 282(52): 37605-37617.

Lorenz, N., Wilson, E.V., Machado, C., Schardl, C., and Tudzynski, P. 2007. Comparison of ergot alkaloid biosynthesis gene clusters in Claviceps species indicate loss of late pathway steps in evolution of C. fusiformis. Applied and Environmental Microbiology 73(22): 7185-7191.

Moon, C.D., Guillaumin, J.-J., Ravel, C., Li, C., Craven, K.D. and Schardl, C.L. 2007. New Neotyphodium endophyte species from the grass tribes Stipeae and Meliceae. Mycologia 99:895-905.

Panavas, T., Stork, J., and Nagy, P. D. 2006. Use of double-stranded RNA templates by the tombusvirus replicase in vitro: Implications for the mechanism of plus-strand initiation. Virology 352(1): 110-120.

Parthasarathy, R., and Palli, S.R. 2007. Developmental and hormonal regulation of midgut remodeling in a lepidopteran insect, Heliothis virescens. Mechanisms of Development 124: 23-34.

Parthasarathy, R., and Palli, S.R. 2007. Stage- and cell-specific expression of ecdysone receptors and ecdysone-induced transcription factors during midgut remodeling in the yellow fever mosquito, Aedes aegypti. Journal of Insect Physiology 53: 216-229.

Schardl, C.L., Grossman, R.B., Nagabhyru, P., Faulkner, J.R., and Mallik, U.P. 2007. Loline alkaloids: currencies of mutualism. Phytochemistry 68: 980-996.

Selosse, M.A., and Schardl, C.L. 2007. Fungal endophytes of grasses: hybrids rescued by vertical transmission? An evolutionary perspective. New Phytologist 173: 452-458.

Sharkey, M.J., Laurenne, N.M., Sharanowski, B., Quicke, D.L.J., and Murray, D. 2006. Revision of the Agathidinae with comparisons of static and dynamic alignments (Hymenoptera: Braconidae). Cladistics 22: 546-567

Sullivan, T.J., Rodstrom, J., Vandop, J., Librizzi, J., Graham, C., Schardl, C.L., and Bultman, T.L. 2007. Symbiont-mediated changes in Lolium arundinaceum inducible defenses: evidence from changes in gene expression and leaf composition. New Phytologist 176: 673-679.

Takahashi, S, Yeo Y., Greenhagen B.T., Song L., Maurina-Brunker J., Rosson R., Noel J.P., and Chappell, J. 2007. Metabolic engineering of sesquiterpene metabolism in yeast. Biotechnology and Bioengineering. 97: 170-181.

Takahashi, S, Yeo Y., Zhao, Y., O'Maille, P.E., Greenhagen, B.T., Noel, J.P. Coates, R.M., and Chappell, J. 2007. Functional characterization of premnaspirodiene oxygenase, a cytochrome P450 catalyzing regio- and stereo-specific hydroxylations of diverse sesquiterpene substrates. Journal of Biological Chemistry 282: 31744-31754.

Timoney, J.F., Qin, A., Muthupalani, S., and Artiushin, S. 2007. Vaccine potential of novel surface exposed and secreted proteins of Streptococcus equi. Vaccine. 25: 5583-5590.

Tiwari, R., Artiushin, S., and Timoney, J.F. 2006. P9, a temperate bacteriophage of Streptococcus equi. In K.S. Sriprakash (ed.) Streptococci - New Insights into an Old Enemy. International Congress Series 1289, Elsevier, The Netherlands. p. 165-168

Tiwari, R., Qin, A., Artiushin, S., and Timoney, J.F. 2007. Se18.9 an anti-phagocytic factor H binding protein of Streptococcus equi. Veterinary Microbiology 121: 105-115.

Venard, C., and Vaillancourt, L. 2007. Colonization of fiber cells by Colletotrichum graminicola in wounded maize stalks. Phytopathogy 97: 438-447.

Venard, C., and Vaillancourt, L. 2007. Penetration and colonization of unwounded maize tissues by the maize anthracnose pathogen Colletotrichum graminicola, and by the related non-pathogen C. sublineolum. Mycologia 99: 368-377.

Mendu V., and Hildebrand, D.F. 2007. Plant Hydroperoxide Lyases and Related Enzymes. Chapt. 24 In: C. Hou (ed.) Biocatalysis and Biotechnology for Functional Foods and Industrial Products. Taylor & Francis, N.Y., p. 399-417.

Wang, E., Dimova, and N., Cambi, F. 2007. PLP/DM20 ratio is regulated by hnRNPH and F and a novel G-rich enhancer in oligodendrocytes. Nucleic Acids Research 35(12): 4164-78.

Wang, R.Y., Kritzman, A. Hershman, D.E., and Ghabrial, S.A. 2006. Aphis glycines as a vector of persistently and nonpersistently transmitted viruses and potential risks for soybean and other crops. Plant Disease 90: 920-926.

Wu, S., Schalk, M., Clark, A., Miles, R.B., Coates, R.M., and Chappell, J. 2006. Redirection of cytosolic or plastidic isoprenoid precursors elevates terpene production in plants. Nature Biotechonlogy 24: 1441-1447.

Wu, S., Schoenbeck, M.A., Greenhagen, B.T., Takahashi, S., Lee, S.B., Coates, R.M. and Chappell, J. 2005. Surrogate splicing for functional analysis of sesquiterpene synthase genes. Plant Physiology 138: 1322-1333.

Xie, J., Wei, D., Jiang, D., Fu, Y., Li, G., Ghabrial, S., and Yi, X. 2006. Characterization of debilitation-associated mycovirus infecting the plant pathogenic fungus Sclerotinia sclerotiorum. Journal of General Virology 87: 241-249.

Yu, K., McCracken, C.T., Jr., Li, R., and Hildebrand, D. 2006. Diacylglycerol acyltransferases from Vernonia and Stokesia prefer substrates with vernolic acid. Lipids 41: 557-566.

Zhao, T., Havens, W.M., and Ghabrial, S.A. 2006. The disease phenotype of virus-infected Helminthosporium victoriae is independent of overexpression of the cellular alcohol oxidase/RNA-binding protein Hv-p68. Phytopathology 96: 326-332.

Zhang, C., and Ghabrial, S.A. 2006. Development of Bean pod mottle virus-based vectors for stable protein expression and sequence-specific virus-induced gene silencing in soybean. Virology 344: 401-411.

Zhang, C., Gu, H., and Ghabrial, S.A. 2007. Molecular characterization of naturally occurring RNA1 recombinants of the comovirus Bean pod mottle virus. Phytopathology 97: 1245-1254.