The 8th Annual

NC State University

Undergraduate Summer Research Symposium

Center for Integrated Fungal Research abstracts

Abstracts are listed in alphabetical order by the last name of the corresponding author.

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Student Author(s):	Brumfield. Leethaniel
Home Institution:	NCSU
Program:	Center for Integrated Fungal Research
College:	CALS
Department(s):	Plant Pathology
Research Mentor(s):	Ralph A. Dean/Plant Pathology Douglas E. Brown/Plant Pathology Joshua K. Sailsbery/Bioinformatics
Title of Presentation:	Bioinformatic Elucidation of Consensus Phosphorylation Motifs Utilizing Inter-Species Functional Data

It is estimated that the world needs to produce 40% more rice by 2030 to feed its more than five billion rice consumers. Fungal disease, particularly that caused by the rice blast fungus Magnaporthe oryzae, is a major factor limiting rice production. The Center for Integrated Fungal Research (CIFR) is actively involved in researching fungal pathogenicity with the intention to control fungal disease, while enhancing the overall safety and protection of the agricultural food supply across the globe. A key to controlling rice blast disease is a better understanding of M. oryzae’s pathogenic mechanisms; an important part of which resides in the ability of cellular signaling molecules (kinases) to phosphorylate a core set of transcription factors (TF) in a direct and controlled manner. Therefore, the more we know about the downstream targets of kinases, their associated pathways, and TF-regulated genes, the more effective controlling pathogenicity efforts will be. Previous pathway and network structure research in S. cerevisiae and H. sapiens may be utilized to better understand cellular signaling in M. oryzae when investigating similar proteins. Large scale protein phosphorylation microarrays can be used to accurately identify functional TF targets of homologous kinases across these species. Potentially phosphorylated binding motifs were identified in these TFs using the Pratt algorithm that detects sequence patterns. These TF phosphorylation motifs were used to examine the shared functionality between homologous kinases. Such motifs may also provide potential chemical targets and aid in developing disease control strategies. In our continued mission to fully understand the transcriptional control of each gene and the targets of each TF involved in controlling infection related development and pathogenicity, future research includes comparing the data compiled from Pratt with other motif-finding programs and eventually composing an open-to-the-public online M. oryzae TF database.

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Student Author(s):	Shows, Angela L.
Home Institution:	Meredith College
Program:	Center for Integrated Fungal Research
College:	CALS
Department(s):	Plant Pathology
Research Mentor(s):	Yeon Yee Oh/Plant Pathology Ralph Dean/Plant Pathology Yeon Yee Oh/Plant Pathology Junhyun Jeon/Plant Pathology Gregory Bernard/Plant Pathology
Title of Presentation:	Ubiqutin-mediated Proetolysis and Magnaporthe oryzae

The SCF complex is a proteasome that works to degrade “trash” proteins that have been ubiqutinated. The SCF complex consists of Skp1, Cul1, Rbx1, and an F-box protein. The protein material broken down by the SCF complex can then be used for other cellular purposes. During spore germination in Magnaporthe oryzae the contents of the spore are degraded and recycled into the developing infection cells, appressorium. MGG_01282 is one of the genes responsible for tagging proteins with ubiquitn. We hypothesized that the deleting this gene would interfere with appressorium formation and reduce virulence. To knockout MGG_01282, a deletion cassette was constructed by PCR containing a gene conferring resistance to hygromycin flanked by boarder fragments of MGG_01282. After screening transformants for hygromycin resistance, DNA was extracted and PCR was done to test for genotypic evidence of mutation. Three gene deletion mutants were found and tested against two ectopic transformants and the 70-15 wild type strain. Phenotypic analysis showed that mutants had consistently underdeveloped spores, low spore concentration, and reduced formation of appressorium. When grown on V8 media, mutants had slower growth rates than that of the ectopic and wild type strains. However, when grown on minimal media, mutant growth rates were only slightly slower than the ectopic strains and both mutant and ectopic strains had faster growth rates than the wild type. A test for pathogenicity against barley plants was attempted, but due to the low spore concentration of the mutants, the test proved to be inconclusive. Further testing will be done to genetically confirm the mutation and assess pathogenicity on host plants.

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