Source: TEXAS A&M UNIVERSITY submitted to
MECHANISMS OF FUNGAL GROWTH AND DEVELOPMENT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1009335
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Mar 29, 2016
Project End Date
Feb 16, 2021
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001
Performing Department
Plant Pathology & Microbiology
Non Technical Summary
Fungi account for as much as 70% of the economic impact of plant diseases. Critical steps in the fungal life style include production of spores (conidia) and germination of these spores to form a thread-like cell called a hyphae. Both cell types are critical to the life cycle of the fungus, and both cell types are unique to fungi. Therefore, an understanding of how the fungus regulates these cell differentiation events will lead to a greater chance of disrupting the disease cycle. The work proposed here will address the molecular and cellular mechanisms utilized by fungi to regulate cell differentiation.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
20640201030100%
Knowledge Area
206 - Basic Plant Biology;

Subject Of Investigation
4020 - Fungi;

Field Of Science
1030 - Cellular biology;
Goals / Objectives
Here we willa) Utilize GFP fused proteins that localize to the cell apex to assess the apical recycling model.A growing catalog of apically localized proteins has been reported in A. nidulans including for example SynA (Tahari-Talesh, 2008); TeaC (Higashitsuji et al. 2009), and DnfA (Schulthaus and Shaw, 2015). Here we will express GFP fusions of apically localized proteins in conditional endocytosis mutants to follow the localization of the proteins when endocytosis is compromised. We anticipate that the localization of each will disperse across the cell in the absence of endocytosis. The dynamics of this alteration of localization will provide insight into the mechanisms of hyphal growth and polarization in fungi.b) Test the Apical Recycling Model through examination of mutants affected in Endocytosis.A Bioinformatic survey of the Aspergillus nidulans genome showed that only 36 proteins contain an NPFxD motif that is diagnostic for endocytosed proteins (Schultzhaus et al. 2015). An examination of one (DnfA; Schultzhaus et al. 2015) demonstrated a clear role of endocytic mechanisms in the dynamics of hyphal growth. Here we will systematically examine the role of other NPFxD containing proteins in hyphal growth. Many of these proteins identified in A. nidulans have no ascribed function associated with them. We anticipate that novel proteins important for hyphal growth will be discovered.c) Complement by over-expression an endocytosis defective mutant of A. nidulans to determine what genes interact with fimbrin during the process of hyphal growthOne phenotype of the fimbrin (endocytosis) mutant is that after multiple rounds of cell cycle progression polarization is recovered even though fimbrin is absent from the endocytic sites in the apical collar of hyphae. We hypothesize that additional actin binding proteins are upregulated to substitute for fimbrin at these sites. To identify these components we will over-express candidate actin binding proteins in the fimbrin mutant and screen for bypass of the polarization defect. Identification of these components will establish further components necessary for the endocytic machinery at the hyphal apex.
Project Methods
Here we willa) Utilize GFP fused proteins that localize to the cell apex to assess the apical recycling model.A growing catalog of apically localized proteins has been reported in A. nidulans including for example SynA (Tahari-Talesh, 2008); TeaC (Higashitsuji et al. 2009), and DnfA (Schulthaus and Shaw, 2015). Here we will express GFP fusions of apically localized proteins in conditional endocytosis mutants to follow the localization of the proteins when endocytosis is compromised. We anticipate that the localization of each will disperse across the cell in the absence of endocytosis. The dynamics of this alteration of localization will provide insight into the mechanisms of hyphal growth and polarization in fungi.b) Test the Apical Recycling Model through examination of mutants affected in Endocytosis.A Bioinformatic survey of the Aspergillus nidulans genome showed that only 36 proteins contain an NPFxD motif that is diagnostic for endocytosed proteins (Schultzhaus et al. 2015). An examination of one (DnfA; Schultzhaus et al. 2015) demonstrated a clear role of endocytic mechanisms in the dynamics of hyphal growth. Here we will systematically examine the role of other NPFxD containing proteins in hyphal growth. Many of these proteins identified in A. nidulans have no ascribed function associated with them. We anticipate that novel proteins important for hyphal growth will be discovered.c) Complement by over-expression an endocytosis defective mutant of A. nidulans to determine what genes interact with fimbrin during the process of hyphal growthOne phenotype of the fimbrin (endocytosis) mutant is that after multiple rounds of cell cycle progression polarization is recovered even though fimbrin is absent from the endocytic sites in the apical collar of hyphae. We hypothesize that additional actin binding proteins are upregulated to substitute for fimbrin at these sites. To identify these components we will over-express candidate actin binding proteins in the fimbrin mutant and screen for bypass of the polarization defect. Identification of these components will establish further components necessary for the endocytic machinery at the hyphal apex.

Progress 03/29/16 to 02/16/21

Outputs
Target Audience:The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will lead to the development of new medicines and pesticides to control Animal and plant disease caused by fungi. I taught anundergraduate course on the impact of fungi on society 5 times. Class enrollment exceeded 500 students and included a high percentage of women, Latinx, and African Americans, which ensured dissemination of my research to underrepresented groups. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student, Dr. Zach Schultzhaus , published 4 papers, defended his dissertation and graduated with his PhD. One graduate student, Dr. Blake Commer published two papers, defended her dissertation and graduated with her PhD. A third graduate student, Mr. Joe Vasselli is begin work to examine the role of endocytosis during hyphal growth and appressorium differentiation in the maize pathogen Colletotrichum graminicola. Two undergraduate students Ellen Kainer and Hope Hancock supported the graduate student's research. Undergraduate students each developed a poster and presented it at a campus wide undergraduate research symposium. My laboratory personnel trained and worked with 10 research personnel from other labs on the use of research microscopes. A graduate student gave a research seminar to the department. How have the results been disseminated to communities of interest?The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Three graduate student presented a departmental seminars and multiple conference talks. I also incorporate my research into my undergraduate and graduate fungal biology class classes that average 100 undergraduates and 10 graduate students per year to total over 500 undergraduates and 50 graduate students during the reporting period. What do you plan to do during the next reporting period to accomplish the goals?Final Report

Impacts
What was accomplished under these goals? 14 peer reviewed publications during the life of this project. a) We utilized GFP fused proteins that localize to the cell apex to assess the apical recycling model. GFP or RFP (mCherry) fusions have been recovered for 14 of the candidate proteins. 9 of them localize to predicted sites of endocytosis and hyphal growth, 5 do not produce detectable levels of fluorescence. Strains are being constructed to co localize these proteins. 2 of these proteins are encoded only by other filamentous fungi making them outstanding candidates for being fungal specific growth factors. b) We Tested the Apical Recycling Model through examination of mutants affected in Endocytosis. The markers described in A, are being assesed using live cell imaging to examine endocytosis during Strains are being constructed to place these markers in endocytic mutants to examine their fate when endocytosis is knocked down. c) No progress to report.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Z. Schultzhaus and B. D. Shaw. 2016. The Flippase DnfB is Cargo of Fimbrin-associated Endocytosis in Aspergillus nidulans, and likely recycles through the late Golgi. Communicative and Integrative Biology. 9:2, e1141843, DOI: 10.1080/19420889.2016.1141843
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Z. Schultzhaus, L. Quintanilla, A. Hilton, and B. D. Shaw. 2016. Live Cell Imaging of Actin Dynamics in the Model Filamentous Fungus Aspergillus nidulans Using Lifeact. Microscopy and Microanalysis.22: 264-274.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: C.-L. Wang and B. D. Shaw. 2016. F-actin localization dynamics during appressorium formation in Colletotrichum graminicola. Mycologia. 108:506-514.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: C.-L. Wang, W.-B. Shim, and B. D. Shaw. 2016. The Colletotrichum graminicola striatin ortholog Str1 is necessary for anastomosis and is a virulence factor. Molecular Plant Pathology.17: 931-942.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: M. R. Mendoza, A. Payne, S. Castillo, M. Crocker, B. D. Shaw, H. B. Scholthof. 2017. Expression of separate proteins in the same plant leaves and cells using two independent virus-based gene vectors. Frontiers in Plant Science 8:1808
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: T. Isakeit, B. Commer, B. D. Shaw, M. Brown and C. Neely. 2017. First Report of Leaf Spot of Barley Caused by Drechslera gigantea in the United States. Plant Disease: 101 1548.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Z. Schultzhaus, W. Zheng, Z. Wang, R. Mourino-Perez and B. D. Shaw. 2017. Phospholipid Flippases DnfA and DnfB Exhibit Differential Dynamics within the A. nidulans Spitzenk�rper. Fungal Genetics and Biology. 99:26-28.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Z. Schultzhaus, T. B. Johnson, and B. D. Shaw. 2017. Clathrin Localization and Dynamics in Aspergillus nidulans. Molecular Microbiology 103: 299-318.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Strycker, Z. Han, B. Commer, B. Shaw, A. Sokolov, and M. Scully. 2019. CARS Spectroscopy of Aspergillus nidulans spores. Scientific Reports. 9(1):1789. doi: 10.1038/s41598-018-37978-6
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Schultzhaus. G. A. Cunningham, R. R. Mouri�o-P�rez, and B. D. Shaw. 2019. The phospholipid flippase DnfD localizes to late Golgi and is involved in asexual differentiation in Aspergillus nidulans. Mycologia. 111(1):13-25. doi: 10.1080/00275514.2018.1543927/
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Blake Commer, Zachary Schultzhaus, Brian D. Shaw. 2000. Localization of NPFxD motif-containing proteins in Aspergillus nidulans. Fungal Genetics and Biology.141 https://doi.org/10.1016/j.fgb.2020.103412.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Blake Commer & Brian D. Shaw. 2020. Current views on endocytosis in filamentous fungi, Mycology https://doi.org/10.1080/21501203.2020.1741471
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Abhinav Bhardwaj, Joseph Vasselli, Matt Lucht, Zhijian Pei, Brian Shaw, Zachary Grassley, Xingjian Wei, Na Zou. 2020. 3D Printing of Biomass-Fungi Composite Material: A Preliminary Study. Manufacturing Letters 24: 96-99. https://doi.org/10.1016/j.mfglet.2020.04.005
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Zehua Han, Benjamin D. Strycker, Blake Commer, Kai Wang, Brian D. Shaw, Marlan O. Scully & Alexei V. Sokolov. 2020. Molecular origin of the Raman signal from Aspergillus nidulans conidia and observation of fluorescence vibrational structure at room temperature. Scientific Reports 10, Article number: 5428. https://doi.org/10.1038/s41598-020-62112-w


Progress 10/01/19 to 09/30/20

Outputs
Target Audience:The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will lead to the development of new medicines and pesticides to control Animal and plant disease caused by fungi. In 2020 I taught one undergraduate course on the impact of fungi on society. Class enrollment was a total of 100 students and included a high percentage of women, latinx, and African Americans, which ensured dissemination of my research to underrepresented groups. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student, Dr. Blake Commer published two paper, listed above, defended her dissertation and graduated with her PhD. A second graduate student, Mr. Joe Vasselli is begin work to examine the role of endocytosis during hyphal growth and appressorium differentiation in the maize pathogen Colletotrichum graminicola. Two undergraduate students Ellen Kainer and Hope Hancock supported the graduate student's research. Undergraduate students each developed a poster and presented it at a campus wide undergraduate research symposium. My laboratory personnel trained and worked with 10 research personnel from other labs on the use of research microscopes. A graduate student gave a research seminar to the department. How have the results been disseminated to communities of interest?The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. A graduate student presented a departmental seminar. I also incorporate my research into my classes that included 100 undergraduates and 10 graduate students What do you plan to do during the next reporting period to accomplish the goals?Graduate student, Mr. Vaselli is working toward his first publication which will the expansion of this field of study into the maize pathogen Colletotrichum graminicola.

Impacts
What was accomplished under these goals? a) Two publications this reporting period relating to objective a. Blake Commer, Zachary Schultzhaus, Brian D. Shaw. 2000. Localization of NPFxD motif-containing proteins in Aspergillus nidulans. Fungal Genetics and Biology.141 https://doi.org/10.1016/j.fgb.2020.103412. Blake Commer & Brian D. Shaw. 2020. Current views on endocytosis in filamentous fungi, Mycologyhttps://doi.org/10.1080/21501203.2020.1741471 b) Test the Apical Recycling Model through examination of mutants affected in Endocytosis. The markers described in A, are being assesed using live cell imaging to examine endocytosis during Strains are being constructed to place these markers in endocytic mutants to examine their fate when endocytosis is knocked down. c) Complement by over-expression an endocytosis defective mutant of A. nidulans to determine what genes interact with fimbrin during the process hyphal growth. No progress to report.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Blake Commer, Zachary Schultzhaus, Brian D. Shaw. 2000. Localization of NPFxD motif-containing proteins in Aspergillus nidulans. Fungal Genetics and Biology.141 https://doi.org/10.1016/j.fgb.2020.103412.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Blake Commer & Brian D. Shaw. 2020. Current views on endocytosis in filamentous fungi, Mycology https://doi.org/10.1080/21501203.2020.1741471
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Abhinav Bhardwaj, Joseph Vasselli, Matt Lucht, Zhijian Pei, Brian Shaw, Zachary Grassley, Xingjian Wei, Na Zou. 2020. 3D Printing of Biomass-Fungi Composite Material: A Preliminary Study. Manufacturing Letters 24: 96-99. https://doi.org/10.1016/j.mfglet.2020.04.005
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Zehua Han, Benjamin D. Strycker, Blake Commer, Kai Wang, Brian D. Shaw, Marlan O. Scully & Alexei V. Sokolov. 2020. Molecular origin of the Raman signal from Aspergillus nidulans conidia and observation of fluorescence vibrational structure at room temperature. Scientific Reports 10, Article number: 5428. https://doi.org/10.1038/s41598-020-62112-w


Progress 10/01/18 to 09/30/19

Outputs
Target Audience:The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will lead to the development of new medicines and pesticides to control Animal and plant disease caused by fungi. Efforts to disseminate my research include: In 2018-2019 I attended two international scientific meetings and presented invited plenary talks at The Fungal Genetics Meeting at Asilomar California, and the 13th National Congress on the Molecular and Cellular Biology of Fungi in Ensenada Mexico. Two presentations relating to my Hatch projected were presented by two graduate students. I estimate that through these presentations my research was disseminated to approximately 1100 scientists. In 2019 I taught one undergraduate course on the impact of fungi on society. Class enrollment was a total of 98 students and included a high percentage of women, latinos, and African Americans, which ensured dissemination of my research to underrepresented groups. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student, Ms. Blake Commer is conducting research in the lab in support of this project during the reporting period. She is preparing her work for publication and it is anticipated that she will complete her PhD in 2020. A second graduate student, Mr. Joe Vasselli is beginning to work to examine the role of endocytosis in appressorium differentiation in the maize pathogen Colletotrichum graminicola. An undergraduate: Reid Thrash supported the graduate student's research. Both graduate students presented their research at scientific meetings (The Fungal Genetics conference). Undergraduate students each developed a poster and presented it at a campus wide undergraduate research symposium. My laboratory personnel trained and worked with 10 research personnel from other labs on the use of research microscopes. A graduate student gave a research seminar to the department. How have the results been disseminated to communities of interest?The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. My students and I have attended scientific meetings including: the Fungal Genetics Conference and the 13th National Congress of the Molecular and Cellular Biology of Fungi in Ensenada Mexico. I was an invited plenary speaker at both meetings. I estimate that more than 1100 total constituents where reached with these efforts. I also incorporated my research into my classes that included 100 undergraduates and 10 graduate students. What do you plan to do during the next reporting period to accomplish the goals?Ms. Blake Commer will continue her characterization of the suite of endocytosis proteins and their role in hyphal growth. Analysis will expand to disruption of the genes to analyze mutant phenotypes. A new graduate student, Mr. Joe Vasselli, was recruited to the project in November 2017. Mr. Vaselli will continue to look at the role of endocytosis in differentiation of appressoria in fungi.

Impacts
What was accomplished under these goals? a) Utilize GFP fused proteins that localize to the cell apex to assess the apical recycling model. GFP or RFP (mCherry) fusions have been recovered for 14 of the candidate proteins. 9 of them localize to predicted sites of endocytosis and hyphal growth, 5 do not produce detectable levels of fluorescence. Strains are being constructed to co localize these proteins. 2 of these proteins are encoded only by other filamentous fungi making them outstanding candidates for being fungal specific growth factors. b) Test the Apical Recycling Model through examination of mutants affected in Endocytosis. The markers described in A, are being assesed using live cell imaging to examine endocytosis during Strains are being constructed to place these markers in endocytic mutants to examine their fate when endocytosis is knocked down. c) Complement by over-expression an endocytosis defective mutant of A. nidulans to determine what genes interact with fimbrin during the process hyphal growth. No progress to report.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Strycker, Z. Han, B. Commer, B. Shaw, A. Sokolov, and M. Scully. 2019. CARS Spectroscopy of Aspergillus nidulans spores. Scientific Reports. 9(1):1789. doi: 10.1038/s41598-018-37978-6
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Schultzhaus. G. A. Cunningham, R. R. Mouri�o-P�rez, and B. D. Shaw. 2019. The phospholipid flippase DnfD localizes to late Golgi and is involved in asexual differentiation in Aspergillus nidulans. Mycologia. 111(1):13-25. doi: 10.1080/00275514.2018.1543927/


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will leadto the development of new medicines and pesticides to control animal and plant disease caused by fungi. Efforts to disseminate my research include: In 2017-2018I attended one international scientific meeting. The International Mycology Congress.One presentationrelating to my Hatch projected was presented by a graduate student. Grad. I estimate that through these presentations my research was disseminated to approximately 100 scientists. In 2018I taught one undergraduate course on the impact of fungi on society. Class enrollment was a total of 100 students and included a high percentage of women, latinos, and African Americans, which ensured dissemination of my research to underrepresented groups. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Twograduate students: Ms. Blake Commer and Mr. Joe Vasselli conducted research in the lab in support of this project during the reporting period. Bothgraduate students presented their research at scientific meetings (one international meetings, sveral on campus). Two undergraduate students participated in summer REU in my lab. Undergraduate students each developed a poster and presented it at a campus wide undergraduate research symposium. My laboratory personnel trained and worked with 12research personnel from other labs on the use of research microscopes. A graduate student gave a research seminar to the department. How have the results been disseminated to communities of interest?The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will leadto the development of new medicines and pesticides to control Animal and plant disease caused by fungi. Efforts to disseminate my research include: In 2015-2016 I attended one international scientific meeting. The Mycological Society of America annual meeting two presentations relating to my Hatch projected were presented by two graduate students. Grad. student Zach Scultzhaus also attended and presented at the Gordon Research conference on Cellular and Molecular Biology of Fungi.. I estimate that through these presentations my research was disseminated to approximately 100 scientists. In 2016 I taught one undergraduate course on the impact of fungi on society. Class enrollment was a total of 150 students and included a high percentage of women, latinos, and African Americans, which ensured dissemination of my research to underrepresented groups. What do you plan to do during the next reporting period to accomplish the goals?A new graduate student (Joe Vasselli) joined the lab. He will continue the characterization of endocytosis during fungal grwoth and development by examining the role of endocytosis in appressorium development. Graduate student Blake Commer will publish a comprehensive assesment of NPFxD motif containing proteins and their role in fungal growth.

Impacts
What was accomplished under these goals? During this reporting period I have two papers accepted.Note these do not yet have volume and page number assigned so I could not report them under outputs. Both will be published in early 2019. B. Strycker, Z. Han, B. Commer, B. Shaw, A. Sokolov, and M. Scully. 2019. CARS Spectroscopy of Aspergillus nidulans spores. Scientific Reports. Z. Schultzhaus. G. A. Cunningham, R. R. Mouriño-Pérez, and B. D. Shaw. 2019The phospholipid flippase DnfD localizes to late Golgi and is involved in asexual differentiation in Aspergillus nidulans. Mycologia. Graduate student Blake Commer gave an invited symposium talk (in my stead) at the Internation Mycology Congress. For this talk she was awarded best graduate student talk for thecongress. To date Commer has GFP tagged 15 proteins and verified that the NPFxD motif is a robust predictor for proteins involve din apical growth in fungi. A new graduate student. Joe Vasselli was recruited to the lab during the reporting period. He will test the role of endocytosis during appressorium formation in the phytopathogenic fungusColletotrichum graminicola.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: Blake Commer, Zach Schultzhaus, Brian D. Shaw. Trafficking of Membrane and Endocytic Cargo Proteins in A. nidulans. 2018. Invited abstract for International Mycology Congress. http://imc11.com/


Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will lead to the development of new medicines and pesticides to control Animal and plant disease caused by fungi. Efforts to disseminate my research include: In 2015-2016 I attended one international scientific meeting. The Mycological Society of America annual meeting two presentations relating to my Hatch projected were presented by two graduate students. Grad. student Zach Scultzhaus also attended and presented at the Gordon Research conference on Cellular and Molecular Biology of Fungi.. I estimate that through these presentations my research was disseminated to approximately 100 scientists. In 2016 I taught one undergraduate course on the impact of fungi on society. Class enrollment was a total of 150 students and included a high percentage of women, latinos, and African Americans, which ensured dissemination of my research to underrepresented groups. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student, Dr. Zach Schultzhaus completed his PhD, while another, Ms. Blake Commer is conductng research in the lab in support of this project during the reporting period. two undergraduates: Mr. William Porter and Ms.. Allison Donovan supported the graduate student's research. Both graduate students presented their research at scientific meetings (two at the Fungal Genetics conference, one at the Mycological Society of America). Undergraduate students each developed a poster and presented it at a campus wide undergraduate research symposium. My laboratory personnel trained and worked with 10 research personnel from other labs on the use of research microscopes. A graduate student gave a research seminar to the department. How have the results been disseminated to communities of interest?The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. My students and I have attended scientific meetings including: the Mycolgogical Society of America, and the Fungal Genetics Conference. I was an invited speaker at the International Fungal Spore Conference in Mexico. Undergraduates presented their research at an Undergraduate Science research fair. I estimate that more than 200 total constituents where reached with these efforts. I also incorporate my research into my classes that included 150 undergraduates and 10 graduate students What do you plan to do during the next reporting period to accomplish the goals?Ms. Blake Commer will continue her characterization of the suite of endocytosis proteins and their role in hyphal growth. Analysis will expand to disruption of the genes to analyze mutant phenotypes. A new graduate student, Mr. Joe Vasseli, was recruited to the project in November 2017. Mr. Vaselli will begin an expansion of the project to look at the role of endocytosis in differentiation of appressoria in fungi.

Impacts
What was accomplished under these goals? a) Utilize GFP fused proteins that localize to the cell apex to assess the apical recycling model. GFP or RFP (mCherry) fusions have been recovered for 11 of the candidate proteins. 7 of them localize to predicted sites of endocytosis and hyphal growth, 4 do not produce detectable levels of fluorescence. Strains are being constructed to co localize these proteins. 2 of these proteins are encoded only by other filamentous fungi making them outstanding candidates for being fungal specific growth factors. b) Test the Apical Recycling Model through examination of mutants affected in Endocytosis. The markers described in A, are being assesed using live cell imaging to examine endocytosis during Strains are being constructed to place these markers in endocytic mutants to examine their fate when endocytosis is knocked down. c) Complement by over-expression an endocytosis defective mutant of A. nidulans to determine what genes interact with fimbrin during the process hyphal growth. No progress to report.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: M. R. Mendoza, A. Payne, S. Castillo, M. Crocker, B. D. Shaw, H. B. Scholthof. 2017. Expression of separate proteins in the same plant leaves and cells using two independent virus-based gene vectors. Frontiers in Plant Science 8:1808
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: T. Isakeit, B. Commer, B. D. Shaw, M. Brown and C. Neely. 2017. First Report of Leaf Spot of Barley Caused by Drechslera gigantea in the United States. Plant Disease: 101 1548.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Z. Schultzhaus, W. Zheng, Z. Wang, R. Mourino-Perez and B. D. Shaw. 2017. Phospholipid Flippases DnfA and DnfB Exhibit Differential Dynamics within the A. nidulans Spitzenk�rper. Fungal Genetics and Biology. 99:26-28.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Z. Schultzhaus, T. B. Johnson, and B. D. Shaw. 2017. Clathrin Localization and Dynamics in Aspergillus nidulans. Molecular Microbiology 103: 299-318.


Progress 03/29/16 to 09/30/16

Outputs
Target Audience:The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. Additionally, the target audience includes undergraduate students and graduate students whom I will mentor in the classroom and in the laboratory. Ultimately, my research will contribute to the scientific knowledge base that will lead to the development of new medicines and pesticides to control Animal and plant disease caused by fungi. Efforts to disseminate my research include: In 2015-2016 I attended one international scientific meeting. The Mycological Society of America annual meeting two presentations relating to my Hatch projected were presented by two graduate students. Grad. student Zach Scultzhaus also attended and presented at the Gordon Research conference on Cellular and Molecular Biology of Fungi.. I estimate that through these presentations my research was disseminated to approximately 100 scientists. In 2016 I taught one undergraduate course on the impact of fungi on society. Class enrollment was a total of 150 students and included a high percentage of women, latinos, and African Americans, which ensured dissemination of my research to underrepresented groups.? Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Three graduate students: Ms. Brigitte Bomer and Mr. Zachary Schultzhaus and Ms. Blake Commer conducted research in the lab in support of this project during the reporting period. Five undergraduates: Ms. Nicole Donhauser, Ms. Grace Cunningham, Ms. Jessie Gwinn, Ms. Elena Edwards and Mr. Stephon Warren supported the graduate student's research. All three graduate students presented their research at scientific meetings (two international meetings, one on campus). Undergraduate students each developed a poster and presented it at a campus wide undergraduate research symposium. My laboratory personnel trained and worked with 10 research personnel from other labs on the use of research microscopes. A graduate student gave a research seminar to the department.? How have the results been disseminated to communities of interest?The direct target audiences for this research project are my colleagues in the fields of fungal developmental biology and fungal genetics. My students and I have attended scientific meetings including: the Mycolgogical Society of America, the Gordon Research Conference on Cellular and Molecular Biology of Fungi, and the Imaging Science Spotlight Series at Texas A&M University. Uudergraduates presented their research at an Undergraduate Science research fair. I estimate that more than 200 total constiuents where reached with these efforts. I also incorporate my research into my classes that included 150 undergraduatesand 10 graduate students. What do you plan to do during the next reporting period to accomplish the goals?A new graduate student, Ms. Blake Commer, was recruited to the project during the last fiscal year. She is beginnning to systematically construct GFP tags of target proteins that contain the NPFxD motif.

Impacts
What was accomplished under these goals? a)Utilize GFP fused proteins that localize to the cell apex to assess the apical recycling model.Two markers have been developed, EcoA-GFP that is a reporter for endocytic sites in hyphal tips and TaoA-GFP that is a reporter for highly polarized sites at hyphal apices. b) Test the Apical Recycling Model through examination of mutants affected in Endocytosis.The markers described in A, are being assesed using live cell imaging to examine endocytosis during hyphal growth. Strains are being constructed to place these markers in endocytic mutants to examine their fate when endocytosis is knocked down. c) Complement by over-expression an endocytosis defective mutant ofA. nidulansto determine what genes interact with fimbrin during the process of hyphal growth. No progress to report.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Z. Schultzhaus and B. D. Shaw. 2016. The Flippase DnfB is Cargo of Fimbrin-associated Endocytosis in Aspergillus nidulans, and likely recycles through the late Golgi. Communicative and Integrative Biology. 9:2, e1141843, DOI: 10.1080/19420889.2016.1141843
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Z. Schultzhaus, L. Quintanilla, A. Hilton, and B. D. Shaw. 2016. Live Cell Imaging of Actin Dynamics in the Model Filamentous Fungus Aspergillus nidulans Using Lifeact. Microscopy and Microanalysis.22: 264-274.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: C.-L. Wang and B. D. Shaw. 2016. F-actin localization dynamics during appressorium formation in Colletotrichum graminicola. Mycologia. 108:506-514.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: C.-L. Wang, W.-B. Shim, and B. D. Shaw. 2016. The Colletotrichum graminicola striatin ortholog Str1 is necessary for anastomosis and is a virulence factor. Molecular Plant Pathology.17: 931-942.