Progress 07/15/07 to 07/14/12
Outputs
OUTPUTS: We have completed or nearly completed all of our objectives for this project; several new projects have spawned from the original Objectives, yielding interesting results and have been lucrative with regard to publication, as outlined below. We completed Objective 1 involving generation of transcriptomic data from the fungus exposed to stress conditions including heat shock, oxidative stress and in planta, to which we added nutrient starvation for a more complete dataset. All conditions were compared to fungus grown in a complete medium (CM). Our dataset was analyzed using four bioinformatic programs, settling on limma (R) for final data analysis. We then prepared lists of genes showing significant fold change differences from each other and fungus grown in CM. Real-time qRT-PCR was used to confirm gene expression and my student has deleted several of these genes; one mutant in particular presents intriguing phenotypes and is being worked upon for a separate publication. The microarray study itself has been published, as described below. Along with a promoter study which is complete, Objective 2 also involved targeted deletions of several genes with predicted stress-related functions in the fungus, including a heat shock-related protein, Hsp104. The Hsp104 mutant generation and characterization is being carried out completely by undergraduate students, hence its slow pace, however I have now trained three undergrads using this system and the project will soon be ready for an all-undergraduate publication targeted for Fungal Genetics and Biology. I intend to complete this project with one work-study undergraduate student, from another funding source. The goal of Objective 3 was to visualize proteins involved in scavenging reactive oxygen species during a susceptible interaction between rice and the rice blast fungus. Our original intent was to look at the transcriptional activator of a glutathione biosynthetic pathway called YAP1, in both the plant and pathogen. The graduate student who received training from this project achieved the following goals: 1) she made fluorescently-tagged fungal lines for both the YAP1 protein as well as its partner protein HYR1; 2) she characterized knock-out mutants of the HYR1 and the YAP1 gene in the fungus; 3) she learned rice transformation; 4) she has several transformed rice lines that express GFP; 5) she recently published work on the HYR1 gene in the fungus in relation to infection. Our results have been disseminated by conference presentations and publications. I have presented talks on our results at three International meetings and was asked to present the HYR1/YAP1 story at the International Fungal Genetics Conference in March 2011 and at the Gordon Conference in June, 2012. My two graduate students receiving training from this grant have presented 8 talks and/or posters at the American Phytopathological Society national and divisional meetings, as well as at the International Rice Blast Conference held in 2010. A former post-doc presented a poster on results from Objective 2 at an APS National meeting in 2008. PARTICIPANTS: Sandra Mathioni began her PhD in my lab in September, 2007. She has completed and published on Objective 1, and assisted with Objective 2 by way of mentoring students and Objective 2.1, examining promoter elements. She is well-versed in generating and analyzing transcriptional data sets using four different analysis programs, including limma, LoX and Ingenuity Pathway Analysis software. She presented her data at four national or regional meetings (two talks and three posters). Many genes of interest emerged from her microarray screen, and she has also spent time working on one of them involved in pathogenicity, and she has made two publications out of this study. She just completed a one-year post-doc at the Noble Research Foundation in Ardmore, OK, and is now just starting as a research associate in the Plant Pathology Department at Federal University of Lavras in Brazil, her home country. Kun Huang, the graduate student working who worked on Objective 3, began her PhD in my lab in February of 2008. She learned and became proficient in real-time qRT-PCR, construct generation and fungal, yeast and plant transformation, confocal and time-lapse microscopy and sequence analysis. She has made two publications from this project, and is working on a third. She is currently a post-doc with Dr. Blake Meyers and Dr. Jeff Caplan at University of Delaware; her project is largely confocal-microscopy and construct-generation based. She was the highest contender for this position, given the skill set she learned from this USDA project fit the needs of the PIs, perfectly. Jessica Cooper has just finished her Masters' Program in Plant Pathology at Virginia Tech. She worked in my lab for three years, first learning standard lab techniques, and then specifically on Objective 2, generating knock-outs of Hsp104 and characterizing their behavior. This led to her interest in obtaining an advanced degree in plant pathology. Matt Sarlo currently works at Fraunhofer, a biotechnology company for plant-based vaccine development. Kishana Williamson worked on Hsp104 mutant characterization at the molecular level in my lab and is now finishing her masters' in public health at George Washington University. Kasia Dinkeloo is a current senior in my lab working and has been working with me for three years. She is doing a senior thesis project on a gene that spun-off from this USDA project, called Tmpl1. One former post-doc was also trained on this grant for ~1.5 years. We have worked closely and strengthened our relationship with the Bio-Imaging Center and its Directors, Kirk Czymmek and Jeff Caplan, as well as a colleague at DuPont, James Sweigard. All of these investigators were advisors on my graduate student's committee, and co-authors on publications. TARGET AUDIENCES: Efforts: This project has been a launching pad for training five undergraduate students; four through direct funding, and one current undergraduate student, Emma Sweeney, who is working for credit on a spin-off of this USDA project, on other stress mechanisms outside of the Hyr1-yAP1 pathway. The five undergrads who have worked on this project over the years have all ended up in scientific fields, including industry and academic. In the case of my current undergraduates, one of them has applied to graduate school in plant pathology, and my most current student will hopefully apply for a scholarship to continue her work in my lab, this summer. This project has been many wonderful things, including being a great training platform for getting undergraduates interested in molecular plant pathology. Not only has this project piqued their interest, but it has retained their interest enough to continue in a scientific profession. One of my former graduate students, Dr. Mathioni, also received an experiential learning experience through doing an internship at DuPont. While she was not paid by this grant during her internship, her time at DuPont did foster a spin-off of this project that I am working on with a visiting graduate student from Brazil. This project studies a gene family of superoxide dismutases, and is in keeping with stress responses in the rice blast fungus. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The outcomes and impacts of our project have had many positive implications for our program, and continue to fuel new questions and projects. Five undergraduates have been/are being trained in molecular biology by working on this project, one of whom just finished her masters' program in Plant Pathology at Virginia Tech, one who now works at DuPont on small RNAs, and one who is finishing her masters' in public health at George Washington University. Funding from this grant has also allowed each of my two former graduate students to gain training in bioinformatics (Objective 1) and various microscopy techniques (Objective 3), which has already served them well in their current positions. The "stressomics" data from Objective 1 is readily available, and will soon also be uploaded to MGOS, a database specific to Magnaporthe information. I have also sent out this data to researchers so they could see how their genes of interest behave under different stresses. Finally, Objective 3 has yielded many new and exciting results, and data from this Objective will form the foundation for future proposals.
Publications
- Mathioni, S.M., Belo, A., Townsend, J.P., Donofrio, N.M. 2011. Getting the most of your fungal microarray data: two cost and time-effective methods. In: Fungal Genomics: Methods and Protocols. ISBN: 978-1-61779-039-3; 280p.
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Progress 07/15/09 to 07/14/10
Outputs
OUTPUTS: We are in the second no-cost extension year of this project on stress responses in the rice blast fungus; we have adhered to our timeline and are finishing the final objective. Several new projects spawned from the original Objectives; these have yielded interesting results and been lucrative with regard to publication (see Outcomes). We have completed Objective 1 involving generation of transcriptomic data from the fungus exposed to stress conditions. All conditions were compared to fungus grown in a complete medium (CM). Our dataset was analyzed using four bioinformatic programs, settling on limma (R) for final analysis. We prepared lists of genes showing significant fold change differences from each other and fungus grown in CM. Real-time qRT-PCR was used to confirm gene expression and my student has deleted several of these genes and generated several publications (see Outcomes). Along with a promoter study which is now complete, Objective 2 involves targeted deletions of several genes with predicted stress-related functions, including a heat shock-related protein, Hsp104. The Hsp104 mutant generation and characterization is being carried out by undergraduate students; the HSP104 mutant, as we have learned from our present studies, impacts expression of chaperone-type genes, and a publication targeted for Fungal Genetics and Biology will be submitted before the end of this grant period and presented at the APS Potomac Division meeting in March. The goal of Objective 3 is to visualize proteins involved in scavenging reactive oxygen species during a susceptible interaction between rice and the rice blast fungus. Our original intent was to look at the transcriptional activator of a glutathione biosynthetic pathway YAP1, in both the plant and pathogen. The graduate student on this objective has achieved the following goals: 1) made fluorescently-tagged fungal lines for both the YAP1 protein as well as its partner protein HYR1; 2) characterized knock-out mutants of the HYR1 and the YAP1 gene in the fungus; 3) learned rice transformation; 4) published 2 peer-reviewed articles on the HYR1 gene in the fungus in relation to infection; 5) preparing her third and final manuscript on HYR1-YAP1 interactions, as well as on generation of ROS during the infection process; 6) obtained genetically transformed rice plants containing a Hyper-AS sensor construct that detects ROS during the infection process, which will be much more useful for finishing studies on the interaction. Our results have been disseminated by conference presentations and publications. I have presented talks on our results at three International meetings and was asked to present the HYR1/YAP1 story at the International Fungal Genetics Conference in March 2011. I have also been asked to present the HYR1 story at 2012 Molecular and Cellular Fungal Biology Gordon Conference. Graduate students receiving training from this grant have presented 8 talks and/or posters at the APS national and divisional meetings, as well as at the International Rice Blast Conference in 2010. A former post-doc presented a poster on results from Objective 2 at an APS National meeting in 2008. PARTICIPANTS: Sandra Mathioni began her PhD in my lab in September, 2007. She has completed and published twice on Objective 1. Through the support of this grant, she has learned every step of the Agilent microarray platform, as well as data analysis using four different microarray analysis programs. Along with Objective 1, she has also worked on Objective 2.1, examination of promoter elements for stress response elements. She has presented her data at four national or regional meetings (two talks and three posters) and had two publications. She has left an excellent legacy of candidate genes, some of which she has already generated knock-out mutations. She has recently graduated and is working at the Noble Foundation as a post-doc. Kun Huang, the graduate student working on Objective 3, has learned and become proficient in real-time qRT-PCR, construct generation and fungal, yeast and plant transformation, confocal and time-lapse microscopy and sequence analysis. Her project on transcriptional activators of oxidative scavenging pathways constantly yields new and exciting directions and two published articles describing HYR1/YAP1 and their interactions. Jessica Cooper is in her first year of a Masters' Program in Plant Pathology at Virginia Tech. She worked in my lab for three years, first learning standard lab techniques, and then specifically on Objective 2, generating knock-outs of Hsp104 and characterizing their behavior. This led to her interest in obtaining an advanced degree in plant pathology. Matt Sarlo is currently working at Fraunhofer, a biotechnology company for plant-based vaccine development. Kishana Williamson was an undergrad who worked on Hsp104 mutant characterization at the molecular level; she has learned RNA extraction techniques, RT-PCR and Southern blots. She is in her first year at George Washington University in a masters' program. Kasia Dinkeloo is a junior in my lab working who has worked on the HSP104 mutant, and is now studying another protein likely to be involved with HYR1/YAP1 signaling, referred to as TmpL. She will do her senior thesis project in my lab, and will present her data at the APS Potomac Division meeting this March. Sean Recce is a senior in my lab who is finishing the HSP104 project. He recently discovered that chaperone-type genes are impacted by presence/absence of HSP104 during recovery from a heat shock. He will present his results at the APS Potomac division meeting, and will also help draft a manuscript to submit before graduation. One former post-doc was trained on this grant for ~1.5 years. We work closely with the Bio-Imaging Center and its Directors, Kirk Czymmek and Jeff Caplan, as well as a colleague at DuPont, James Sweigard. These investigators are advisors on my graduate student's committee, and co-authors on publications. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Five undergraduates and two graduate students have been/are being trained in molecular biology, confocal microscopy, and bioinformatics through working on this project. The "stressomics" data from Objective 1 is readily available. Objective 3 has yielded many new and exciting results, and data from this Objective will form the foundation for future proposals. Participants Sandra Mathioni began her PhD in my lab in September, 2007. She has completed and published twice on Objective 1. Through the support of this grant, she has learned every step of the Agilent microarray platform, as well as data analysis using four different microarray analysis programs. Along with Objective 1, she has also worked on Objective 2.1, examination of promoter elements for stress response elements. She has presented her data at four national or regional meetings (two talks and three posters) and had two publications. She has left an excellent legacy of candidate genes, some of which she has already generated knock-out mutations. She has recently graduated and is working at the Noble Foundation as a post-doc. Kun Huang, the graduate student working on Objective 3, has learned and become proficient in real-time qRT-PCR, construct generation and fungal, yeast and plant transformation, confocal and time-lapse microscopy and sequence analysis. Her project on transcriptional activators of oxidative scavenging pathways constantly yields new and exciting directions and two published articles describing HYR1/YAP1 and their interactions. Jessica Cooper is in her first year of a Masters' Program in Plant Pathology at Virginia Tech. She worked in my lab for three years, first learning standard lab techniques, and then specifically on Objective 2, generating knock-outs of Hsp104 and characterizing their behavior. This led to her interest in obtaining an advanced degree in plant pathology. Matt Sarlo is currently working at Fraunhofer, a biotechnology company for plant-based vaccine development. Kishana Williamson was an undergrad who worked on Hsp104 mutant characterization at the molecular level; she has learned RNA extraction techniques, RT-PCR and Southern blots. She is in her first year at George Washington University in a masters' program. Kasia Dinkeloo is a junior in my lab working who has worked on the HSP104 mutant, and is now studying another protein likely to be involved with HYR1/YAP1 signaling, referred to as TmpL. She will do her senior thesis project in my lab, and will present her data at the APS Potomac Division meeting this March. Sean Recce is a senior in my lab who is finishing the HSP104 project. He recently discovered that chaperone-type genes are impacted by presence/absence of HSP104 during recovery from a heat shock. He will present his results at the APS Potomac division meeting, and will also help draft a manuscript to submit before graduation. One former post-doc was trained on this grant for ~1.5 years. We work closely with the Bio-Imaging Center and its Directors, Kirk Czymmek and Jeff Caplan, as well as a colleague at DuPont, James Sweigard. These investigators are advisors on my graduate student's committee, and co-authors on publications.
Publications
- Huang, K., Caplan, J., Sweigard, JA., Czymmek, KJ and Donofrio, NM. 2011. HYR1-mediated detoxification of reactive oxygen species is required for full virulence in the rice blast fungus. PLoS Pathog 7(4): e1001335. doi:10.1371/journal.ppat.1001335.
- Mathioni, SM., Belo, A., Rizzo, CJ., Dean, RA and Donofrio, NM. 2011. Transcriptome profiling of the rice blast fungus during invasive plant infection and in vitro stresses. BMC Genomics 12:49. doi:10.1186/1471-2164-12-49.
- Huang, K., Caplan, J., Sweigard, JA., Czymmek, KJ and Donofrio, NM. 2011. Suppression of plant-generated reactive oxygen species is required for successful infection by the rice blast fungus. Virulence 2, 559-562.
- Mathioni, SM., Belo, A., Townsend, JP. and Donofrio, NM. (2011) Getting the most out of your fungal microarray data: two cost and time- effective methods. In JR Xu, BH Bluhm (Eds.) Methods in Molecular Biology. Volume 722, pages 29. Springer Science, USA.
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Progress 07/15/08 to 07/14/09
Outputs
OUTPUTS: We are in a one year no-cost extension of this project outlined in our grant on stress responses in the rice blast pathogen; we have mostly adhered to our timeline and completed several Objectives. Several new projects have spawned from the original Objectives; these have yielded interesting results and been lucrative with regard to publication, as outlined below. We have completed Objective 1 involving generation of transcriptomic data from the fungus exposed to stress conditions including heat shock, oxidative stress and in planta, to which we added nutrient starvation for a more complete dataset. All conditions were compared to fungus grown in a complete medium (CM). Our dataset has been analyzed using four bioinformatic programs, settling on limma (R) for final data analysis. We then prepared lists of genes showing significant fold change differences from each other and fungus grown in CM. Real-time qRT-PCR was used to confirm gene expression and my student has deleted several of these genes; one mutant in particular presents intriguing phenotypes and is being worked upon for a separate publication. The microarray study itself has been published, as described below. Along with a promoter study which is now complete, Objective 2 also involves targeted deletions of several genes with predicted stress-related functions in the fungus, including a heat shock-related protein, Hsp104. The Hsp104 mutant generation and characterization is being carried out completely by undergraduate students, hence its slow pace, however I have now trained three undergrads using this system and the project will soon be ready for an all-undergraduate publication targeted for Fungal Genetics and Biology. The goal of Objective 3 is to visualize proteins involved in scavenging reactive oxygen species during a susceptible interaction between rice and the rice blast fungus. Our original intent was to look at the transcriptional activator of a glutathione biosynthetic pathway called YAP1, in both the plant and pathogen. The graduate student receiving training from this project has achieved the following goals: 1) she has made fluorescently-tagged fungal lines for both the YAP1 protein as well as its partner protein HYR1; 2) she has characterized knock-out mutants of the HYR1 and the YAP1 gene in the fungus; 3) she has learned rice transformation; 4) she has obtained all vectors needed for rice transformation and generated the constructs; 5) she has recently published work on the HYR1 gene in the fungus in relation to infection. Our results have been disseminated by conference presentations and publications. I have presented talks on our results at three International meetings and was asked to present the HYR1/YAP1 story at the International Fungal Genetics Conference in March 2011. My two graduate students receiving training from this grant have presented 7 talks and/or posters at the American Phytopathological Society national and divisional meetings, as well as at the International Rice Blast Conference held in 2010. A former post-doc presented a poster on results from Objective 2 at an APS National meeting in 2008. PARTICIPANTS: Sandra Mathioni began her PhD in my lab in September, 2007. She has completed and published on Objective 1. Through the support of this grant, she has learned every step of the Agilent microarray platform, as well as data analysis using four different microarray analysis programs, including limma, and Ingenuity Pathway Analysis software. Along with analyzing her new data, she has also worked on Objective 2.1, examination of promoter elements for stress response elements. She has presented her data at four national or regional meetings (two talks and three posters). Many genes of interest have emerged from her microarray screen, and she has also spent time working on one of them involved in pathogenicity; our goal is to submit this project as a publication before the end of summer 2011. Kun Huang, the graduate student working on Objective 3, began her PhD in my lab in February of 2008. She has learned and become proficient in real-time qRT-PCR, construct generation and fungal, yeast and plant transformation, confocal and time-lapse microscopy and sequence analysis. Her project on transcriptional activators of oxidative scavenging pathways constantly yields new and exciting directions and a manuscript describing HYR1/YAP1 and their interactions is in process. Jessica Cooper is in her first year of a Masters' Program in Plant Pathology at Virginia Tech. She worked in my lab for three years, first learning standard lab techniques, and then specifically on Objective 2, generating knock-outs of Hsp104 and characterizing their behavior. This led to her interest in obtaining an advanced degree in plant pathology. Matt Sarlo is currently working at Fraunhofer, a biotechnology company for plant-based vaccine development. Kishana Williamson is a senior working on Hsp104 mutant characterization at the molecular level; she has learned RNA extraction techniques, RT-PCR and Southern blots. She is characterizing mutant phenotypes when placed under extreme heat conditions. Kasia Dinkeloo is a current sophomore in my lab working together with Kishana, along with performing all pathogenicity assays with the Hsp104 mutant. One former post-doc was also trained on this grant for ~1.5 years. We have worked closely and strengthened our relationship with the Bio-Imaging Center and its Directors, Kirk Czymmek and Jeff Caplan, as well as a colleague at DuPont, James Sweigard. All of these investigators are advisors on my graduate student's committee, and co-authors on publications. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The outcomes and impacts of our project are constantly being realized. Four undergraduates have been/are being trained in molecular biology by working on this project, one of whom is starting her first year of a masters' program in Plant Pathology at Virginia Tech. Funding from this grant has also allowed each of my two graduate students to gain training in bioinformatics (Objective 1) and various microscopy techniques (Objective 3), which I have no doubt will serve them well when they apply for post-doctoral positions. My graduate student working on Objective 3 is an expert in confocal techniques, as well as efficiently generating translational fusion constructs with reporter genes. The "stressomics" data from Objective 1 is readily available, and will soon also be uploaded to MGOS, a database specific to Magnaporthe information. I have also sent out this data to researchers so they could see how their genes of interest behave under different stresses. Finally, Objective 3 has yielded many new and exciting results, and data from this Objective will form the foundation for future proposals.
Publications
- Huang, K., Caplan, J., Sweigard, JA., Czymmek, KJ and Donofrio, NM. 2011. HYR1-mediated detoxification of reactive oxygen species is required for full virulence in the rice blast fungus. PLoS Pathogens, In Press.
- Mathioni, SM., Belo, A., Rizzo, CJ., Dean, RA and Donofrio, NM. 2011. Transcriptome profiling of the rice blast fungus during invasive plant infection and in vitro stresses. BMC Genomics 12:49. doi:10.1186/1471-2164-12-49.
- Mathioni, S.M., Belo, A., Townsend, J.P., Donofrio, N.M. 2009. Getting the most of your fungal microarray data: two cost and time-effective methods. In: Fungal Genomics: Methods and Protocols. ISBN: 978-1-61779-039-3; 280p.
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Progress 07/15/07 to 07/14/08
Outputs
OUTPUTS: Two years into the project outlined in our grant on stress responses in the rice blast pathogen, we have mostly adhered to our timeline and completed several Objectives, as outlined below. We are on-target to continue following our timeline. We have completed Objective 1 involving generation of transcriptomic data from the fungus exposed to stress-inducing conditions including heat shock, oxidative stress and in planta, to which we added nutrient starvation for a more complete dataset. All conditions were compared to fungus grown in a complete medium (CM). Our dataset has been analyzed using four bioinformatic programs, settling on limma for final data analysis. From this data, we have prepared lists of genes showing significant fold change differences from each other and fungus grown in CM. Real-time qRT-PCR has been used to confirm gene expression and my student has deleted several of these genes; one mutant in particular presents intriguing phenotypes and will become a separate publication. We are completing Objective 2 of our grant, involving a promoter study of predicted fungal stress genes, as well as genes mined from Objective 1. The graduate student who also carried out Objective 1 has performed promoter searches for stress-responsive elements and is currently performing a second run with a more complete dataset. Objective 2 also involves targeted deletions of several genes with predicted stress-related functions in the fungus, including an activator of the major chaperone Hsp90 called AHA1, and another heat shock-related protein, Hsp104. The Aha1 mutant shows virulence reduction when spores are first exposed to elevated temperatures. The Hsp104 mutants were recently obtained and we are preparing for phenotype assays. The goal of Objective 3 is to visualize genes involved in scavenging reactive oxygen species during a susceptible interaction between rice and the rice blast fungus. We are looking at the transcriptional activator of a glutathione biosynthetic pathway called YAP1, in both the plant and pathogen. The graduate student has generated all transgenic fungal lines required for this Objective, and is starting microscopy. The next step of this project is to generate constructs for plant transformation, and transform rice with the rice homolog of the YAP1 gene. My student is beginning the plant work now, having had training in rice transformation last winter. This project yielded an intriguing related side project; YAP1, in other organisms, interacts with an oxidative species sensor gene, HYR1. She has knocked this gene out in the fungus and found a reduced virulence phenotype; when she stains for reactive oxygen species, she finds that the mutant is unable to break down plant-generated ROS. We are finishing some experiments and my student has a working draft of a manuscript. Our results have been disseminated by means of conference presentations. I have presented talks on our results at two International meetings. My graduate student has presented 4 talks and/or posters at the American Phytopathological Society meetings (microarray) and a former post-doc presented a poster on results from Objective 2 at an APS meeting. PARTICIPANTS: Sandra Mathioni began her PhD in my lab in September, 2007. She has completed Objective 1, and is finishing several final experiments in preparation for publication. Through the support of this grant, she has, in a period of six months, not only completed Objective 1, but learned every step of the Agilent microarray platform, as well as data analysis using four different microarray analysis programs. Along with analyzing her new data, she has also worked on Objective 2.1, examination of promoter elements for stress response elements. She has presented her data at four national or regional meetings (one talk and three posters). Kun Huang, the graduate student working on Objective 3, began her PhD in my lab in February of 2008. Through support from this grant, she has learned and become proficient in real-time qRT-PCR, construct generation and fungal transformation, confocal microscopy and sequence analysis. Her project on transcriptional activators of oxidative scavenging pathways has yielded new and exciting projects that we are currently focused on with a manuscript in process. Two undergraduates have been trained on this grant, along with one former post-doctoral researcher. One of the undergraduates is continuing on with me for her third year of research in my lab. She has been working on Objective 2.2, deleting heat-stress related genes from the genome. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The outcomes and impacts of our project are constantly being realized. Two undergraduates have been trained in molecular biology by working on this project, one of whom will be starting her third year of research with me this semester. Funding from this grant has also allowed each of my two graduate students to gain training in bioinformatics (Objective 1) and various microscopy techniques (Objective 3), which I have no doubt will serve them well when they apply for post-doctoral positions. My graduate student working on Objective 3 is virtually an expert in confocal techniques, as well as efficiently generating translational fusion constructs with reporter genes. I have polled the Magnaporthe community with regard to the "stressome" transcription data from Objective 1, and they are excited to have the data in-hand. I have already sent out some of this data to various researchers so they could see how their genes of interest behave under different stress conditions. With the completion of our current data set using limma for analysis, we feel confident that this is a complete and robust dataset. My graduate student has a working draft of a manuscript in hand, and we are awaiting on several final experiments before submitting. The "stressome" transcriptomics and the HYR1/Yap1 manuscripts will be submitted for publication before the end of 2009.
Publications
- No publications reported this period
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