Michele Kiyoko Nishiguchi - US grants

This award will support research at the Laboratory of Ecological and Evolutionary Genetics in the Department of Biology at New Mexico State University. A number of investigators will make use of the DNA sequencer including: 1) Dr. Michele Nishiguchi (PI), who studies the evolution and speciation of symbionts in a squid-luminous bacterium model system and is interested in the evolution of infection mechanisms in related pathogenic bacteria of the genus Vibrio, which are responsible for the radiation and spread of infectious disease. 2) Dr. Peter Houde (co-PI), whose research program focuses on the systematics of birds, with an emphasis in the molecular systematics of the avian order Gruiformes, an ill-defined group within avian phylogenetics. 3) Dr. Daniel Howard (co-PI), whose group is interested in species formation, the process responsible for the diversity of life on earth using a model system of closely related ground crickets. 4) Dr. Brook Milligan (co-PI), whose research interests include developing statistical methods to disentangle genetic and environmental factors affecting quantitative traits thereby identifying the genetic architecture of important medical, physiological, or ecological traits of natural populations. 5) Dr. Kevin Oshima (co-PI), whose research focuses primarily on characterizing and examining environmental factors that influence the diversity, ecology and evolution of fish rhabdoviruses. 6) Dr. Geoffrey Smith (co-PI), whose primarily interests include investigating the relationships in the microbial community between metabolic function and phylogenetic structure of biodegradative microorganisms as they respond to chlorinated pollutant exposure.
This award will be used to purchase an Applied Biosystems (ABI) 377XL automated sequencer. In addition to sequencing, this machine can also run microsatellite gels (used in population genetics for identifying specific DNA fragments from individual specimens), as well as AFLP (amplified fragment length polymorphism) techniques used in genome mapping in areas such as agricultural and evolutionary biology. The ABI system does not require any radioactive labeling, enabling a radiation-free environment in the laboratory.
The faculty of the Laboratory of Ecological and Evolutionary Genetics have major interests in the evolution and radiation of organisms with respect to environmental and ecological influences. The ABI 377XL will contribute to the research and teaching of numerous graduate and undergraduate students who are pursuing careers in those fields.

The mutualistic association between sepiolid squids (Mollusca: Cephalopoda) and their luminous bacterial symbionts (Genus Vibrio) provides a versatile model system to study population biology and speciation among animal-bacterial partnerships. Although the squid light organs are open to the seawater, only two Vibrio species stay inside the organ, indicating adaptation and evolution of specificity. Using state-of-the-art techniques involving fluorescent microscopy, molecular biology, and gene expression, this project will examine whether environmental processes or the squid hosts determine the architecture of these interactions. Central to this study will be determining how different the squid-Vibrio pairs are among geographically isolated populations. A key element of this proposal is the several international and collaborative efforts that have been established by the Nishiguchi laboratory for this work.

Results from this proposal will be shared (through publications, meetings, web sites, courses) with both the research and public communities. This study will provide unique research opportunities for students, building on programs for under-represented minorities at New Mexico State University. NMSU is the only Carnegie I research institution ranked by the National Science Foundation as one that serves significant numbers of two ethnic minority student populations, Hispanics and Native Americans. This research will lead to a better understanding of speciation and evolution of bacterial-animal relationships and provide information about the transmission of bacteria in the environment.

This award is for the purchase of a transmission electron microscope, a dedicated digital camera for image acquisition and a room temperature ultramicrotome for the multi-user Electron Microscopy Laboratory at New Mexico State University. These new set of instruments will significantly improve the research and student training on campus and will be used by a group of fifteen investigators from colleges of Arts and Sciences, Agriculture and Home Economics, and Engineering. This research group includes senior investigators with well established research projects as well as junior faculty members who will benefit enormously from the instruments for their career development. The following research projects will make use of the acquired instruments: (1) plant-virus interaction in Arabidopsis thaliana, (2) evolutionary ecology of animal bacterial mutualisms, (3) investigation of assembly of Hepatitis C nucleocapsid-like particles and morphology of hepatocytes cultured on a rotary tissue culture system, (4) effect of vancomycin and house cleaners on cell wall morphology of house-cleaner tolerant Staphylococcus aureus mutants, (5) study of mechanosensory hair cell differentiation during organogenesis. Other projects include structural study of symbiotic fungi in native desert plants, membrane lipid endocytosis by Giardia lamblia, fuel cell technology, mine tailing remediation, subcellular localization of viral replication processes and storage protein bodies in plants, interaction of pathogenic fungus Cercospora beticola with sugar beet and safflower, interaction of soil-borne pathogens with economically important crops in Southern New Mexico, immunolocalization of various Zein proteins in alfalfa, investigation of regulation of the actinomysin and microtubule cytoskeletons for proper coordination of mitosis and cytokinesis in animal cells, and influence of environmental, insect, and herbicide stresses on crop and weed productivity. The newly acquired instruments will be made available to all potential users from various other departments across campus and they will be invaluable for the lab's strong teaching mission and local community outreach activities.

[unreadable] DESCRIPTION (provided by applicant): The mutualistic association between sepiolid squids (Mollusca: Cephalopoda) and their Vibrio symbionts is an experimentally tractable model to study the evolution of animal and bacterial associations at various levels of interaction (genetics, physiology, population ecology). Since symbiotic bacteria are environmentally transmitted to new hosts with every generation, it provides an opportunity to resolve how the ecology of the free-living symbiont affects the architecture of bacterial-host interactions. Bacteria encounter potentially conflicting selective pressures, competing with one another to colonize and persist in the sepiolid light organ, but also competing for scarce resources in the environment outside the squid. A hierarchy of colonization exists, where particular V. fischeri strains out-compete other V. fischeri strains for colonization of their native squid partner, and abiotic factors such as water and temperature can affect the fitness of different strains. Genetic factors that contribute to intra-strain variations in fitness have not been elucidated, and neither have the genetic factors that likely contribute to intra-strain fitness in the environment. The following project will examine how genes that are highly-expressed exclusively in sea water or expressed exclusively in the light organ contribute to bacterial fitness. The present proposal is an extension of the current work in my laboratory, and focuses on four specific goals: [unreadable] i) Characterize environmentally-relevant phenotypes of strains with null mutations in highly-expressed, variable genes. [unreadable] ii) Determine whether a mutant strain complemented with the wild-type operon from the same strain is fit for environmental persistence (free-living) than the same strain complemented with the operon from a different strain. [unreadable] iii) Examine whether symbiotic strains with specific variable loci complemented with the wild-type operon or gene from the same strain are fit for host colonization than the same strain complemented with the operon/gene from a different strain. [unreadable] iv) Measure whether symbiotic genes are found in pathogenicity islands through whole genome comparisons, and demonstrate positive selection of these genes compared to normal housekeeping genes. [unreadable] [unreadable] [unreadable]

Mutualistic associations between bobtail squids and their luminescent bacterial symbionts (genus Vibrio) make excellent models to study the evolution of animal-acterial associations. Since symbiotic bacteria are transmitted through the surrounding seawater to new hosts with every tion, it provides a uniqe opportunity to resolve how the ecology of free-living bacteria effects the make-up of these squid-luminous bacterial partnerships, particularly with respect to factors such as salinity and temperature. Since Vibrio bacteria are quite easily adaptable to changing environmental conditions, this may lead to the possibility of range expansion of these bacteria to different squid populations, and may eventually lead to the evolution of new species of Vibrio. This proposal will examine both free-living and symbiotic vibrios under a variety of conditions in the laboratory (salinity, temperature) to determine if abiotic factors contribute to the distribution of Vibrio among wide ranging host populations. Simultaneously, monitor genetic changes of various Vibrio bacteria in different squid host species will be monitered in the lab to determine if the host has any influence upon Vibrio evolution. These experiments will be completed via growth studies of the bacteria in culture or in juvenile squids that are raised in the laboratory. Clearly, there is a need to understand how quickly organisms such as bacteria can adapt to different environments; with the noticeable change in global climate, it will be a benefit to determine if these changes will increase or decrease biodiversity on a much larger scale. The expected outcome of this research is that adaptation occurs at a much higher frequency due to fluctuations in the environment, which then allows successful colonization of different host species, providing a new trajectory for bacterial speciation to occur. Results from this proposal will be disseminated through publications, teaching, web sites, and outreach to have a greater impact for both the public and research communities.

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).

This Major Research Instrumentation-Recovery and Reinvestment (MRI-R2) award funds the purchase of a confocal microscope to support a diverse array of research and teaching activities at New Mexico State University. The instrument will be housed in NMSU's centralized microscopy facilities and administered by the Electron Microscopy Laboratory to improve current and future research and training for the university and surrounding community. The new confocal microscope will support research in cellular and developmental biology, neurobiology, animal/microbial interactions, plant molecular biology, nanotechnology and chemical biology. The projects will take advantage of the microscope's ability to perform tandem scanning for live cell imaging, a spectral detector to separate overlapping spectra, which will be essential for applications requiring non-traditional excitation/emission spectra. The new system substantially broadens the research capacities of faculty members and of graduate and undergraduate students, enhancing careers in multiple scientific fields. The new instrumentation also offers significant opportunities for research training and education. The 18 participating investigators mentor over 100 graduate and undergraduate students, (60% female, 57% underrepresented minorities), and these numbers reflect a history of commitment to the recruitment and training of women and other underrepresented groups. The system will be used in support of educational and outreach activities including the continued offering of 2 courses, the introduction of confocal microscopy in other existing Biology courses and demonstrations for STEM programs aimed at recruiting and enhancing the research skills of women and underrepresented minority students and faculty at NMSU and throughout the state of New Mexico. The opportunity for students to use state-of the- art fluorescent imaging and analytical tools will profoundly impact their research projects, rendering them more competitive in gaining entry to top graduate programs or obtaining top postdoctoral positions. Data from the new system will be disseminated by publication in peer-reviewed journals, and by student and faculty presentations are regional or national meetings.

PROJECT SUMMARY/ABSTRACT The New Mexico State University RISE Program is diversifying the community of health scientists by supporting PhD degree completion by NMSU students who are members of groups underrepresented in biomedical/biobehavioral/bioengineering research. NMSU is able to achieve this goal because of the institution's unique status as a doctoral, research-intensive (NIH-INBRE), land grant, minority (Hispanic) serving university with a diverse student population and a cadre of accomplished mentors. Key outcomes of the seventeen-year NMSU RISE research education program are: 80.1% retention to science careers after NMSU PhD completion; 93.4% retention to degree completion by NMSU PhD students; student-authored publications; achievement of student independent support; national recognition for student research excellence; and student entry to postdoctoral positions in R1 institutions. This application requests funds to continue to train twelve RISE to the Postdoctorate (RTP) graduate scholars yearly under RISE Option III. The project aims are: RTP/PhD will enable completion of a PhD by 90% of the PhD Scholars and their subsequent entry to postdoctoral positions. Students in the NMSU RISE Program are prepared for the next step of their career through mentored research experiences, professional development activities, formal courses, scientific workshops that develop cutting edge quantitative and technical skills, and through the guided expansion of their research social network. Three developmental activities will provide core skills to prepare students for success in different scientific fields and careers: Activity 1, LEAD IDEA (LEADership in InterDisciplinary Exploration and Advancement) Program, develops the professional skills that maximize research productivity, funding success, and degree completion in 5 years; Activity 2, START-UP (Student Training and Research through United Partnerships) Program provides scientific and technical workshops and enhanced student off-site research training by establishing a formal network with R1 university department heads, program directors, center leaders, and renown faculty mentors; Activity 3, Bioscience TRAIN (Team Research And INnovations) Program provides a new set of activities that emphasize critical thinking, multidisciplinary research and communication skills as well as team leadership through independent interdepartmental research projects supported in the form of mini grants. The program will also include a Students-to-Students Technical Skills Workshop component. RTP graduate scholars are guided toward career independence through preparation of individualized development plans and electronic professional portfolios for yearly review, and through submission of fellowship and internship applications. RTP student activities are institutionalized through course offerings and dissemination in university-wide supported workshops. Students participate in formal education in Responsible Conduct in Research (RCR) that is reinforced by faculty mentor practices and educational activities developed in partnership with colleagues in the START-UP network. Assessment and evaluation of program practices and outcomes are integrated into the design and continual refinement of programmatic elements.