Progress 10/01/06 to 09/30/09
Outputs
OUTPUTS: During the three years of the project, the results of the study were published in peer-reviewed journals; the publications were cited by many papers; the results were presented at regional (11), national (6), and international (9) conferences; discussed in seminars at 9 universities in the US, Taiwan, Japan, and India; discussed in several courses at UNH; presented at teacher workshops (4), and disseminated to public in several lectures in the US and abroad. The activities have included training undergraduate students in designing and doing experiments, teaching, mentoring, presentations at conferences, demonstration sites, field days, symposia, workshops, and training of high school teachers. The study also resulted in making contacts with the industry for consulting. Curricular changes for introductory biology lab included the development of a manual to include experience with plant cloning and tissue culture. Some examples include: Subhash C. Minocha September 13, 2007 - The functional genome of the polyamine metabolic pathway - The polyaminome Purdue University, W. Laffayette, IN. Subhash C. Minocha - November 2, 2007 - The functional genome of the polyamine metabolic pathway - The polyaminome; Joint seminar at the International Center for Genetic Engineering and Biotechnology; Plant Genome Research Institute and Jawaharlal Nehru University, New Delhi, India Subhash C. Minocha - November 9, 2007 - The functional genome of the polyamine metabolic pathway - The polyaminome; Delhi University, South Campus, New Delhi Subhash C. Minocha Biotechnology and Genetic Engineering: Should We Play God Exeter Rotary Subhash C. Minocha - November 19th, 2007; 12:15-1:30, The Inn of Exeter, Exeter, NH Subhash C. Minocha - From the Polyaminome to Decaffeinated Tea and Sticky Tomatoes - New Biology Department Inaugural Seminar - February 8, 2008 Subhash C. Minocha - March 7, 2008 North Bengal University - Regional Workshop on Bioinformatics Subhash C. Minocha - Regulation of the Expression of Genes of the Polyamine Metabolic Pathway: "The Polyaminome" Subhash C. Minocha - February 29, 2008. Punjab University, Chandigarh, India - Department of Botany and the center for Environmental Study Seminar - Functional Genomics of the Polyamine Pathway: The "Polyaminome" Subhash C. Minocha - March 4, 2008. Delhi University, Delhi - National Refresher Course for College Teachers - Phytoremediation Subhash C. Minocha - March 21, 2008. Growing Barrels of Wine on the Vine - The World of Plant Biotechnology. Active Retirement Association. The products also included DNA sequences that have been submitted to the GenBank database. PARTICIPANTS: Participants Sridev Mohapatra - Ph.D. student - 3 years Rajtilak Majumdar - Ph. D. student - 3 years Michelle Serapiglia - MS student - 2 years Vasavi Challa - MS student - 2 years Smita (Singh) Cherry - MS student - 3 years Lin Shao - Ph. D. student - 1 year Undergraduate students - more than 20 High school students - More than 80 High school teachers - more than 100 Scientific community and general public General public Partner Organizations: Dr. Rakesh Minocha (Co-PI and Co-Author) and her group - USDA Forest Service, NRS, Durham, NH Dr. Autar K. Mattoo (Co-PI and Co-Author), USDA/ARS, Beltsville, MD Dr. Gopi Podilla and his group, University of Alabama, Huntsville, AL Dr. Lucas Li, University of Illinois Biotechnology Center, Urbana, IL Dr. Avtar K. Handa, Purdue University, W. Lafayette, IN Dr. Barbara Moffatt, University of Waterloo, Ontario, Canada Dr. Margret Sauter, Christian-Albrechts-Universitaet, Kiel, Germany Training and Professional Development Six graduate students, four international visiting scientists, two Postdoctoral Research Scientists, three research technicians, several undergraduate students, and high school teachers. TARGET AUDIENCES: Scientific community, graduate and undergraduate students, high school teachers, high school students and general public within the US and internationally PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The results of the study helped us understand the regulation of plant metabolism (particularly that of polyamines) in response to genetic manipulation; a commonly used approach to genetic improvement of agricultural crops and forest trees. The results suggested that genetic manipulation of a single step in a metabolic pathway has consequences that go beyond that pathway. Thus the products of plant genetic engineering, particularly where metabolism related to stress response or nutritional improvement have been targeted, need to be carefully analyzed for unintended consequences on other parts of metabolism. The results can be useful in modifying the metabolism of amino acids and polyamines to produce low-polyamine plant foods for use as specialty diet for patients undergoing chemotherapy using polyamine biosynthesis inhibitors (there are several studies involving polyamine metabolism as a target for chemotherapy for cancer and parasitic infections), and to produce plants that are tolerant of abiotic stress because polyamines play a critical role in abiotic stress response of plants. It was also found that increased polyamine biosynthesis results in increased uptake and utilization of both carbon and nitrogen, which would result in increased growth of trees thus increasing biomass production for energy or other uses of forest products. The use of polyamines as markers of stress in forest trees can help us in developing forest management practices to alleviate and/or minimize damage to forest productivity as a consequence of environmental stress and climate change. The publications, presentations and participation in conferences and classrooms resulted in the training of several young scientists, and public at large. A major outreach activity in which the results and techniques of the project were discussed was Project SMART (Science and Mathematics Achievement through Research training), a summer program aimed at highly talented high school students, including a large proportion of under-represented and minority students. Likewise, presentation of workshops for high school teachers would allow the project results to reach beyond the scientific community to the future generation of scientists. More than 20 undergraduate students were trained in the techniques of plant molecular biology and genetic engineering and about the applications and implications (moral, ethical, environmental, economic, and legal) of plant biotechnology and genetic engineering. The project also allowed us to develop new collaborations with the University of Alabama, the University of Illinois, USDA Forest Service, USDA/ARS, and private industry in the US and Canada.
Publications
- Mattoo, A.K., Minocha, S.C., Minocha, R., Handa, A.K. (2009) Polyamines and cellular metabolism in plants: transgenic approaches reveal different responses to diamine putrescine versus higher polyamines spermidine and spermine.Amino Acids DOI 10.1007/s00726-009-0399-4
- Mohapatra, S., Minocha, R. Minocha, S.C. (2009) Transgenic Manipulation of a Single Polyamine in Poplar Cells Affects the Accumulation of all Amino Acids. Amino Acids DOI 10.1007/s00726-009-0322-z.
- Majumdar, R., Minocha, S.C. (2009) The SAMDC and SPDS gene family: Their tissue-specific expression and response to abiotic stress treatments; Annual Meeting of the Northeastern Section of American Society of Plant Biologists, Plattsburgh, NY; May 1-2, 2009, Abs.
- Birt, A., Minocha, S.C. (2009) Regulation of Expression of the Polyamine Pathway Genes: What does a Promoter Look Like Annual Meeting of the Northeastern Section of American Society of Plant Biologists, Plattsburgh, NY; May 1-2, 2009, Abs.
- Birt, A., Minocha, S.C. (2009) Regulation of Expression of the Polyamine Pathway Genes: What does a Promoter Look Like Annual UNH URC; April 25, 2009, Abs.
- 11 other Abstracts published during 2006-2008
- Minocha, S.C. (2008) So that the Plants can Stand Tall even under Stress: Advancements in Research, DNA may help. Amar Ujala, Chandigarh, Tuesday November 18, 2008. Popular article in a newspaper.
- Minocha, R., Minocha, S.C. et al. (2008) Species-Specific Elevation Dependent Response of Foliar Physiology of Trees to Calcium Supplementation at Hubbard Brook Experimental Forest, NH, USA; 93rd Annual Meeting of the Ecological Society of America, Milwaukee, WI Aug 3-8, 2008, Abs.
- Minocha, S.C. and R. Minocha (2008)Abiotic Stress in Plants: Response, Perception, Detection and Amelioration Indian Society for Plant Physiology. Golden Jubilee Conference; Challenges and Emerging Strategies for Improving Plant Productivity, New Delhi, India (Nov. 12-14, 2008, Abs.
- Minocha, R. and S.C. Minocha. (2008)Climate Change and Forest Productivity: Nitrogen and Calcium: Indian Society for Plant Physiology. Golden Jubilee Conference; Challenges and Emerging Strategies for Improving Plant Productivity, New Delhi, India (Nov. 12-14, 2008), Abs.
|
Progress 10/01/07 to 09/30/08
Outputs
OUTPUTS: Experiments were performed to study the regulation of polyamine metabolism in poplar cells. A detailed metabolomic analysis of two cell lines differing in putrescine was done; the results showed several key metabolites changing in response to genetic manipulation of putrescine. Data were analyzed and used for writing thesis/manuscripts/presentations at national and international conferences. Three Ph.D. students, one M.S. student, and 5 undergraduate studnets were mentored and trained in the techniques of molecular biology and biochemistry. Results of this research were presented at national and international conferences. A summer workshop was conducted for 32 inner-city, under-represented, minority groups of high achool students. The material was included in teaching undergraduate and graduate level courses. New sequences were added to the database of expressed genes in poplar. A collaboration was established with the Center for Metabolomics, University of Illinois. One Ph.D. student submitted his thesis and graduated. PARTICIPANTS: Sridev Mohapatra, Ph.D. student UNH Smita Singh, MS student, UNH Carolyn Madden, undergraduate student, UNH Sara Espinal, undergraduate student, UNH Tania DiVirgilio, undergraduate student, UNH Gabriella martinez, Postdoctoral research fellow, UNH Rakesh Minocha, Research Scientist, USDA Forest Service Stephanie Long, Research Technician, USDA Forest Service Sridevi Ganapathi, Postdoctoral visiting scientist from India P. Thangavel, Postdoctoral visiting scientist from India Christie Williams, Research scientist, Purdue University TARGET AUDIENCES: National and internations scientific community for peer-reviewed articles and presentations as listed. UNH graduate students UNH undergraduate students General public for public presentations High school students from New York/New Jersey area PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The scientific community gained from a better understanding of the effect of genetic manipulation of a single metabolite on the overall metabolism of plant cells, particulalry on the oxidative stress in cells. A better understanding of total nitrogen metabolism was gained through the analysis of the metabolome and individual amino acids. Methods developed as a part of the project were used by USDA-ARS, Beltsville, MD and others for analysis of polyamines. The summer workshop helped enhance the interest and aptitude in agricultural sciences for 32 students from inner-city high schools, who will make informed choices in taking advantage of the opportunities for higher education in agriculture-related fields. These students reflected a change in attitude towards agricultural biotechnology and learned the practical skills of plant cloning and genetic engineering. This knowledge should lead to their active participation in decision-making RE the benefits and potential risks associated with agricultural biotechnology.
Publications
- Serapiglia, MJ, Minocha, R., Minocha, SC. 2008. Changes in polyamines, inorganic ions and glutamine synthetase activity in response to nitrogen availability and form in red spruce (Picea rubens). Tree Physiol. 28: 1793-1803.
- Mohapatra, S., Minocha, R. Long L. Minocha, S.C. 2008 Putrescine overproduction negatively impacts the oxidative state of poplar cells in culture. Plant Physiol. Biochem. doi:10.1016/j.plaphy.2008.12.007.
- Williams, C., Subramanyam, S., Sardesai, N., Minocha, S. 2008. Virulent Hessian fly larvae trigger increased wheat polyamine levels; Plant and Animal Genome Conference; January 8-13, 2008; San Diego, CA.
- Minocha, S. 2008 Regulation of the Expression of Genes of the Polyamine Metabolic Pathway: "The Polyaminome". Regional Workshop on Bioinformatics - North Bengal University, Siliguri, India. March 7-9, 2008
- Minocha, R. et al. 2008. Elevation dependent sensitivity of northern hardwoods to Ca-depletion at Hubbard Brook Experimental Forest, NH, USA Tree Physiol. (Under revision)
- Mohapatra, S. 2008. Understanding Polyamine Metabolism through Transgenic Manipulation in Poplar Suspension Cultures. Ph.D. Thesis Submitted to UNH, May, 2008.
- Presentations 2008
- Subhash C. Minocha - 2008. From the Polyaminome to Decaffeinated Tea and Sticky Tomatoes - New Biology Department Inaugural Seminar - February 8, 2008
- Subhash C. Minocha - 2008. Punjab University, Chandigarh, India - Department of Botany and the center for Environmental Study Seminar - Functional Genomics of the Polyamine Pathway: The "Polyaminome". February 2008.
- Minocha, S. 2008. "Bioremediation and Phytoremediation: A Solution to Pollution" National Refresher Course in Environmental Studies for College Teachers, Institute of Life Long Learning, Academic Research Centre, University of Delhi; March 4, 2008 **
- Subhash C. Minocha - 2008 - Round table Discussion (2 sessions) on Biotechnology and Nanotechnology at the National Junior Science and Humanities Symposium for high school students, Orlando, FL May 1, 2008
- Subhash C. Minocha - 2008 - Teachers Workshop on Biotechnology: Solution to our Environmental Problems; at the National JSHS Symposium, Orlando, FL; May 3, 2008
|
Progress 10/01/06 to 09/30/07
Outputs
OUTPUTS: The major goals of the proposed project are to comprehensively investigate the impact of genetic manipulation of a single step in PA metabolism on the related metabolic pathways, and to study the response of cells with elevated PAs to abiotic stress. The effect of enhanced putrescine metabolism on the oxidative state of poplar (Populus nigra x maximowiczii) cells was compared in two cell lines; one transformed with a β-glucuronidase (GUS) gene (used as control) and the other with a mouse ornithine decarboxylase (mODC) gene, which resulted in over-production of putrescine and a consequent enhancement in its catabolism. Activities of several key enzymes of the Reactive Oxygen Species (ROS) scavenging machinery were measured over a seven day culture cycle. Cellular contents of the common reductant glutathione along with amino acids proline and glutamate were also measured. The activities of glutathione reductase (EC 1.6.4.2) and monodehydroascorbate reductase (EC 1.6.5.4)
increased concomitant with enhanced putrescine metabolism, and the activity of ascorbate reductase (EC 1.11.1.11) remained mostly unchanged. Reduced glutathione was significantly lower in cells with high putrescine. While proline content increased on some days, glutamate was lower on all seven days of culture in the high putrescine cells as opposed to the control cells. There was no significant change in the mitochondrial activity; however, enhanced putrescine metabolism was associated with increased membrane damage in the cells. There was an increased influx of Ca accompanied by a concomitant efflux of K in the high putrescine cells. We conclude that, whereas increase in putrescine accumulation may have a protective role against ROS in plants, enhanced turnover of putrescine actually makes plant cells vulnerable to increased oxidative damage.
PARTICIPANTS: Individual who worked on the Project: Andrew Page, Rakesh Minocha, Sridev Mohapatra, Rajtilak Majumdar, Smita Singh, Carolyn Madden, Leland Czeke, Gopi Podila, Carolyn Madden, Autar K. Mattoo Partner Organizations and Collaborators: USDA Forest Service, Northern research station, Durham, NH USDA ARS, Beltsville, Maryland University of Alabama, huntsville Alabama Dr. Andrew Page Received tarining as Post doc Sridev Mohapatra, Rajtilak Majumdar, Smita Singh were trained as graduate students Carolyn Madden was an undergraduate student
TARGET AUDIENCES: Faculty and staff at varrious national and international research organizations - Formal lectures and discussions; workshops Graduate and undergraduate students - hands-on research experince, discussions and formal classroom teaching High school students - hands on experience and lectures during a month-long summer Institute - Project SMART General Public - public lectures on Biotechnology
PROJECT MODIFICATIONS: None
Impacts
The results demonstrate clearly that genetic manipualtion of a single step in a metabolic pathway has far reaching effects on the cellular metabolism; some of these effects can be benefiacial and others can be detrimental to the cell. A clear understanding of these effects will help us design better plants with modified nutrition as well as those that can tolerate abiotic stress without compromising their intrinsic economic value. Genetic manipulation of the polyamine pathway has demonstrated that a thorough analysis of the metabolic effects of modifying a single step must be undertaken to assess the overall gains in the quality of the product. Presentations: April 28, 2007. Genetic manipulation of polaymines and stress response in plants - Institute of Agricultural Biotechnology; Academia Sinica, Taiwan. May 2, 2007. Genetic manipulation of a metabolic pathway - the case of polyamines. Department of Biological Sciences, Tokyo University, Tokyo, Japan September 13,
2007 - The functional genome of the polyamine metabolic pathway - The polyaminome November 2, 2007 - The functional genome of the polyamine metabolic pathway - The polyaminome: Joint seminar at the International Center for Genetic Engineering and Biotechnology; Plant Genome Research Institute and Jawaharlal Nehru University, New Delhi, India November 9, 2007 - The functional genome of the polyamine metabolic pathway - The polyaminome; Delhi University, South Campus, New Delhi November 19th, 2007 - Biotechnology and Genetic Engineering: Should We Play God? Exeter Rotary Club, Exeter, NH Students and Post-docs: DR. Andrew Page _ Post doc completed 4 years on the project and moved on to a job in industry Sepetember, 2007 Two high school students (Lucy and Zoey) and an undergraduate student received training in the techniques of gene transfer. two graduate students (Smita Singh and Sridev Mohapatra worked for their Ph.D. thesis on this project. Courses Labs etc. - The material produced in
the project was used for discussion and hands-on experience for Project SMART (a Summer Institute in Biotechnology for High school students) and for Cell Culture (PBIO 751/851).
Publications
- Andrew F. Page, Sridev Mohapatra, Rakesh Minocha and Subhash C. Minocha (2007). The effects of genetic manipulation of putrescine biosynthesis on transcription and activities of the other polyamine biosynthetic enzymes. Physiol. Plant. 129: 707-724
- Minocha, S.C. and A.F. Page. 2006. Microbes and their contributions to plant biotechnology. In R.C. Ray (ed), Microbial Biotechnology in Agriculture and Aquaculture, Vol. 2.pp. 1-24.
- A. Page, R. Minocha, S. Minocha. Global impact of genetic manipulation of polyamines on the cells transcriptome. American Society of Plant Biologists and Botanical Society of America Joint Meeting. Chicago, July 7-11, 2007.
- S. Mohapatra, R. Minocha, S.C. Minocha Putrescine overproduction changes the oxidative state of poplar cells in culture and aids in aluminum tolerance. American Society of Plant Biologists and Botanical Society of America Joint Meeting. Chicago, July 7-11, 2007.
- Rajtilak Majumdar, Jeff Mitchell, Scott Sanders, Subhash Minocha. Analysis of the expression of key polyamine biosynthetic genes in Arabidopsis. American Society of Plant Biologists and Botanical Society of America Joint Meeting. Chicago, July 7-11, 2007.
- Rakesh Minocha, Stephanie Long, P. Thangavel, Subhash C. Minocha, Christopher Eagar and Charles T. Driscoll Monitoring Remediation of Physiological Stress in Trees after Calcium Addition at Hubbard Brook Experimental Forest, NH, USA. Ecological Society of America; August, 6-11, 2007- San Jose, CA.
- S. Smita, R. Minocha, S.C. Minocha Enhanced polyamine metabolism and sulfur amino acids in poplar cells. American Society of Plant Biologists and Botanical Society of America Joint Meeting. Chicago, July 7-11, 2007.
- S. Minocha, A., L. Czeke G. Podila, R. Minocha. Global Impact of Genetic Manipulation of Putrescine on the Cell's Transcriptome. International Congress on Biogenic Amines: Biological and Clinical Perspectives October 17-21, 2007 Catania (Sicily), ITALY
- R. Minocha, S. Long, W.C. Shortle, K. Smith, P. Wargo, J. Aber, S. Minocha, F. A. Bazzaz, G. Lawrence, C. Driscoll, T. Fahey, C. Eagar. Calcium-putrescine interaction at cellular to organismal levels: Using foliar putrescine as an indicator of calcium depletion and stress in forest trees. International Congress on Biogenic Amines: Biological and Clinical Perspectives. October 17-21, 2007 Catania (Sicily), ITALY
|