The Effects of Heat Stress on Microgametophyte Viability in Arabidopsis thaliana (Blog #2)

Over the summer months, I have sought out to investigate and gain a better understanding of how an increase in global temperatures will affect overall plant growth and reproduction. It is undeniable that global climate change is occurring, however if we are the contributing to the rise in temperatures we must decide as a whole to use current technologies to mitigate our impact.

The literature shows a strong correlation between increased temperatures and an inability for many organisms to tolerate the increase, but what about our economically valuable plants?  Our agricultural plants?  Cultivated ornamentals?  What about the grasslands and deciduous forests?  Plants do not have the option to leave and area when it becomes inhospitable.  Our research efforts are concentrated on showing morphologically and physiologically what can happen to these organisms in the face of climate change.

One of the results of my literature searches and readings have helped me to determine the specific protocols I will use to help answer my research question.    For the sake of reproducibility, I have selected more than one protocol to target each of my parameters: pollen size, pollen count, pollen morphology, and pollen viability. We have received in all chemicals and reagents in order to go forward with the selected protocols, I put together an extensive timeline for the project, from seed germination to data collection, and have begun planting.

As with any research project, I knew that I was bound to face a few obstacles. The purpose of research and experimentation is trial and error. It is called research, after all!  I have been able to apply my coursework into a real-life laboratory setting. Between the courses I have taken at Pace and my work as a Laboratory Technician for the Department of Biology, I feel better prepared to work within time constraints and with my ability to juggle and multitask.   Finding protocols that were going to help me answer my specific research questions proved to be a bigger challenge than expected, but I was also familiarizing myself more the current scientific literature in the process.  Working with live organisms also presents a unique set of challenges!  I have learned many valuable lessons from this experience that include time management and ‘how to do science.’

As I have advanced throughout my college career and working on this research project, which I will continue into the next academic year, I have decided that working in a laboratory setting is something that is very interesting to me.  As I continued to spend more time in the lab this summer I realized I would like to continue with this type of work in my academics and hopefully someday as a fulltime career.  After I graduate, I am considering graduate work in Ecology or Environmental Science. I really enjoy the independence that comes with working in a laboratory setting and I feel confident that I could pursue a career in the same type of work environment. Ultimately, Professor Kipp and I would like to have our work published and I would also like to gain the experience of presenting my work at appropriate conferences.   .

The Effects of Heat Stress on Microgametophyte Viability in Arabidopsis thaliana

Environmental and ecological changes due to temperature increase, such as those associated with global climate change, have been well documented.  Since temperature fluctuations are greater on a regional rather than global scale, research quantifying plant responses to heat stress has become a principal objective for environmental and ecological biologists.  The objective of this research is to gain a better understanding of how an increase in global temperatures will affect overall plant growth and reproduction.  To that end, Professor Kipp and I will subject Arabidopsis thaliana, a commonly used lab rat, to slightly elevated temperatures.  We will compare these test plants to plants grown optimally.  In this experiment, we are going to monitor how heat affects the plant’s microgametophyte–pollen.  We will document pollen development and morphology, test pollen viability, and record overall pollen numbers.   We will use statistical tools to analyze our data in order to draw conclusions.  We hope to publish our research findings so that the data can be used to project the effects of heat stress in agricultural and other economically valuable plants.  Thus, the results of this research can be applied to the fields of agronomy, ecology, and plant physiology, as well as to quantify future consequences of global warming and help give insight into long-term stewardship and conservation of crop infrastructure.

My research began with searching for and reading scientific literature relevant to my experimental questions: “Does heat affect pollen production and maturation?”  And if it does, How?”  “Does it affect the ability of the plant to reproduce?”  “Are pollen numbers affected?”  “Is the morphology and overall size of pollen altered?” “Are pollen produced but their viability compromised?”

By reading journal articles, I have begun to learn about current research and methodologies, as well as how to search for and dissect peer-reviewed literature.  Professor Kipp has delivered several PowerPoint Presentations in order for me to understand the basics of plant biology and climate change and to help me gain more from each paper we read.  I have collected various protocols from books and the journal articles that I will use to carry out the experiments and eventually generate data.  I am in the process of becoming more familiar with the protocols and laboratory equipment I will need in order to carry out the experiments successfully. One goal is to become more adept at literature reviews and using laboratory equipment.

Once I have established a comfort level with the literature, equipment, and selected protocols, I will begin growing two sets of plants: one set of plants will be grown at 23°C during the day and 23°C at night.  This will be my control set of plants as A. thaliana grows best at 23°C.  The second set of plants will be grown at 29°C during the day and 23°C at night.  This second set of plants will serve as the experimental set as they will be subjected to heat stress during the day.  Both sets of plants will receive equal water, fertilizer, 12 hours of daylight, and 12 hours of night.  We will grow the plants in growth chambers so temperature and humidity, photoperiods, and all experimental parameters can be accurately controlled.

Once the anthers develop (site of pollen production), I will begin collecting pollen from both the control set and the heat-stressed set of plants.  The control set will provide the base line in order to compare any differences in pollen between these optimally gown plants and the heat-stressed plants.  Carrying out this research will not only help with my overall skills in a research laboratory, but it also will help with my communication skills, written and verbal, thru blogging, keeping a lab notebook, reading scientific literature, and presenting our work at several conferences and poster sessions.  This is another personal goal, to increase my scientific literacy.  I am learning to ask good questions in the laboratory and trying to find answers to those questions by developing sound experimental protocols, which will help me with my analytical skills.  Once the experiments begin, I hope to learn how to effectively troubleshoot in the laboratory.  The learning process that comes with performing this research has already proven to be worthwhile, and I look forward to continuing my research throughout the summer with Professor Kipp.