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Center for Plant Cell Biology



REU 2002


REU Students and their Summer 2002 Experiences

Please click on the following student links to see student photos and read about their Summer 2002 experiences working in CEPCEB laboratories.

Eduardo Cen
Marjannie Eloi
Susan Garity
Courtney Hamada
Mattie Irion
Jennifer Jones
Derrick Sergeant
Isha Wallace

Eduardo  EDUARDO CEN
Zhenbiao Yang Lab

I have been working with pollen tubes from transgenic Arabidopsis thaliana plants as a model to study a G protein-mediated signaling pathway that acts as a switch for an array of physiological effects in the cell. The genetic transformation technology and molecular techniques I have been learning are valuable information and will most likely prove very useful in my goals as a research scientist. The experience of working in Plant Cell Biology has shown me that good science can be applied to a variety of systems and models; and conversely, one model can be used in a variety of applications of good science.

Overall, my experience here has been very rewarding.

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Marjannie MARJANNIE ELOI
Elizabeth Bray Lab

I am currently working in Dr. Elizabeth Bray's laboratory at UC Riverside. The focus of study in this laboratory is on the effects of plants under water-deficit stress. An important response of plants to this stress is the accumulation of the plant hormone, abscisic acid (ABA). The goal of my project this summer is to determine how other plant hormones interact with the accumulation of ABA under water-deficit stress. Two plant hormones, ethylene and auxin, are being examined to determine if there is any significant interaction. I am growing Columbia wild-type Arabidopsis seedlings on plates containing precursors of ethylene and auxin. After three days of incubation, I am exposing these seedlings to different water-deficit stress levels. I am measuring ABA and proline levels using an ABA radioimmuniassay and proline assay. I will determine if any of these hormones affect the accumulations of ABA or proline under water-deficit stress.

This is my first research experience. I am enjoying my time and learning a lot of valuable information. I really enjoy the hands-on experience of working on and running my own experiments. I really like this program because I have my own project to work on. I now have a much clearer view of what research is really like. The program is great and I am very happy and thankful that I was selected to be one of the eight students in this program. I feel my experience here will stay with me for a lifetime.

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Susan

SUSAN GARITY
Linda Walling Lab

The project that I am working on is to identify genes regulated by a phloem-feeding insect, the silverleaf whitefly (Bernisia argentifolii) using gene trap/enhancer trap technology. The project is two-fold. Objective 1 required us to grow wild-type Arabidopsis plants in insecticide-free soil and infest them with silverleaf whiteflies. We then collected tissue from these plants and also our controls at 6 different time points over a course of 36 days. After all 6 time point tissues have been collected, we then can extract RNA from both our control and infested leaves and run an agarose gel. After this is completed, we need to transfer the RNA to a membrane and hybridize it with two different gene probes; PDF1.2 defensin and BGL2, β-glucanase. These genes are known to act exclusively in one of two defense-response pathways; the jasmonic acid-dependent and salicylic acid-dependent. The last part of this objective would be to autoradiograph signals to see the increases in RNAs during infestation. This experiment will identify if WF feeding induces any of the known defense-signaling pathways. The second objective of my project is screening a part of the Arabidopsis gene-trap (GT) and enhancer trap (ET) line available here at UCR. After growing these lines in insecticide-free soil under long-day conditions, we infested each flat with 750 wilverleaf whiteflies and changed them to short-day conditions. Non-infested controls of the Arabidopsis lines were also grown. After 21 days of infestation, we then harvested the developmentally-matched tissue from both the infested plants and our controls. After taking down the screen, the leaves were histochemically stained to identify any GUS activity. Microscopy was done to determine any GUS staining patterns, which were recorded in a GT/ET datasheet. After selecting two lines that I thought were good candidates based on their staining patterns, I then re-grew the two lines and extracted Genomic DNA from two-week old seedlings. TAIL PCR will be done to identify the gene driving reporter gene expression. A database search will let me know if the gene I have isolated is novel or has been previously characterized. Both objectives will help in determining how plants respond to phloem-feeding insects.

Over the course of this internship I have been able to gain knowledge of many different lab methods and techniques. I have been able to see for myself what goes on in a research laboratory and have the opportunity to do hands-on experiments on my own project. The skills and knowledge that I have acquired over the past weeks are priceless. I have learned so much from my PI, Dr. Linda Walling; mentor, Sonia Zarate; and other lab members. It has been an awesome experience and the guidance, instruction and encouragement have been amazing. This truly will be a summer that I will never forget, and the skills and techniques that I have learned over the past few weeks will surely be put to use in the future. I am very thankful for being able to participate in such a great internship as an undergraduate.

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COURTNEY HAMADA
Elizabeth Lord Lab


This summer I have been working on determining which chemicals disrupt pollen tube adhesion. The chemical genetics experiment consists of creating an artifical matrix of pectin and SCA and adding to it chemicals as well as pre-germinated pollen tubes, and seeing the effect it has on the adhesion of the pollen tubes to the matrix. Before this summer, I had never worked in a lab before. I was a little intimidated at first; however, that disappeared once my time in the lab began. Every day I learn new and interesting information. My time here always seems to just fly by. I also could not ask for a better enrionment to work in. My mentor, Jean-Claude Mollet, has to be the most patient and understanding person I have ever encountered. If I do not understand something at first, he has at least five other ways he can explain it to me. Also, the graduate students (Sunran Kim and Juan Dong), Kathie Eckard, the other undergraduate student (Kimberly Tan), and of course, Dr. Lord, have all helped me immensely. The experience I have gained working in Dr. Lord's lab is invaluable. I know it will be extremely useful, not only in the rest of my studies, but also in my future plans. I want to go to medical school and possibly be involved in research. I am very grateful for having this opportunity made available to me.

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MATTIE IRION
Shou-Wei Ding Lab

In the seven weeks doing research in the Ding lab, I have had the opportunity to participate in extensive laboratory procedures directly correlated with the cutting-edge research presently being studied involving RNA silencing and the involvement of miRNAs in gene regulation. It was a privilege to be involved in the review process of several papers submitted for possible publication in this growing area of genomics -- a responsibility that Dr. Ding normally reserves for graduate and post doctorate participants. I feel blessed to have been mentored by Hong Wei Li, a post doctorate participant in the lab. He was incredibly patient and determined to expose me to the most valuable lab techniques and indispensable information in genomics that I am certain to be ahead of my peers and that I am certain to carry over into medical school. This type of experience and exposure is an "invaluable opportunity" for any undergraduate, which many must wait to get once in graduate school. I am honored to have been selected as one of the first eight participants of this NSF-funded research program, and I would like to recognize and thank all those that participated from CEPCEB.

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JENNIFER JONES
Julia Bailey-Serres Lab

The objective of my project in the Bailey-Serres lab is to determine whether Arabidopsis thaliana RopGAP genes (encoding Rho like protein of plants, GTPase Activating Proteins) are differentially expressed in response to stress induced by hypoxia, cold and DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea, an inhibitor of photosynthetic electron transport).

Airica Baxter-Burrel, a graduate student in Dr. Bailey-Serres' lab, has demonstrated that RopGTPases participate in transduction of signals that regulate gene expression in response to hypoxia (Baxter-Burrel et al. 2002). Part of this includes the differential regulation of one RopGAP gene (RopGAP4) in response to oxygen deprivation. However, it is not known whether any other Arabidopsis RopGAP genes (RopGAP 1,2,3,5 or 6) are involved in the hypoxic response or whether RopGAP genes are involved in other plant stress responses. This is being tested using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR, a method used to determine the abundance of an mRNA transcript).

Another question being addressed is whether DCMU causes oxidative stress that induces the same Rop signaling pathway involved in transduction of hypoxia signals. This can be determined by probing differential regulation of other components of the Rop signaling pathway: ADH (Alcohol Dehydrogenase), GUS (β-glucuronidase) and H2O2, in addition to the RopGAP4 gene. To do this, RT-PCR, ADH activity assays, GUS activity assays and H2O2 measurements are being performed using both, tissue from wild-type Arabidopsis and tissue from a mutant in which the RopGAP4 gene is disrupted with a GUS reporter gene.

At the time I came to the REU program the Bailey-Serres lab had protocols for hypoxia and cold stress induction in Arabidopsis, but not for DCMU stress induction. One of my tasks was to develop a protocol for treating Arabidopsis seedlings with DCMU and determining the degree of DCMU inhibition of photosynthetic electron transport in Arabidopsis seedlings using a fluorescence induction assay. This employed my previous experience using fluorescence assays to screen cyanobacterial mutants for perturbed oxygen evolving complexes.

I am appreciative of Dr. Bailey-Serres' sensitivity to my interests in allowing me to integrate my previous work using fluorescence assays with this project by allowing me to investigate the effect of DCMU on gene expression. She and other members of her lab have done a fine job teaching and answering my questions. This and the wide applicability of the techniques I am learning in this project make this an experience extremely worthwhile.

Baxter-Burrell AY, Yang Z, Springer PS, and Bailey-Serres J (2002) RopGAP4-Dependent Rop GTPase Rheostat Control of Arabidopsis Oxygen Deprivation Tolerance. Science 296: 2026-2028

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DERRICK SERGEANT
Eugene Nothnagel Lab

My ultimate goal is to become a college or university science professor. I already knew that I enjoyed teaching, having had a lot of experience in that area. But a science professor must also do research. I had very little experience doing full-time work in research before entering this program. I am happy to say that the experience has been extremely valuable and enjoyable. The instruction I have received on laboratory and research techniques are sure to serve me well in the future. My research focused on some of the components of plant cell walls and what happens when their normal function is interrupted. I had a wonderful time investigating this question. What I enjoyed most of all was the freedom to be able to eventually set up my own experiments and be responsible for my own data collection and results evaluation. I have only good things to say about the ten weeks I spent here.

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ISHA WALLACE
Natasha Raikhel Lab

I've been researching genes involved in the biosynthesis of plant cell walls. I use molecular techniques to study the expression of two genes that seem to be related. I also use computer analysis programs to compare homologues of the two genes in other organisms.

Working in a research laboratory has been a great experience. I get to do most of my research on my own and there is always some one to ask for help when I need. it. This experience is not intimidating, as I initially thought. Everyone in the lab is friendly and very supportive.

I enjoyed the experience so much that now I realize more than ever that I really want a career in research. (I also realized that I like working with plants.) I learned a lot about scientific research this summer. I don't think I could have learned so much anywhere else.

I thank Dr. Raikhel for an opportunity that I will never forget.

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Center for Plant Cell Biology
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E-mail: genomics@ucr.edu

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