Proposal Requirement 5 Example: Chemical Plant Biology

Writer: Swasti Singhai

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Proposed Solution This study plans on using the traditional liquid chromatography-mass spectrometry to detect OPFR due to its uses in both plant research and OPFR detection in animal models. In the case that the compounds are insufficiently separated, we will use a tandem mass spectrometer to increase the sensitivity of separation.  Arabidopsis thaliana is the first plant we will be detecting and monitoring due to its adoption as a model organism in plant biology. This plant is easy to work with due to its short generation time, small size, and rapid seed production through self-pollination [1]. In the event that OPFR is insufficiently detected in a large, natural sample size, we will manually increase the exposure of OPFR to the plants by growing Arabidopsis thaliana and/or glycine max in a laboratory setting with soil containing ~0.3 μM of TPHP (1 type of OPFR). This will help determine whether or not OPFR can penetrate plants through the soil, an important discovery in regions with high OPFR concentrations in sediment [2]. While keeping the amounts of temperature, light, water, and nutrients constant, one control group will consist of little to no OPFR concentration in the soil and plant. The rest of the plants will be separated in groups based on OPFR concentration. All groups will be regularly observed and tested for OPFR concentration using LC-MS over the course of a few months. The concentrations will be graphed, and at the end of the study, these graphs will be examined to identify the plants’ retention of OPFR. The majority of this study depends on two factors: the detectability of OPFR and the measurability of OPFRs’ effects in plants. Moreover, the previous research showing evidence of human consumption of OPFR and the structural similarities with organophosphate pesticides strongly indicates the above possibility. Preferably, a physical change eventually occurs in the plants, and the specific mechanism of action (a certain biochemical/biophysical process that is affected by the contaminant) is identified through the symptoms. Regardless of whether this occurs or not, groups with OPFR and the control group then will both be tested for known carcinogens supported by the state of California, such as TDCIPP and TCEP, which are chlorinated OPFRs, using LC-MS/MS coupled with electrospray ionization in order to increase the sensitivity of our results [3]. Multiple trials will have to be conducted to ensure that the conclusions are due solely to OPFR’s effect.  Then, the same process will be repeated in glycine max, the soybean plant. Due to the soybean plant’s widespread use in the food industry and its role as a dietary staple in many countries, this study is crucial to assess the potential toxic exposure. As shown in the steps of our study, our solution to many of the possible insufficiencies with the techniques used is repetition and variation. By varying the species and repeating the study with >50 samples, this study aims to be as accurate and efficient as possible in advancing our understanding of the toxicity of OPFRs. 

Analysis The author clearly goes over the study’s procedure, including the technology, model organisms, and methods of cultivation. With that, the author talks about not only the potential insufficiencies, but also possible solutions. This can help the researcher plan ahead, and if applying for a grant, the information would be useful to the sponsor as well. It’s important to note that this section does not cover technology too much in-depth, because the entire previous section is devoted to current technology/insufficiencies. The author also clearly described the procedure in chronological order. A diagram may have also helped the reader understand, and the author also could have gone more in detail with implications regarding costs and accessibility (if the model the research requires is extremely rare, it would be important to note that as an implication). Otherwise, the author does clearly go through the procedure, implications, and how to solve them (if you're interested in reading the technology part of this piece, you can check out the previous post I wrote regarding this topic). 

References [1] Koornneef, Maarten, and David Meinke. “The Development of Arabidopsis as a Model Plant.” The Plant Journal, vol. 61, no. 6, 2010, pp. 909–921., doi:10.1111/j.1365-313x.2009.04086.x. [2] Cao, Shuxia, et al. “Levels and Distributions of Organophosphate Flame Retardants and Plasticizers in Sediment from Taihu Lake, China.” Environmental Toxicology and Chemistry, vol. 31, no. 7, 2012, pp. 1478–1484., doi:10.1002/etc.1872. [3] Dodson, Robin E., et al. “After the PBDE Phase-Out: A Broad Suite of Flame Retardants in Repeat House Dust Samples from California.” Environmental Science & Technology, vol. 46, no. 24, 2012, pp. 13056–13066., doi:10.1021/es303879n.