Thursday, October 3, 2019
Impact of Increased Temperature on Delosperma Cooperi Pollen
Impact of Increased Temperature on Delosperma Cooperi Pollen Eunice Oh The Impact of Increased Temperature due to Global Warming on Pollen Germination ofà Delospermaà Cooperi Introduction: There is an ongoing crisis that is beginning to influence ecosystems throughout the world,à which may lead to largeà scaleà natural disasters due to the rise in temperature from global warming. According to NASAââ¬â¢s Goddard Institute for Space studies,à 0.8à °Cà have increased around the world since 1880. In addition, the rise in temperature is pervasive andà isà increasing at a faster rate in the last two decades (SITE1). This warming phenomenon can disturb ecosystemsà andà lead to extinction in extreme cases.à Such ecosystems are dependent on plant growth and proliferation to sustain itself.à Therefore, an experiment toà observeà theà effects ofà a significantà rise in temperature on pollen germination was conducted to predict the adaptability ofà Delopspermaà cooperi,à aà common species ofà iceplantà grown around the world, toà this phenomenonà . Tà Delosperma cooperià (trailing iceplant) was compared toà Tulbaghia violaceaà (society garlic)à to obtain a broader view of how different plants from the same environment would react to a distinct change in temperature.à An increase of 10à °C was chosen as the variable to performà analysis with the Q10à temperature coefficient.à Pollen is a fine powder that contains microgametophytes of seed plants and produces male gametes. When pollination occurs, the pollen grainà germinates and a tube is producedà as a conduit to transport the male gametes from the stigma to the pistilsà of the ovule in flowering plantsà (SITE2).à In nature, germination occursà when the stigma is hydrated fromà waterà sources (e.g. rain)à . can also be inducedà in vitroà usingà aà germination media andà theà hanging drop method (SITE 3).à Three replicates were observed the wereà analyzedà withà statisticsà toà measure the significance of the variable à (via a T-test, and Dixon Q). à The plantââ¬â¢s temperature dependence was quantified with the Q10à temperature coefficient. It was predictedà that the increase in temperature would result inà a significant improvement ofà pollen germination rateà and longer pollen tubes than the controlà due toà Delospermaà cooperiââ¬â¢sà adaptive traits (quote). Materials and Methods: Germination ofà Delospermaà cooperià was induced in basic germination media, composed ofà 1mM KCl, 0.1mM CaCl2, 1.6mM H3BO3, 10% glucose,and distilled water. Standard labà equipments wereà used: lightà microscope,à gardenà gaskets, depression slides, slides warmer, petri dish,à and micropipettes. The light microscope was used under the 10x objective to track the germination process and measure the elongation of pollen tubes. Toà accommodateà forà a large sample volume (50à µLà transferred using micropipettes), garden gaskets were employed to extend the capacity of the depression slides.à Aà slides warmerà was used to maintain the high temperature environment (37à °Cà )à andà wetà petrià dishes wereà utilized as germination chambers. The hanging drop method consistsà of several steps. à A gasket was placed on top of the slide in order to create an area for the hanging drop to be intact with the cover slideà and held together with grease. The slides were placed in the humidityà chamber toà allow germination andà prevent drying. Two sets of the hanging drops were prepared, one for the higher temperature (37à °C),à and another for theà positiveà controlà (27à °Cà ). The negative control was prepared by observing the pollen without any germination media. Statistical analysis methodology: Theà germinationà elongation rates were recordedà by sampling fiveà pollenà tubes from each slide in 30 minutesà intervals, up to 150 minutes.à This data was analyzedà using biostatistics.à Aà Dixonà Q test was performed to identify and remove outliers.à Theà Dixon Q testà was calculated using the equation, Q= (gap)/(range). The gap refers to the absolute difference between the outlier and the closest number to the outlierà and the range is simply between the smallest and largest valuesà (CITE). After the elimination of outliers from the Dixon Q test, a student T-Testà (with a 95% confidence interval)à was performed to determine whether the variables were statistically significant in the difference of their elongation ratesà using P valuesà (SITE).à Finally, aà Q10à value was determined from the mean ofà elongationà rates.à It was calculated by using the following equation: Q10à = (R2/R1)10/(T2-T1).à Q10à is a unit-less measur ement thatà quantifyà the change of a biological systemà due to temperature change. Results: The purpose of the experiment was toà measure theà elongation rates after every 30 minute interval, 32 points of data were obtained and analyzed.à Overall, the elongation rateà ofà Delosperma cooperifor the highà temperature variable was as much as threeà times fasterà compared to the controlà temperatureà (0.686à à µm/min vs.à 0.278à µm/min)à in trial three. The percent germination wasà alsoà noticeably better for theà high temperature variableà versus the control, whereà it wasà approximately 60% compared to 20%à after 120à minutes from initiation. From the list of data, theà Dixon Q-test result indicated the data point 0.780à µm/min of the higher temperature control as an outlierà with a 95% confidence level. The mean elongation rate for the room temperature was 0.314à µm/min and 0.454à µÃ m/min for the higher temperature control. The student T-Testà yieldedà a P value of 0.0447, which indicatesà that the result is statistically significant at a 95% confidence interval.à Theà Q10à temperature coefficientà forà Delosperma cooperià was calculated to beà 3.59, categorized as a temperature dependent biological system. Figure 1.à The graph shows the averageà elongationà rates ofà Delospermaà cooperià at two differentà temperatures. The tubule elongation rate wasà 0.314à µm/minà for the control andà 0.454à µm/minà for the variable. Error bars denote one standard deviationà (0.152à µm/minà andà 0.177à µm/min, respectively)à above and below the mean. Figureà 2. The graph shows the average elongation rates ofà Tulbaghiaà Violaceaà at two differentà temperatures. The tubule elongation rate wasà 17.4à µm/min for the control andà 3.00à µm/min for the variable. Error bars denote one standard deviation (1.95à µm/minà andà 0.279à µm/min, respectively)à above and below the mean. Discussion: The results appear to support the hypothesis, whereà Delospermaà cooperià was positively affected by the increasedà temperatureà by approximatelyà a 0.140à µm/minà and 40% germinationà improvement.à The result shows that the higher temperature yielded in an improvement in both percentage germination and pollen tube length growthà at a significant level (P10à value is higher than 2. Q10à is a unit-less measurement that establish a temperature coefficientà that correlates a systemââ¬â¢s change to temperature differenceà (of 10à °C)à (SITE 4) In addition,à theà higher percentage germination was observed from the higher temperature controlà correspond to an articleà in whichà Delosperma cooperià is more adapted to a higher temperature environmentà due toà increased metabolic rate under temperature stressà (SITE 5). The results ofà Delopsermaà cooperià were compared withà Tulbaghiaà violaceaà and suggest that the increased temperature had the opposite effect onà Tulbaghiaà violacea, whereà pollen germination percentage and pollen tube growth were more effective in the room temperature control.à Tulbaghia violaceaà is known to be better suited in the colder environmentà while highà temperaturesà restrict their germination (SITE 6). However, the data was determined to be not significantly significant.à (P>0.6).à A possible future experiment includes testing a greater variety of indigenousà flower pollensà under more temperature variances. The experiment provided a glimpse into how certain plants would respond toà the consequences ofà globalà warmingà and more studies are needed for a more comprehensive overview.à References Leistner, O. A. (ed.). 2000.à Seed plants of southern Africa: families and genera. Strelitzia10. National Botanical Institute, Pretoria. Mozaffar Ebrahim Edmund John Pool (2010). The effect ofà Tulbaghiaviolaceaà extracts on testosterone secretion by testicular cell cultures.à Journal ofà Ethnopharmacologyà 132(1): 359ââ¬â361 Reyes, A.B.,à Pendergast, J.S., andà Yamazaki, S. 2008. Mammalian peripheral circadian oscillators are temperature compensated. J.Biol. Rhythms 23: 95-98. ââ¬Å"Global Warming Facts.â⬠2007. National Geographic.à http://news.nationalgeographic.com/news/2004/12/1206_041206_global_warming.html Raven, Peter H.; Ray F. Evert, Susan E. Eichhorn (2005).Biology of Plants, 7th Edition. New York: W.H. Freeman and Company Publishers. pp.504ââ¬â508. Pfahler PL (1981).In vitro germination characteristics of maize pollen to detect biological activity of environmental pollutants. Health Perspect.37: 125ââ¬â32. Reyes, A.B.,Pendergast, J.S., and Yamazaki, S. 2008. Mammalian peripheral circadian oscillators are temperature compensated. J.Biol. Rhythms 23: 95-98. Rinnanà R, Steinke M,à McGenityà T, Loreto F. Plant volatiles in extreme terrestrial and marine environments.Plant Cell Environ. 2014 Mar 7. http://autocite.durkmed.com/
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