Pteridophytes are vascular plants that do not utilize seeds for reproduction. Instead, they switch between a gametophyte and sporophyte process to flourish. There are ways to expedite the growth process by using fertilizers that contain nutrients for the plant. In this lab the, the purpose was to see what the effect of adding 1% ammonium nitrate on the C-fern spores germination. For this experiment, two petri dishes with agar gel and C-fern spores spread out on top and with the 1% ammonium nitrate only being added to one petri dish. The samples were then placed in a isolated area of the room for 6 weeks to be incubated. Over the 6 weeks, it was observed that the experimental petri dish, which had the 1% ammonium nitrate, actually germinated at a much slower pace compared to the control. It was concluded that that the addition of the 1% ammonium nitrate actually had a negative affect on the spores because the excess nitrogen slowed down the C-fern germination process.
Nitrogen is a vital part of the development and germination. Almost 80% of the earth’s atmosphere is made up of nitrogen gas. However, plants have to undergo nitrogen fixation to absorb the nitrogen because they are unable to in their natural form. The atmospheric nitrogen is converted to ammonium, or nitrate, by diazotrophic bacteria that are symbiotic to plants because regular nitrogen gas is useless to the plant (Boundless). The level of nitrogen that is absorbed by the plant has a great affect on the growth of the plant. There was a study done on how variations on the amounts of nitrogen absorbed by Foliage plants would hinder the growth (Steinkamp). For nine months, seven different plants were given different fertilizers with varying amounts of nitrate and ammonium in the fertilizer. The growth data was recorded every three months to compare the results. The results showed that plants that were given an average amount to nitrate and ammonia yielded the best results with regards to height, plant quality, and the color. The other samples expressed decent results however; the color and height of the plants were not up to par. There is another study as well that yielded the same result at the previous experiment. The University of Minnesota conducted a research on Purple Lupines by giving 296 field plots different amounts of nitrogen (Sciencedaily). This experiment was conducted within an experimental plot where the conditions were regulated such as light. After six years, the scientist concluded that the plants that were given less nitrogen led to higher levels of carbon dioxide in the atmosphere. This means that plants who are deprived of their nutrients lack in performance (photosynthesis) and show a lack of physical characteristics.
In this lab, the purpose was to observe how ammonia nitrate affects spore germination on Ceratoptris Ferns. Every week, the sample of Ceratoptris Ferns, also known as C-Ferns, was observed to see if the addition of ammonium nitrate increases, decreases, or has no affect on the germination rate of Ceratoptris Ferns. It was hypothesized that if the Ceratoptris Ferns are provided the ammonium nitrate, the germination rate will be slightly faster compared the average germination rate. The null hypothesis for this experiment was the average number of days for germination to take place would be about the same for the control and experimental test. Observing the different environments that the Ceratoptris Fern are grown in lets us have a better understanding of how the ammonium nitrate affected the germination process.
For this experiment, Ceratoptris Ferns were used as the subjects. There were two petri dishes obtained, one labeled control and one labeled experimental. The control petri dish had a medium of agarose gel and the experimental group had agarose gel but it also had the 1% ammonia nitrate. When the handling the petri dishes with the agarose, be sure to not damage the gel solution. A paper clip, Bunsen burner, and pipet were used for the next step. Using the aseptic technique, the paper clip was bent straight. After putting on safety goggles, the Bunsen burner was turned on and the paper clip was waved over the open flame to sterilize it. After that, the C-Fern spores were transferred into a solution using a pipet. Then three drops were transferred to each petri dish. Using the paper clip that is sterile, gently slide it across the agarose gel at the bottom of the petri dish making sure not to disrupt the gel itself. The experimental dish will need an extra ingredient, which is the 1% ammonium nitrate. Both the petri dishes where then covered with the covers provided and tape the edges. The dishes were then placed under florescent lights at the designated location at room temperature. The samples were observed every week during class to see how the germination compared to the control group. Both dishes were seen under a microscope.
Table 1: The table shows the average spore count for both petri dishes.
Group Number Number of spores in Control group Number of spores in Experimental group
1 93 64
2 104 105
3 114 26
4 116 47
5 169 62
6 99 34
Average 115.8 67.6
Chart 1: The chart shows a the average number of spores germinated over the 6 week period for both samples.
Figure 1: The equations show the sample variance of the control and experimental samples.
Control : = 754.97
Experimental: = 793.1
Figure 2: The calculations show the homoscedasticity of the control and experimental variance.
= = 1.05 *Degrees of Freedom= n-1=5
* 1.05 <5.0503
Figure 3: The figure below reflects the results of the t- test to determine the condition of the null hypothesis.
*Degrees of Freedom= (n1 +n2) – 2= 10
= = 3.02
The data was recorded after a 6-week period for the spores to properly germinate. After the 6 weeks, the number of spores was counted by all the groups and averaged with the other section. In table 1, the average spore count for all the groups after the data was pooled from all the sections and averaged. Also in the first table is the average number of spores from all 6 groups (Table 1). Those values were then used to make the chart to reflect the average number of spores germinated for both samples. The height of the bar shows which sample had the higher spore count and a quick visual comparison of which yielded the best results. The error bars on the chart were derived from the standard deviation of the data (Chart 1). With the information until this point, the sample variance was calculated for both samples (Figure 1). The values calculated for the variance were then used in the F-test to calculate the homoscedasticity. The α- value used was .05 and the degree of freedom was 5 since there were 6 groups. The F-test value was 1.05 and when compared to the critical value of 5.0503 to see if it supported the null hypothesis (Figure 2). For Welch’s T-test, the α- value was .05 again but the degree of freedom was 10. Welch’s T-test was calculated using the sample variance values to see if it also supported the null hypothesis (Figure 3).
According to the data obtained from this experiment, it is observed that the C-fern culture without the 1% ammonium nitrate germinated at a much faster rate compared to the experimental inoculated with the 1% ammonium nitrate. The control group had an average spore count of 115.8 compared to the experimental culture, which had an average spore count of 67.6. From the data presented, it is observed that the 1% ammonium nitrate held back the germination of the spores. The F-test value was 1.05, which is way below the accepted critical value. The variance of both cultures was relative so this prevents us from rejecting the null hypothesis. The F-test, calculated using Welch’s T-test, yielded 3.02. The critical value was 1.812, the degree of freedom was 10, and the alpha value is .05. The calculated value was a little higher than the critical value, which led to the rejection of the null hypothesis meaning the ammonium nitrate, impeded the germination of the spores.
Although nitrogen, phosphorus, and potassium are all necessary for germination of spores, the perfect amount needs to be utilized. If an excess of one of these occurs, the process slow down (Brazilian). The excess of the nitrogen throws off the process because the culture has to make up for the lack in the other nutrients since there’s abundance in nitrogen (Home).
There are a number of possible errors that could’ve occurred during this experiment possibly rejecting the null hypothesis. The inoculation of spores could be a factor because if they weren’t done properly and spread out, only one area was affected by it then. The petri dishes could’ve been contaminated resulting slower germination. This experiment could’ve been executed better if the petri dishes were checked daily instead of weekly to get a better understanding of how the spores are coming along and what changes everyday.
This experiment is a prime way to understand the germination process of C-fern spores. Also it helps give a better understanding of how nitrogen affects the culture through the addition of the 1% ammonium nitrate. . For the future, this lab should be conducted for a longer period because the first couple weeks there was no germination and by the time the culture was getting to its prime, they were tossed out.
“Brazilian Journal of Botany – Germination of Spores and Growth of Gametophytes and Sporophytes of Rumohra Adiantiformis (Forst.) Ching (Dryopteridaceae) after Spore Cryogenic Storage.”Brazilian Journal of Botany – Germination of Spores and Growth of Gametophytes and Sporophytes of Rumohra Adiantiformis (Forst.) Ching (Dryopteridaceae) after Spore Cryogenic Storage. N.p., n.d. Web. 20 Nov. 2014.
“Higher Carbon Dioxide, Lack Of Nitrogen Limit Plant Growth.” ScienceDaily. ScienceDaily, n.d. Web. 15 Oct. 2014.
“The Nitrogen Cycle – Boundless Open Textbook.” Boundless. N.p., n.d. Web. 15 Oct. 2014.
Steinkamp, K. “Nitrogen Form Affects Foliage Plant Growth A Summary of Research at CFREC Apopka.” Https://mrec.ifas.ufl.edu. CFREC, n.d. Web. 15 Oct. 2014.
“What Do Ferns Require to Grow & Germinate?” Home Guides. N.p., n.d. Web. 19 Nov. 2014.
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