The Role of Phytochrome in Germination of Lettuce Seeds Formal Lab Report

Bio 102 Section 09 Lisa Ivester

Title: The Role of Phytochrome in Germination of Lettuce Seeds

Abstract: Phytochrome regulates many responses to light by a red - far red conversion mechanism. The purpose of the experiment is to determine the role of phytochrome in lettuce seed germination. Light control is very important for the plant. The hypotheses tested were that germination will occur in far red light if the variety of seed is concept and also that germination will occur in red light if the variety of seed is two starr. Dark light is used as a control while results are checked in green light. The seeds are exposed to the light for 15 minutes and then checked on the second day after the initial experiment and again on the eighth day after. Results are as follows; 9 lettuce seeds germinated under white light, 4 under far red light, 1 under the red light, and no seeds germinated under the dark control. Also, the concept lettuce seeds had a higher rate of germination than the two starr lettuce seed. These results of the experiment supported the hypotheses.

Introduction: Light is critical in a plants environment for much of the plants growth and development.(Roux 1984) Photomorphogenesis, which is growth and development in a plant, begins with light absorption by a pigment in the plant, called phytochrome. (Roux 1984) Phytochrome regulates responses to light in many different types of plants. (Campbell et al. 2009) It is known to be involved in germination and flowering response of long-day plants. (Casal 1993) It was discovered by studying seed germination. (Campbell 2009) The USDA discovered the specific light spectrum for light induced

lettuce seed germination in the 1930s. (Campbell et al. 2009) The red light wavelength of 660 nm has the most effect on the germination. Far red light, at 730 nm, inhibits germination. (Campbell et al. 2009) The photoreceptors that caused the opposing effects of the two lights are phytochromes. A phytochrome has two subunits. In these subunits, a polypeptide component is joined to a no polypeptide chromophores by a covalent bond. Chromophores are the light absorbing part of the subunit.(Campbell et al. 2009) The chromophore is photoreversible. The pigment has been proved to be a chromo protein that exists in two forms that, by light, are interconvertable. (Kendrick and Spruit 1973) Phytochromes interconvert between two forms when responding to red and far red light. (Sparace 2010) The two isomeric forms are Pr and Pfr. Pr being the isomer form that absorbs red light, and Pfr the isomer form that absorbs far red light. (Campbell et al. 2009) Pfr triggers developmental responses to light in plants. A prime example would be, in a lettuce seed, Pr exposed to red light is converted to Pfr. This conversion triggers the responses in cells that lead to germination. Likewise, when the red light seeds are exposed to far red light, the germination response is inhibited. (Sparace 2010, Campbell et al. 2009) In nature, plants synthesize phytochrome as Pr. In complete darkness, the pigment will remain mostly in Pr form, which is why germination does not occur without light exposure. Sunlight contains red and far red light, but Pfr converts faster than Pr, meaning there is more Pfr and germination is triggered. (Campbell et al. 2009) The experiment was conducted to test the role of phytochrome in lettuce seed germination. The study was performed to provide further insight into lettuce seed

germination under the effects of different lighting. It was performed to determine how different types of lettuce seeds react to the different lighting also. It was hypothesized that if the lettuce seed variety was concept then germination will occur in response to far red light. The null hypothesis is that germination will not occur in the seeds exposed to other types of light. It was also hypothesized that if the lettuce seed variety was two starr then germination will occur in response to red light(but not far-red light). The null hypothesis fro this second variety is that if it is two starr, then the germination will not occur in response to red light.

Materials and Methods: Materials used in the experiment to test the role of phytochrome in lettuce seed germination are as follows; green safelight, eight petri dishes, approximately one hundred lettuce seeds, filter paper, water and foil. A dark cabinet for storage is also needed to conduct this experiment. (Sparace, 2010) The experiment was conducted in a dark room, using a green safelight to see. Eight petri dishes were labeled according to the seed variety and type of light treatment. Four were labeled concept seed variety and the other four were labeled two starr seed variety. The date of the experiment is also labeled on the petri dishes. Next, filter paper was placed in each dish. Two to three droppers full of water were added to the petri dishes. The filter paper was thoroughly moist, but water was not accumulating in the dish. (Sparace, 2010) Twenty five lettuce seeds was added to each of the four petri dishes. Next, the seeds in the petri dishes were exposed to the light regimens. One of each kind of lettuce seed was exposed to one type of light. For example, one petri dish containing

concept seeds and one petri dish containing two starr seeds were exposed to white light. Likewise, one petri dish containing concept seeds and one dish containing two starr were exposed to red light, and so on for the two remaining light regimens. Table 1 in the results section illustrates the light regimens. The dishes were wrapped in foil immediately after being exposed to light. The seeds were stored on a level surface in a dark cabinet. (Sparace, 2010) After forty eight hours, the seeds were checked for germination. It is crucial to check the seeds under a green light because seeds will germinate if checked under white light. The total number of germinated seeds were counted in each variety. (Sparace,2010) The percent germination for each treatment is calculated based on the results. The data was put into a table for better visibility and organization. Results: Table 1: Seed Exposure to Light
Dark control Time of seed exposure Ongoing exposure: kept in dark. Red light 15 minutes Far red light 15 minutes White light 15 minutes

Table 1 illustrates the light regimens used in the experiment and the amount of time the seeds were exposed to the light.

Table 2: Concept Seed Germination Records

Date 2-3-11 2-8-11 Dark control 0/25 concept 0/25 concept Red light 0/25 concept 0/25 concept Far red light 1/25 concept 2/25 concept White Light 2/25 concept 7/25 concept

Table 2 illustrates the results of the concept seed variety on the dates checked. On February 3, only one seed exposed to far red light had germinated, while two of the seeds exposed to white light had germinated. On February 8, two seeds exposed to far red light had germinated, and seven of the concept seeds exposed to white light had germinated.

Table 3: Two Starr Seed Germination Records

Date 2-3-11 2-8-11 Dark Control 0/25 0/25 Red light 0/25 1/25 Far red light 0/25 0/25 White light 1/25 2/25

Table 3 illustrated the results of the two starr seed variety on the dates checked. On February 3, one of the two starr seeds that had been exposed to white light had germinated. By the second check, on February 8, one two starr seed that was exposed to red light had germinated and two that were exposed to white light had germinated.

Table 3:Percent Germination Overall

Dark Control Concept 0% Red light 0% Far red light 8% White light 28%

Two starr

0%

4%

8%

8%

Table 3 shows the results in Tables 2 and 3 as percentages of seed germination.

Discussion: After the first reading, on February 3, 2011, which is two days after the experiment took place, on February 1, 2011, only the far red and white light treatments had produced any results. 1 seed of the concept variety that was exposed to far red light had germinated. 2 concept variety seeds and 1 two starr variety seed has germinated under white light. These results supported both of the hypotheses. The results after the second reading, on February 8, 2011 were that 9 seeds germinated under white light. 4 seeds were germinated under far red light, 1 under red light, and none in the dark. The seeds that were exposed to far red light did not germinate because the far red light, wavelengths 730 nm, inhibit seed germination. The Pfr in the seeds were converted to Pr, which inhibited the germinating response. The last light that the seed is exposed to determines the response of the germination. (Campbell et al. 2009) The seeds that were exposed to the red light germinated more so because the red light increases germination percentage. The Pr in the seeds were converted back to Pfr, which triggered the plant response to germinate. The wavelength for red light is 660 nm. The plants were only read on two dates because after the 8th of February, the seeds were exposed to white light, which resulted in total germination of all seeds. The results were not 100% accurate. To be more accurate, more seeds could have been tested. Also, stricter light settings should be used to be sure the seeds are not exposed to white light etc. Future experiments could add other factors into the experiment such as humidity control or temperature control. Another option for future experiments is to used more

seed varieties. Literature Cited: Campbell, Neil A. et al. 2009. Biology, 8th Edition. Custom for Tri County Technical College (Boston, MS. Pearson Education.) Pg 836 - 837.

Casal, J. 1993. Novel effects of phytochrome status on reproductive shoot growth in Triticum aestivum L. New Phytologist. Vol. 123: 45 - 51.

Kendrick, R.E. and Spruit, C.J.P. 1973. Phytochrome Properties and the Environment. Plant Physiology. Vol. 52: 327 - 331.

Molecular

Roux, Stanley J. January 1984. Ca2+ and Phytochrome Action in Plants. BioScience. Vol. 34: 25 - 29.

Sparace, K. 2010. Biology 102 Laboratory Manual. (Tri-County Technical College Press, Pendleton, SC.) pg 64 - 68.