Plant Physiology Experiment Using Tomato Plant Lab Report

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PLANT PHYSIOLOGY Enquiry Class project: Measurement of the Light Dependence of Photosynthesis: Light response curves, photochemical efficiency and light compensation points of C3 and C4 Plants Photosynthesis is the most important energy-capturing process in both aquatic and terrestrial ecosystems. As the name suggests, photosynthesis is a process by which light is converted to chemical energy and is used to synthesize compounds within the organism. The green pigment chlorophyll is the central light-absorbing pigment that makes this energy capture possible. Chlorophyll, along with various other accessory pigments, is located in chloroplasts within the cells of the green portions of a plant. The leaves are the main organs of the plant housing chloroplasts and they are very well designed to capture the maximum amount of light, as you will see when you examine their structure in detail. The light energy captured by the chloroplast pigments is converted into chemical energy in the form of ATP and NADPH2. These compounds can then be used to convert CO2 molecules into sugar molecules, which are used to feed the plant and us. In the overall process of photosynthesis CO2 molecules are absorbed by the leaf and O2 molecules are released as a waste product of the process, as seen in the equation below: 6 CO2 + 12 H2O C6H12O6 + 6 O2 + 6 H2O Thus the rate of photosynthesis can be determined by measuring the rate of either O2 production or CO2 uptake. The purpose of this experiment is to demonstrate that leaves produce O2 during photosynthesis and to measure photosynthetic rate by measuring the rate at which O2 is evolved from a leaf at various light intensities or wavelengths. In addition the results will show that the rate of photosynthesis increases with light intensity until a light saturation point is reached. At that point, photosynthetic rate is limited either by the ability of the leaf to convert the light energy it absorbs to chemical energy, or by the supply of some other factor required for photosynthesis, such as CO2. Materials Leaves of a C3 and C 4 species of higher plant Qubit Systems photosynthesis laboratory package with oxygen sensor Plant Physiology Laboratory Exercise: L a b 2 : Mineral Nutrition in Tomato Plants Introduction: At least 17 mineral elements are considered essential for plant growth. Plants acquire the majority of these elements from the soil solution by absorption through the roots. Elements may also enter plants via the stomata in gaseous states. Elements entering plants as gases may not only include carbon as CO2, but also sulfur as gaseous SO2 and nitrogen as gaseous NO2 or NH3. Other elements such as chlorine may also enter the plant via the atmospheric route carried on dust particles or water droplets. In the case of essential micronutrients (those elements required in relatively small amounts; < 0.1 mg/g dry wt tissue), the atmospheric pathway could possibly supply most if not all of a plant's requirements for that element. The required macronutrients (those elements required in relatively large amounts; usually considerably more than 1 mg/g dry wt of tissue), except for carbon and oxygen, typically cannot be supplied in sufficient quantity through the atmosphere to meet a plant's requirements for those elements. Low levels or the absence of a particular essential element will result in specific deficiency symptoms in a given plant species. These deficiency symptoms can be useful for determining what role(s) these elements play in the physiology of the plant. Colors, organ or tissue shapes, or excreted products are examples of symptoms that may be clues about these essential roles. Whether the deficiency symptoms first appear on the youngest or oldest portions of the plant also can provide clues about the mobility and thus the identity of the element. Symptoms that first appear on older portions of the plant such as fully expanded leaves are indicative of a mobile element. Plants can mobilize some elements that are in short supply and transport them from older parts to actively growing (younger) regions of the plant. Deficiency symptoms that first appear in younger regions of the plant are indicative of non-mobile elements. A plant cannot remobilize these elements when they are in short supply and therefore requires a constant supply of these elements to actively growing regions of the plant. Deficiency symptoms can best be studied using a water culture system (hydroponics), where the presence or absence of essential mineral elements can be controlled precisely. In addition, hydroponics allows the experimenter to observe deficiency symptoms that occur on the roots, which cannot be observed easily in soil-grown plants. This experiment is designed to demonstrate the deficiency symptoms caused by the deletion of specific macro- or micro-nutrients from the hydroponic solution. Students will also be given a chance to exercise their diagnostic skills by identifying nutrient deficiencies in plants given to them as unknowns. Materials You will use 4-6 week old tomato plants (Lycopersicon esculentum) for all tests. TOMATO (SOLANUM LYCOPERSICUM): A MODEL FRUIT-BEARING CROP INTRODUCTION Tomato (Solanum lycopersicum) is one of the most important vegetable plants in the world. It originated in western South America, and domestication is thought to have occurred in Central America. Because of its importance as food, tomato has been bred to improve productivity, fruit quality, and resistance to biotic and abiotic stresses. Tomato has been widely used not only as food, but also as research material. The tomato plant has many interesting features such as fleshy fruit, a sympodial shoot, and compound leaves, which other model plants (e.g., rice and Arabidopsis) do not have. Most of these traits are agronomically important and cannot be studied using other model plant systems. There are 13 recognized wild tomato species that display a great variety of phenotypes and can be crossed with the cultivated tomato. These wild tomatoes are important for breeding, as sources of desirable traits, and for evolutionary studies. Current progress on the tomato genome sequencing project has generated useful information to help in the study of tomato. In addition, the tomato belongs to the extremely large family Solanaceae and is closely related to many commercially important plants such as potato, eggplant, peppers, tobacco, and petunias. Knowledge obtained from studies conducted on tomato can be easily applied to these plants, which makes tomato important research material. Because of these facts, tomato serves as a model organism for the family Solanaceae and, specifically, for fleshy-fruited plants. PLANTING TOMATOES: Tomato seed germinates fairly quickly, within 5-10 days. The plants also develop fast, so seeds can be started 6-8 weeks before you intend to transplant outdoors. Keep in mind that the warmer your seedlings are kept and the more light they are given, the faster they will grow. You don't want them to get so large indoors that you have to keep moving them to larger p Preparing the Pots for Planting Dampen the Potting Soil: It's easier to dampen the potting mix before you put it the containers. Add some water and work it through. Keep adding water until the mix stays compressed in your hand, but is not dripping wet. It should break apart when you poke it with your finger. Fill the Pots: Fill your containers and gently firm the soil so that it is about an inch from the top. Use your label to make a 1/4 inch furrow in the planting mix. Sprinkle 2–3 seeds into the furrow and cover them with a sprinkling of potting mix. Gently firm the mix down, so the seeds make good contact with the soil. You can spray the surface with water, if it doesn’t feel moist enough. Be Patient: At this point, you should place your containers somewhere warm and check them daily to make sure the soil is moist - not wet - and watch for germination. Putting containers inside a plastic bag can create a mini greenhouse. Remove the bag when the seedlings emerge. Caring for seedling Once your tomato seedling has true leaves, it's time to start feeding it. Any good liquid fertilizer can be used once a week. Dilute it to half the label recommended dose. Light is critical now. Keep your tomato seedlings close to your grow lights and rotate the plants if they seem to be growing or leaning I one direction. Tomato stems grow sturdier if they are tossed about by the wind. You can simulate this indoors by putting a fan on your plants for an hour a day or simply running your hand across them each time you pass them. When the tomato seedlings are 2--3 inches tall and have a couple of sets of true leaves, it's time to pot them up or move them into larger pots of their own. Potting Up Tomato Seedlings You should transplant individual tomato seedlings into bigger pots, to continue growing stronger indoors. Three to four inch containers are good for seedlings this size. You may need to move them to larger pots later, if you can't move them outdoors. Fill the new pots with moist potting mix, just as you did when you started the seeds. If more than 1 seed germinated in your containers, you will need to thin them. Either gently jiggle entangled roots apart or simply snips off unwanted seedlings at soil level. This ensures that you won't damage the seedling you want to keep. Plant the tomato seedling in the new pot, a little deeper than it was in its original container. If it is tall and leggy, you can plant it right up to its top most leaves. Firm the soil gently around the seedling. Materials: - Walz Imaging-PAM M-Series MINI Chlorophyll Fluorescence System Background: Chlorophyll Fluorescence is a method used to determine photosynthetic activity. Chlorophyll Fluorescence utilizes many parameters to obtain a conclusion regarding the rate of photosynthetic activity in a plant. A few observable parameters that are useful for analysis include, but are not limited to Electron Transport Rate, maximal Fluorescence yield, and Effective Photosystem II yield. The Walz Imaging PAM MINI provides all this information in an effective way. Light Curve Analysis Light Curve Analysis is done by exposing a plant leaf to increasing light intensities over a short interval of time (20 seconds) and instant light-response curves are recorded. This method can be criticized because photosynthesis will most likely not be in steady state, but instant light curves can nevertheless provide reliable information about pivotal points of photosynthesis and can provide essential details regarding the intrinsic properties of the plant leaves being measured. The Light curve responses that we obtained were performed using the Walz Imaging-PAM MSeries MINI system. The leaf was exposed to increasing intensities of actinic light over a 2 minute interval in 6 steps, where each step followed each other after 20 seconds. The range of actinic light was measured based on Photosynthetically Active Radiation (PAR) values that ranged from 0µmol m⁻² s⁻¹ to 231µmol m⁻² s⁻¹. The Imaging system provides clear information regarding the fluorescence yield parameters and Electron Transport Rate under the different lighting conditions. Electron Transport Rate There is a positive linear relationship between the Electron Transport Rate (ETR) in Photosystem II and the net photosynthetic rate of a plant leaf. The parameter ETR has been shown to correlate well with linear electron flow calculated on the basis of O2 evolution rates (Flexas et al. 1999). As we are well aware of the important role that the electron transport chain plays in the process of photosynthesis in plants, the knowledge of the ETR under different lighting conditions can help us understand the photosynthetic capacity resident in a plant leaf. Maximal Fluorescence Yield Chlorophyll fluorescence can be defined as the red to far-red light emitted by photosynthetic tissues/organisms when illuminated by light of approximately 400–700 nm (photosynthetically active radiation or PAR) (Kalaji.et al. 2016). Actinic Light at different intensities is flashed on a leaf and the chlorophyll in the leaf supplies a light emission in response. Analysis of the maximum amount emitted under the given conditions is useful in providing knowledge of the growth rate capacity resident in the leaves at the time that the light curve analysis was administered. Effective Photosystem II Yield Photosystem II (PSII) is a specialized protein complex that uses light energy to drive the transfer of electrons from water to plastoquinone, resulting in the production of oxygen and the release of reduced plastoquinone into the photosynthetic membrane. The key components of the PSII complex include a peripheral antenna system that employs chlorophyll and other pigment molecules to absorb light, a reaction centre at the core of the complex that is the site of the initial electron transfer reactions, an Mn4OxCa cluster that catalyses water oxidation and a binding pocket for the reduction of plastoquinone. PSII is the sole source of oxygen production in all oxygenic photosynthetic organisms, which include plants, algae and cyanobacteria. In these organisms, PSII operates in series with other protein complexes, including the PSI reaction centre, to produce the reduced form of nicotenamide–adenine dinucleotide phosphate (NADPH) and adenosine triphosphate (ATP), which is used in the Calvin–Benson cycle to produce carbohydrates from carbon dioxide. The sensitivity of PSII activity to abiotic and biotic factors has made this parameter an observable and essential one for understanding the photosynthetic mechanisms in a plant. The yield recorded is the operating efficiency of PSII photochemistry, Fq′/Fm′ (Genty et al., 1989), is calculated from (Fm′–F′)/Fm′ (and does not require a darkadapted measurement). This parameter is often termed ϕPSII or ΔF/Fm′ (Maxwell and Johnson, 2000; Baker, 2008). It gives the proportion of absorbed light that is actually used in PSII photochemistry (Genty et al., 1992) and can therefore be used to estimate the rate of electron transport through PSII with knowledge of light absorptance by the leaf and photosystems. Procedure: 1. 2. 3. 4. Select a leaf from around the middle of the stem Attach to the Imaging-PAM system with the leaf holder. Utilizing the computer software, run Light Curve Analysis. Repeat for the remaining plant species/treatments. Data: PAR vs ETR PAR: MJ 0.1 MJ 0.25 0 2 32 80 137 231 MJ 0.5 0.0 0.4 2.0 0.0 0.0 0.0 0.5 0.5 5.2 12.5 5.6 0.0 MJ 0.75 Unknown 0.1 Unknown 0.25 Unknown 0.5 Unknown 0.75 SA 0.1 SA 0.25 SA 0.5 SA 0.75 References: 1. Flexas J, Escalona JM, Medrano H. Water stress induces different levels of photosynthesis and electron transport rate regulation in grapevines. Plant Cell Environ. 1999;22:39–48. doi: 10.1046/j.1365-3040.1999.00371.x. 2. Kalaji HM, Schansker G, Brestic M, et al. Frequently asked questions about chlorophyll fluorescence, the sequel [published correction appears in Photosynth Res. 2017 Apr;132(1):67-68]. Photosynth Res. 2016;132(1):13–66. doi:10.1007/s11120-016-0318-y 3. Genty B Goulas Y Dimon B Peltier G Briantais JM Moya I . 1992. Modulation of efficiency of primary conversion in leaves, mechanisms involved at PS2. In: Murata N, ed. Research in photosynthesis, Vol. IV: Proceedings of IXth International Congress on Photosynthesis . Nagoya, JapanAugust 30–September 4, 603–610. 4. Maxwell K Johnson GN . 2000. Chlorophyll fluorescence—a practical guide. Journal of Experimental Botany 51, 659–668. 5. Baker NR . 2008. Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annual Review of Plant Biology 59, 89–113. 6. Genty B Goulas Y Dimon B Peltier G Briantais JM Moya I . 1992. Modulation of efficiency of primary conversion in leaves, mechanisms involved at PS2. In: Murata N, ed. Research in photosynthesis, Vol. IV: Proceedings of IXth International Congress on Photosynthesis . Nagoya, JapanAugust 30–September 4, 603–610. Soaking Before the tomato seeds were planted in soil they first soaked in their respective elicitor for 24 hours. A total of 192 tomato Meyer seeds were soaked in petri dishes, 16 seeds soaked in MJ 0.1, 16 seeds soaked in MJ 0.25, 16 seeds soaked in MJ 0.50 and 16 seeds soaked MJ 0.75. This same process was done for Salicylic acid and the unknown elicitor with the same varying concentration of 0.1, 0.25, 0.50 and 0.75. Planting February 19, 2019 After the seeds were soaked for 24 hours, twelve 12inch planting pot were then labeled as MJ 0.1, MJ 0.25, MJ 0.50, MJ 0.75, SA 0.1, SA 0.25, SA 0.50, SA 0.75, Unk 0.1, Unk 0.25, Unk 0.50 and Ukn 0.75; the numbers correspond to the concentration. The 12in pots were then filled with planting soil then thoroughly soaked with water for 10-15 seconds. Four holes were then made in the soil of pot along the and with equal space; roughly 1/4-1/2 inch deep. The soaked seeds were place onto a paper towel to soak the excess liquid, then planted into their respective labeled pots and left in the Bowie State University greenhouse with controlled environment. Maintenance 8 weeks Two weeks after planting treatment began, 1000L of each elicitor was poured into their respective pot and respective concentration. A 500ml graduated cylinder was used for treatment and were thoroughly rinsed out after each treatment; this was done twice a week until April 30th, 2019. Photosynthetic Activity Analysis After the 10-week experiment of varying elicitor treatments the plant were then analyzed for photosynthetic activity with the Walz Imaging-Pam M-Series Mini Chlorophyll Fluorescence System. This system will be used to provide quantitative analysis for the Electron Transport Rate, maximum Fluorescent yield and effective Photosystem II yield. A leaf was selected (remained attached) from all twelve of the treated plants, then clamped to the Imaging-PAM system, the leaf was then scanned with Light Curve Analysis for 2 minutes and varying parameter. All data collect was recorded and is presented in the tables and figures below. Result: Table 1 Photosynthetically Active Radiation vs Electron Transport Rates PAR: 0 2 32 80 MJ [0.10] 0.0 0.6 4.4 5.1 MJ [0.25] 0.0 0.6 1.0 0.0 MJ [0.50] 0.0 0.7 3.9 0.0 MJ [0.75] 0.0 0.6 2.2 0.0 Unknown 0.5 0.5 3.1 0.0 [0.10] Unknown 0.0 0.6 3.6 0.0 [0.25] Unknown 0.0 0.7 4.5 2.1 [0.50] Unknown 0.3 0.4 1.4 0.0 [0.75] SA [0.10] 0.4 0.4 1.2 0.0 SA [0.25] 0.5 0.5 3.4 0.0 137 0.0 0.0 0.0 0.0 0.0 231 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 SA [0.50] SA [0.75] 0.5 0.3 0.5 0.4 2.9 1.4 0.0 0.0 Table 2 Photosynthetically Active Radiation vs Photosystem II PAR: 0 2 32 80 MJ [0.10] 0.472 0.493 0.318 0.149 MJ [0.25] 0.537 0.483 0.076 0.0 MJ [0.50] 0.543 0.536 0.278 0.0 MJ [0.75] 0.486 0.470 0.162 0.0 Unknown 0.383 0.378 0.221 0.0 [0.10] Unknown 0.494 0.485 0.257 0.0 [0.25] Unknown 0.599 0.548 0.322 0.061 [0.50] Unknown 0.450 0.475 0.096 0.0 [0.75] SA [0.10] 0.305 0.305 0.087 0.0 SA [0.25] 0.426 0.428 0.244 0.0 SA [0.50] 0.392 0.379 0.211 0.0 SA [0.75] 0.258 0.278 0.103 0.0 Table 3 Photosynthetically Active Radiation vs Fluorescent Yield PAR: 0 2 32 80 MJ [0.10] 0.1755 0.1711 0.1265 0.0824 MJ [0.25] 0.2828 0.2618 0.1554 0.1353 MJ [0.50] 0.4824 0.4750 0.3172 0.2446 MJ [0.75] 0.6142 0.6029 0.3510 0.2804 Unknown 0.6574 0.6525 0.4892 0.4343 [0.10] Unknown 0.4544 0.4475 0.3765 0.2980 [0.25] Unknown 0.4515 0.4426 0.2907 0.2255 [0.50] Unknown 0.5618 0.5510 0.4456 0.3520 [0.75] SA [0.10] 0.6574 0.6471 0.4490 0.4034 SA [0.25] 0.5426 0.5377 0.4475 0.3917 SA [0.50] 0.5176 0.5265 0.4407 0.3299 SA [0.75] 0.4912 0.4887 0.4377 0.3314 0.0 0.0 0.0 0.0 137 0.0 0.0 0.0 0.0 0.0 231 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 137 0.0721 0.1265 0.2206 0.2569 0.3858 231 0.0691 0.1191 0.1931 0.2431 0.3211 0.2680 0.2328 0.2064 0.1936 0.3069 0.2701 0.3539 0.3515 0.2843 0.3059 0.3142 0.3206 0.2613 0.3279 Table 4. Final Height of Tomato Plants after 8 weeks of elicitor treatment Elicitor Final Height (cm) MJ [0.10] MJ [0.25] MJ [0.50] MJ [0.75] Unknown [0.10] Unknown [0.25] Unknown [0.50] Unknown [0.75] SA [0.10] SA [0.25] SA [0.50] SA [0.75] 55.88 66.04 94.06 91.77 91.44 101.6 81.28 76.2 83.82 93.98 92.71 88.9 Figure 1. Graph of Maximum ETR in varying concentration of MJ, SA and Unknown Photosynthetically Active Radiation vs Maxima Electron Transport Rates 6 5 4 [0.10] 3 [0.25] [0.50] 2 [0.75] 1 0 MJ SA Unknown Fig 1. In this graph the maximum ETR were plotted for each concentration for the three elicitors during the imaging of PAM in the two-minute period. Figure 2. Graph of Maximum Photosystem II in varying concentration of MJ, SA and Unknown Photosynthetically Active Radiation vs Photosystem II 0.7 0.6 0.5 [0.10] 0.4 [0.25] 0.3 [0.50] 0.2 [0.75] 0.1 0 MJ SA Unknown Fig 2. In this graph the maximum Photosystem II were plotted for each concentration for the three elicitors during the imaging of PAM in the two-minute period. Figure 3. Graph of Maximum Fluorescent Yields in varying concentration of MJ, SA and Unknown Photosynthetically Active Radiation vs Fluorescent Yield 0.7 0.6 0.5 0.1 0.4 0.25 0.3 0.5 0.2 0.75 0.1 0 MJ SA Unknown Fig 3. In this graph the maximum Fluorescent Yields were plotted for each concentration for the three elicitors during the imaging of PAM in the two-minute period. Figure 4. Final Growth Measurement of Tomato Plants at 10 Weeks Final Growth Measurements of Varying Ellicitor Treatments 120 Length in Centimeters 100 80 0.1 0.25 60 0.5 0.75 40 20 0 MJ SA Unknown Fig 4. These are the measurement of the tallest plants in each of the experimental pots which were taken April 30th, 2019
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Running Head: PLANT PHYSIOLOGY EXPERIMENT

Lab Report: Plant Physiology Experiment Using Tomato Plant

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PLANT PHYSIOLOGY

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Background
The tomato stands chief among the few vegetables plants which are developed as
greenhouse crops. In its way of life under glass, particularly in the northern expresses, the
questing of adequate light for its best advancement and most elevated efficiency emerges and
winds up intense. As explained by Foca, Oancea, and Condurache, (2004), the light of the
common day, amid the winters months, give off an impression of being deficient as for its span
and furthermore to its standard force.
Certainly photosynthesis is the most significant energy catching procedure in both
terrestrial and aquatic environments which conditions the production and behavior of plants. By
definition, photosynthesis is a procedure by which light is changed over to chemical energy
which is used in synthesizing compounds within the organism. The light-absorbing pigment
which enables the capturing of energy is referred as chlorophyll. Alongside other adornment
pigments, Chlorophyll is situated in chloroplasts inside the cells of the green parts of a plant.
The leaves are the primary organs of the plant which carries the chloroplasts and they are
very much intended to catch the greatest amount of light. The light energy caught by the
chloroplast shades is changed over into compound energy as NADPH2 and ATP (Fan et al.,
2013). These mixes would then be able to be utilized to change over CO2 particles into
molecules of sugar, which are used in feeding us and the plant. In the general procedure of
photosynthesis CO2 particles are consumed by the leaf and O2 atoms are discharged as a waste
result of the procedure, as found in the underneath equation:
6 CO2 + 12 H2O

C6H12O6 + 6 O2 + 6 H2O

In determining the photosynthetic activity, Chlorophyll Fluorescence is used. This is
technique uses numerous parameters to get a conclusion with respect to the rate of
photosynthetic action in a plant. A couple of discernible parameters that are helpful for
investigation incorporate...


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