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Germination Experiment

• Hypothesis: I will place seeds in regular potting soil. I am testing the presence of a light source (growth lights, natural light coming through a window, and absence of light in a dark cupboard). There will be no difference in the germination rate of each pot of seeds with or without a light source. My germination will be measured as soon as the cotyledons are present. Growth will be measured immediately after the cotyledons are present. Seeds placed under the growth light will germinate much faster than those placed in the window sill and the dark cupboard.

Experimental Design
• Unknown number of seeds per pot due to size. • Regular Potting Soil used. • Pot #1 placed under growth light. • Pot #2 placed in window sill. • Pot #3 placed in dark cupboard. • Each will be watered as needed.

Germination Results
• See separate Germination Log (Pages 1-5). • Independent Variable: Light Sources used: growth light, window sill, and dark cupboard (absence of light). • Dependent Variable: Germination Rate • The seeds placed in the window sill germinated much faster than those placed under the growth light. No germination occurred with the seeds placed in the absence of light. • Conclusion: My experiment shows tobacco seeds germinate faster in natural light than under a growth light or in darkness.

• Hypothesis: Now that I have moved pot #3 from the dark cupboard to under the growth light, I will test to see if the seeds in pot #3 will germinate now with a presence of light. There will be no change in the germination of pot #3 from the dark cupboard to the growth light. Seeds placed under the growth light will not germinate due to the fact that they may have rotted in the wet soil when a source of light was not present in the dark cupboard. I am also looking at growth rate between pot #1 still under the growth light and pot #2 in the window sill. Both have germinated. There will be no difference in the growth of the two plants. I predict that there will be considerably more growth of the plant in pot #2 in the window sill as it was the first plant to germinate.

Second Experiment Results
• See separate germination and growth log (Pages 5-8) • Independent Variable: Light Sources used: growth light (pot #3 and #1), and window sill • Dependent Variable: Germination/Growth of previously germinated plants • The seeds previously placed in the dark cupboard germinated within 5 days, the fastest germination of all pots in the presence of light and pot #2 placed in the window had considerably more growth than pot #1 under the growth light. True leaves were present 28-30 days after germination in all pots. • Conclusion: This experiment shows light is required for tobacco seeds to germinate and that previously germinated seeds grow better in natural light compared to under a growth light.

Experimental Design
• No new seeds were planted in this experiment. • The soil content was not changed. Still using regular potting soil. • Water as needed.

Pot #1

Pot #2

Pot #3

Identification DNA Extraction and PCR
• Extraction
– Chloroplast, Nucleus, and Mitochondrial DNA were extracted – Tissue was used from several leaves – The plant was alive at the time of DNA extraction

• PCR
– Primers used:
• Forward: B49317 • Reverse: A49855

– Chloroplast DNA was amplified

Identification Gel Electrophoresis

Pos 1

2

3 Neg

Pos 5

6

67 Neg JHN

7

8

10

Neg

• Lane 7 held my sample • The lane right after lane 10 held my tables negative control • I was unsuccessful in amplifying
– May be due to the fact that tobacco leaves are very hairy (many trichomes) and therefore DNA was not successfully extracted.

Identification Results
• DNA sequence used in blasting:
tggctcacaa aattgaattt agagaaggcc ctgctttgcc cgttcttgat caactggttg aagatgtgag tttactccct taccaaatct atcttataca tttgctttta attatctctt tattactatt ttttattatt gataatgaga atccgatgaa gcttatttag tttagggatt 1

• Taxonomic Lineage:
Kingdom Plantae – Plants Subkingdom Tracheobionta – Vascular plants Superdivision Spermatophyta – Seed plants Division Magnoliophyta – Flowering plants Class Magnoliopsida – Dicotyledons Subclass Asteridae Order Solanales Family Solanaceae – Potato family Genus Nicotiana L. – tobacco Species Nicotiana tabacum L. – cultivated tobacco2

• Latin Name: Nicotiana tabacum • Common Name: Tobacco

http://plants.usda.gov/java/profi le?symbol=NITA&photoID=nita_ 001_avp.jpg

• Historic use

Ethnobotany

– Tobacco first growing in the Americas in 6,000 B.C. – It was first used for medicinal and religious practices by American Indians. – Used to dress wounds, chewing tobacco was thought to cure a toothache. – Christopher Columbus given tobacco as a gift by American Indians in 1492, tobacco then taken to Europe. – Alkaloid nicotine isolated in 1820’s5. – The leaves are dried and used to make cigarettes, cigars, snuff and chewing tobacco4. – Over 60% of total nicotine is found in the leaves (although, nicotine is found in all parts of the plant)3.
• This would explain why the leaves are the only part of the plant which are dried.

• Current Usage: You got a light? SMOKING!
• Alkaloid Nicotine (this is very addictive)
– Synthesized in the roots and transported to the leaves, where it is stored in vacuoles6.

– Affects the nervous system and brain receptors. • Increase in blood pressure/heart rate3. • Some of the more than 7,000 Chemicals in tobacco smoke: – Cyanide – Benzene – Formaldehyde – Methanol (wood alcohol) – Acetylene (the fuel used in welding torches) – Ammonia7

http://www.clker.com/clip art-2767.html

• Smoking causes CANCER?!
– Smoking accounts for at least 30% of all cancer deaths in the United States and 80% of lung cancer deaths. Smoking also causes cancers of the nasopharynx (upper throat), nasal cavity, paranasal sinuses, lip, larynx (voice box), mouth, pharynx (throat), esophagus (swallowing tube), and bladder. It also has been linked to the development of cancers of the pancreas, cervix, ovary, colorectum, kidney, stomach, and some types of leukemia3.

• Tobacco was first smoked by Native American Indians for religious purposes5. • As of 2011, Nearly 1 in 5 Americans were smoking tobacco and remains the leading cause of preventable death in the United States8. • In Michigan… – As of May 1, 2010 tobacco use was outlawed in public places (restaurants, bars, hotels, etc.)9. – No one under the age of 18 is allowed to purchase, use, or have possession of tobacco or tobacco products.

• At Aquinas…
– Smoke free campus except for designated areas.

• Tobacco is a major U.S. industry. In 1998, consumers spent an estimated $59.3 billion on tobacco products, chiefly on cigarettes ($55.7 billion)10.

• I have never used a tobacco product, but know people who have. Having an addiction to nicotine seems annoying. People are always having to leave a conversation or activity to have a cigarette. SUCH A WASTE OF YOUR LIFE!
http://www.istockp hoto.com/stockphoto-50907cigaretteburning.php

Conclusion
• Tobacco is a very slow germinating and growing plant. My experiment showed tobacco seeds will not germinate in the absence of light. Seeds are very small, frequent watering is essential. • Possible sources of errors:
– Planting depth (no direction on how to plant seeds). – One plant was drowned when lab overheated causing death. – Plants may have been shaded by towering neighboring plants in competition for light. – Watering as needed rather than a certain amount and schedule.

Seed Facts: -Seeds develop in pods -Pods are dried and then crushed to remove the seeds. -One acre of tobacco can yield 300 pounds of seeds!11

11: http://science-in-farming.library4farming.org/Seeds_1/PRODUCTION-OFSEEDS/Tobacco-Seed.html

http://takhs000.blogspot.com/2011/08/tobacco.html

http://www.icr.org/article/4182/

Does anyone have any BURNING questions?