Part VI


Applications of Biotechnology:

Plant Biotechnology



REFERENCES


Biotechnology in Plants


Below is a list of references you may find helpful in understanding the topic. If you need a copy of the item, call your local library or state university library. Michigan State University telephone number is (517) 353-8700. They can arrange for Interlibrary Loan if the item is not in their collection.


Beardsley, Timothy M. Green thumbs; doing agricultural genetics in the marketplace.             Scientific American, April 1990 v262 n4 p24(1). IAC Coll. 54B0505.

 

Benbrook, C.M. & Moses, P.B. (1986, November/December). Engineering crops to resist herbicides. Technology Review, pp.55+.


Biotechnology could begin green revolution in the fields. (1988, June 14). Durham Sun, n.p.


Bridgen, Mark (1986). Do it yourself cloning. Greenhouse Grower, May 1986:43-47.

 

Carey, J. (1986, November/December). Brave new world of super-plants. International Wildlife, pp.16-18.

 

Chilton, M.D. (1983). A vector for introducing new genes into plants. Scientific American, 48(6), 50-59.

 

Collins,G.B. and Genovesi, A.D.(1982). Another culture and its application to crop improvements. Application of Plant Cell Tissue Culture to Agriculture and Industry. D.T. Tomes et al, eds. Plant Cell Culture Centre, University of Guelph. Guelph, Ontario, Canada.

 

Cowen, R.C. New wave crops headed for the farms. (1988, June 1). Christian Science Monitor, pp. 16-17

 

Crawford, Mark. Biotech companies lobby for federal regulation. (marketing and testing agricultural biotechnology products). Science, May 4, 1990 v248 n4955 p546(2).

 

Dixon, R.A., ed. (1985). Plant Cell a Practical Approach. IRL Press, Oxford- Washington, D.C.

 

Dodds, John H. and Roberts, Korin W. Experiments in Plant Tissue Culture, 2nd edition. London: Cambridge University Press, 1985.


Gene may protect beans from weevils. (1988, April 19). New York Times, p.28.


Goodwin, P.B. And Adisarwanto, T. (1980) Propagation of potato by shoot tip culture in petri dishes. Potato Res. 23:445-448.

 

Gould, F. Evolutionary biology and genetically engineered crops. (1988, January). BioScience, 38(1), n.p.


Griffs, John L. Jr., Hennen, Gary and Oglesby, Raymond P. (1980). Establishing            tissue-cultured plants in soil. Proc. Int. Prop. Soc, 33: 618-622.

 

Kyte, Lydiane. Plants From Test Tubes, An Introduction to Micropropagation. Portland, OR: Timber Press, 1983.

 

McClean, Phillip. "Plant Biotechnology," prepared for Encyclopedia of Food Science and Technology, 1991.


McCullouch, Steven M. and Briggs, Bruce A. (1982). Preparation of plants for              micropropagation. Proc. Int. Plant Prop. Soc. 32:297-303.

 

Millstein, Marc. NTGargiulo sees future ripe for green tomatoes. (tomato wholesaler and distributor). Supermarket News, August 6, 1990 v40 n32 p40(1).

 

Murashige, T. (1974). Plant propagation through tissue cultures. Ann. Rev. Plant Physiol. 25: 135-166.

 

Plant biotechnology to play key role in agriculture. (1986, May 26). European Chemical News, 46(1), 15.

 

Sommer, Harry E., and Caldas, Linda S., (1981). In vitro methods applied to forest trees. Plant tissue culture, methods and applications in agriculture. Trevor A. Thorpe, ed., Academic Press.

 

Stamets, Paul and Chilton, J.S. (1983). The mushroom cultivator, a practical guide to growing mushrooms at home. Agarikon Press, Olympia, WA.

 

Tomes, D.T., Ellis, B.E., Harney, P.M., Kasha, K.J., and Peterson, R.L., editors. (1982). Application of Plant Cell and Tissue Culture to Agriculture and Industry. Plant Cell Culture Centre: University of Guelph, Guelph, Ontario, Canada.

 

Vincent, Gary. The fruits of summer testing; 1988 biotech field trails bring borer-resistant corn one step closer. Successful Farming, February 15, 1989 v87 n3 p34B(2). IAC Coll. 43T0985 and 48L2641.

 

Wetherrell, D.F. (1982) Introduction to in vitro propagation. Avery Pub. Group Inc., Wayne, N.J.


Wilkins, Malcom B. (1984). Advanced Plant Physiology: Pitman Publishing.

 

Wisconsin Fast Plants Manual. University of Wisconsin, Madison, WI. Distributed by Carolina Biological Supply, 1989.

 

Zimmerman, R.H., Griesbach, R.J., Hammerschlag, F.A. and Lawson, R.H. editors,(1986). "Tissue Cultures as a Plant Production System for Horticultural Crops." USDA. Martinus Nijhoff Pub.


Biotechnology Applications


Below is a list of references you may find helpful in understanding the topic. If you need a copy of the item, call your local library or state university library. Michigan State University telephone number is (517) 353-8700. They can arrange for Interlibrary Loan if the item is not in their collection.

 

Anderson, Walter Truett. Food without farms: the biotech revolution in agriculture. (artificial food production). The Futurist, Jan-Feb 1990 v24 n1 p16(6). IAC Coll. 49P2231 and 53D2703.

 

Brosten, Dennis and Simmonds, Brenda. The evolution of biotech revolution. Agricultural Age, Oct 1989 v33 n9 p6(4).

 

Clary, Mike. Using biotechnology to create tastier, longer-lasting veggie. (cover story - brand name produce: FreshWorld Inc. to introduce designer carrots and celery sticks). Adweek's Marketing Week, Oct 30 1989 v30 n44 p28(1).

 

Darrow, Edward E., editor. The Science Workbook. Columbus, OH: The Ohio State University Press, 1985.


Gasser, Charles S. and Fraley, Robert T. "Genetically Engineered Plants For Crop      Improvement." Science Vol. 244, June 16, 1989.

 

Gibson, W. David. Biotech companies eye future harvests. (Farm Chemicals '90 supplement). Chemical Marketing Reporter, May 21,1990 v237 n21 pSR20(2).

 

Harlander, Susan K. and Labuza, Theodore P. Biotechnology in Food Processing. Park Ridge, NJ: Noyes Publications, 1986.


Of the Earth: Agriculture and the New Biology. St. Louis: Monsanto Company, 1987.

 

Olson, Steve: Biotechnology: An Industry Comes of Age. Washington, DC: National Academy Press, 1986.

 

Rauber, Paul. Better nature through chemistry: biotech's breakthroughs: hardball tomatoes, blue roses and potatoes poisons can't kill. Is this the best we can do? Sierra. July -August 1991 v76 n4 p.32(3). IAC Coll. 60J0773


Rensberger, B. Plant gene fights herbicide. (1988, October 31). The Washington Post, p. A3.

 

Schmitz, Tom. New bacterium takes the bite out of frost. (FrostBan , Frost Technology Corp.). Journal of Commerce and Commercial, Jan 17, 1991 v387 n27397 p7A(1).


What is Biotechnology? Washington, DC: Industrial Biotechnology Assoc., 1989.

 

Wheeler, D. Agricultural scientists step up efforts to find biological weapons against crop pests grown resistant to chemical insecticides. (1988, August). Chronicle of Higher Education, 34(47), pp. A4, A5, A10.



TITLE OF BIOTECHNOLOGY ACTIVITY:

 

A.    Jumping Genes in Corn

(Adapted from "A Sourcebook of Biotechnology Activities", North Carolina Biotechnology Center.)

 

OBJECTIVES:

 

Upon completion of this activity, students should be able to:

 

                        1.         Use Mendelian principles to explain certain aspects of the determination of kernel color and morphology in corn (Activity I).

 

                        2.         Use non-Mendelian principles to explain further the determination of kernel color (Activity II).

 

                        3.         Define transposon and describe how transposons (jumping genes) can affect kernel color and morphology (Activity III).

 

                        4.         Explain how transposons are used to insert foreign genetic material into plants (Activity III).

 

                        5.         Appreciate that there are various levels at which a scientific problem can be understood.

 

                        6.         Appreciate that the hereditary material of a cell is a dynamic, constantly changing system, of which gene movements and interactions are an important part.

 

                        7.         Recognize and understand the following words:

 

Activity I: embryo, gametes, alleles, Mendelian genetics, pericarp, aleurone layer, endosperm, Punnett square

 

Activity II: epistasis, locus, multiple alleles

 

Activity III: transposon, dissociation, autonomous, translocation

 

MATERIALS:

 

per lab team

▾ Ears of the following commercially available genetic corn (expressed as a ratio of kernels). At least one ear is needed per group:

 

Activity I:

 

▾ 1 yellow starchy: 1 yellow sweet

▾ 3 purple starchy: 1 yellow starchy

▾ 9 purple starchy: 3 purple sweet:

3 yellow starchy: 1 yellow sweet

 

Activity II:

 

▾ 9 purple: 3 red: 4 white

▾ 12 purple: 3 yellow: 1 white

▾ 13 yellow: 3 purple

▾ 2 purple: 1 yellow: 1 white

▾ 9 red: 7 white

▾ 9 purple: 3 red: 3 yellow: 1 white

▾ 3 yellow: 1 purple

▾ 9 yellow: 3 white: 4 purple

 

Activity III:

 

▾ "Indian corn"

 

BACKGROUND:

 

Introduction

 

For the first part of this century, geneticists believed that genes were static with regard to their loci or positions on chromosomes. However, in 1947 Barbara McClintock proposed that genes could indeed move within and between chromosomes. McClintock kept careful records of genetic crosses in corn. From her data, she concluded that there must be genetic elements that could move to and be inserted at the loci for color in corn kernels. These fragments of DNA are now called transposons. Although her work initially did not have much impact on genetic theory, it was later corroborated by findings of similar phenomena in bacteria and mammalian retroviruses. She received the Nobel prize for her work in 1983.

 

In order to get a gene from one kind of cell to be expressed in a different kind of cell, frequently the gene must be inserted in the recipient cell's chromosome. Transposons are a useful tool for genetic engineering in plants. Like certain viruses, they provide the mechanism for inserting foreign DNA into a host cell's chromosome. They have been used in research laboratories in the sometime complex series of steps required to obtain a plant that expresses a gene from another organism.

 

This lesson is composed of three activities, each building on or extending what was learned in the previous one. Activity I involves simple Mendelian inheritance, Activity II deals with the complex genetic mechanisms involving gene interaction mechanisms involving gene interaction and Activity III introduces the concept of how transposons (jumping genes) can affect the color of "Indian" corn kernels.

 

ACTIVITY I:

 

Simple Inheritance

 

Each kernel on an ear of corn is a potential corn plant. It contains an embryo resulting from the union of two gametes, one (the egg) contributed by the female plant part and the other (the sperm) contributed by the male plant part.

 

The structure of a corn kernel is illustrated below. It includes a thin outer layer, the pericarp; a thin underlying aleurone, which is the outermost layer of the endosperm; the remainder the endosperm, which contains the kernel's stored food; and the embryo. Each of the three outer layers may be pigmented or not, with the presence or absence of pigmentation being under the control of several different genes.

 

The chemical form of food stored in the endosperm affects the appearance of the kernel. This is under genetic control, with the Su gene for starchy (smooth and rounded appearance) being dominant to the su gene for sweet (wrinkled and raisin-like).

 

Each of the three outer layers may be pigmented or not. Pigmentation in the aleurone layer is under control of a number of genes. In Activity I, you will be concerned with only one of these genes, the R locus. R is dominant to r and causes a purple or red color; the r allele does not cause any pigmentation of the aleurone.

 

ACTIVITY II:

 

Complex Inheritance

 

Pigmentation in the aleurone layer is actually under the control of at least three different genes, R, Pr and C, each of which may occur in a number of different forms, or alleles. At the R locus, allele R produces a red pigment and/or allows a purple color to be present in the aleurone; R is dominant to r which does not produce a pigment and does not allow the expression of a purple color. Thus, the expression of a second set of alleles at a different locus, the Pr locus, varies with whether the R or r allele is present. Pr produces purple color, while pr contributes no color to the aleurone. Pr is dominant to pr. In order for the Pr gene to be expressed, there must be at least one R allele present. If there are at least one R gene and Pr gene (i.e., a Pr_ R_ genotype) then the kernel can be purple; a genotype of prpr R_ can produce a reed aleurone; prpr rr contributes no color to the aleurone. The relationship between the R and Pr loci is an example of epistasis, a condition that results when genes at one locus affect the expression of genes at another locus.

 

Note that we said the aleurone can be purple. Aleurone color is affected by yet another locus, C, which provides an example of both epistasis and multiple alleles. In order for the aleurone to be pigmented, there must be a least one C allele at the C locus, i.e., CC or Cc. A cc individual will lack aleurone pigmentation. Besides C and c, there is another allele C1; it is dominant to C and prevents any color from being expressed in the aleurone. For instance, a PrPr RR C1C individual would lack aleurone coloration. A locus with more than two alleles is said to be under the control of multiple alleles.

 

Another locus that is involved in determining the color of corn kernels is the Y locus. This is a case of simple dominance, with Y_ producing yellow endosperm and yy producing white endosperm. The color of the endosperm is often not visible because of the overlying purple or red pigments in the aleurone.

 

ACTIVITY III

 

Jumping Genes and Inheritance

 

For the first part of this century geneticists believed that genes were static with regard to their positions on chromosomes. Work first reported by Barbara McClintock in 1947, however, revealed that genes could indeed move within and between chromosomes. McClintock kept careful records of genetic crosses in corn and observed the color changes on individual kernels of ears of corn over numbers of generations. From her data, she concluded that there must be genetic elements that could move to and be inserted at the loci for color in corn kernels. These fragments of DNA are now called transponsons or transposable genetic elements. When she published her unorthodox work in the early 1950's it was discounted as impossible because it did not support the hypothesis under which all other geneticists were operating, that is, that the position of genes was fixed. Her work was later corroborated by findings of similar phenomena in bacteria, yeast, fruit flies and mammalian retroviruses. She received the Nobel prize for her work in 1983.

 

Specifically, McClintock showed that there was a genetic element which could move to and be inserted at the C locus. Insertion of this Ds element (for dissociation, so named because it was frequently a site of chromosome breakage, however; rather this required the presence of another element, called Ac for Activator. If the Ds element (in the presence of the Ac element) moved from the C locus during the development of a kernel, all cells resulting from that cell would be purple. Each cell in which such a translocation took place could eventually result in a purple spot on an otherwise yellow or white kernel of corn.

 

The P locus was another found to affect kernel color. It was found that the Ac element could insert itself at the P locus, disrupting thereby the production of a red-orange pigment in the pericarp. Translocation to and from the locus several times during kernel development resulted in the red-orange swirls characteristic of many kernels on "Indian corn."

 

Subsequent studies in corn have revealed other "families" of transposons analogous to the Ac-Ds family. Each family contains element that, like Ac, can effect their own movement and that of other elements in the family from one site to another. Such elements are called autonomous. In addition, each family generally contains nonautomonous elements that, like Ds, can only move in the presence of an autonomous member of the same family. In many cases nonautonomous elements appear to be derived from autonomous elements be deletion of DNA sequences.

 

PROCEDURE:

 

Teacher Preparation

Order the corn.

 

Teaching Tips

 

These three activities should be taught several days apart rather than on consecutive days to allow students to assimilate the information. This strategy will also provide a unifying theme for other components of a genetics unit:


 

Before Activity I:

 

                                    1.         Review simple dominance, Mendel's laws and Mendelian ratios with the students.

 

                                    2.         Briefly describe the status and basic structure of corn kernels using information from the student pages.

 

                                    3.         Use the table below for the class discussion.


 

R/r

Su/Su

  purple/starchy

R _

Su _

  purple/sweet

R _

susu

  yellow/starchy

rr

Su _

  yellow/sweet

rr

susu


 

Before Activity II:

 

                                    1.         Review the results of Activity I.

 

                                    2.         Describe the more complex mechanisms underlying the inheritance of kernel morphology using information from the student pages.


 

PrPr

R/r

C/CI/c

Y/y

  Purple

Pr _

R _

CC or Cc

_ _

  Red

prpr

R _

CC or Cc

_ _

  Yellow

prpr

rr

_ _

yy

  White

_ _

_ _

CI_or cc

yy

  White

prpr

rr

_ _

yy

 

Before Activity III:

 

                                    1.         Review the results of Activity II.

 

                                    2.         Describe transposons in corn and how they may affect kernel color using information from the student pages.

 

                                    3.         It's recommended that students begin each of these three activities by working in groups, then work individually to ensure that each understands the concepts involved. (The number of ears available may dictate that students work in groups).

 

Teaching Procedure:

 

(See student pages for detailed protocol.)

 

                                    1.         Have students do Background Reading.

 

                                    2.         Have students work through the steps of Activity I and present their conclusions to the class.

 

                                    3.         Have students work through the steps of Activity II and present their conclusions to the class.

 

                                    4.         Have students work through the steps of Activity III and present their conclusions to the class.

 

                                    5.         Have students answer the Questions.

 

Activity I:

 

Simple Inheritance

 

                                    1.         Obtain an ear of corn for your group.

 

                                    2.         Using slides of the ears themselves, point out examples of purple vs. yellow, and starchy vs. sweet kernels.

 

                                    3.         Purple color is caused by pigmented cells in the aleurone. It is under control of a pair of alleles, R which allows the color to be expressed and r which does not allow expression. R is dominant for r. Smooth, round kernels (called "starchy") result from the presence of a dominant gene Su, whereas wrinkled kernels (called "sweet") have the genotype susu.

 

                                    4.         Determine the genotypes of the parents that produced your ear of corn. Hint: Count the number of kernels of each phenotype, then make a Punnett square and "reason backward" to obtain the parents' genotypes. In the interest of time you can count only a few rows rather than the whole ear.

 

                                    5.         Choose a spokesperson (if working in a group) to present your results in a whole class discussion.

 

ACTIVITY II:

 

Complex Inheritance

 

                                    1.         Obtain an ear of corn for your group.

 

                                    2.         Make sure you can distinguish purple, red, yellow and white kernels.

 

                                    3.         Use the information in the Background Reading to determine with your group the possible genotypes of purple, red, yellow and white kernels.

 

                                    4.         Determine the genotypes of the parents that produced your ear of corn. Two hints that may be of use to you are:

 

                        Hint 1:           Look for examples of familiar Mendelian ratios. For instance, an ear that has 9 purple: 3 red: 4 yellow has a non-yellow:yellow ratio of 3:1. Such a ratio could have resulted from crossing two heterozygous parents Rr X Rr. In addition, the ratio among the non-yellow kernels is 3 purple: 1 red, suggesting parental genotypes of Prpr Rr X Prpr Rr. The fact that all the kernels lacking aleurone pigment are yellow suggests that at least one parent was homozygous for the Y gene, i.e. Prpr Rr YY S Prpr Rr _ _ . These parental genotypes account for the 9:3:4 ratio provided at least one parent was homozygous for C and neither had C1, giving one possible answer of Prpr Rr YY CC X Prpr Rr yy CC. What are some other "correct" answers?

                        Hint 2:           As an alternative, it may be helpful to make a Punnett square and "reason backward" to obtain the parents' genotypes.

 

                                    5.         Choose a spokesperson (if working in a group) to present your results in a whole class discussion.

 

ACTIVITY III:

 

Jumping Genes and Inheritance

 

                                    1.         Obtain an ear of "Indian corn" for your group.

 

                                    2.         Ask each group to look for examples of:

●kernels with purple or white spots

                                                          kernels with red-orange swirls resulting from translocations to/from the P locus.

                                                          Other color patterns that might be explained by similar genetic mechanisms.

 

                                    3.         Use the information in the Background Reading to determine how:

                                                          purple or white spots result from translocations to/from the C locus.

                                                          red-orange swirls result from translocations to/from the P locus.

 

                                    4.         Choose a spokesperson (if working in a group) to present your results in a whole class discussion.

 

QUESTION(S) FOR INVESTIGATION:

 

                        1.         Assume a corn kernel had the genotype Prpr Rr YY Cc. What color would its endosperm be? What color would the kernel appear to be? Explain your answers.





 

                        2.         Assume that an ear of corn is produced by the following cross:

 

Prpr RR Yy CC X prpr RR yy CC1




 

                        3.         Assume that the phenotypic ratio on an ear of corn is 9 purple: 7 white. What were the genotypes of the parents?





 

                        4.         How do you think the C1 gene prevents pigment from occurring in the aleurone even when the Pr, R and C genes are present?





 

                        5.         How might transposons be used one day in genetic engineering?




 

ANSWERS TO QUESTION(S) FOR INVESTIGATION

 

Activity II Procedure Answer

 

Some other possibilities include:

 

● Prpr Rr YY CC X Prpr Rr Yy cc

● Prpr Rr yy CC X Prpr Rr YY Cc

● Prpr RR YY Cc X Prpr RR yy Cc

 

Answers

 

                                    1.         Its endosperm would be yellow, but the color would be masked by the purple in the overlying aleurone. The kernel would appear purple.

                                    2.         1 purple: 1 red: 1 yellow: 1 white

                                    3.         PrPr Rr yy Cc X PrPr Rr yy Cc

                                    4.         Answers will vary depending in part on the students' background in genetics. Some might suggest that the CI gene codes for a protein that destroys purple and red pigment in aleurone cells. Others might suggest that the CI gene product somehow prevents the Pr and R gene products from being produced, perhaps by binding to the chromosome and preventing transcription. Still others might suggest that the C gene codes for a product that is necessary for the production of purple and red pigment, and that the CI product prevents the C product from being made. There are also other possibilities.

                                    5.         Genetic engineering usually involves getting a cell to express foreign genetic material. Appropriate expression can depend on inserting the genetic material at a specific chromosome locus. Transponsons provide a means for controlling with precision where it is inserted.

 

RESOURCES:

 

Audio-visuals

 

"The Biological Revolution: 100 years of Science at Cold Spring Harbor" contains a short piece on McClintock's work. It is published by Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, NY 11724. Also available from: Cabisco Biotechnology, Burlington, NC 27215; 800-334-5551 or 900-632-1231 (NC only).

 

Printed Materials

 

Federoff, N.V. (1984). Transposable genetic elements in maize. Scientific American, 250(6), 85-98.

 

Sprague, G. & Dudley, J.W. (1988). Corn and corn improvement (3rd ed.). Madison, WI: American Society of Agronomy.

 

Computer Software

 

Have your students design and write a computer program that simulates the mechanisms of inheritance in corn. Such a program could be modeled after the CATLAB software developed by Judith Kinnear and published by Conduit, The University of Iowa, Oakdale Campus, Iowa City, Iowa 52242.



TITLE OF BIOTECHNOLOGY ACTIVITY:


B.    Hydroponic Greenhouse Systems

(Adapted from "Biotechnology", by Hard, Harris and Kedenburg, NWCCC, St. Martins College, Lacey, WA.)

 

OBJECTIVE(S):

 

Upon completion of this activity, students should be able to:

 

                        1.         Enhance and improve design and problem solving in hydroponics.

 

                        2.         Describe the biological systems and control mechanisms involved in hydroponic gardening.

 

                        3.         Make informed career choices regarding hydroponics.

 

                        4.         Explain the impacts of technological advances in agriculture.

 

                        5.         Introduce, implement and enhance the use of computers and electronic sensors as a control mechanism in gardening.

 

                        6.         Reinforce the mathematical concepts of measurement, percentage calculation, fractions and liquid measurement.

 

                        7.         Strengthen science concepts of characteristics of plants, pH level and testing, measurements of light, moisture and temperature.

 

MATERIALS AND/OR EQUIPMENT:

Many supplies may be found locally in garden and nursery supply stores, and Creative Learning Systems sells hydroponic supplies.

                                  Seeds for the desired crop or crops

                                  Commercial liquid nutrient solutions

                                  Water

                                  Sunlight

                                  Sand, gravel, vermiculite, or styrofoam pellets

                                  2 Liter clear pop bottles (preferably with a removable base section)

                                  A Plastic Dishpan or a deepbowl

                                  One gallon milk jugs to hold nutrient solutions

 

TIME REQUIRED FOR ACTIVITY:

 

Continuous process

 

BACKGROUND MATERIAL:

 

Hydroponic gardening is expected to increase in scope and quantity as we move away from labor intensive farming methods and into a more technologically intensive system of producing agricultural goods. In addition, home of the future are expected to contain small scale hydroponic gardens for the production of fresh vegetables right in the kitchen, and the entrepreneurial possibilities of marketing hydroponic systems directly to restaurants remain unexplored. This activity focuses on basic hydroponic gardening techniques and activities to be completed in the technology lab.

It is recommended that crops such as radishes, which grow quickly and lettuces that require little support above the root be tried, although more enterprising students may wish to grow cucumber or tomatoes.

 

Students will participate in alternative methods of growing food. The hydroponic method will be introduced as one using minimal amounts of both water and nutrients. A prototype of a hydroponic greenhouse will be constructed and used to demonstrate the effectiveness and simplicity of this type of agriculture. Emphasis should be placed on how technology can be adapted to the environment.

 

This use of two liter bottles also reinforces recycling efforts and shows that we often throw away items that may be useful if we change our manner of thinking about their uses.

 

PROCEDURES:

 

Two liter pop bottles offer a cheap and readily available structure to use as a greenhouse. Two liter bottles may be used in three ways.

 

A drain down system may be used in which the roots of the plants are supported by an aggregate of vermiculite, or small pellets of styrofoam. The nutrient is poured through the aggregate, or the roots are set into the water and allowed to hang suspended gently from the aggregate into the solution of nutrients. The aggregate serves the purpose of holding the plant upright. In some cases the aggregate may need to be supported above the liquid by a screen or mesh. The solution is reused and periodically poured through the aggregate to replenish the water supply in the aggregate.

 

In a wick system, the plant is suspended as in the drain down system, and a cotton wick is placed in the solution to carry the liquid to the plant. Cut the bottle in half and using the neck as a container for the aggregate. Support for the roots is easily accomplished. A cotton wicking system is used to deliver necessary water and nutrients to the plants roots. A section of another bottle can also be replaced over the neck, and used as the removable "greenhouse," or use the removable base of a bottle for a container.

 

In a water culture, the plant is held in a cotton or styrofoam disc that fits in the bottle, and the roots allowed to hang into a solution of nutrients. The remaining section of the bottle is used as a container for the nutrient liquid. The nutrient solution will need to be replenished from time to time. The neck of the bottle is not used in this case.

 

In all cases, it is better to use too little liquid fertilizer than too much. A teaspoon of fertilizer to a cup of water is a good ratio.

 

DATA AND SUMMARY ANALYSIS:

 

The advantage of hydroponic systems is that several problems are eliminated.

 

                                    1.         Weeds are not a factor, since the built in controls eliminate them entirely.

                                    2.         The root system is smaller, so more plants may be grown in a given area.

                                    3.         Plants do not need soil, only the nutrients that are dissolved in it.

                                    4.         The nutrients are reused, while in soil gardening these nutrients are lost.

                                    5.         You can grow plants in areas and climates that normally will not support agriculture.

                                    6.         Labor costs are less as hydroponics is not as labor intensive as other forms of agriculture.

                                    7.         Soil fertilizers are made from oil. Everyday, 200,000 barrels of oil are made into fertilizer.

 

Getting started in hydroponics could be an activity in which an entire class can get involved. After sprouting, the activity can be left alone while the hydroponic system is chosen and made ready. Once implanted in a system, watering and feeding can be put on a schedule until the crops mature, and are ready for harvest. If the entire project can be put aside and out of the way, the teacher can move the class into other activities. Hoisting the entire classes small greenhouse up to a skylight gets it completely out of the way. Incorporate design activities into the unit for a storage system for the bottles and plants and let the students themselves solve the problem of safe storage.

 

IDEA(S) FOR ADDITIONAL ACTIVITIES:

 

                        1.         Hydroponic growing of plants using pop bottles. (National Geographic World Magazine, National Geographic Society, July 1968) (A well illustrated, simple explanation of a hydroponic system.)

                        2.         Use of a greenhouse on a larger, more permanent scale and scope.

                        3.         Computerized electronic controls of sensors to control lighting, nutrients, pumps, water.

                        4.         Experimentation with various crops.

                        5.         Experimentation with various nutrient solutions.

                        6.         Future impacts on society. Widespread hydroponics in the kitchen? Design and develop a home based hydroponics system.

 

RESOURCES:

 

Circular #844, Illinois Cooperative Extension Service, The University of Illinois at Urbana-Champaign.

 

Dikerman, Alexandra and John, Discovering Hydroponic Gardening.

 

Gericke, W.F. The Complete Guide to Soilless Gardening, Prentice Hall, New Your, 1940.

 

Harris, Dudley, Hydroponics, Growing Plants Without Soil.

 

Johnsen, Jan, Gardening Without Soil.

 

Kramer, Jack, Gardens Without Soil.

 

Kenyon, Stewart, Hydroponics For the Home Gardener.

 

National Geographic World, National Geographic Society, July 1988

 

Nicholls, Richard E., Beginning Hydroponics

 

Douglas, James Sholto, Beginners Guide to Hydroponics.

 

Douglas James Sholto, Advanced Guide to Hydroponics.

 

Saunby, T., Soilless Culture.

 

Sullivan, George, Hydroponics-Growing Plants Without Soil.




TITLE OF BIOTECHNOLOGY ACTIVITY:

 

C.    Cytoplasmic Inheritance of Leaf Variegation

(Adapted from "Biotechnology in Agriculture", MAVCC, Stillwater, OK.)

 

OBJECTIVES:

 

Review objectives in Wisconsin Fast Plants™ Cytoplasmic Inheritance Kit # 15-8786K available from Carolina Biological Supply Company.

 

MATERIALS AND OR EQUIPMENT:

 

                                  Wisconsin Fast Plants™ Cytoplasmic Inheritance Kit # 15-8786K

 

TIME REQUIRED FOR ACTIVITY:

 

45 - 60 days

 

BACKGROUND MATERIAL:

 

Plants with variegated leaves have a pattern of white markings on their leaves as opposed to solid green leaves. This is an inherited trait carried by genes in chloroplasts located in the cell cytoplasm. Growing 2 generations of plants with this genetic trait will allow you to see how variegation is expressed and the frequency with which it appears. Interestingly, triazine resistance is also a cytoplasmic gene and is presented similar to variegation.

 

PROCEDURE:

 

                        1.         Refer to the kit's written materials, and review the background material, student worksheet, page 1. Ask your instructor to explain any information you do not understand.

 

                        2.         Adjust your light system so that the lights are 2" above the plants. Continuous lighting is required.

Note:The light system you built in Job Sheet 2 will need to be adjusted for this experiment. Bypass the timer because no dark period is needed. Check with your instructor for help. Place plants on a platform that can be raised and supported by bricks, blocks, or other means. An alternate light system that is adjustable may also be used.

 

                        3.         Review the new terms on page 1s of the worksheet.

 

                        4.         Review the objectives and obtain the materials needed to complete this laboratory activity.

 

                        5.         Answer the pre-lab questions on page 2s and review these with your instructor before proceeding.

 

                        6.         Record beginning information in your laboratory notebook as described in Unit 1, "Introduction to Biotechnology."

 

                        7.         Read the planting and watering instructions for Rapid Cycling Brassica.

 

                        8.         Plant the variegated plant seeds and record the planting date in your notebook.

 

                        9.         Experiment day 1: 3 to 5 days after planting variegated seeds, plant the wild type seed and record the date.

 

                        10.       Experiment day 4 to 5: Thin plants according to instructions on student worksheet page 3s. Record your observations.

 

                        11.       Experiment day 10 to 14: Place barriers as shown on page 3s. Record observations.

 

                        12.       Experiment day 14 to 18: Pollinate plants following the directions on page 4s. Record observations.

 

                        13.       Experiment day 18 to 40: Continue growing plants to maturity following the "Growing Instructions." Record your observations during this time period.

 

                        14.       Experiment day 45: Collect seeds from your plants following directions on page 4s. Record date and observations.

 

                        15.       Plant second generation seeds and record day 1 information.

 

                        16.       Count plants according to type. Follow directions on page 4s and record information in your laboratory notebook.

 

DATA AND SUMMARY ANALYSIS:

 

                        17.       Complete discussion questions on page 5s and review these with your instructor.

 

                        18.       Make your conclusions and record these in your laboratory notebook.

 

                        19.       Write a laboratory report and turn it in for evaluation. Follow the laboratory report instructions in Unit 1.

 

QUESTION(S) FOR INVESTIGATION:

 

See questions in kit.




TITLE OF BIOTECHNOLOGY ACTIVITY:


D.    Micropropagation of Dry Bean Shoot Tips

(Adapted from "Biotechnology in Agriculture", MAVCC, Stillwater, OK.)

 

OBJECTIVES:

 

Upon completion of this activity, the student should be able to:

 

                        1.         Demonstrate proper micropropagation techniques of any dry bean shoot tips.

 

MATERIALS:

 

                                  8 dry bean seeds (pinto beans or seeds for green beans - garden varieties)

                                  4 petri dishes, each containing 25 ml MS5 media

                                  2 petri dishes, each containing 25 ml B5 media

                                  1 half-pint wide mouth jar containing 75ml MS media

                                  500 ml sterile distilled water

                                  200 ml 80% ethanol for use in a mist spray bottle

                                  200 ml 40% household bleach (80 ml household laundry bleach and 120 ml water)

 

Note: Above three items have sufficient quantity for about 30 experimental set-ups.

 

                                  Fine tip thumb forceps

                                  Scalpel with #11 blade

                                  Masking tape or parafilm

                                  Felt tip laundry marker

                                  Still air chamber

                                  Fluorescent light set up for culturing material

 

TIME REQUIRED FOR ACTIVITY:

 

3 - 5 days

 

BACKGROUND MATERIALS:

 

Introduction: Micropropagation is use of an actively growing piece of plant tissue to develop an entire plant. In this case, a small piece of actively growing stem (shoot) tissue will be used. Gelatin-like media that contains nutrients to support plant growth and hormones to stimulate development of plant structures will be used. Changing hormones and concentrations stimulate development of different plant structures as plantlets are moved to different media. Cleanliness to prevent contamination is important.

 

PROCEDURE:

 

Follow the steps and check each as you complete it.

 

                                    1.         Complete beginning entries in your laboratory notebook.

 

                                    2.         Disinfect your seed by soaking up to 200 seeds in 200 mil of 40% household bleach for 5 minutes.

 

                                    3.         Pour off the household bleach and rinse the seeds 3 times using sterile distilled water.

 

                                    4.         Set up the still air box and spray down the interior with 80% ethanol and allow it to air dry. Place equipment and materials inside the box. Spray each with ethanol as you put it in the box.



Figure 1. Equipment Set Up For Bean Germination.


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                                    5.         Transfer the beans directly from the sterile water rinse and place 4 on the media in each of 2 petri dishes. Label the dish with the date, your name, bean germination, and MS5 media. Seal the petri dishes with tape or parafilm.

 

                                    6.         Germinate the seeds for 7 days under the culture lights or until the emerging root is about 1/2" long. Discard seeds that are contaminated. Record observations in the laboratory notebook.

 

                                    7.         Set up and disinfect the still air box and equipment as in step 4. Include the germinated bean dishes and two petri dishes with MS 5 media.

 

                                    8.         Dissect out the shoot tip. Use the inside of the lid of the bean germination dish as a cutting surface. Carefully place 4 tips with the cut edge down on each of the petri dishes. Seal and label your cultures.


Figure 2 Process For Dissecting Out Shoot Tip.


pg170.gif




 

                                    9.         Place the shoot tip culture under the grow lights for two weeks or until two leaves are developed and expanded to about 1/4 inch. Record observations.

 

                                    10.       Prepare the work area for transferring the shoots to B5 media for root development. Select the most vigorous shot for transfer and cut it as near the base as possible. Transfer the shoot to the center of the B5 dish placing the cut surface firmly on the media.



Figure 3 Transferring Shoots



pg171.gif


 

                                    11.       Seal and label the petri dish and culture under the grow lights for 1 week or until roots start developing. Record observations.

 

                                    12.       Mature the plantlet by transferring it to the media in the widemouth jar. Transfer the entire plantlet and place the root securely on the media surface. Seal and label the jar.

 

                                    13.       Culture the plantlets under the grow lights for about 3 weeks or until they reach 1 to 1 1/2" in height. Record observations.

 

                                    14.       Transfer the plantlets to sterile potting soil. Use clean procedure but not aseptic technique. Remove the agar and plant from the jar and gently work the agar off the roots by careful massage in warm water. Pot each plant in a 6" pot or 2 quart milk carton base. Keep the soil moist using 1/2 strength liquid plant fertilizer. Keep plants in a box and cover with clear plastic to prevent excessive moisture loss.

 

                                    15.       After three days begin removing the plastic by opening a small section and enlarging it over several days until all is removed.

 

                                    16.       Grow plant to maturity under greenhouse conditions.

 

DATA AND SUMMARY ANALYSIS:

 

Record your conclusions in your laboratory notebook.

 

Write a laboratory report and turn it in for evaluation.




TITLE OF BIOTECHNOLOGY ACTIVITY:

 

E.    Tissue Culture of Cauliflower

(Adapted from Biotechnology in Agriculture", MAVCC, Stillwater, OK.)

 

OBJECTIVES:

 

Follow objectives in cauliflower tissue culture kit

 

MATERIALS AND/OR EQUIPMENT:

 

Cauliflower Tissue Culture kit available from Carolina Biological supply Company is required. Similar kits may be substituted.

 

TIME REQUIRED FOR ACTIVITY:

 

Follow suggested time in kit

 

BACKGROUND MATERIALS:

 

Tissue culture is the process of developing undifferentiated tissue (callus) from a plant followed by development of nearly identical plants (clones) from the callus. This process depends on nutrient support and hormone treatment for the various stages of development. This technology is practiced commercially as well as for research activities. Applications of tissue culture have potential in horticulture, floriculture, and crop systems.

 

PROCEDURE:

 

Directions: Follow the procedures in the cauliflower tissue culture kit manual. Check each step off on the checklist as you complete it.

 

                        1.         Set up the work area with a still air box or laminar flow hood.

 

                        2.         Set up culture growth area with controlled lighting.

 

                        3.         Review aseptic technique.

 

                        4.         Obtain the materials and equipment you need from the instructor. Review the function and use of all items with your instructor.

 

                        5.         Record the introductory information and purpose for this activity in your laboratory notebook.

 

                        6.         Prepare explants or obtain them from the instructor after completing the non-sterile procedures.

 

                        7.         Complete the sterile procedures.

 

                        8.         Grow cultures for 1 to 2 weeks in growth area. Record observations in the laboratory notebook.

 

                        9.         Transfer plant material to shoot initiation media following the subculturing procedure.

 

                        10.       Record observations during the shoot development stage.

 

                        11.       After 4 weeks, transfer shoots to a rooting medium.

 

                        12.       Record observations during the rooting stage.

 

                        13.       After roots are well developed, transfer the plantlets to soil.

 

                        14.       After acclimatizing the plants, they can be grown to maturity in a greenhouse or transplanted outdoors.

 

DATA AND SUMMARY ANALYSIS:

 

                        15.       Record all observations during the acclimatizing and transplant phase. Complete your conclusions in the laboratory notebook.

 

                        16.       Write a laboratory report and turn it in for evaluation.




TITLE OF BIOTECHNOLOGY ACTIVITY:


F.    Diagnostic Testing (ph, nitrates, aflatoxin)

 

OBJECTIVES:

 

Upon completion of this activity, the students should be able to:

 

                        1.         Demonstrate a simple, convenient and economical method of accurately testing for nitrates, pH and Triazine using the AGRI-SCREEN test kits produced by Neogen Corporation.

 

MATERIALS AND/OR EQUIPMENT:

 

                                  Nitrate Field Screening Kit (I.D. No. 80)

                                  pH Field Screening Kit (I.D. No.81)

                                  Triazine Resistance Field Screening Kit (I.D. No. 82)

 

TIME REQUIRED FOR ACTIVITY:

 

Two class periods

 

BACKGROUND MATERIAL:

 

Nitrate Test

While the majority of water in this county is in good shape, there are some problem areas-wells that are contaminated by groundwater from poorly placed septic tanks, cracked sewer pipes, leaking landfills, animal wastes, or the improper use of fertilizer. While the test itself will not improve your water, it certainly can give you the homeowner and the livestock feeder, a feeling of confidence to know that your water tests less than the federal standards allow-10 parts per million of nitrate nitrogen or 44 parts per million of nitrate. Keeping low levels of nitrate in water improves feed efficiency for livestock and poultry and ensures better general health for both humans and animals. If your water tests higher than the federal standards, you can then take corrective steps.

 

Now you can know the nitrate levels of your tap, well, pond or tile drainage water instantly with the new technology provided by AGRI-SCREEN Nitrate TEST kit. Each test kit includes materials to conduct three tests-a test tube, indicator strip, and colored chart for measuring nitrate levels. Each individual indicator strip is hermetically sealed in aluminum to keep the air from damaging the strip and to provide an accurate reading at the time of testing water. The AGRI-SCREEN Nitrate TEST is accurate, instant, and easy to use and read.

 

pH

Check pH, the chemical measure of acidity or alkalinity, on-the-spot for aiding crop and livestock management. The new AGRI-SCREEN pH TEST is an accurate, rapid method of measuring pH in soil and water.

 

On the pH scale of 0 to 14, most crops perform best near neutral (pH=7). Use AGRI-SCREEN to check for soil acidity, indicated by low Ph, and the need for lime. Alkaline soil, with high Ph, may require special reclamation for the best crop performance. Whether you are growing rhododendrons or azaleas, or fields of peas and potatoes-you can expect the best results when your Ph is balanced for the different needs of your individual crops.

 

Alkaline poultry litter or livestock bedding can produce ammonia gas and promote the growth of disease organisms. Check the need for acidification with AGRI-SCREEN. A simple test can increase animal productivity.

 

Measure the pH of the water supply you use for preparing pesticide tank mixes. Many combinations are pH sensitive. Acid or alkaline water may reduce pesticide effectiveness, and simple measures are available for adjusting pH.

 

The clogging of water lines and irrigation systems is often caused by water pH problems. AGRI-SCREEN will show you whether water pH treatment is needed.

 

AGRI-SCREEN pH tests are accurate and easy to use and read. They can be used in the field, greenhouse, backyard, barnyard or almost anywhere for accurate results in five minutes or less.

 

Triazine

Populations of some weed species have developed resistance to triazine herbicides. Now you can determine before spraying whether your weeds are resistant to Atrazine or other triazines. Weeds shown to have resistance include lambsquarter, pigweed, ragweed, horseweed and many other common weeds.

 

AGRI-SCREEN Triazine Plant Sensitivity Test offers rapid detection using a floating leaf disc technique pioneered by Drs. Donald Penner and James J. Kells, Michigan State University. Each kit contains materials to conduct three tests. It contains tubes of Atrazine solution, a leaf disc cutter, and a syringe to withdraw air from the leaf tissue. Under normal growing conditions (sunlight or plant growth light), a user can obtain accurate results in about 15 minutes.

 

If you are planning to use Atrazine or another triazine, this test can save you both time and money. It is a convenient, economical method of quickly and accurately testing weeds on site before you apply herbicides.

 

Test suspect weeds for resistance to these Triazine herbicides:

 

                                              Atrazine                                            Prometryne

                                              Simazine                                           Cyanazine

                                              Metribuzin


Use AGRI-SCREEN

Triazine Plant Sensitivity Test on:

 

                                              Weeds that survive pre-emergence application

                                              Weeds in late summer to select next year's herbicide

 

Regular use aids in better, more efficient management of herbicidal programs while eliminating the time and costs for agent conducted tests. Easy to use and read AGRI-SCREEN Triazine Plant Sensitivity Test checks suspect weeds on site with results in about 15 minutes.

 

PROCEDURE:

 

                        1.         Follow instructions as specified in the directions of each of the following kits:

 

                                              Nitrate Field Screening Kit (I.D. No. 80)

                                              pH Field Screening Kit (I.D. No.81)

                                              Triazine Resistance Field Screening Kit (I.D. No. 82)

 

                        2.         Nitrate Field Screening Kit:

Supplies available for 3 tests

Compares sample to 0-250 ppm standards

                                    Contains:        3 nitrate water test strips

Collection tube

Color comparison chart

 

                        3.         pH Field Screening Kit:

Supplies available for 5 tests

                                    Contains:        5 pH test strips

Collection/Preparation containers

Color comparison chart from pH 2 to 9

 

                        4.         Triazine Resistance Field Screen Kit:

Supplies available for 3 tests

                                    Contains:        Activating reagent

Collection/Preparation containers

Leaf punch

Vacuum syringe

 

QUESTION(S) FOR INVESTIGATION:

 

                        1.         How can each of the testing kits benefit you or your community?

 

IDEA(S) FOR ADDITIONAL ACTIVITIES:

 

                                  The testing of pH, nitrates or triazine can be used not only as a class laboratory assignment, but also as a BOAC community service project.

 

REFERENCES:

 

Neogen Corporation 620 Lesher Place, Lansing, Michigan 48912

1-800-234-5333