Plant engineering

1. The perpetual search for improvement Plant engineering

2. Salute to Norman E. Borlaug Father of “Green Revolution” Iowa native and Nobel Laureate (1970) used Biotechnology and modern farming practices to combat world hunger. Dr. Borlaug’s development of high-yield and disease-resistant wheat varieties bore results in Mexico, Pakistan and India that stretched the imagination of viable agriculture in developing countries.  Recently, Borlaug worked to apply farming practices and methods of increasing food production to Asia and Africa, and he has continued to advocate for the use of biotechnology to combat world famine.  Established the World Food Prize in 1987 and expanded this with the help of John Ruan of Des Moines in 1990. Died September, 2009 at the age of 95.

3. Plant cloning Leaf or root cutting from a plant grown into a new plant (vegetative propagation) – cloning – same genetic makeup as the donor plant. Vegetative propagation: cutting forms a mass of non-specialized cells – callus Callus grows, divides and forms various specialized cells (roots, stems) new plant.

4. Cloning approaches Leaf cuttings Stem of a leaf in soil new plant, e.g. African violets, snake plants, gloxinia, and begonias. Some plant leaves, when cut from their stems and placed flat on top of the soil, develop roots at particular intervals along the cut veins of the leaf. The parent sprouts several baby plants, all growing vertically from the leaf at various points along the veins, e.g.begonia and geranium. Stem cuttings Piece of stem inserted in soil horizontally or vertically. Roots develop downward from stem  new plant from top. Stem with bud cut from plant,  new plant from bud, e.g: begonia, gardenia, christmas cactus, lantana, and impatiens. Budding Common in propagation of many fruit trees. A bud, which is an undeveloped branch, leaf or flower protruding from the stem of the plant, cut from parent, placed into a notch made into the parent stem, wrapped in place and allowed to grow until it can be removed from the parent and allowed to grow in soil on its own. Plant division Many flowers, such as the commonly found daylily, hostas, certain orchids, and ferns, are cultivated through the division of their thick roots. Runners Wild strawberry plants, e.g., put out horizontal stems that touch the soil. These develop roots from which another identical strawberry plant will grow. Grafting Cutting from plant attached to a piece of the root, or to the rooted stem of another plant. The pieces become united, and grow as one plant. Desirable properties of each plant can be mated to produce a new hybrid plant without producing hybrid seeds, e.g. producing dwarf fruit and oriental trees for home landscaping

5. Meristem cloning

6. Genes and Chromosomes This diagram shows a gene in relation to the double helix structure of DNA and to a chromosome (right). The chromosome is X-shaped because it is dividing. Introns are regions often found in eukaryote genes that are removed in the splicing process (after the DNA is transcribed into RNA): Only the exons encode the protein. This diagram labels a region of only 50 or so bases as a gene. In reality, most genes are hundreds of times larger. The total complement of genes in an organism or cell is known as its genome, which may be stored on one or more chromosomes; the region of the chromosome at which a particular gene is located is called its locus.

7. Mendelian inheritance Crossing between two pea plants heterozygous for purple (B, dominant) and white (b, recessive) blossoms

8. DNA chemistry The chemical structure of a four-base fragment of a DNA double helix

9. Mutations DNA replication is extremely accurate, with an error rate per site of around 1 in 10−6 to 10−10 in eukaryotes. Rare, spontaneous alterations in the base sequence of a particular gene arise from a number of sources, such as errors in DNA replication and the aftermath of DNA damage. These errors are called mutations. The cell contains many DNA repair mechanisms for preventing mutations and maintaining the integrity of the genome; however, in some cases—such as breaks in both DNA strands of a chromosome — repairing physical damage to the molecule is a higher priority than producing an exact copy. Due to the degeneracy of the genetic code, some mutations in protein-coding genes are silent, or produce no change in the amino acid sequence of the protein for which they code; for example, the codons UCU and UUC both code for serine, so the U↔C mutation has no effect on the protein. Mutations that do have phenotypic effects are most often neutral or deleterious to the organism, but sometimes they confer benefits to the organism's fitness. Mutations propagated to the next generation lead to variations within the population of a species. Variants of a single gene are known as alleles, and differences in alleles may give rise to differences in traits. Although it is rare for the variants in a single gene to have clearly distinguishable phenotypic effects, certain well-defined traits are in fact controlled by single genetic loci. A gene's most common allele is called the wild type allele, and rare alleles are called mutants. However, this does not imply that the wild-type allele is the ancestor from which the mutants are descended.

10. Biopharmaceuticals from transgenic plants • Glycoproteins can be made (bacteria like E. coli cannot do this) • Virtually unlimited amounts can be grown in the field rather than in expensive fermentation tanks • There is no danger from using mammalian cells and tissue culture medium that might be contaminated with infectious agents. • Purification is often easier Corn is the most popular plant for these purposes, but tobacco, tomatoes, potatoes, and rice are also being used.

11. Biotech basics For centuries, humankind has made improvements to crop plants through selective breeding and hybridization — the controlled pollination of plants. Traditional plant breeding involves the crossing of hundreds or thousands of genes, whereas plant biotechnology allows for the transfer of only one or a few desirable genes. This more precise science allows plant breeders to develop crops with specific beneficial traits and without undesirable traits. Many of these beneficial traits in new plant varieties fight plant pests —that can be devastating. Others provide quality improvements, such as tastier fruits and vegetables; processing advantages, such as tomatoes with higher solids content; and nutrition enhancements, such as oil seeds that produce oils with lower saturated fat content. Crop improvements like these can help provide an abundant, healthful food supply and protect our environment for future generations.

12. Plant Biotechnology Myths & Facts • There are no biotech food products currently on the market. • Biotech foods are unsafe to eat. • Biotech foods are not regulated or tested. • Meat, milk and eggs from livestock and poultry fed biotech feed products are not safe. • Organic or conventional crops are more nutritious or safer than biotech crops. • Biotech foods taste different than foods made from conventional crops. • The United States does not require labeling of biotech foods. • Biotech foods and crops have been rejected by consumers. • The United States is the only country growing and consuming biotech crops. • Biotech crops are harmful to monarch butterflies. • Biotechnology is only being applied to a few crop varieties. • The "pipeline" of biotech plants products is dried up - no new products being developed • Biotech crops harm the environment. • Biotech foods can't feed the world. • Biotech crops increase food allergies. • Using biotechnology to improve plants is not natural. • Biotech companies won't disclose where field trials of biotech crops are being grown • Biotech crops harm the environment. • Biotech crops will cause "superweeds" to develop. • The only people who benefit from biotech plants are the agricultural companies

13. Animal Biotechnology Myths & Facts • Only humans, not animals can benefit from medical biotechnology. • Biotech and cloned animals are still years away - science fiction. • Pets do not benefit from biotechnology at all. • Biotech and cloned animals are different from normal animals. • Wild animals cannot benefit from cloning technology. • Biotech will cause disease outbreaks such as avian flu, mad cow disease and West Nile virus. • Organ transplants from animals are an unreal fantasy. • We are just exploiting animals by applying biotechnology to them. • Meat, milk and egg products from biotech animals are unsafe to eat. • Biotech animals suffer more pain or distress than conventional animals. • Cloned animals have higher death rates than conventionally-bred animals. • Cloned animals are not as healthy as non-cloned animals. • Animal cloning is not safe. • If biotech animals or fish escape into the wild, they will endanger wild animals and environment. • Animals are misused in research. • The famous cloned sheep Dolly did not lead a full and healthy life.

14. Myth: Biotech foods are unsafe to eat Fact: The Food and Drug Administration (FDA) has determined that biotech foods and crops are as safe as their non-biotech counterparts. The American Medical Association, the American Dietetic Association, and the U.S. National Academy of Sciences have also declared biotech foods safe for human and animal consumption. In addition, since being introduced to U.S. markets in 1996, not a single person or animal has become sick from eating biotech foods. Other international groups that have concluded biotech foods and crops are safe are The United Nations Food and Agriculture Organization, the World Health Organization, the International Council for Science, the French Food Agency, and the British Medical Association. The European Food Safety Authority (EFSA) has also found several biotech varieties to be safe for human and animal consumption.

15. Myth: no biotech food products marketed Fact: 70+% of processed foods on grocery store shelves contain ingredients and oils from biotech crops. The first biotech crop, a tomato improved through biotechnology, was sold in 1994. The first biotech commodity crops - an insect resistant variety of corn - were grown and sold in 1996. Today, the most popular biotech crops are corn, soybean, cotton and canola.

16. C5 Plum pox resistant plums Plums that have been genetically engineered to be resistant to the plum pox virus

17. Tobacco mosaic virus resistance Tomato plants infected with tobacco mosaic virus (which attacks tomato plants as well as tobacco). The plants in the back row carry an introduced gene conferring resistance to the virus. The resistant plants produced three times as much fruit as the sensitive plants (front row) and the same as control plants. (Courtesy Monsanto Company.)

18. Pesticide resistance Effect of the herbicide bromoxynil on tobacco plants transformed with a bacterial gene of which a product breaks down bromoxynil (top row) and control plants (bottom row). "Spray blank" plants were treated with the same spray mixture as the others except the bromoxynil was left out. (Courtesy of Calgene, Davis, CA.)

19. Bt Corn Bt corn is a variant of maize, genetically altered to express the bacterial Bt toxin, which is poisonous to insect pests. The pest is the European Corn Borer.

20. Bt Corn: Cont’d • Expressing the toxin achieved by inser-ting a gene from the lepidoptera pathogen Bacillus thuringiensis into the corn genome. This gene codes for a toxin causing the formation of pores in the larval digestive tract. These pores allow naturally occurring enteric bacteria such as E. coli and Entero-bacter to enter the hemocoel where they multiply and cause sepsis (Broderick et al., PNAS 2006). Contrary to the common notion that Bt toxin kills the larvae by starvation. • Bt176 varieties were voluntarily with-drawn, in 2001, from the list of approved varieties by the USEPA after finding to have little or no Bt expression in the ears and not found to be effective against second generation corn borers. (Current status of Bt Corn Hybrids, 2005) • Effect on non-target invertebrates depends on the standard of comparison

21. Preventing Bt resistance in pests • By law, farmers in the United States who plant Bt corn must plant non-Bt corn nearby. The non-modified fields provide a location to harbor pests. These refuges slow the evolution of resistance to the pests to the Bt pesticide. Doing so enables an area of the landscape where wild type pests will not be immediately killed. • It is anticipated that resistance to Bt will evolve in the form of a recessive allele in the pest. Because of this, a pest that gains resistance will have an incredibly higher fitness than the wild type pest in the Bt corn fields. If the resistant pest is feeding in the non-Bt corn nearby, the resistance is neutral and offers no advantage to the pest over any non-resistant pest. • Ensuring that there are at least some breeding pests nearby that are not resistant, increases the chance that resistant pests will choose to mate with a nonresistant one. Since the gene is recessive, all offspring will be heterozygous, and the offspring from that mating will not be resistant to Bt and therefore no longer a threat. Using this method scientists and farmers hope to keep the number of resistant genes very low, and utilize genetic drift to ensure that any resistance that does emerge does not spread.

22. The effect of Bt corn on monarchs • A small, preliminary study done at Cornell University, and reported as a note in Nature in June 1999, indicated that monarch butterflies under laboratory conditions might be harmed by eating pollen from Bt corn plants. That experiment used a small number of caterpillars and gave them no choice about avoiding eating leaves that had been treated with a thick layer of Bt corn pollen. It did not attempt to duplicate real world environmental conditions.

23. The effect of Bt corn on monarchs • There is no significant risk to monarch butterflies from environmental exposure to Bt corn, according to research conducted by a group of scientists coordinated by the Agricultural Research Service (ARS), U.S. Department of Agriculture. This research was published in the Proceedings of the National Academy of Sciences (PNAS).he studies in this project showed that monarch caterpillars have to be exposed to pollen levels greater than 1,000 grains/cm2 to show toxic effects. • Caterpillars were found to be present on milkweed during the one to two weeks that pollen is shed by corn, but corn pollen levels on milkweed leaves were found to average only about 170 pollen grains/cm2 in corn fields. http://www.pnas.org/cgi/content/full/211297698v1

24. Bt Cotton Saves 3.46 M lb raw material- Conserves 1.48 Mgal fuel oil- Eliminates 2.16 M lb industrial waste - Transports and stores 416,000 gal insecticide less- Conserves 604k gal fuel oil - 1.04 million lb insecticide less 2.5 fewer applications per acre- 416k fewer insecticide containers- Saves 41,250 10-h workdays- Eliminates 2,150 10-h days of aerial application- Conserves 2.41 Mgal fuel and 93.7 Mgal water -Accrues $168 million in economic benefits from lower production costs and increased cotton yield - Reduces pesticide exposure risk- Preserves beneficial insect populations- Creates wildlife benefits- Gives cotton producers more time for family and community activities- Gives cotton producers peace of mind Produces fiber equivalent to that found in all consumer products derived from cotton

25. Golden rice • Progress in production of transgenic cereals for developing countries • Achievement has been the introduction of genes that produce beta-carotene – the precursor of vitamin A – in the rice grain. • Beta-carotene is present in the leaves of the rice plant, but conventional plant breeding has been unable to put it into the grain. • Dr. Ingo Potrykus of the Swiss Institute of Plant Sciences in Zurich, with Rockefeller funding, transferred one bacterial and two daffodil genes. • The transgenic rice grain has a light golden-yellow color and contains sufficient beta-carotene to meet human vitamin A requirements from rice alone. • Potrykis has also added a gene from the French bean to rice that increases its iron content over threefold.

26. The StarLink corn controversy • StarLink variety of Bt corn patented by Aventis (acquired by Bayer AG, 2002), for use in animal feed. • U.S. regulatory authorities permitted StarLink seed but not to be grown for human consumption. • This restriction was based on the possibility of an allergic reaction to the Bt protein used in StarLink. • StarLink corn was subsequently found in food. Taco Bell taco shells was particularly well publicized 1 • A public relations disaster for Aventis and the biotechnology industry as a whole! Sales of StarLink seed were discontinued. Aventis voluntarily withdrew the registration for Starlink varieties in 20002 • 28 people reported apparent allergic reactions. However, the US Centers for Disease Control studied the blood of these individuals and concluded there was no evidence of hypersensitivity to Bt protein 3 • 5 weeks after the FDA/CDC declared StarLink safe, based on blood tests, advisers to the EPA - including some of the leading food allergists, released a thorough critique of the FDA's allergy test and other aspects of the StarLink investigation. They said the research to declare StarLink safe had many shortcomings including, lack of adequate controls, not sensitive enough and failed to follow standard protocols that helped prevent false interpretations. (FIFRA Sci. Advisory Panel Report #2001-09, July ‘01) • Aid sent by the UN and the US to Central African nations also contained some StarLink corn. The nations involved refused to accept the aid. • The southern portion of the U.S. Corn Belt planted the greatest amount of StarLink corn. It is this portion of the U.S. where corn borer damage creates the greatest economic loss to farmers. • The US corn supply has been monitored for the presence of the Starlink Bt proteins since 2001. No positive samples have been found since 2004, showing that it was possible to withdraw this GM crop without leaving traces in the environment once it has been used in the field 4

27. Ethanol Contribution to Energy and Environmental GoalsAlexander E. Farrell, Richard J. Plevin, Brian T. Turner, Andrew D. Jones, Michael O'Hare, Daniel M. Kammen SCIENCE    VOL 311 27 JANUARY 2006 www.sciencemag.org http://rael.berkeley.edu/ebamm

28. Genetically modified salmon • Armed with a gene from the ocean pout -- the new salmon, which originally hails from the Atlantic, grows twice as fast as its less endowed peers out at sea. • The Food and Drug Administration has concluded that the new salmon is safe to eat and safe for the environment, suggesting that approval is likely at a hearing planned for later this month. The decision would make the fish the first genetically modified animal allowed for human consumption. The issue has raised questions about impacts on the environment, evolution and the future of food.

29. Genetically modified sugar beets • Roundup resistant • Challenged in court September, 2010

30. Proteins produced by transgenic crops • Human growth hormone with the gene inserted into the • chloroplast DNA of tobacco plants. • Humanized antibodies against such infectious agents as • HIV • respiratory syncytial virus (RSV) • sperm (a possible contraceptive) • herpes simplex virus, HSV, the cause of "cold sores" • Protein antigens to be used in vaccines • An example: patient-specific antilymphoma (a cancer) vaccines. B-cell lymphomas are clones of malignant B cells expressing on their surface a unique antibody molecule. Making tobacco plants transgenic for the RNA of the variable (unique) regions of this antibody enables them to produce the corresponding protein. This can then be incorporated into a vaccine in the hopes (early trials look promising) of boosting the patient's immune system — especially the cell-mediated branch — to combat the cancer. • Other useful proteins like lysozyme and trypsin http://images.google.com/imgres?imgurl=http://olegvolk.net/olegv/macro/plants/tomato.JPG&imgrefurl=http://kikoshouse.blogspot.com/2006/03/science-saturday-iv-oh-you-sweet-thing.html&usg=__8rqaLks4T2a8o8Wi3FcOOjH7XQE=&h=750&w=707&sz=155&hl=en&start=8&um=1&tbnid=GUU8gNWaI4XS2M:&tbnh=141&tbnw=133&prev=/images%3Fq%3Dtransgenic%2Bplants%26hl%3Den%26rlz%3D1T4RNWE_enUS312US312%26sa%3DX%26um%3D1

31. Agrobacteriumtumefaciens – t-DNA Invades damaged plant sites Releases transfer DNA (t-DNA) T-DNA invades plant cells, is incorporated into the genome Infected cells grow rapidly to form galls The infected cells produce food for the bacteria The invasion is DNA transfer between kingdoms This is exploited to introduce desirable genes This produces transgenic plants Salt tolerant rice Golden rice

32. Transgenetics with A. tumefaciens

33. Exposure to engineered Agrobacteria

34. Reducing the rate of insect resistance development The rate of resistance development to transgenic plants can be reduced by either expressing proteins with different modes of action, or binding characteristics in one plant (gene pyramiding) or by expressing proteins with different modes of action or binding characteristics in separate plants and then using these plants either in a mosaic or sequential fashion

35. Food Prices Worldwide food prices have also increased considerably. But biofuels only consume roughly 4% of the world's grains. USDA Secretary Schafer stated: last year "Higher oil prices affect much more than just the cost of driving; they are actually one of the major factors behind higher food costs." The USDA and the White House Council of Economic Advisors have stated that increased corn demand is only responsible for "3 percent of the more than 40 percent increase we have seen in world food prices this year." Ethanol is made from "dent" or yellow "feed" corn. People accuse ethanol of increasing the price of beer, pasta and tortillas, which is curious because none of those products are made from "dent" corn. Major food companies are profiting nicely from the food price crisis: Land O'Lakes (earnings up 16%), Kraft Foods (earnings up 21%), Sara Lee (net income up 55%), and General Mills (profits up 61% from 2007).

36. Galls: maple trees Galls on a maple leaf

37. Galls: Rose Rose bedeguar gall

38. Galls: OAK Oak artichoke gall caused by a wasp

39. Galls: Oak Marble Galls Cross-section (above) Oak marble galls, one with a gall wasp exit hole and another with Phoma gallorum fungal attack Green stage

40. Galls: Lime Nail Gall Caused by a mite, Eriophyes tiliae, of 0.2 mm long

41. Bacterial galls Pseudomonas syringae causes bacterial gall on oleander and olive. ...Gall bacteria reproduce in fissured or galled bark and are spread by ...

42. Bacterial galls: rose Agrobacterium tumefaciens manages to survive in the rhizosphere on materials that leak from roots. It infects wounded host plant stems and roots.

43. Fungal galls Fungus stem gall or "poop gall" on choke cherry (Prunus virginiana) in Montana. The swollen stems are caused by the fungus Dibotryon morbosum

44. Giant fungi and trees 1. The mycelium of some forest fungi can extend enormous distances. A single individual of Armillaria bulbosa permeates >30 ac forest soil in northern Michigan and may be one of the world's largest living organisms. Some scientists speculate that it was spawned by a single spore thousands of years ago. Another Armillaria in Washington was recently found to consist of a subterranean mycelial network with erect, above-ground mushrooms covering > 1000 ac 2. These fungal monstrosities are rivaled in total size and mass by a 106 acre, 6,000 ton stand of genetically identical quaking aspen in the Rocky Mountains. The aspen clone is connected by a common root system, and has literally climbed over mountains and across meadows. Any discussion of massive clonal colonies should also include the conjoined polyps of coral reefs. However, the question still remains: Do these clonal colonies qualify as a single individual, as in the 1200 ton General Sherman tree of California's Sequoia National Park?

45. Giant fungus: oregon Mycologists investigated if Armillaria could be killing trees dying in the Malheur National Forest in the Blue Mountains of eastern Oregon and were astonished. This most recent find was estimated to cover over 2,200 acres (890 hectares) and be at least 2,400 years old, possibly older.