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June 2010 Updates

Paper Industry Tests Genetically Altered Trees

By Mitch Stacy
Associated Press
June 7, 2010

TAMPA, Fla. - The commercial paper industry's plans to plant forests of genetically altered eucalyptus trees in seven Southern states have generated more cries from critics worried that such a large introduction of a bioengineered nonnative plant could throw natural ecosystems out of whack.

ArborGen, a biotechnology venture affiliated with three large paper companies, got U.S. Department of Agriculture approval last month for field trials involving as many as 250,000 trees planted at 29 sites during the next few years. Much smaller lots of the genetically altered trees have been growing in some of the states for years.

Australian eucalyptus trees grow faster than native hardwoods and produce high-quality pulp perfect for paper production, but thus far, they have been able to thrive only in very warm climates. South Carolina-based ArborGen genetically altered the trees to withstand freezing temperatures, and the idea with the test forests is to see how far north they can now be grown.

The test sites will cover a total of about 300 acres in Florida, South Carolina, Texas, Alabama, Mississippi, Georgia and Louisiana.

While genetically engineered crops such as corn and soybeans have become common, ArborGen's experiment marks the first large planting of designer trees in the United States. The company says plantations of hearty, faster-growing eucalyptus could produce more timber in a smaller area and allow conservation of natural forests.

But critics say that despite the USDA's assurance that the trees pose no environmental threat, not enough is known about their effect on natural surroundings.

"We have many reservations about it," said Neil J. Carman, a biologist who serves on the Sierra Club's genetic engineering committee. "We don't think the scientific evidence is in yet that says this is a good idea."

Anne Petermann, executive director of the activist group Global Justice Ecology Project, said eucalyptus trees are invasive, require vast amounts of water that could reduce groundwater levels, and increase the wildfire risk because they are so flammable.

"This is quite a dangerous tree to be mass planting," Petermann said.

But ArborGen CEO Barbara Wells said eucalyptus trees have not proven invasive in dozens of tropical countries where they are widely grown on plantations. Also, ArborGen genetically modified the trees to limit their ability to disperse seed and spread.

Although the new field trials will significantly increase the number of genetically engineered trees being grown, Wells called it "very confined research."

"The total is 300 acres, but when you're doing tree research, that really is very small acreage," she said, noting that about 20,000 acres of genetically unaltered eucalyptus trees are already grown in central and southern Florida for production of wood chips and mulch. The new test forests will show whether the genetically altered trees can thrive farther north in Florida, where freezing temperatures can occur in the winter.

Donald Rockwood, a professor emeritus in the University of Florida's School of Forest Resources and Conservation, has worked for about 30 years on developing eucalyptus trees that thrive in Florida. He uses traditional breeding techniques, not genetic modification.

The genetically unaltered trees growing in controlled plantations in Florida have not proven invasive, are relatively efficient users of water and are no more flammable than other hardwoods, said Rockwood, who was hired by ArborGen to do a report on eucalyptus trees' invasiveness because of his experience working with them at the university.

Still, Rockwood said, introduction of any genetically altered species poses risks. For example, the gene that makes the trees resistant to cold could be transferred to surrounding plants, allowing them to spread farther north than nature intended.

"It certainly needs to be done carefully, it needs to be regulated and there needs to be a period of well-defined observations," Rockwood said.

The ArborGen trees will be planted in seven counties throughout Florida, four counties each in South Carolina and Texas, two each in Alabama and Mississippi and single counties in Georgia and Louisiana. Rockwood said they can grow about 25 feet per year and be ready to harvest in less than three years.


 

Growing Chemicals

By Melody Voith
Chemical and Engineering News
June 7, 2010

New biotech traits enable fuel and polymer production in crops


When Jeremy Johnson and R. Michael Raab founded Agrivida in 2005, they were still finishing their Ph.D.s at Massachusetts Institute of Technology. Although both are chemical engineers, they based their start-up not on chemical plant technology, but on a technology for making chemicals in plants.

The work, Johnson says, is a sort of homecoming for a kid from Kansas who grew up working on a small dairy farm. And it is farmers who would benefit from Agrivida's products. The company is developing specialized crops that manufacture their own enzymes to more easily convert their cellulose into sugars for ethanol production.

Agrivida is one of a number of plant biotechnology firms that are hoping to leapfrog current technologies. Making today's biobased fuels and materials involves growing crops and then using physical and chemical means to extract sugars that can be fermented into the desired product. The firm's goal is to make plants do more of the manufacturing work.

For example, biobased plastics firm Metabolix is growing polyhydroxyalkanoate (PHA) resins in switchgrass, and the firm sees a future in biorefineries based on such modified crops. Other companies are forming partnerships in an attempt to engineer chemicals inside crops.

Syngenta is collaborating with research firm Edenspace Systems to develop easier-to-process plants for fuel production. Plant biotech start-up Ceres has worked with Rohm and Haas, now Dow Chemical, to determine whether energy crops could simultaneously produce methacrylate monomers. And Mendel Biotechnology has a partnership with BP to improve dedicated energy crops to speed reactions that break down cellulose and lignin.

Turning crops such as corn, switchgrass, miscanthus, and sorghum into living chemical factories holds the promise of lowering processing costs for renewable fuels and chemicals, proponents contend. Industry analysts agree that the plant biotechnology developments are showing early successes, and they confirm there would be strong demand for the plant-made products.

But for biobased technologies to displace petroleum, farmers will need to gamble on new, risky crops planted on vast acreages. And the new traits do not address questions that face the broader push for cellulose-based fuels and chemicals, including how processors will access and efficiently transport cellulosic feedstocks (C&EN, April 27, 2009, page 10).

It will be a few years before farmers have the chance to plant the new chemical-making crops. In the meantime, research and development of new traits is moving at a brisk pace in the labs of Agrivida and Metabolix, both located near MIT in Cambridge, Mass.

At first glance, the activities at Agrivida's headquarters look like those at any other analytical lab. Scientists watch over automated machines pipetting small amounts of liquid for screening tests, while biological samples grow in stacks of petri dishes. But a visitor might note a curious-looking plant in the corner. The 4-foot-tall sorghum specimen is a source of genetic material for a team of DNA-transfer experts.

Johnson explains that the researchers are working to modify plant genes to produce a specialized enzyme that breaks down the cellulose in the cell wall. In addition to the enzyme, Agrivida will insert a switch, made from a protein segment, that activates the enzyme under specific conditions.

"Cell-wall-degrading enzymes, though desirable for sugar production, are bad for plant growth," Johnson points out. With the addition of the enzyme alone, plants soon become limp and die. To preserve plant growth, the company is working with enzyme-arresting protein segments called inteins. "An intein is a peptide sequence within a protein that has the ability to cleave itself out and reconnect the rest of the sequence. That way there is no negative impact on the plant when it is inactive," Johnson says.

The switch can be designed to splice out under different conditions, such as by heating a crop after it is harvested. Then the activated enzymes would begin to degrade the cell wall and make sugar from the cellulose.

Johnson's team is working with wild-type enzymes and a selection of the 400 known inteins to find a pair that combines low initial enzymatic activity with high sugar production after the switch is triggered. The result for a cellulosic ethanol maker would be to reduce the need for expensive added enzymes, which can cost up to 70 cents per gal of ethanol. In addition, Johnson says, the sugar could be obtained with less mechanical, thermal, or chemical processing of the plant biomass.

The plants are not yet commercially viable. But Johnson says lab tests show that the modified samples produce a larger amount of sugar than control plants in cases where no enzymes are added, as well as when some or a lot of enzymes are added.

In contrast to Agrivida's focus on sugar, Metabolix is working to produce plastics and chemicals in plants. The company currently uses sugar as a feedstock to manufacture its biodegradable plastic, a PHA resin called Mirel, through microbial fermentation. Its joint venture with Archer Daniels Midland recently started producing Mirel at a 110 million-lb-per-year facility in Clinton, Iowa.

In March, Metabolix told investors it had successfully produced PHA inside switchgrass plants at a dry weight concentration of 6%. The company can also produce PHA in oil seeds and sugarcane. "Our goal is to get to commercially viable crops in field trials within two years," says Metabolix Chief Executive Officer Richard Eno. By then, he tells C&EN, the firm will have increased PHA content to a level where producing it inside of plants will be more cost-competitive than fermentation.

To get the most value from the plastic-producing crops, Eno says, they could be processed in a biorefinery. "We would harvest the plants, recover the PHA, and convert it to either polymers or industrial chemicals," he says. The rest of the plant biomass, he adds, could be used for cellulosic ethanol, gasification, or for fuel in the refinery.

Metabolix is already finding a market for the relatively costly PHA made by fermentation, according to Laurence Alexander, chemicals analyst at the investment firm Jefferies & Co. He told investors recently that with recent Food & Drug Administration approval for Mirel in food contact applications, the firm's addressable market size is about 4 billion lb per year. "Indicated customer interest to date should suffice to sell out the first commercial plant and justify expansions," he predicted.

But as other firms enter the biobased plastics market, Metabolix will need to compete on price. "Right now, PHA can withstand a certain premium in consumer products because it is biobased and biodegradable," says Samhitha Udupa, biobased materials analyst at Lux Research. "But that won't last more than five to seven years." Udupa says engineered crops may help get the price down, but she questions the scale the company would have to achieve to gain a sizable share of the polymers market.

All of the plant-based technologies face the same problem of reaching significant scale, Udupa says. "In recent years, venture capitalists and governments have been investing heavily in technology to convert biomass or sugar to chemicals or fuel. We've focused on how to get there but not on how much stuff goes into the front end." The challenge, she says, is growing and accessing the large amount of biomass that would be needed to displace a meaningful amount of petroleum.

At Mendel, Donald M. Panter, senior vice president for bioenergy seeds, is optimistic that farmers will want to plant perennial grasses modified for energy production because they can "make use of secondary land without using inputs or taking away from the food and fiber supply chain." According to the Department of Agriculture, 38 million acres of idled cropland is available for growing energy crops, if farmers feel they would make a profit.

Large agriculture businesses such as seed firm Monsanto and ethanol giant Poet would make good partners for the biotech firms, Udupa says, because they already have supply chains and distribution channels in place. Johnson agrees, and says Agrivida is looking to enter licensing deals with a large seed company or processor.

Plant biotech executives insist that the industry as a whole is developing more efficient technologies and processes that will make growing, collecting, and processing biomass for energy and chemicals profitable. But for now, the promise of large-scale cellulosic fuel production has yet to be fulfilled, and that concerns Johnson. "For our model to be truly successful, we'll be somewhat dependent on the success of others," he says.

 

Bayer Must Pay Dow $5M in Fees in Corn Patent Loss

By Mike Cherney
Law 360, USA
June 9, 2010

Bayer BioScience NV has been ordered to pay $4.9 million in attorneys? fees to Dow AgroSciences LLC following a ruling that four Bayer patents for genetically modified corn were invalid because of inequitable conduct before the U.S. Patent and Trademark Office.

Judge James A. Beaty Jr. of the U.S. District Court for the Middle District of North Carolina issued the ruling Tuesday, another blow to Bayer, which was ordered by a federal court in Missouri to pay nearly $8.4 million in attorneys? fees to Monsanto Co. in a similar dispute.

The current suit was originally brought by Aventis CropScience NV, which alleged Dow infringed four patents. The roles were reversed in Missouri, with Monsanto bringing suit and seeking a declaration that the Bayer patents were invalid. Bayer acquired Aventis CropScience in 2002.

The North Carolina suit was eventually stayed pending the outcome of the Missouri suit. In Missouri, the court granted summary judgment to Monsanto, finding the patents were invalid, but the U.S. Court of Appeals for the Federal Circuit reversed the decision and remanded the case for trial.

At trial, however, it was determined again the patents were invalid, and the judge determined the case was exceptional, allowing Monsanto to seek attorneys? fees from Bayer. The findings were once more appealed to the Federal Circuit, which affirmed them.

The stay was lifted in the North Carolina case in December 2008. Because the Federal Circuit upheld the trial court?s findings that the patents were invalid and the case exceptional, Bayer agreed it could not argue otherwise in the North Carolina action.

Bayer did contend, however, that Dow?s attorneys? fees request of just over $5 million should be denied, or at least reduced, because the amount was unreasonable. It also claimed that it had already been severely punished in the Monsanto case by being forced to pay fees there.

Judge Beaty, however, was not persuaded by Bayer?s arguments.

?The court?s decision to award attorneys? fees and costs does not hinge on whether a party has been sufficiently ?punished? in another proceeding, but instead, focuses on the issue of whether in the court?s discretion, [Dow] should be reimbursed for the cost of defending against a suit which Bayer has conceded was based upon invalid and unenforceable patents,? the judge said.

?The court notes that Bayer initiated these proceedings against [Dow] in this forum, seeking to exploit the protections afforded by the United States patent laws for patents that it improperly procured based upon its own inequitable conduct,? he added.

Ultimately, the judge awarded $4.9 million to Dow, trimming about $190,000 from the initial fees request to accommodate a reasonable rate for travel time and correct partners? hourly billing rates.

A second defendant in the case, Pioneer Hi-Bred International Inc., previously settled its fees request with Bayer.

The patents-in-suit are U.S. Patent Numbers 5,254,799; 5,767,372; 6,107,546; and 5,545,565.

Dow is represented by Orrick Herrington & Sutcliffe LLP and Smith Moore Leatherwood LLP.

Bayer is represented by Womble Carlyle Sandridge & Rice.

The case is Aventis CropScience NV v. Pioneer Hi-Bred International Inc. et al., case number 00-463, in the U.S. District Court for the Middle District of North Carolina.

To see court documents and read more articles on this subject, visit Law360.com.

 

Dawn of the Frankenfish

The Economist
June 10, 2010

The Belgian blue is an ugly but tasty cow that has 40% more muscle than it should have. It is the product of random mutation followed by selective breeding - as, indeed, are all domesticated creatures. But where an old art has led, a new one may follow. By understanding which genetic changes have been consolidated in the Belgian blue, it may be possible to design and build similar versions of other species using genetic engineering as a short-cut. That is precisely what Terry Bradley, a fish biologist at the University of Rhode Island, is trying to do. Instead of cattle, he is doing it in trout. His is one of two projects that may soon put the first biotech animals on the dinner table.

Belgian blues are so big because their genes for a protein called myostatin, a hormone that regulates muscle growth, do not work properly. Dr Bradley has launched a four-pronged attack on the myostatin in his trout. First, he has introduced a gene that turns out a stunted version of the myostatin receptor, the molecule that sits in the surface membrane of muscle cells and receives the message to stop growing. The stunted receptor does not pass the message on properly. He has also added two genes for non-functional variants of myostatin. These churn out proteins which bind to the receptors, swamping and diluting the effect of functional myostatin molecules. Finally, he has added a gene that causes overproduction of another protein, follistatin. This binds to myostatin and renders it inoperative.

The upshot of all this tinkering is a trout that has twice the abdominal muscle mass of its traditional counterparts. Moreover, this muscle is low in fat, like that of its bovine counterparts. That, and the fact that the animal's other organs are unaffected, means it does not take twice as much food to grow a fish to maturity.

The genetic engineers at Aqua Bounty, a company based in Waltham, Massachusetts, have taken a different route using a different species. They are trying to grow supersize salmon by tinkering with the genes for growth hormone. Two snippets of DNA are involved. One, taken from a relative of the cod called the ocean pout, promotes the activity of the gene that encodes growth hormone. The other, taken from a chinook salmon, is a version of the growth-hormone gene itself. Unmodified salmon undergo a period of restricted growth when they are young. Together these two pieces of DNA produce growth hormone during that lull, abolishing it. The result is a fish that reaches marketable size in 18-24 months, as opposed to 30 months for the normal variety.

It is one thing to make such fish, of course. It is quite another to get them to market. First, it is necessary to receive the approval of the regulators. In America the regulator in question is the Food and Drug Administration, which Aqua Bounty says it has been petitioning for more than a decade and which published guidelines for approving genetically engineered animals in 2009. Aqua Bounty has now filed its remaining studies for approval, and hopes to hear the result this year. Dr Bradley has not yet applied for approval.

It seems unlikely that either of the new procedures will yield something that is unsafe to eat. But what happens if the creatures escape and start breeding in the wild? For that to be a problem, the modified fish would have to be better at surviving and reproducing than those honed by millions of years of natural selection. On the face of it, this seems unlikely, because the characteristics that have been engineered into them are ones designed to make them into better food, rather than lean, mean breeding machines.

But there is a chink in this argument. As Mark Abrahams, a biologist at Memorial University in Newfoundland, points out, it is not just the fish that have been modified by man, but also the environment in which they could escape. Many of the creatures that eat salmon and trout, such as bears and some birds, have had their ranks thinned by human activity. Dr Abrahams thinks it possible that fast-growing salmon could displace the natural sort in places where predators are rare.

Aqua Bounty is addressing such concerns by subjecting developing eggs to high pressures. This alters their complement of chromosomes, giving them three sets per cell instead of the usual two. Such "triploid" fish are perfectly viable, but they are sterile. Only a small, sequestered breeding stock is allowed to remain diploid. The company claims a 99% success rate with its pressurising technique which, according to John Buchanan, its research director, meets the FDA's requirements. As for the trout, Dr Bradley says his fish have enough trouble breeding on their own for it to be unlikely that they would do well in the wild. To get them to lay eggs and produce milt (seminal fluid) you have to squeeze them by hand. But he says his fish could also be made triploid if necessary.

Whether people will actually want to buy or eat the new fish is another question - though they buy the meat of Belgian blue cattle at a premium. If people will pay extra for meat from a monstrosity like the Belgian blue, anything is possible.

 

A Potato Remade for Industry Has Some Swedes Frowning

By John Tagliabue
NY Times
June 10, 2010

SKARA, Sweden - Johan Bergstrom, a blond and boyish man of 31, who farms here with his father, reached into the dark, soft soil and extricated a tennis-ball-size potato, holding it gently so as not to snap off any of a half-dozen white shoots that were growing out of the potato's eyes. He advised against tasting the potato, whose dulcet name Amflora belies its harsh flavor, a result of genetic jiggling that has made it almost pure starch.

The potato, the first genetically engineered organism to be allowed in the European Union in more than a decade, was planted on 16 acres of land on the fringes of this town in southwestern Sweden, after a quarter-century of bureaucratic wrangling.

Although inedible, Amflora is a kind of miracle potato on two counts: for one, there is its starch content, which makes it precious to the starch industry, a major employer in Sweden; and then there is its feisty resilience in surviving some 25 years of tests, regulations, rules, ordinances and applications for approval by both Sweden and the European Union, of which Sweden is a member.

While not grown as a food crop, the Amflora potato is giving many people in this region of rolling hills, broad lakes and small farms a bad case of indigestion.

Though genetically engineered crops like corn, cotton or soybeans are common enough in the United States, they remain a rarity in Europe, where public resistance is high. The European Union takes the position that the long-term effects of genetic engineering on the environment and on plant and animal life cannot yet be known with scientific certainty, and so urges extreme circumspection. In few places is that caution as much in evidence as in Skara.

"I generally don't like modified potatoes, carrots, what have you," said Bengt Uilsun, 74, a veterinarian, interrupting his shopping one recent Friday morning. "Perhaps it's not unhealthy for humans now, but it may be unhealthy over the long term for other creatures."

Still, the Amflora is something of the pride of Sweden.

Development began in the mid-1980s, at the beginning of the revolution in biotech foods. A Swedish farmers' cooperative, Lyckeby, one of Europe's biggest starch producers, was searching for potatoes with high starch content to supply the starches it sells for manufacturing paper, textile finishes, glues and other products. "Genetic engineering was first emerging," said Kristofer Vamling, 51, managing director of Plant Science Sweden, a company that grew out of the original research efforts. "We thought this could perhaps be something for the new engineering."

But then, in 1998, the European Union imposed an indefinite moratorium on approval of genetically modified organisms, and no one at Plant Science knew when it would end. "I heard every year: 'Next year,' " Mr. Vamling said.

The moratorium was finally lifted in 2004, but it was another six years before the bureaucrats in Brussels, perhaps concerned about falling too far behind in biotech, gave the green light for planting. None too soon for Mr. Bergstrom.

Since 2004, when he finished agricultural college, Mr. Bergstrom has run a 590-acre farm just north of Skara, raising wheat, rye, barley and other crops. His family has farmed the lands here since the 1660s.

In 2006 a neighbor asked whether he wanted to try the new Amflora potato. "He asked if I was interested, we talked about it," Mr. Bergstrom said. "It's one more leg to stand on."

Holding one potato, he said, "I cannot tell the difference."

He is aware of the controversy.

"You need both sides," he said. "But the debate has gone the wrong way, and that's bad."

"I don't see any risk, or very low risk," he said. "There are so many papers to fill out; if only everyone did the same inspecting we do."

Just a few miles west of here, Anders Lunneryd, 47, disputes that. Working the 425-acre farm his grandfather bought in 1942, he grows wheat, oats, barley and a variety of other crops, but like an increasing number of farmers hereabout, he has done so organically for the past 10 years. When spraying his fields in the past with insecticides or weed killer, he explained, he often came too close to the village, and people would complain bitterly.

"I'd stop immediately, I'd tell them, if I could afford to," he said. "After all, I'm the one getting sick from all the chemicals."

As demand for organic crops soared, he switched to organic farming. "We have increased demand all the time," he said. "People are asking for it." Now about 7,000 acres of land in the area are organically cultivated, he said.

He objects to genetically modified foods, for their complexity and the control they give to big corporations. The genetic codes, he said, "are like a piano keyboard, but going four times around the planet earth, and now you're going to play that piano?"

"And it's even more complex," he said, "because you're playing in an orchestra."

He compares biotech crops in farming to performance-enhancing drugs in sports. "In the short run, it enhances your performance," he said. "In the long run, you get sick from it."

The bees that Ann-Charlotte Berntsson keeps along with angora rabbits and deer on her 90-acre farm are far upwind from Mr. Bergstrom's potato fields, but still she worries. The bee is a particularly sensitive animal, and in recent years entire bee colonies have been known to suddenly collapse, though no such significant cases have been reported in Sweden. Still, some experts speculate that poisons from biotech plants may be one cause.

"Modern agriculture is one of the bee's biggest threats, and we are also the farmer's best friend," she said. "The organic farmer is our best friend."

Not everyone in Skara shares her concern. "It's not a big issue," said Tomas Ek, 43, a local radio reporter, as he strode into a market on the main street. "People are more interested in the price of a product, and its fat content," he said, than whether it is genetically engineered.

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