| Good Intentions and Engineering Organisms that Kill Wheat
 by Elaine Ingham, Oregon State University
 <www.soilfoodweb.com>
 
 A genetically engineered Klebsiella-planticola had devastating
 effects on wheat plants while in the same kind of units, same incubator,
 the parent bacteria did not result in the death of the wheat plants.
 
 Consider that the parent species of bacteria grows in the root systems of
 every plant that has been assessed for Klebsiella's presence. The
 bacterium also grows on and decomposes plant litter material. It is a very
 common soil organism. It is a fairly aggressive soil organism that lives
 on exudates produced by the roots of every plant that grows in soil.
 This bacterium was chosen for those very reasons to be engineered:
 aggressive growth on plant residues.
 
 Field burning of plant residues to prevent disease is a serious cause of
 air pollution throughout the US. In Oregon, people have been killed
 because the cloud from burning fields drifted across the highways and
 caused massive multi-car crashes. A different way was needed to get
 rid of crop residues. If we had an organism that could decompose the
 plant material and produce alcohol from it; then we'd have a win-win
 situation. A sellable product and get rid of plant residues without
 burning. We could add it to gasoline. We could cook with it. We could
 drink grass wine-although whether that would taste very good is
 anyone's guess. Regardless, there are many uses for alcohol.
 
 So, genes were taken out of another bacterium, and put into
 Klebsiella-planticola in the right place to result in alcohol
 production. Once that was done, the plan was to rake the plant residue
 from the fields, gather it into containers, and allow it to be
 decomposed by Klebsiella-planticola. But, Klebsiella would produce
 alcohol, which it normally does not do. The alcohol production would
 be performed in a bucket in the barn. But what would you do with the
 sludge left at the bottom of the bucket once the plant material was
 decomposed? Think about a wine barrel or beer barrel after the wine
 or beer has been produced? There is a good thick layer of sludge left
 at the bottom. After Klebsiella-planticola has decomposed plant
 material, the sludge left at the bottom would be high in nitrogen and
 phosphorus and sulfur and magnesium and calcium-all of those
 materials that make a perfectly wonderful fertilizer. This material
 could be spread as a fertilizer then, and there wouldn't be a waste
 product in this system at all. A win-win-win situation.
 
 But my colleagues and I asked the question: What is the effect of the
 sludge when put on fields? Would it contain live Klebsiella-planticola
 engineered to produce alcohol? Yes, it would. Once the sludge was spread
 it onto fields in the form of fertilizer, would the
 Klebsiella-planticola get into root systems? Would it have an effect
 on ecological balance; on the biological integrity of the ecosystem; or on
 the agricultural soil that the fertilizer would be spread on?
 
 One of the experiments that Michael Holmes did for his Ph.D. work was to
 bring typical agricultural soil into the lab, sieve it so it was nice and
 uniform, and place it in small containers. We tested it to make sure it
 had not lost any of the typical soil organisms, and indeed, we found
 a very typical soil food web present in the soil. We divided up the
 soil into pint-size Mason jars, added a sterile wheat seedling in
 every jar, and made certain that each jar was the same as all the
 jars.
 
 Into a third of the jars we just added water. Into another third of the
 jars, the not-engineered Klebsiella-planticola, the parent organism,
 was added. Into a final third of the jars, the genetically engineered
 microorganism was added.
 
 The wheat plants grew quite well in the Mason jars in the laboratory
 incubator, until about a week after we started the experiment. We came
 into the laboratory one morning, opened up the incubator and went,
 "Oh my God, some of the plants are dead. What's gone wrong? What did
 we do wrong?" We started removing the Mason jars from the incubator.
 When we were done splitting up the Mason jars, we found that every
 one of the genetically engineered plants in the Mason jars was dead.
 Wheat with the parent bacterium, the normal bacterium, was alive and
 growing well. Wheat plants in the water-only treatment were alive and
 growing well.
 
 From that experiment, we might suspect that there's a problem with this
 genetically engineered microorganism. The logical extrapolation from this
 experiment is to suggest that it is possible to make a genetically
 engineered microorganism that would kill all terrestrial plants. Since
 Klebsiella-planticola is in the root system of all terrestrial plants,
 presumably all terrestrial plants would be at risk.
 
 So what does Klebsiella-planticola do in root systems? The parent
 bacterium makes a slime layer that helps it stick to the plant's roots.
 The engineered bacterium makes about 17 parts per million alcohol.
 What is the level of alcohol that is toxic to roots? About one part
 per million. The engineered bacterium makes the plants drunk, and
 kills them.
 
 But I am not trying to say that all genetically engineered organisms are
 technological terrors. What I am saying is that we have to test each and
 every genetically engineered organism and make sure that it really does
 not have unexpected, unpredicted effects.
 
 They have to be tested in something that approximates a real world
 situation. I've worked with folks in the Environmental Protection Agency
 (EPA) and I know the tests the EPA performs on organisms. They often begin
 their tests with "sterile soil." But if it's sterile, then it's not really
 soil. Soil implies living organisms present. If you use "sterile soil" and
 add a genetically engineered organism to that sterile material, are you
 likely to see the effects of that organism on the way nutrients are
 cycled, or on the other organisms in that system? No, you're not
 likely to. So it's probably no surprise that no ecological effects
 are found when they test genetically engineered organisms in sterile
 soil. They really need to put together testing systems, which assess
 the effects of the test organism on all of the organisms present in
 soil.
 
 What do we mean, organism-wise, when we talk about soil? Agricultural
 soil should have 600 million bacteria in a teaspoon. There should be
 approximately three miles of fungal hyphae in a teaspoon of soil. There
 should be 10,000 protozoa and 20 to 30 beneficial nematodes in a teaspoon
 of soil. No root-feeding nematodes. If there are root feeding
 nematodes, that's an indicator of a sick soil.
 
 There should be roughly 200,000 microarthropods in a square meter of soil
 to a 10-inch depth. All these organisms should be there in a healthy soil.
 If those conditions are present in an agricultural soil, there will be
 adequate disease suppression so that it is not necessary to apply
 fungicides, bactericides, or nematicides. There should be 40 to 80% of the
 root system of the plants colonized by mycorrhizal fungi, which will
 protect those roots against disease.
 
 What happens when you apply the most fungicides and pesticides to soil?
 In every single case where we have looked at foodweb effects of
 pesticides, there are non-target organism effects, and usually very
 detrimental effects. The sets of beneficial organisms that suppress
 disease are reduced. Organisms that cycle nitrogen from
 plant-not-available forms into plant-available forms are killed.
 Organisms that retain nitrogen, phosphorus, sulfur, magnesium,
 calcium, etc. are killed. Organisms that retain nutrients in the soil
 are killed. Once retention is destroyed, where do those nutrients go?
 They end up in our drinking water; or end up in our ground water. You
 and I as taxpayers have to pay in order to clean up that water so we
 can drink it.
 
 Wouldn't it be much wiser to keep those organisms present in the soil so
 those nutrients would be retained and become available to the next crop of
 plants instead of ending up in our drinking water where we have to pay in
 order to have clean drinking water? How do you do that? You get the
 organisms back into the soil. If you grow the proper number and types of
 bacteria, fungi, protozoa, nematodes and microarthropods, mycorrhizal
 fungi in the root systems of the plants, you can do away with
 pesticides. It's been done. We can reduce significantly the amount of
 fertilizer that goes into that soil. In experiments that have been
 done all over the country, all over the world, inorganic fertilizer
 inputs have been reduced, or are not added at all, without reduction
 in plant growth. Where green manure or legumes are not available,
 approximately 40 pounds of nitrogen fertilizer, once every four
 years, are still necessary.
 
 Let's talk about why today's conventional agricultural systems require
 such massive inputs of pesticides and fertilizers. When a healthy soil is
 first plowed out of native grassland, for example, the disease-suppressive
 bacteria and fungi, protozoa and nematodes are present. For the first 5 to
 15 years after plowing native grassland you don't have to use any
 pesticides. No fertilizers are required because there is natural nutrient
 cycling, natural nitrogen retention, and disease suppression. As you plow
 that soil, you start to kill the beneficial organisms, you lose the
 organic matter, and you lose the food to feed the beneficial
 organisms. After about 10 to 15 years, if you're not adding back
 adequate plant residue to feed those organisms, you lose them, and
 start having significant disease problems. Then you either leave that
 land and farm elsewhere, or in the US, we used fertilizers to keep
 yields high. As more and more of the organisms were killed by the
 salt effect of the fertilizers, and the constant plowing mined out
 more and more of the organic matter, starving the beneficial
 organisms to death, disease became a serious problem. And we started using
 more and more pesticide to knock the disease back.
 
 In California, around 1955, those disease problems became so severe that
 they thought they would lose agricultural production. So the University of
 California came up with a better way to kill those disease-causing
 organisms. It's called methyl bromide. This chemical kills disease-causing
 organisms-but it also kills everything else. There is very little natural
 disease suppression going on in agricultural soils in California.
 
 How many organisms are left in strawberry fields that have been
 methyl-bromided 2 to 3 times a year for the last 14 years? There are no
 microarthropods left. There are no beneficial nematodes left; only root
 feeding nematodes. And there is nobody to control root-feeding nematodes
 in those soils. How many protozoa are left in that soil? None. You
 cannot cycle nutrients. There is nobody home to make nitrogen
 plant-available. So what do you have to do? You have to add
 fertilizer. We force ourselves to have to add fertilizer. We have no
 other choice if you're going to grow those plants in those soils.
 
 How many fungi do you have left in that soil? No beneficial fungi-they're
 all disease-causing. How many bacteria are left? All are gone, except for
 100 per gram of soil. We should have 600 million per teaspoon in that
 soil; we have 100 left. There is nothing left to retain nitrogen in
 those soils, nothing. So you apply fertilizer. What happens to the
 fertilizer? Whatever fertilizer contacts the roots of the plants is
 indeed taken up; the rest of it flushes through the soil into the
 ground water, into the river. Take Santa Maria River in California as
 an example. This land has had methyl bromide applied 2 to 3 times a
 year for the last 14 years or more. Fertilizer is applied as
 sidedress when strawberries are planted. About two weeks later, the
 river goes up to around 150 parts per million nitrates. What is the
 toxic level for nitrate for humans? Ten parts per million nitrates is
 what the EPA tells us. It used to be three parts million but that
 evel was increased. Can you drink that water in the river in the Santa
 Maria valley? Not unless you'd want to die. You would destroy your kidneys
 pretty fast if you drank that water. It is high in nitrate. It is so toxic
 that you can't even put that water back on the plants. The high nitrate
 burns the plants.
 
 We have a simple solution for this problem. Get the right kind of
 organisms, the right numbers of organisms, back in the soil and let them
 start performing their functions again. Put food for the organisms back
 into the soil; put the organisms back into the soil. It's that
 simple. Send us your soil samples and we can tell you whether you
 have that food web in your soil.
 
 How are you going to fix that set of organisms it if you don't have a
 healthy foodweb? We can help you with that question. We can indeed move
 towards that time when we really don't need pesticides anymore; where you
 only apply fertilizer once every four years and in very small amounts. We
 can move to a sustainable agriculture. It takes time and effort, but it is
 possible.
 
 This article is adapted from the presentation the author gave on July
 18, 1998 at the First Grassroots Gathering on Biodevastation: Genetic
 Engineering.
 
 See also: Holmes, M.T., Ingham, E.R., Doyle, J.D., & Hendricks, C.W.
 (1998). Effects of Klebsiella-planticola SDF20 on soil biota and
 wheat growth in sandy soil. Applied Soil Ecology, 326, 1-12.
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