Klebsiella planticola--The Gene-Altered Monster

That Almost Got Away

The Deadly Genetically Engineered Bacteria that Almost Got Away: A

Cautionary Tale



Web Note: In the early 1990s a European genetic engineering company was

preparing to field test and then commercialize on a major scale a

genetically engineered soil bacteria called Klebsiella planticola. The

bacteria had been tested--as it turns out in a careless and very

unscientific mannner--by scientists working for the biotech industry and

was believed to be safe for the environment. Fortunately a team of

independent scientists, headed by Dr. Elaine Ingham of Oregon State

University, decided to run their own tests on the gene-altered Klebsiella

planticola. What they discovered was not only startling, but terrifying--

the biotech industry had created a biological monster--a genetically

engineered microorganism that would kill all terrestrial plants. After

Ingham's expose, of course the gene-altered Klebsiella planticola was never

commercialized. But as Ingham points out, the lack of pre-market safety

testing of other genetically altered organisms virtually guarantees that

future biological monsters will be released into the environment. Moreover

it's not only genetic engineering that poses a mortal threat to our soil

ecology, the soil food web, as Ingham calls it. Chemical-intensive

agriculture is slowly but surely poisoning our soil and our drinking water

as well.



This article orginally appeared in the Green Party publication

Synthesis/Regeneration 18 (Winter 1999)

Ecological Balance and Biological Integrity

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.

