HORT 250 - Biotechnology in Agriculture

Lecture 16 - Transgenic plants with resistance to insects (continued)

 

Last time we discussed two strategies that have been used to develop transgenic plants with resistance to insects.

Note that there are some important differences between these two strategies. Bt toxin is normally lethal to the insect that ingests this protein. In contrast, the protease inhibitors and alpha-amylase inhibitors are not lethal but will merely slow down the growth, maturation and life cycle of the insect. From studies done in other systems, the selection pressure that is applied by the Bt toxin is more likely to result in the evolution of insects with resistance to this toxin. In contrast, resistance is less likely to develop to a control method that merely slows down development.

Both of these methods have shown promise and commercial products have been developed that incorporate the Bt toxin in potatoes, cotton and corn. What are the economics of using these Bt crops? I don't know the details of this for corn, but the cost of hybrid corn seed with a Bt toxin gene is a little higher than regular corn hybrids. Growers must therefore weigh this against the cost of other measures to control European corn borer (insecticides), or the yield loss that might result from not using any insecticide, with a non-Bt hybrid.

For cotton, the economics are a little more complicated. Bt cotton seed costs about the same as regular cotton seed. However, the grower must pay Monsanto a licensing fee of $32 per acre to plant Bt cotton. In 1996, approximately 1.8 million acres of cotton were planted to Bt cotton, generating revenues of about $60 million dollars for Monsanto! The normal cost for insect control in cotton might be between $100 and $200 per acre, to the best of my knowledge. Remember that cotton accounts for the largest fraction of insecticide use in the U.S. If the Bt cotton yields the same as conventional cotton, then there is a clear advantage to the Bt cotton purely on the basis of production costs. Even if there is a need for one or two applications of insecticides at about $10 per acre per spray, the Bt cotton is likely to come out ahead.

This is a fairly simple analysis, but it gives an indication as to the economic advantages of using a genetic/biotechnological solution to controlling insect pests. In addition to these cost considerations, there are a number of other advantages to using this method (expressing proteins in plants to control insects) that are discussed below, followed by a discussion of the major problem, development of insects with resistance to these control measures.

What additional benefits are likely to come from these biotechnology methods to control insect pests?

  1. Reduced application of chemical pesticides.
  2. Potentially dangerous pesticides may be eliminated.
  3. Efficacy of the treatment is independent of the weather. For example, the Bt toxin that is expressed in the plant cannot be washed off by rain.
  4. These controls are in place throughout the growing season. If the Bt toxin is expressed throughout the growing plant, then the toxin will be ingested by the insect whenever it feeds on the plant.
  5. Treatment can be designed to be truly systemic, reaching all parts of the plant that pesticides may not penetrate. It may be difficult for insecticides to get into developing cotton bolls for purely physical reasons. But the Bt toxin can be expressed specifically in these tissues.
  6. The treatment can be designed to be in place when the insect is at its most sensitive stage of its life cycle, when it is easier to control. For example, some insects are easier to control at a young larval stage. It may not be possible to spray when the insects are at that stage, or the insects may develop throughout the course of the growing season. The Bt toxin can be designed to be expressed throughout the season, so that the feeding insects will always take up the toxin. This benefit is similar to the previous two above.
  7. The only insects that are affected are those that actually feed on the plant, because the plant is the only source of the insecticide. "Beneficial" insects and others that do not feed on the crop are not affected.
  8. The active agent is usually a biodegradable protein, so there is little potential for ground water contamination or other environmental problems.
  9. There is widespread consumer acceptance of this technology, compared to the use of pesticides.
  10. There are no human toxicology problems with Bt toxin proteins. Even though the toxins are integrated into the plasma membrane in insects, they are not toxic to humans and are simply digested as part of the diet.

What are the potential problems with these biotechnology methods to control insect pests?

The most important concern is that insects will develop resistance to these Bt toxins. It has been argued by some that this is unlikely to occur, because Bt spores have been used over the past 20 years and there have been essentially no reports of insect resistance developing against these controls. However, there are a number of important differences between using Bt spores as insecticides and expressing a Bt toxin in transgenic plants.

Currently, Bt crops typically express only a single Bt toxin protein, at relatively high levels, throughout the growing season of that plant. Under these conditions, it is much more likely that insects will develop resistance to Bt crops than they will to Bt spores. First, there is high selection pressure when a population of any organism is exposed continuously to a toxin. This is compared to the transient exposure that insect populations face with Bt spores. Second, insects need develop resistance only to a single toxin expressed in the plant, compared to the suite of toxins in spores. Simultaneous development of several different resistance mechanisms is highly unlikely.

Therefore, in spite of the fact that few examples of resistance to Bt spores have been reported, there is good reason to expect that insects will quickly develop resistance to Bt toxin in plants. Already there are some results pointing in this direction.

  1. Laboratory strains of insects have been selected that show resistance to individual Bt toxins. Resistance has been shown to be caused by at least two different mechanisms. Insects may lack the protease in their gut that is responsible for cleaving the protoxin into the active toxin molecule. Mutations in the receptor protein that normally mediate incorporation of the toxin into the membrane can prevent the toxin from being incorporated into the gut cell membrane.
  2. Studies of insects in the field where Bt crops are growing. Insects that are targeted for control by Bt may be developing resistance, but there may be other reasons for finding them in these fields (just passing through, or a new arrival in that field).
  3. Recent genetic studies indicate that alleles for Bt resistance are already present at a fairly high level in insect populations. By crossing insects captured in fields of Bt crops with lab strains that are already resistant to Bt toxin, it is possible to estimate the frequency of genes for resistance to Bt toxins in the wild insect populations. These studies indicate that Bt resistance alleles are present at a frequency of around 1 in a 1,000 (Proc. Natl. Acad. Sci. 94:3519-3523). This does not mean that 1 out of every 1,000 insects is resistant, because resistance is a recessive genetic trait. Therefore, the frequency of resistant individuals may be closer to 1 in a million.

These and other results clearly demonstrate that the genetic potential to develop resistance to Bt toxins exists in insect populations. I think it is widely accepted that insects will develop resistance to this control method, just like they have developed resistance to most other insecticides.

How can the potential problem of insects developing resistance to Bt toxins be countered? Many of the companies that are involved in this area of research and development are very concerned about this problem. A couple of strategies have been proposed to delay or prevent the appearance of resistance.

The first is to plant a mixture of Bt and non-Bt varieties, not interspersed but in adjacent blocks. The non-Bt blocks are known as refuges. My understanding is that the current recommendation to growers is to plant 20% non-Bt corn which can be treated with another insecticide, or 5% non-Bt corn which cannot be treated with any insect control. These provide refuges for insects that are not resistant to the Bt toxin.

To discuss how refuges can delay the development of insect resistance, first we will discuss the likely genetics of this trait. Resistance is recessive, i.e. individuals that are r/r will be resistant, while sensitive individuals can be either S/S or S/r. (S and r are different alleles at the same locus.) If resistance to a single Bt toxin is a dominant trait, there is little that can be done to prevent this spreading rapidly through an insect population. The refuge model also assumes that Bt-resistant individuals, and likely the heterozygotes, are at a selective disadvantage under normal (non-Bt) conditions compared to the sensitive insects. The refuge model works like this, at least in theory:

This strategy is based on population genetics and is sound in theory. However, it does rely on grower compliance. Will a farmer willingly plant some of his/her crops with a variety that is susceptible to insects? Maybe he/she will rely on the neighbor to be the one to plant the non-Bt varieties. It should be emphasized that this strategy is not going to prevent the development of insect resistance to Bt toxins, it is designed merely to slow down the inevitable spread of resistance.

Are there alternatives that are being considered to enhance the long term usefulness of Bt toxins in plants? This is an important question because Bt toxins are a valuable biological resource, one of the few examples of highly active insecticidal proteins. It is in no one's interest to see this resource lost quickly by using it injudiciously in the first generation of transgenic crops. What alternatives are being considered?

In the future it may be possible to design genes that can encode novel proteins to perform specific functions, such as kill or inhibit the growth of insects. But our current understanding of protein structure and function do not yet make this sort of rational design of proteins possible.

Two final points to consider with regard to transgenic plants with resistance to insects:

  1. While organic farmers generate only a small fraction of agricultural production, should there be any concern that they might lose one of the few organic methods for insect control that is available to them? If insects develop resistance to Bt crops, organic farmers may find that Bt spores are no longer able to control insect pests on their crops.
  2. If a similar investment had been made designing non-chemical control methods as has been spent on the development of chemical pesticides, would there be a number of alternatives to control insects in crops?

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