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INTERNATIONAL BIOSAFETY TRAINING:A TECHNICAL REPORTBy JEROLET C. SAHAGUN,Project Evaluation Officer, FPA

PART II. HIGHLIGHTS OF THE TRAININGThe biochemical basis and development of Bacillus thuringiensis (Bt)Plant Incorporated Protectant (PIP)

In studying the characteristics of the source of Bt strains, the biochemistryand the early attempts to improve the microorganisms through geneticengineering will provide a context on how to deal with the environmentalbiosafety issues associated with using Bt crops.Bt grows rapidly and reproduces asexually by simple cell division (a.k.avegetative growth). Bt cells produces a spore that only germinates whenconditions is favorable again. During sporulation, it produces a crystallineproteinaceous inclusion called the parasporal body. When certain insectincidentally ingest the sporulated Bt cells, with their parasporal body, thealkaline midgut (i.e insect digestive tract) solubilizes the crystallineparasporal body releasing protein toxin known as delta-endotoxin. Theendotoxins are protoxins that must be cleaved by the insect midgutproteases into the molecular from that eventually kills the insect due tobacterial septicemia.Inconsistency of result of using Bt products that were deployed in agricultureand forestry before 1970, led the scientist in discovering a new isolate of Btwhich was found to be 200 more times more active against pest targeted bythe existing Bt products. This new isolate represents the new subspecies, andwas called

kurstaki

and was given the appellation HD-1. The

Kurstaki Bt

isolates were found to be variably pathogenic to differentspecies of Order Lepidoptera. Also another Bt strain toxic to primitive flies of the Order Diptera (mosquitoes and blackflies) was isolated and namedsubspecies

israelensis.

Today there are approximately 280 Bt strains that have been isolated frominsects, foliage and grain dusts. These strains are differentiated by thecharacteristics of the crystalline protein, its gene sequence and its spectrumof insecticidal activity. The various strains that contain one or more differentendotoxins that are designated by the acronym Cry which means crystallineprotein. The Bt crops were developed in order to address the limitations of Btinsecticides which has very limited coverage and found only to be pathogenicto larvae. In addition to this, the Bt insecticides quickly breakdown when

exposed to sunlight so that farmers apply it repeatedly spending more timeand money. The different strains of Bt produce different Cry proteins that are toxic to avariety of insect. Bt toxin works when an insect ingested the protoxins, thistoxin is solubized in high pH and I is activated when it cleaved to the insect’smidgut by the gut proteases. Later the insect will die due to bacterialsepticemia. Bt toxin is practically non-toxic to human because human beingshas gut low in pH and has no gut receptors. The Bt crops has Bt protein in all parts of the plant. It means insecticide iswithin the plant in high dose and will not easily degrade even plants areexposed to sunlight. The economic issue is only in the cost of seeds.Another GM crop is the Roundup Ready crops which are geneticallyengineered to tolerate the herbicide Roundup. It carries a version of 5-enolpyruvylshikimate-3-phospahate synthase (EPSPS) from agrobacteriumresistant to glyphosate inhibition. It is produced by monsato Company. Itwas developed in order to address problem on weeds. Using the RR crops,the farmers could use the glyphosate as a post-emergence herbicide againstmost broadleaf and cereal weeds. The farmers could spray their fields toeliminate weeds without damaging the crop. There are a lot of RoundupReady crops that are already in commercialization stage.

Insect Resistance Management

In insect resistance management, human factors are important. It dependson the strength of enforcement and the question of whether there arepenalties for those who violate the agreement of having a 20% refuge infield. According to Dr. difonzo, an entomologist in MSU, the challenges of working with transgenic crops is that it requires higher level of growereducation and it is currently very controversial.Dr. Edward J. Grafius (Department of Entomology, MSU) had discussed the Theoretical basis for pest resistance management. According to him thepesticide resistance serves as a model for resistance management fortransgenic crops. Currently, pest resistance to pesticides is one of the mostimportant problems in worldwide agriculture. Some scientists think thatresistance to genetically engineered crops may become an equally importantissue.Resistance means tolerance to pesticides, tolerance to geneticallyengineered crops or host plant resistance. According to Dr. Grafius, pesticideresistance is the heritable ability of the pest to withstand a pesticidetreatment that would normally be lethal and it is not an acquired trait.Organisms have gain resistance to pesticides due to the evolution of anorganism in response to selection pressure or mortality which includepreexisting adaptation to toxins (e.g. plant defensive chemicals), naturalvariability and mutations. Currently the good agricultural practices (GAP) hasbeen employed to manage the pesticide resistance by using the integrated

Pest Management Techniques (Diverse Mortality Factors), chemicalmanagement, refuges for susceptible genotypes and resistance monitoring. The parallels in the management of transgenic crop resistance are thefollowing:

•

Alternate chemicals = different resistance genes in differentvarieties

•

Tank mixes = gene pyramiding

•

High dose use of insecticides = high gene expression

•

Refuges = crops without the resistance genes or weed hosts of the pestBased on the report of Dr. Grafius, it is concluded that in order to managetransgenic crop resistance the following should be generally employed by thegrowers:

•

Use multiple control strategies, including Bt varieties, croprotation, standard insecticides, biological control, etc.

•

Leave refuge of non-Bt crop within normal dispersal distance foryour pest

–Require by law if necessary

–Use crop rotation to increase gene flow

•

Monitor Bt fields for surviving individualsIt was also suggested that additional management factors and inplaceresistance management before a problem arise are necessary to effectivelymanage the resistance.

The Gene Flow from GE Crops to wild relatives

The principles involved in Gene flow from GE crops to its wild relatives werediscussed by Dr Jim Hancock of MSU-Department of Horticulture. Accordingto Dr. Hancock, the different concerns about the environmental risk of GEcrops are the non-target effects of novel toxins; development of resistance inpests, transgene will alter crop fitness and competitive ability making it moreinvasive; pollen flow from GE crop will alter native species fitness; andcompetitive ability making it more invasive. The overall concern is that GEcrops and related species will become more invasive and reduce biodiversity.Based on history, successful crop and wild hybridizations are common mostespecially if crops grown adjacent to wild relatives. Several factors that canslow rates of gene flow are breeding system (selfing vs. outcrossing), agentof pollination (wind vs. insect), type of seed dispersal and flowering time.Several studies show that all crops have relatives somewhere or anywhere inthe world and there will be a great possibility of gene to escape if relative isgrowing nearby. Biosafety issues related to gene flow will focus on the threemajor factors to determine the environmental risk of GE crops such aspresence of compatible relative, invasiveness of crop or native relative and