Designer Genes May Quell Fungus
The researchers designed genes to cue wheat plants to make enzymes
called chiti nase and glucanase. These are the types of enzymes Fusarium
manufactures when it needs to tear down some of its cell walls to grow
and expand. Cell walls of Fusarium are made up, in part, of compounds
called chitin and glucan, which the enzymes break down. This process
occurs over and over again in the growing tip of the fungus's microscopic,
root like structure, called a hypha.
A hypha is formed when a Fusarium sporeperhaps carried
by wind or rainlands on a wheat floret and germinates. The hypha's
mission is to reach the developing kernel, where a storehouse of energy-rich
starch compounds awaits.
In rebuilding wheat plants to make cell-wall-degrading chitinase and
glucanase, the scientists intend to disrupt the hypha's orderly progress
toward the food source inside the wheat plant. The strategy is to overwhelm
the fungus with chitinase and glucanase that it didn't make and can't
control.
Scientists have known about the role of chitinase and glucanase for
years. But Okubara, Blechl, Hohn, and Berka are the first to use pieces
of Fusarium chitinase and glucanase genes for making unique,
antifungal genes. To mimic the microbe's own enzyme-making machinery
as closely as possible, the researchers borrowed gene segments from
a F. graminearum relative called F. venenatum. Its genes
had been copied earlier into a "library" maintained by Novo
Nordisk. That made this cousin a convenient source of the needed material.
Tooling Wheat To Make Crucial Enzymes
The scientists already knew that engineering a wheat plant to make
chitinase or glucanase was unlikely to hurt the plant itself. That's
because there's little or no chitin or glucan in wheat cell walls.
Okubara, Hohn, and Marcie Moore, who is with ARS at Peoria, have extensively
rebuilt some of the F. venenatum genes. They have, for instance,
filled in missing pieces of the original F. venenatum chitinase-forming
gene. Okubara also outfitted the genes with a custom-built promoter.
This on-off switch would enable a wheat plant to better use the newly
redesigned genes.
At Albany, Blechl moved the lab-built genes into wheat embryos. She
started by excising the embryos from immature wheat kernels. Then she
placed the embryos into petri dishes positioned in the line of fire
of a gene gun, or bioblaster. Blechl ferried the genes into the embryos
on gold particles propelled by the gene gun.
Later, Blechl tested the target embryos to see whether they had taken
up the genes. If they had, she nurtured them into healthy greenhouse
plants. Blechl then shipped seeds from the plants to colleague Ruth
Dill-Macky in St. Paul, Minnesota, for a rigorous program of screening.
Dill-Macky, a plant pathologist with the University of Minnesota, is
part of a team that evaluates promising new wheat plants for resistance
to Fusarium head blight. The indoor and outdoor tests pinpoint new lines
that may someday replace current varieties that have been clobbered
by the fungus in the past.
Pyramiding: A Strategy To Strengthen Defense
Preliminary results with the novel genes have been mixed. "But
that's not unusual for biotech experiments," comments Blechl. "Our
priority is to make sure the plants are using, or expressing, the new
genes to the greatest extent possible. We don't know yet if we've reached
that point."
In addition, bundling these genes with others is likely a key to packing
a more powerful punch against Fusarium head blightan approach
known as pyramiding. To fend off Fusarium, plants of the future
might be loaded with a multiplicity of antifungal genes, for what's
called multiple-gene resistance. The plant and this pathogen have co-evolved
in such a way that wheat plants probably turn on many genes to survive
attack.
Yet wheat harbors no single gene that enables the plants to eliminate
the fungus. "Following nature's lead," explains Blechl, "we're
continuing to develop an array of genes. Some are from organisms other
than Fusarium. We think that carefully developed combinations
of unique genes may provide the ultimate protection against this destructive
fungus."
The research is funded in part by the North American Millers' Association
and the U.S. Wheat and Barley Scab Initiative.By Marcia
Wood, Agricultural Research Service Information Staff.
This research is part of Plant Biological and Molecular Processes,
an ARS National Program (#302) described on the World Wide Web at http://www.nps.ars.usda.gov.
For more information on U.S. Patent Application Serial No. 09/649,747,
"Nucleic Acid Sequences Encoding Cell-Wall Degrading Enzymes and
Their Use To Engineer Resistance to Fusarium and Other Pathogens,
contact Patricia A. Okubara, USDA-ARS
Root Disease and Biological Control Research Unit, P.O. Box 646430,
Washington State University, Pullman,
WA 99164-6430; phone (509) 335-7824, fax (509) 335-7674, or Ann
E. Blechl, USDA-ARS Crop
Improvement and Utilization Research Unit, Western Regional Research
Center, 800 Buchanan St., Albany, CA 94710; phone (510) 559-5716, fax
(510) 559-5777.
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