Purdue University scientists Natalia Dudareva and Joseph Lynch have been searching for a
way to increase a plant’s production of phenylalanine, a compound important for
plant survival and used by humans in flavors, fragrances, biofuels,
insecticides and pharmaceuticals. Their work led to discovery last year of a
previously unknown metabolic pathway that they thought could be engineered to
allow plants to produce more phenylalanine than they do on their own.
A genetic modification that should have ramped up phenylalanine production
led to an unexpected reduction of the compound. This setback, however,
illuminated a hidden connection between phenylalanine biosynthesis and the
plant hormone auxin, which has implications for not just amino acid metabolism,
but also our understanding of growth and development.
“For many years, we didn’t know how fluctuations through these pathways were
regulated and interconnected with plant hormones and other compounds,” said
Dudareva, a distinguished professor of biochemistry and member of Purdue’s Center for Plant Biology, whose findings were published in
Nature Chemical Biology. “We found a cross-talk with auxin, which may explain
why plants don’t use this second pathway and create larger amounts of
phenylalanine.”
Plants use phenylalanine as building blocks for compounds to attract
pollinators, for defense, reproduction, growth and development. While
sufficient for those purposes, the amounts are small for human uses.
Phenylalanine production happens mainly in plastids, the small organelles
such as chloroplasts. But Dudareva, Lynch, who is a Purdue research scientist,
and graduate student Yichun Qian discovered that plants also can produce
phenylalanine in cytoplasm and may be able to make larger quantities there.
The scientists grew petunias to maturity, and then induced production of an
enzyme that would increase phenylalanine production in the cytosol.
“It worked beautifully. We got a threefold increase in phenylalanine
synthesis,” Lynch said.
Then they integrated a gene into the petunia genome that would increase
production of the same enzyme, which should have yielded similar results.
Instead, phenylalanine production increased slightly in the cytosol, but
dropped significantly in the plastids. That led to an overall decrease in
phenylalanine production.
That’s because both phenylalanine and auxin, a plant hormone necessary for plant
growth, can use a compound called phenylpyruvate as a substrate for
biosynthesis. By producing more phenylalanine in the cytosol, phenylpyruvate
increased in that compartment and created more auxin.
Slight variations in plant hormones can cause significant developmental
problems. In this case, the increase in auxin led to the production of fewer
plastids and a drop in phenylalanine output.
“Our strategy for creating more phenylalanine won’t work. We kind of hit a
dead end because of the unexpected cross-talk with auxin,” Lynch said. “We will
continue to try to increase phenylalanine, but we’ll work through the plastid
pathway and try to overcome the bottlenecks that limit production there.”
Dudareva said the findings not only show how phenylalanine and auxin are
linked, but offer a suggestion about why plants have the less-often-used
cytosolic pathway at all.
Plants likely produce enough phenylalanine through the tightly regulated
plastid pathway and don’t produce more so as not to throw off the auxin balance.
But when a plant is injured and needs more phenylalanine for defense or to
heal, the cytosolic pathway can kick into gear to provide what’s needed.
“It looks like the pathway is used by plants as a first response to stress
or damage,” Dudareva said. “This is important to know because initially it
wasn’t clear if plants used this pathway at all for phenylalanine
biosynthesis.”
The National Science Foundation and the U.S. Department of Agriculture
National Institute for Food and Agriculture supported this research.
This story was written by Brian Wallheimer.
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