1. Algae
at
the
Nexus
of
21st
Century
Food,
Energy
and
Water
Needs?
George
A.
Oyler
MD
PhD
October
17,
2011
Big
Red
is
The
New
Green

2. BioEnergy
ENERGY
Agriculture
Environmental
Products
Enhancement
FOOD
WATER

3. Why
Algae
for
Biofuels?
Algae
have
great
potenAal
in
biofuels
and
GHG
abatement
• Algae
have
extremely
high
producJvity.
• Algae
photosyntheJc
and
sequester
CO2.
• Algae
do
not
compete
with
food
crops.
• Algae
do
not
require
farm
land.
• Algae
can
remediate
water.
• Use
of
algae
for
wastewater
treatment
offers
huge
value
added
potenJal.
Caveats:
•
Algae
biofuels
are
not
yet
economically
viable.
•
Major
breakthroughs
in
both
engineering
and
biology
are
required.

7. World
human
populaJon
vs
Jme
(human
history)

12. •
• Midwest corn
production for
biofuels has lead
Hypoxic to record size of
Zone Dead Zone this
summer.

16. Hypoxic zones are a world problem
Oxygen-depleted
coastal zones
Persistent
Annual
Episodic
Periodic

19. Algae can be combined with animal agriculture, AD, and
Aquaculture - an integrated industry for Nebraska and Europe
Algae
producJon
Aquaculture
producJon

21. Algae industry can be phased in with high
value products
Therapeutics

23. Highlights
of
Algal
Research
at
The
University
of
Nebraska-­‐Lincoln
Presenter:
Don
Weeks

26. Transcriptomic analysis of expression of TAG
biosynthesis genes under N deprivation

28. Raman spectra of algae on a gold surface

29. Raman image of N-depleted algae
Non-­‐starved
algae
(+N)
N-­‐depleted
algae
(-­‐N)
Triglycerides

30. TALEN (TAL Effector Nuclease) technology
PotenAal
to
knockout
the
acAvity
of
any
gene
in
any
organism
and
the
potenAal
to
replace
that
gene
with
an
improved
version
inal
F k
N-­‐Terminal
A
G
G
T
A
C
T
C
G
A
A
T
C
C
T
N N N oN
G
C-­‐Term
N Nucle
I
TAL
Domain
TAL a
DN Nse
D
N
N omainN oma
in
NI
NN
NN
NG
NI
HD
NG
HD
NN
NI
NI
NG
HD
HD
NG
NN
N N N N N N N T A G G T A C T C G A A T C C T G N N N N N N N N N N N N N N N N N N N N N C T C C C A T A T G A A G C G T A N N N N N N N
N N N N N N N A T C C A T G A G C T T A G G A C N N N N N N N N N N N N N N N N N N N N N G A G G G T A T A C T T C G C A T N N N N N N N
HD
NG
NI
NG
NN
NN
NN
NI
NI
HD
NN
HD
NG
NG
HD
NI
N omainN DNmaN
e
D
N
o ins TAL
Domain
TAL a
C-­‐Term
N Nucle
I
N N N oN A
C
G
G
G
A
T
A
T
C
T
C
T
G
A
C
N-­‐Terminal
ina k
Target
Gene
DNA
Sequence
F l
Double
Strand
DNA
Break
N N N N N
N N N N N N NN N N N N N
G A A T C C T G N N N N N N N N N N NN N N C T C C C
A T A T G A A
N T A G G
T A C T C C N N N N N N N N N N G G C G T A N N
N N N N N N T T A G G A A G G G T A
T A C T T C G N N N N N
A T G A G C C A T N N N N
N A T C C N N N
N N N N N N
Error-­‐prone
NonHomologous
End-­‐Joining
Repair
N N N N N N NN N N N N N N T A G G T A C T C G A A T C C T G N N N N N N N N N N N N N N C T C C C A T A T G A A G C G T A N N N N N N N N N N N N N N
N N N N N N NN N N N N N N A T C C A T G A G C T T A G G A C N N N N N N N N N N N N N N G A G G G T A T A C T T C G C A T N N N N N N N N N N N N N N
DeleJons,
InserJons,
Frame-­‐shi
MutaJons
=
Gene
Knockout
Proven
in
Yeast
Now
tesAng
in
algae

31. Greenhouse
Algal
Growth
Facility
George
Oyler

32. UNL’s
University
and
Commercial
Partners
in
Algae
R&D
(ParJal
List,
U.S.
Only)
Johns
Hopkins
University
CAB-­‐COMM
Collaborators
University
of
California,
San
Diego
Iowa
State
University
University
of
Nebraska-­‐Lincoln
University
of
Maryland
Rutgers
University
(UMBC,
UMBI,
UMD)
Scripps
InsJtuJon
of
Oceanography
University
of
California,
Davis
New
Mexico
State
University
Sapphire
Energy
University
of
California
–
Berkeley
Life
Technologies
Life
Cycle
Associates
University
of
California
–
LA
University
of
Delaware
Other
Commercial
Partners
SynapJc
Research
CGC
(Clean
Green),
LLC
Los
Alamos
NaJonal
Laboratory
Prime
BioSoluJons
Sandia
NaJonal
Laboratory
NEW
Nebraska
Start-­‐up
NanoGreen
Technologies,
LLC

33. VIRUSES AND ALGAE:
THE GOOD AND THE BAD
James L. Van Etten
University of Nebraska
Food
and
Fuel
Conference
–
University
of
Nebraska
Oct.
17
–
18,
2011

34. Viruses Can Serve 3 Functions
One bad and two good
• 1. Pathogens - bad
• 2. Enzymes to degrade cell walls for
harvesting lipids – good
• 3. Provide genetic elements for altering
algae – e.g. promoter & enhancer
elements - good

35. PBCV-1 Replication in Chlorella NC64A
(1-3 min) (3-5 min)
100 nm 100 nm
(3-5 hr) (6-8 hr)
500 nm 1 µm

36. Virus Attaching to & Infecting an Algal Cell

37. + Virus - Virus

38. An
Algal
Virus
in
Nature
750 nm

39. Selected Viruses Infecting Unicellular Algae
Size Genome Genome
Virus Host Principal references
(nm) type size (kb)
Chaetoceros
CnRNAV 31 ssRNA - Y. Shirai (pers. comm.)
neogracilis
Chaetoceros
CsNIV 38 (ss+ds)DNA 6 Nagasaki et al. (2005c)
salsugineum
Chlorella
PBCV-1 190 dsDNA 330 Van Etten (2003)
variabilis
Micromonas
MpRHAV 120 dsRNA 25.5 Brussaard C et a. (2004)
pusilla
Mizumoto et al. (2007), Nagasaki
Heterocapsa
HcRNAV 30 ssRNA 4.4 et al. (2004b, 2005a, 2006),
circularisquama
Tomaru et al. (2004a)

40. Host Cell Wall Degradation Using Chlorella-
Virus- Enzymes
Control Enzyme treated

41. Viral Elements for Genetically Engineering
Algae
• > Viral promoters elements work very
well in plants and bacteria – never
tested in algae
• > Viral enhancer element works very
well in plants – never tested in algae

42. Summary
Viruses Can Serve 3 Functions
• 1.
Pathogens
• 2.
Enzymes
to
degrade
cell
walls
for
harvesAng
lipids
• 3.
Provide
geneAc
elements
for
altering
algae
–
e.g.,
promoter
&
enhancer
elements