On the move to field (5 min)
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On the move to field (5 min) Lukáš Spíchal Laboratory of Growth Regulators, IEB AS CR & UP Olomouc, Czech Republic Centre of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic 1,2 1 2
description
On the move to field (5 min). Lukáš Spíchal Laboratory of Growth Regulators , IEB AS CR & UP Olomouc, Czech Republic Centre of the Region Haná for Biotechnological and Agricultural Research , Olomouc, Czech Republic. 1,2. 1. 2. Acknowledgements. - PowerPoint PPT Presentation
Transcript of On the move to field (5 min)
On the move to field (5 min)
Lukáš SpíchalLaboratory of Growth Regulators, IEB AS CR & UP Olomouc, Czech Republic
Centre of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic
1,2
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Laboratory of Growth Regulators, IEB AS CR & UP Olomouc :Miroslav StrnadMarek ZatloukalLucie SzüčováVáclav MikKarel DoležalMarkéta GemrotováJana Klásková Jaroslav NislerJiří GrúzOndřej NovákJaromír MikulíkRadoslav Koprna
Department of Biochemistry, PřF UP Olomouc:Petr GaluszkaDavid Kopečný
Laboratory of Genome Integrity, UP Olomouc & BioPatterns, Ltd., Olomouc:Martin Mistrík
Institute of Biology/Applied Genetics, FU Berlin:Tomáš WernerThomas Schmülling
Acknowledgements
Grant Support: MSM 6198959216, LC06034, ED0007/01/01, GA522/08/0920
organic synthesis
biochemistry/biology
analytical chemistryfield experiments
boss
Development of CK-derived PGRs
Organic synthesis
Testing in CK bioassays
Interaction with CK metabolism/perception
In vivo applications (temporal/spatial specific)
In vivo applications in field conditions
SAR
PhenotypeGene profilingHormone levels
Testing of toxicity
Testing of toxicity
TAR
GE
TED
SY
NTH
ES
IS
What to change to gain…
NHN
NN
NH
BA
NHN
NN
NH
R1
R2
NH
NHN
NN
R1
R2
NCC
C
N
NC
C
N
NC
CC
CC
C
C
H
H
H
H
H
H
H
HH
H
H
BA
Cytokininantagonist
CKX inhibitor
Spíchal et al., 2009Nisler et al., 2010
Zatloukal et al., 2008Spíchal et al., submitted
Development of plant growth regulators at Laboratory of Growth Regulators(http://rustreg.upol.cz/)
Inhibitors of cytokinin degradation (INCYDE)
Inhibitors of cytokinin signalling (PI-55)
Inhibitors of cytokinin biosynthesis
Cotton defoliants
Stimulators of legume nodulation (N2-INCYDE)
Vigour enhancing compounds and anti-senescence activity
development of fluorescent cytokinins
development of cytokinin derivatives for affinity purification of CK binding proteins
Modulation of cytokinin degradation
Ashikari et al., 2005
Bartrina et al., 2011
Petr Galuszka – lecture
INCYDEINhibitor of CYtokinin DEgradation
INhibitor of CYtokinin oxidase/DEhydrogenase
INCYDE
INCYDE rescues WT phenotype of CKX overproducing plants
INCYDE action
INCYDE inhibits CKX activity √Can we phenocopy the ckx mutants by INCYDE application?
Targeted INCYDE application on Arabidopsis WT plants
- INCYDE + INCYDE
Targeted INCYDE application on Arabidopsis WT plants
INCYDE as a growth regulator for modulation of traits of crops
winter rapeseed 2009-2010
Branches p
er stem
Siliques
per branch
Total yiel
d
Biomass (
DW, shoot)
Biomass (
DW, root)
020406080
100120140160
10 µM50 µM
% o
f con
trol
INCYDE
winter rapeseed 2010-2011
10% increase in field yeild
Field trials
winter rapeseed 2009-2010, 2010-2011
winter wheat 2009-2010, 2010-2011
barley 2010, 2011
maize 2010, 2011
rice 2010, 2011
different regions
different phenological phasesdifferent doses
different types of application
control 28dpi infected
INCYDE negatively affects Verticillium longisporum proliferation
7dpi
14dp
i21
dpi
28dp
i7d
pi14
dpi
21dp
i28
dpi
0
10
20
30
40
50
60
70
80
control infected
leaf
are
a in
cm
²
control infected0
1
2
3
4
5
6
7
8
9
10
Col-0 + DMSO necrotic
Col-0 + INCYDE necrotic
Col-0 + DMSO chlorotic
Col-0 + INCYDE chlorotic
chlo
rotic
& n
ecro
tic le
aves
/ pl
ant
28dp
i
Col-0 +10µM INCYDE
Col-0 +DMSO
Col-0 + DMSOCol-0 + 10 µM INCYDE
A B
C D
E
F G
*** ***
*****
Col-0 + DMSO
Col-0 + INCYDE
Col-0 inf. + DMSO
Col-0 inf. + INCYDE
0.00
5.00
10.00
15.00
20.00
25.00
30.00
pg
fung
al D
NA/1
08 a
ctin
28d
pi
Col-0 + DMSOCol-0 + 10 µM INCYDE
**n.d. n.d.
T. Teichmann, unpublished
INCYDE improves functional properties of edible plants
Functional foods can be defined as those providing health benefits beyond basic nutrition and include whole, fortified, enriched or enhanced foods which have a potentially beneficial effect on health when consumed as part of a varied diet on a regular basis at effective levels (Hasler CM, 2000).
Shikimate pathway hydroxybenzoic acids (HBA)
L- Phe
trans-cinnamic acid
hydroxycinnamic acids (HCA)
PAL
Deikmann et al., 1995
INCYDE improves functional properties of edible plants
ORAC - radical scavengingTPC - phenolic content
Grúz et al., unpublished
HBA - hydroxybenzoic acids HCA - hydroxycinnamic acids
INCYDE is not cyto- and genotoxic
Cytotoxicity:
- epithelial and endothelial fibroblasts:NEGATIVE
In vivo oral toxicity:
MTD > 400 mg/kgPharmacokinetics - elimination half life ~ 2 h
Genotoxicity:
- Ames test - NEGATIVE- DNA damage - NEGATIVE
Conclusion
In the rigth time the right dose of INCYDE application
improves functional properties of edible plants
protects against biotic stress - strongly reduces symptom development and negatively affects Verticillium proliferation
has positive effect on quantitative traits of plants (increased size and number of flowers, fruits and seeds; increased biomass)
INCYDE is not cytotoxic, genotoxic and shows very high tolerance in oral toxicity in vivo studies
Inhibitors of cytokinin signalling
Riefler et al., 2006
Nishimura et al., 2004Higuchi et al., 2004
Inhibitors of cytokinin signalling – PI-55, LGR-991
PI-55 stimulates root branching and increases number of lateral roots
PI-55 accelaretes seed germination
PI-55 reduces binding of the natural ligand to cytokinin receptors
PI-55 antagonizes cytokinin effects in bioassays with different plant species
PI-55 protects development of seedlings grown on contaminated soils
Development of compounds stimulating nodulation and nitrogen fixation of legumes
Regulation of perception and metabolism of cytokinins, important plant hormones, was recently shown to be very important during the nodule formation of legumes (Gonzalez-Rizzo et al., 2006; Murray et al., 2007; Tirichine et al., 2007; Lohar et al., 2004). Preliminary data shows that application of the tested compounds to the roots of pea leads to increment in nodule number and subsequent higher rate of nitrogen fixation. Importantly, in comparison to classical cytokinins the applications of our substances has no negative effects on root and shoot growth and development.
24 h pulsing + 10 days
Control (DMSO)
Anti-senescent activity and post-harvest storage
T-2 (50 µM) T-4 (50 µM)
Shimon Meir, Volcani Center, Israel
New cotton defoliants derived from cytokinins
control
control
TDZ (250 µM)
LGR-951 (500 µM)
Defoliation of cotton leaves
0
10
20
30
40
50
60
70
80
90
100
0 2 4 6 8 10 12
Days
Defo
liatio
n (%
) LGR-2210 (500)
LGR-1042 (500)
LGR-951 (500)
LGR-950 (500)
TDZ (250)
TDZ (500)
• Dropp 4 SC, Free Fall 50 WP, Leafless 4 F, Ginstar 1.5 EC
• GROSSMANN, K. (1990) Induction of Leaf Abscission in Cotton Is a Common Effect of Urea-and Adenine-Type Cytokinins. Plant Physiol. 234-237.
contain thidiazuron as active compound
Berlin 2009
