Factsheet 20 fire resistance of timber frame wall constructions pl 2
Expression profiles of the late blight resistance gene …...Expression profiles of the late blight...
Transcript of Expression profiles of the late blight resistance gene …...Expression profiles of the late blight...
Expression profiles of the late blight resistance gene Rpi-phu1
Śliwka J, Świątek M, Tomczyńska I, Stefańczyk E, Chmielarz M & Zimnoch-Guzowska E
Plant Breeding and Acclimatization Institute – National Research Institute, Młochów, Poland
Rpi-mcq1
Rpi-edn2
Rpi-dlc1
R8
I
VIII VII
VI V IV III II
XII XI X IX
R1
R3a R3b R5
R6 R7 R9 R10
R11 Rpi-sto2
Rpi-pcs1
Rpi-vnt1.1-1.3
R2 R2-like
Rpi-blb3
Rpi-abpt
Rpi-snk1.1-1.2
Rpi-edn1.1
Rpi-hjt1.1-1.3
Rpi-mcd1
Rpi-dmsf1
Rpi-bst1
Rpi-blb2
Rpi-ver1
Rber/Rpi-ber/Rpi-
ber1
Rpi-ber2
Rpi-blb1/RB
Rpi-bt1
Rpi-sto1
Rpi-pta1
Rpi-ver1-8
Rpi1
Rpi-avl1
Rpi-cap1
Rpi-qum1
Rpi-edn3
Rpi-pur1
Rpi-vnt2
Hot spots of R genes for resistance to P. infestans on potato genetic map
Solanum phureja
Solanum stenotomum
http://www.cgn.wageningen-ur.nl/pgr/collections/crops/potato/species.htm
Diploid , 2 EBN
Identification of the Rpi-phu1 gene on chromosome IX of potato genome
Rpi-phu1
GP129***
CP135***
GP39***
B7***
GP94***
S1D11*
2,7 cM
12,3 cM
6,4 cM
3,4 cM
4,9 cM
21,7 cM
IX
PR PS resistant susceptible progeny
Markers significantly linked with the resistance: * - p <0.05;*** - p<0.001
(U Mann-Whitney test)
okaNBS-Hae
Rpi-oka1.1
GP39
Rpi-phu1
2.7 cM
2.7 cM
2.7 cM
13.4 cM
TG551
okaNBS-Hae
4.0 cM
8.7 cM
2.7 cM
GP94
GP129
CP135
B7
S1D11
Rpi-phu1
(n=148)
1.3 cM
2.0 cM
T0156
TG591b
5.4 cM
TG186 [R]
TG35
S2h4 [R]
8.7 cM
1.3 cM
3.3 cM
0.16 cM
Rpi-oka1.1
(n=1213)
SokaM2.9L
TG186
C2_At2g47590
S. okadae = S. venturii
73
6 R
74
7 S
A6
18
+ -
Rpi-phu1 = Rpi-vnt1.1
Why are we interested in expression?
LB symptoms on Rpi-phu1 plants late in the season
Isolation of P. infestans
The isolate is not virulent on Rpi-phu1 plants in the lab
Hypotheses: 1.Isolate lost virulence in vitro 2.Rpi-phu1 gene is not (sufficiently) expressed due to:
•Genetic background •Plant age •Day length
?
Material
P. infestans
• MP324 avirulent on Rpi-phu1 plants
• MP1162 isolated from Rpi-phu1 plants but avirulent in lab tests
• EC1 described as virulent on Rpi-vnt1.1 plants
Plant material
diploid (2x)
tetraploid (4x)
Śliw
ka e
t a
l., 2
01
2
Methods
• Plants grown in climatic chambers • High relative humidity75-90% • temperature
– 16˚C night – 18˚C day
• Sampled on 0, 1, 3 and 5th day • Spray inoculation
Methods
• RNA isolation • Spectrum Plant Total RNA Kit, Sigma
• DNase I, Sigma
• RT-PCR • Maxima First Strand cDNA Synthesis Kit for RT-qPCR, Fermentas
• real-time PCR
• SYBR Green
• Relative expression
• α-tubulin
Relative expression level of Rpi-phu1 after inoculation with different P. infestans isolates
• P. infestans • MP324
• MP1162
• EC1
• Plants
• test: 04-IX-21 with Rpi-phu1 gene
• control: Craigs Royal
MP324
MP1162
EC1
Infection
04-IX-21 Craigs Royal
None of the P. infestans isolates was able to infect Rpi-phu1 plants efficiently
Potato genetic background
Potato genetic background day by day
Plant material
diploid (2x)
tetraploid (4x)
Śliw
ka e
t a
l., 2
01
2
Donor genome
12.5% 6.25% 3.12%
Plant age and Rpi-phu1 gene expression
• Gene Rpi-phu1 was expressed in all plants
• All plants were resistant
phu6 (Rpi-phu1) 3 6 9 12 14 weeks
Plant age
Plant age, day by day
Daylegth and Rpi-phu1 gene expression
• P. infestans: MP324
• Plants
• test: 04-IX-21 with Rpi-phu1
• control: Craigs Royal
• Warunki
• Short day (8h)
• Long day (16h)
Day length and Rpi-phu1 gene expression
Short day Long day
Re
lati
ve e
xpre
ssio
n
Summary
• Compatible interaction is a challenge in case of Rpi-phu1 • Rpi-phu1 expression is different in 2x and 4x potatoes
(donor genome?) and upregulated 1 day post inoculation (2x)
• In 3-week-old plant Rpi-phu1 expression is lower but it is enhanced after contact with pathogen
• No good explanation for the field observation so far • Day length? • Interaction of the above factors? • Avr-vnt1 may be switched off
Acknowledgements
J. Jones
The Sainsbury Laboratory, UK
Financed by:
LIDER/06/82/L-1/09/NCBiR/2010