Pentland Ivory. (Nc:Ny:Nz) cv. Desiree. (nc:Ny:nz). PVYN-Wi symptoms in potato differentials. Non-inoculated upper leaves. 11. Inoculated local. Leaves. (no HR) ...
Trends in PVY population changes in Poland Zhimin Yin, Zofia Wójcik, Krystyna Michalak, Mirosława Chrzanowska, Renata Lebecka, Ewa Zimnoch-Guzowska Młochów Research Center Plant Breeding and Acclimatization Institute National Research Institute, Poland
16th triennial meeting of the Virology Section of EAPR Ljubljana, Slovenia, May 31 – June 3, 2016
1
Outlines 1. Infection rate of PVY on tested potato
samples in Poland from 2001-2012. 2. Changes of PVY population in Poland from
1980 -2014. 3. Structure of the PVY population in Poland
from 2008-2012. 2
Potato samples used for evaluation of PVY infection rate and PVY population structure in 2008-2012
Location checked
Number of forms tested
Total # of tubers tested for PVY infection rate
26
64 cvs
11 729
Total # of PVY infected tubers tested for population structure
3 737 3
PVY map (2008-2012)
4
Infection rate of PVY on tested potato samples from production % tubers infected by PVY
70,0 60,0 50,0 40,0 30,0
20,0 10,0 0,0 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 5
Methods for identification and differentiation of isolates in PVY population studies Serological test: polyclonal antibody for all PVY strain (1980-1990) monoclonal cocktail antibody for PVYall (1994-) monoclonal antibody specific for PVYN (1994-) monoclonal antibody specific for PVYO (2010-)
Biological test: potato cvs. King Edward, Desiree, Pentland Ivory, tobacco cv. Samsun (Singh et al. 2008) Multiplex RT-PCR for PVY strain typing: Rigotti and Gugerli 2007 Lorenzen et al. 2006 Chikh Ali et al. 2010 6
Changes in PVY population tested on tobacco bait plants at Młochów, IHAR-PIB, 1980 - 2014 100% 90% 80% 70% 60%
Mix PVY NTN PVY N-Wi PVY O
50%
40% 30% 20% 10% 2014
2012
2010
2008
2004
2003
2002
2001
2000
1998
1996
1994
1992
1990
1988
1986
1984
1982
1980
0% 7
Structure of PVY population on potato samples from 2001 to 2012 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0%
Mix PVY NTN PVY N-Wi
PVY O
2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 8
Structure of PVY population on potato samples from 2001 to 2012 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0%
Mix PVY NTN PVY N-Wi
PVY O
2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 9
PVY symptoms in Nicotiana tabacum Healthy
VN PVYN-Wi
VCl PVYO
VN PVYNTN
10
PVYN-Wi symptoms in potato differentials cv. King Edward (Nc:ny:nz)
cv. Desiree (nc:Ny:nz)
cv. Pentland Ivory (Nc:Ny:Nz)
Inoculated local Leaves (no HR)
Non-inoculated upper leaves
11
PVYNTN symptoms in potato differentials cv. King Edward (Nc:ny:nz)
cv. Desiree (nc:Ny:nz)
cv. Pentland Ivory (Nc:Ny:Nz)
Inoculated local Leaves (no HR)
Non-inoculated upper Leaves
12
PVYO symptoms in potato differentials cv. King Edward
cv. Desiree
(Nc:ny:nz)
cv. Pentland Ivory (Nc:Ny:Nz)
(nc:Ny:nz)
Inoculated local leaves HR ?
HR
HR
Non-inoculated upper leaves SHR HR - Hypersensitive resistance SHR - Systemic hypersensitive resistance
SHR 13
PVY strain typing by multiplex RT-PCR according to Lorenzen et al. 2006 PVYNTN
PVYN:O
-689 bp -452 bp
-181 bp
14
14
PVY strain typing by multiplex RT-PCR according to Chikh Ali et al. 2010 PVYNTN (A) PVYNW (B)
-1307 bp
-853 bp -633 bp -441 bp
15 = 1307 bp + 441 bp PVYNTN (B)
15
PVY with VCl tested by multiplex RT-PCR PVYN:O PVYO
PVYO
PVYO
PVYN W (B) with an additional RJ as in SYRIII
-853 bp
-689 bp
-532 bp
-660 bp -530bp -440 bp
-441 bp -267 bp
-180 bp
Rigotti and Gugerli 2007
-278 bp
-181 bp
Lorenzen et al. 2006 VCl: vein clearing
Chikh Ali et al. 2010 16
16
Summary of PVY strain types in potato Potato differentials
PVY-specific resistance genes
PVY strain group reaction
PVYC
PVYO
PVYN
PVYZ
PVYE
King Edward
HR
s
s
s
s
Nc:ny:nz
Desiree
s
HR
s
s
s
Nc:Ny:nz
Pentland Ivory
HR
HR
s
HR
s
Nc:Ny:Nz
Tobacco cv. Samsun
s
s
VN
s
s
# PVY isolates
0
2
23
0
4
ELISA
O+
N+ (15) O+ (8)
O+
Multiplex RT-PCR
PVYO
PVYNTN (15) PVYN-Wi (8)
PVYN-Wi
Sum=29
+: positive. HR: Hypersensitive resistance response (localized or systemic). S: susceptible (systemic infection, no necrosis). VN: local and systemic veinal necrosis
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Conclusions
1. PVY infection rate on ware potato samples tested
from 2008-2012 ranged from 30-60%.
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Conclusions 2. The two main types of the recombinant strain, PVYN-Wi and recombinant PVYNTN, predominant in
the PVY infected ware potatoes. The ratio between them is variable.
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Conclusions 3. One PVYO isolate induced HR and systemic HR, leaf deformation and mosaic in cvs. Desiree and Pentland Ivory. However, systemic spreading of PVY which confirmed by ELISA suggested overcoming Ny, Nc and Nz. This may arouse discussion about the criteria to assess hypersensitive resistance (HR) response and to distinguish PVYO and PVYE in such biological tests. 20
Acknowledgements • Prof. dr hab. Mirosława Chrzanowska • Dr Zhimin Yin • MSc. Zofia Wójcik • MSc. Krystyna Michalak • Dr hab. Renata Lebecka • Prof. dr hab. Ewa Zimnoch-Guzowska
• IHAR-PIB, Młochów, Poland
This research is supported by Ministry of Agriculture and Rural Development of Poland Grant: PW 3.1 . 21