(XAP) in the ornamental Prunus lauroceracus (cherry laurel)
(XAP) in the ornamental Prunus lauroceracus (cherry laurel)
(XAP) in the ornamental Prunus lauroceracus (cherry laurel)
Create successful ePaper yourself
Turn your PDF publications into a flip-book with our unique Google optimized e-Paper software.
Invasion of X. a. pv pruni<br />
<strong>in</strong> <strong>the</strong> <strong>ornamental</strong> <strong>cherry</strong><br />
<strong>laurel</strong> <strong>in</strong> NL<br />
24 May 2012<br />
Maria Bergsma-Vlami<br />
National Reference Centre<br />
Food and Consumer Product Safety<br />
Authority, NL
2<br />
Outl<strong>in</strong>e<br />
1. Xanthomonas arboricola pv. pruni<br />
* organism and distribution<br />
* situation <strong>in</strong> NL <strong>in</strong> 2009, 2010 and 2011<br />
* symptoms on <strong>cherry</strong> <strong>laurel</strong><br />
2. Molecular typ<strong>in</strong>g of Dutch X.a. pv. pruni isolates<br />
3. Epidemiology of X.a. pv. pruni <strong>in</strong> NL<br />
4. Conclusions and future plans<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Xanthomonas arboricola pv. pruni<br />
• Quaranta<strong>in</strong>e status <strong>in</strong> <strong>the</strong> EU<br />
(Council Directive 2000/29/EG)<br />
and EPPO (IIA2 status)<br />
• Highly host specific: just one plant<br />
genus<br />
3<br />
Major Hosts Common name<br />
<strong>Prunus</strong> armeniaca apricot<br />
<strong>Prunus</strong> domestica european plum<br />
<strong>Prunus</strong> persica peach<br />
M<strong>in</strong>or Hosts<br />
<strong>Prunus</strong> avium sweet <strong>cherry</strong><br />
<strong>Prunus</strong> cerasus sour <strong>cherry</strong><br />
<strong>Prunus</strong> dulcis sweet almond<br />
<strong>Prunus</strong> salic<strong>in</strong>a japanese plum<br />
Incidental Hosts<br />
<strong>Prunus</strong> <strong>lauroceracus</strong> <strong>cherry</strong> <strong>laurel</strong><br />
<strong>Prunus</strong> davidiana ch<strong>in</strong>ese wild peach<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012<br />
(Source EPPO-PQR)
Distribution map of X. a pv pruni (Source EPPO-PQR)<br />
4<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Earlier <strong>in</strong>cidents of X.a. pv. pruni <strong>in</strong> Europe<br />
5<br />
• 1995: <strong>in</strong> France <strong>in</strong> peach, later <strong>in</strong> plum, nectar<strong>in</strong>e and apricot.<br />
• 2005: <strong>in</strong> Switzerland <strong>in</strong> apricot and <strong>in</strong> Japanese plum.<br />
• 2006: <strong>in</strong> Italy <strong>in</strong> <strong>Prunus</strong> laurocerasus.<br />
• 2008: <strong>in</strong> Spa<strong>in</strong> <strong>in</strong> plum and peach.<br />
• 2009: <strong>in</strong> Spa<strong>in</strong> <strong>in</strong> almond, plum and peach.<br />
• 2009: <strong>in</strong> <strong>the</strong> Ne<strong>the</strong>rlands <strong>in</strong> <strong>Prunus</strong> laurocerasus.<br />
• 2010: <strong>in</strong> Germany <strong>in</strong> <strong>Prunus</strong> laurocerasus.<br />
• 2010: <strong>in</strong> Italy aga<strong>in</strong> <strong>in</strong> <strong>Prunus</strong> laurocerasus.<br />
• 2011: <strong>in</strong> Spa<strong>in</strong> <strong>in</strong> <strong>Prunus</strong> sp.<br />
(Source: EPPO Report<strong>in</strong>g Service/EU Notifications)<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Situation <strong>in</strong> <strong>the</strong> Ne<strong>the</strong>rlands - numbers<br />
6<br />
Year Number X. a. pv pruni<br />
2009 43<br />
2010 68<br />
2011 63<br />
Total number 174<br />
(Source: NPPO)<br />
Plant species Cultivar # of positive samples<br />
<strong>Prunus</strong> laurocerasus rotundifolia 43<br />
<strong>Prunus</strong> laurocerasus otto luyken 39<br />
<strong>Prunus</strong> laurocerasus etna 34<br />
<strong>Prunus</strong> laurocerasus novita 32<br />
<strong>Prunus</strong> laurocerasus caucasica 10<br />
<strong>Prunus</strong> laurocerasus herbergii 6<br />
<strong>Prunus</strong> laurocerasus anbri 2<br />
<strong>Prunus</strong> laurocerasus <strong>cherry</strong> brandy 2<br />
<strong>Prunus</strong> laurocerasus polster 2<br />
<strong>Prunus</strong> lusitanica angustifolia 1<br />
<strong>Prunus</strong> laurocerasus a<strong>the</strong>ne 1<br />
<strong>Prunus</strong> pumila depressa 1<br />
<strong>Prunus</strong> laurocerasus gajo 1<br />
<strong>Prunus</strong> laurocerasus gruner teppich 1<br />
<strong>Prunus</strong> laurocerasus mano 1<br />
<strong>Prunus</strong> laurocerasus mischeana 1<br />
<strong>Prunus</strong> laurocerasus mount venon 1<br />
<strong>Prunus</strong> laurocerasus olympus 1<br />
<strong>Prunus</strong> laurocerasus reynvaanii 1<br />
<strong>Prunus</strong> laurocerasus zabe liana 1<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Symptoms on <strong>cherry</strong> <strong>laurel</strong><br />
Bacterial leaf spot and shot hole (necrotic brown centre with dist<strong>in</strong>ct chlorotic marg<strong>in</strong>)<br />
7<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
8<br />
Confirmation: X.a. pv. pruni<br />
EPPO PM 7/64, 2006<br />
Isolates characterized by:<br />
•Katabolic biochemical test range<br />
•Fatty Acid Analysis (FAME)<br />
•Immunofluorescence (IF)<br />
•Pathogenicity: on peach plants (cultivars ‘Peregr<strong>in</strong>e’ and ‘Vaes<br />
Oogst’) and on detached leaves of P. laurocerasus cv.‘Novita’.<br />
Tjou-Tam-S<strong>in</strong> N.N.A., van de Bilt J.L.J., Bergsma-Vlami M., Koenraadt H., Westerhof J., van<br />
Doorn J., Pham K.T.K. and W.S. Mart<strong>in</strong> 2012. First Report of Xanthomonas arboricola pv.<br />
pruni <strong>in</strong> <strong>ornamental</strong> <strong>Prunus</strong> <strong>lauroceracus</strong> <strong>in</strong> <strong>the</strong> Ne<strong>the</strong>rlands, Plant Disease 96 (5), p 759<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Molecular typ<strong>in</strong>g of X.a. pv. pruni isolates<br />
• No knowledge of epidemiology on <strong>cherry</strong> <strong>laurel</strong><br />
• By molecular typ<strong>in</strong>g ga<strong>in</strong> <strong>in</strong>formation on pathogen spread,<br />
variation and relatedness.<br />
• Assess efficacy of typ<strong>in</strong>g techniques for X.a. pv. pruni<br />
9<br />
Four typ<strong>in</strong>g methodologies applied as epidemiological tools for<br />
discrim<strong>in</strong>ation of isolates: VNTR, AFLP, BOX-PCR and gyrB.<br />
Bergsma-Vlami M., Mart<strong>in</strong> W., Koenraadt H., Teunissen H., Pothier J.F., Duffy B. and J.<br />
van Doorn 2012. Molecular typ<strong>in</strong>g of Dutch isolates of X. a pv. pruni isolated from<br />
<strong>ornamental</strong> <strong>cherry</strong> <strong>laurel</strong>, accepted JPP.<br />
Bergsma-Vlami M., Pham K., Koenraadt H., Teunissen H., Pothier J.F., Duffy B. and J. van<br />
Doorn. Epidemiology of X. a pv. pruni isolated from <strong>ornamental</strong> <strong>cherry</strong> <strong>laurel</strong> <strong>in</strong> <strong>the</strong><br />
Ne<strong>the</strong>rlands, <strong>in</strong> preparation.<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
10<br />
What is VNTR analysis?<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Results from VNTR analysis on NL isolates<br />
• 6 VNTR loci selected for f<strong>in</strong>al analysis<br />
• Some loci very discrim<strong>in</strong>ative (<strong>XAP</strong> 1)<br />
Loci with medium/high diversity Loci with low or no diversity<br />
• NL isolates show dist<strong>in</strong>ct variation at several VNTR Loci<br />
11<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012<br />
Source: PPO-Lisse
Number location/year comb<strong>in</strong>ation Cultivar<br />
12<br />
1 A-2009 Novita<br />
2 A-2010-1 Novita<br />
3 A-2010-2 Otto Luyken<br />
4 B-2009 Novita<br />
5 B-2011 Rotundifolia<br />
6 C-2010-1<br />
7 C-2010-2 Etna<br />
8 D-2010 Etna<br />
9 D-2011 Otto Luyken<br />
10 E-2010 Otto Luyken<br />
11 E-2011-1 Otto Luyken<br />
12 E-2011-2 A<strong>the</strong>ne<br />
13 F-2010 Otto Luyken<br />
14 F-2011 Otto Luyken<br />
15 G-2009 Novita<br />
16 G-2011 Novita<br />
17 H-2010 Rotundifolia<br />
18 H-2011 Novita<br />
19 I-2009 Otto Luyken<br />
20 I-2010 Otto Luyken<br />
21 J-2010 Rotundifolia<br />
22 J-2011 Rotundifolia<br />
23 K-2009-1 div. cultivars<br />
24 K-2009-2 Rotundifolia<br />
25 K-2009-2 Etna<br />
26 K-2011<br />
27 L-2009 Etna<br />
28 L-2010-1 Otto Luyken<br />
29 L-2010-2 Otto Luyken<br />
30 L-2010-2 Etna<br />
31 M-2010 Rotundifolia<br />
32 M-2011 Rotundifolia<br />
33 N-2009-1 Otto Luyken<br />
34 N-2009-2 Caucasica<br />
35 N-2009-2 Novita<br />
2ond <strong>in</strong>troduction <strong>in</strong> 2011<br />
<strong>in</strong> location B<br />
S<strong>in</strong>gle <strong>in</strong>troduction <strong>in</strong><br />
locations E and F<br />
<strong>XAP</strong> 1<br />
S<strong>in</strong>gle <strong>in</strong>troduction <strong>in</strong><br />
location D <strong>in</strong> 2010 and 2011<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012<br />
Source: PPO-Lisse
Results from AFLP analysis on NL isolates<br />
Source: Naktu<strong>in</strong>bouw<br />
13<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
14<br />
AFLP data<br />
5<br />
100<br />
10<br />
100<br />
100<br />
15<br />
Source: Naktu<strong>in</strong>bouw<br />
98<br />
20<br />
100<br />
25<br />
99<br />
30<br />
97<br />
35<br />
LMG 12703 X.a. pv. juglandis<br />
LMG 8659 X.a. pv. coryl<strong>in</strong>a<br />
LMG 12141 X.a. pv. populi<br />
40<br />
45<br />
50<br />
55<br />
60<br />
65<br />
70<br />
75<br />
80<br />
85<br />
90<br />
85<br />
86<br />
87<br />
95<br />
83<br />
80<br />
74<br />
100<br />
51<br />
AFLP Nummer<br />
165<br />
185<br />
167<br />
168<br />
169<br />
171<br />
44<br />
45<br />
60<br />
128<br />
178<br />
181<br />
190<br />
166<br />
100<br />
170<br />
172<br />
163<br />
164<br />
93<br />
98<br />
112<br />
116<br />
117<br />
160<br />
173<br />
174<br />
77<br />
162<br />
199<br />
158<br />
175<br />
201<br />
200<br />
Orig<strong>in</strong>ele isolaataanduid<strong>in</strong>g<br />
IVIA 2647.3-1<br />
IVIA 2826-8<br />
IVIA 2832-10b<br />
IVIA 2832-30a<br />
IVIA 3467-1<br />
NAK-1959<br />
NAK-1960<br />
NAK-2351<br />
NAK-2678<br />
IVIA 2826-1<br />
NAK-2459<br />
IVIA 3161-2 col 1<br />
IVIA 3487-1 col 1<br />
IVIA 2626-1<br />
IVIA 2647.1-2<br />
NAK-2377<br />
NAK-2457<br />
NAK-2604<br />
NAK-2608<br />
NAK-2609<br />
NAK-2634<br />
NAK-2659<br />
NAK-2757<br />
Q20<br />
NAK-2632<br />
Q24<br />
Q21<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012<br />
NBC nr<br />
NBC-3897<br />
NBC-3938<br />
NBC-3899<br />
NBC-3900<br />
NBC-3901<br />
NBC-3903<br />
NBC-1959<br />
NBC-1960<br />
NBC-2351<br />
NBC-2678<br />
NBC-3931<br />
NBC-3934<br />
NBC-3943<br />
NBC-3898<br />
NBC-2459<br />
NBC-3902<br />
NBC-3904<br />
NBC-3895<br />
NBC-3896<br />
NBC-2377<br />
NBC-2457<br />
NBC-2604<br />
NBC-2608<br />
NBC-2609<br />
NBC-2634<br />
NBC-3923<br />
NBC-3924<br />
NBC-2659<br />
NBC-2757<br />
NBC-2632<br />
NBC-3925<br />
orig<strong>in</strong>ele code<br />
04512863<br />
cantaena<br />
PD-04804516<br />
PD-04678011<br />
PD-04803337<br />
AM-328<br />
PD-04677975<br />
PD-04458975<br />
AM-343<br />
AM-344<br />
PD-04802772<br />
LMG 12703<br />
LMG 8659<br />
PD-04709631<br />
PD-05018496<br />
PD-04742888<br />
LMG 12141
Results from BOX/ERIC-PCR on NL isolates<br />
15<br />
Atypical f<strong>in</strong>gerpr<strong>in</strong>t<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012<br />
PC<br />
M<br />
(Source: Naktu<strong>in</strong>bouw)<br />
BOX- and ERIC-PCR among more than 120 X.a. pv. pruni isolates<br />
showed almost no isolate-specific, or unique profiles.<br />
Extra<br />
band <strong>in</strong><br />
PC<br />
Eric<br />
Box
Results from gyrB analysis on NL isolates<br />
gyrB phylogeny (Park<strong>in</strong>son et al., 2007) confirmed lack of unique profiles<br />
with 99,6% homology to ICMP51 X.a .pv. pruni reference stra<strong>in</strong><br />
• Young J.M. et al., 2010. Plant Pathol. 59 (2), 270-281.<br />
• Young J.M. et al., 2008. Syst. Appl. Microbiol. 31 (5), 366-377.<br />
16<br />
BOX-PCR<br />
gyr B<br />
sample 1 new pattern 99,3 % with GQ183105 X arboricola stra<strong>in</strong> ICMP 8452 *<br />
sample 2 new pattern 99,6 % with EU498984 X a pv celebensis stra<strong>in</strong> ICMP 1488<br />
sample 3 new pattern 99,0 % with EU498984 X a pv celebensis stra<strong>in</strong> ICMP 1488<br />
sample 4 new pattern 99,7 % with GQ183099 X. dyei stra<strong>in</strong> ICMP 2374 **<br />
sample 5 new pattern 99,0 % with GQ183106 X arboricola stra<strong>in</strong> ICMP 4939 ***<br />
sample 6 new pattern 99,6 % with EU498984 X a pv celebensis stra<strong>in</strong> ICMP 1488<br />
sample 7 new pattern 99,6 % with EU498984 X a pv celebensis stra<strong>in</strong> ICMP 1488<br />
* Magnolia sp.<br />
** Aralia sp, New Zealand, Young et al., 2010<br />
*** Mahonia lomariifolia<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
17<br />
Conclusions<br />
• VNTR analysis is highly discrim<strong>in</strong>ative among X.a. pv pruni isolates.<br />
• Use of VNTR for epidemiological studies, i.e. track and trace<br />
applications.<br />
• There is one cluster of X.a. pv pruni isolates with relatively little<br />
variation found <strong>in</strong> AFLP, BOX/ERIC-PCR and gyrB analysis.<br />
• In diagnostics, use of one of AFLP, BOX/ERIC-PCR and gyrB analysis<br />
is strongly recommended <strong>in</strong> cases of isolates belong<strong>in</strong>g to <strong>the</strong><br />
outgroup (no X.a. pv pruni) avoid<strong>in</strong>g false positive results.<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Plans for 2013<br />
• Whole genome sequenc<strong>in</strong>g of representative Dutch isolates <strong>in</strong> <strong>the</strong><br />
outgroup, <strong>in</strong> collaboration with Agroscope Chang<strong>in</strong>s-Wädenswil<br />
Research Station ACW, CH<br />
18<br />
gyr B<br />
sample 1 99,3 % with GQ183105 X arboricola stra<strong>in</strong> ICMP 8452 *<br />
sample 2 99,6 % with EU498984 X a pv celebensis stra<strong>in</strong> ICMP 1488<br />
sample 4 99,7 % with GQ183099 X. dyei stra<strong>in</strong> ICMP 2374 **<br />
sample 5 99,0 % with GQ183106 X arboricola stra<strong>in</strong> ICMP 4939 ***<br />
• Comparative genomics to identify potential hits <strong>in</strong> <strong>the</strong> X. arboricola<br />
pv. pruni genome sequenced, as well as <strong>the</strong> X. arboricola pv.<br />
juglandis and <strong>the</strong> X. arboricola pv. fragariae stra<strong>in</strong>s sequenced so<br />
far.<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Acknowledgements<br />
• Inspection Service Naktu<strong>in</strong>bouw<br />
19<br />
Harrie Koenraadt, Hedwich Teunissen<br />
• Applied Plant Research, Wagen<strong>in</strong>gen University & Research Centre<br />
Joop van Doorn, Khanh Pham, Wendy Mart<strong>in</strong>s<br />
• Agroscope Chang<strong>in</strong>s-Wädenswil Research Station ACW, CH<br />
Brion Duffy, Joël Pothier, Andreas Bülhmann<br />
• National Reference Centre<br />
Leon Tjou-Tam-S<strong>in</strong>, Jeroen van de Bilt, Bart v/d Vossenberg<br />
QBOL-EPPO Conference, Haarlem, NL<br />
24 May 2012
Change language