Fusarium dry rot on potatoes in Norway:
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1 Fusarium dry rot on potatoes in Norway: Identification of pathogenic Fusarium spp. and implementation of molecular diagnostic methods Pia Heltoft Jensen May Bente Brurberg Arne Hermansen
2 Improved quality of Norwegian fruits, potatoes and vegetables after long-and short-term storage ( ) The project is mainly funded by "Fondet for forskningsvgift på landbruksprodukter" (65%) and "Jordbruksavtalen" (35%), with private partners contributing with around 10%. The partners involved in the final 10% is (highest contributions first): AL Gartnerhallen, Tine/Fellesjuice, Bama Gruppen AS, Findus Norge AS, HOFF SA, Graminor AS, NordGrønt, Norgesgrønt, Maarud AS and Kims Norge AS.
3 Main tasks 1. Find out which Fusarium spp. currently causes dry rot problems in Norway 2. Develop a multiplex real-time PCR detection assay to detect the most important Fusarium species and Boeremia foveata (gangrene).
4 1. Fusarium spp. causing dry rot in potatoes in Norway Why: Better understanding of the Fusarium dry rot problem and thereby make basis for a better control strategy towards the disease. How: Potato samples collected from different regions in Norway 3 seasons
5 2010 Potato samples collected from most important potato growing regions in Norway Østfold Akershus Vestfold Hedmark Oppland Agder Rogaland Nord-Trøndelag Troms
6 Varieties Asterix Laila Saturna Beate Mandel Sava Berber Ostara Secura Bruse Oleva Solist Folva Peik Troll Guløye Pimpernell Van Gogh Innovator Polaris Kerrs Pink Rutt
7 Procedure Wounding Incubation 6 weeks 10 degrees C high relative humidity Description of symptoms
8 Symptom descriptions on tuber Volume of rotted area (mm) Width Depth Surface Mycelium Cavity
9 Procedure Isolation on agar (PDA) from diseased tubers Description of mycelium on agar Colour Size (72 hours in darkness, room temp) Transfer to other agar medium If in doubt -> inoculation trials
10 Inoculation trials Growth on PDA for 5 days Potato plugs (5mmx10mm) removed Replaced with PDA plugs with mycelial growth Incubation in dark room, 13 degrees C After 4 weeks we describe and measure symptoms on tubers Morfological determination of Fusarium spp.
11 Procedure Identification of Fusarium spp. Morfological determination Determination by PCR-test and sequencing
12 Results % of tubers developed rots Penicillium Mucor Fusarium Boeremia (Phoma) Cylindrocarbon 49 % of developed rots are Fusarium
13 Results We found five species of Fusarium: 1. F. coeruleum 2. F. avenaceum 3. F. culmorum 4. F. cerealis 5. F. sambucinum
14 Results Frequencies (%) of Fusarium spp. isolated from commercial potato tubers 2010 Fusarium coeruleum 64 Fusarium avenaceum 31 Fusarium culmorum 4 Fusarium sambucinum 1 Fusarium cerealis 1 Total number of isolates 137 Total number of tubers tested 5400
15 Variety Fusarium spp. Percent of total Asterix cul, ave,coe 24,8 Beate ave, coe 2,2 Berber - - Bruse - - Folva ave, coe, cul, cer 8,8 Gulløye coe, ave 22,6 Innovator - - Kerrs Pink coe, ave 1,5 Laila ave 3,6 Mandel ave, coe 2,2 Oleva ave 13,1 Ostara - - Peik ave, sam 2,2 Pimpernell - - Polaris sam, cul 1,5 Rutt coe 12,4 Saturna ave, coe 5,1 Sava - - Secura - - Solist - - Troll coe 0,7 Van Gogh - - Total 100,7
16 0 Fylke Fusarium spp. Percent of total Østfold coe 0,7 Vestfold ave 1,5 Hedmark ave, coe 8,8 Oppland ave, sam 6,6 Agder coe, cul, sam 13,8 Rogaland ave, coe, cul, cer 7,3 Nord-Trøndelag ave, coe 36,5 Troms ave, coe, cul 24,8 Total 100
17 2. PCR and multiplexing Use of already established Real-time PCR-method Further development into multiplexing Development of primers for important Fusarium spp.
18 2. PCR and multiplexing Detection of more Fusarium spp. in the same test Detection of Boeremia spp. (Phoma) in the same reaction
19 Already established PCR methods Already established PCR methods for: F. avenaceum F. culmorum Boeremia/Phoma
20 Design of primers Developing primers for: F. coeruleum Primer Express program version 2.0 (Applied Biosystems, Foster City, CA, USA) ITS region (ITS 1-4) By sequencing we found that our F. coeruleum is very different from other F. coeruleum
21 Design of primers for F. coeruleum Test of primers (sybr green) Real time PCR Waiting for good results Test of probes Hope to get a good test that can be used together with other tests in multiplexing
22 Thank you
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