ORIGINAL ARTICLE
The influence of long-term storage in liquid nitrogen on survival and pathogenicity of Phytophthora infestans isolates
 
More details
Hide details
1
Plant Breeding and Acclimatization Institute – National Research Institute Młochów Research Centre, Platanowa 19, 05-831 Młochów, Poland
CORRESPONDING AUTHOR
Jadwiga Śliwka
Plant Breeding and Acclimatization Institute – National Research Institute Młochów Research Centre, Platanowa 19, 05-831 Młochów, Poland
Submission date: 2012-04-04
Acceptance date: 2012-10-24
 
Journal of Plant Protection Research 2012;52(4):479–485
KEYWORDS
TOPICS
ABSTRACT
A set of 14 aggressive Phythophthora infestans (Mont.) de Bary isolates with a wide range of virulence, was stored for eight years (2002–2010) in liquid nitrogen at –196°C (209 samples) and under paraffin oil at 7°C (70 test tubes). The survival rate of samples stored in liquid nitrogen was scored as 88%, 45 days after thawing. The revived isolates were passaged through potato tissues four times and tested for virulence on a set of 11 Black’s differentials. The mean number of virulence factors per isolate, of these P. infestans isolates evaluated before storage, was 7.4. The isolates stored in liquid nitrogen and under paraffin oil and passaged four times through potato tissues showed a similar mean number of virulence factors per isolate, 7.3 and 6.9, respectively. Isolates stored under paraffin oil showed no expression of factor avr5 , but expression of this factor occurred in six isolates after liquid nitrogen treatment. The initial expression of this factor occurred in four isolates. Before storage, the average aggressiveness of the tested isolates was assessed as 1.7, on a 1–9 scale, where 1 means the most aggressive. After storage and after four subsequent passages through potato tissues, the mean aggressiveness of isolates stored in liquid nitrogen and under paraffin oil reached the level of 1.5 and 2.1, respectively. The mean aggressiveness was not significantly different from the level of initial assessment.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
REFERENCES (22)
1.
Andrivon D., Avendaño-Corcoles J., Cameron A.M., Carnegie S.F., Cooke L.R., Corbière R., Detourne D., Dowley L.J., Evans D., Forisekova K., Griffin D.G., Hannukkala A., Lees A.K., Lebecka R., Niepold F., Polgar Z., Shaw D.S., Thompson J., Trognitz B., van Raaij H.M.G., Zimnoch-Guzowska E. 2011. Stability and variability of virulence of Phytophthora infestans assessed in a ring test across European laboratories. Plant Pathol. 60 (3): 556–565.
 
2.
Caten C.E., Jinks J.L. 1968. Spontaneous variability of single isolates of Phytophthora infestans. I. Cultural variation. Can. J. Bot. 46 (1): 329–348.
 
3.
Dahnen H., Staub Th., Schwinn F.J. 1983. Technique for long-term preservation of phytopathogenic fungi in liquid nitrogen. Phytopathology 73 (2): 241–246.
 
4.
Haas B.J., Kamoun S., Zody M.C., Jiang R.H., Handsaker R.E., Cano L.M., Grabherr M., Kodira C.D., Raffaele S., Torto-Alalibo T., Bozkurt T.O., Ah-Fong A.M., Alvarado L., Anderson V.L., Armstrong M.R., Avrova A., Baxter L., Beynon J., Boevink P.C., Bollmann S.R., Bos J.I., Bulone V., Cai G., Cakir C., Carrington J.C., Chawner M., Conti L., Costanzo S., Ewan R., Fahlgren N., Fischbach M.A., Fugelstad J., Gilroy E.M., Gnerre S., Green P.J., Grenville-Briggs L.J., Griffith J., Grünwald N.J., Horn K., Horner N.R., Hu C.H., Huitema E., Jeong D.H., Jones A.M., Jones J.D., Jones R.W., Karlsson E.K., Kunjeti S.G., Lamour K., Liu Z., Ma L., Maclean D., Chibucos M.C., McDonald H., McWalters J., Meijer H.J., Morgan W., Morris P.F., Munro C.A., O’Neill K., Ospina-Giraldo M., Pinzón A., Pritchard L., Ramsahoye B., Ren Q., Restrepo S., Roy S., Sadanandom A., Savidor A., Schornack S., Schwartz D.C., Schumann U.D., Schwessinger B., Seyer L., Sharpe T., Silvar C., Song J., Studholme D.J., Sykes S., Thines M., van de Vondervoort P.J., Phuntumart V., Wawra S., Weide R., Win J., Young C., Zhou S., Fry W., Meyers B.C., van West P., Ristaino J., Govers F., Birch P.R., Whisson S.C., Judelson H.S., Nusbaum C. 2009. Genome sequence and analysis of the Irish potato famine pathogen Phytophthora infestans. Nature 461 (7262): 393–398.
 
5.
Hodgson W.A., Sharma K.P. 1967. Restoration of virulence of stored cultures of Phytophthora infestans. Can. J. Plant Sci. 47 (4): 447–449.
 
6.
Lapwood D.H. 1963. Potato haulm resistance to Phytophthora infestans. IV. Laboratory and field estimates compared, and further field analyses. Ann. Appl. Biol. 51 (1): 17–28.
 
7.
Meryman H.T. 1956. Mechanics of freezing in living cell and tissues. Science 124 (1): 515–521.
 
8.
Nagai T., Ideno A., Tsuge M., Oyanagi C., Oniki M., Kita K., Horita M., Aoki T., Kobayashi T., Tsuchiya K. 2000. Preservation of fungi in an atmosphere over liquid nitrogen after uncontrolled freezing. Microbiol. Cult. Coll. 16 (1): 71–74.
 
9.
Pietkiewicz J. 1972. Investigations on the resistance of potatoes to Phytophthora infestans de By. on detached leaves. Biul. Inst. Ziemn. 9 (1): 17–32.
 
10.
Sobkowiak S., Zarzycka H. 1999. The influence of methods of preparation of Phytophthora infestans cultures to liquid nitrogen freezing on their survival. “Biodiversity in European Plant Pathology at the turn of the centuries”. X General Assembly of the PPS Members, Poznań, 7–9.09.1999, 133 pp.
 
11.
Sobkowiak S., Zarzycka H., Zimnoch-Guzowska E. 2004a. Effect of deep freezing of Phytophthora infestans cultures on their survival and pathogenicity. Plant Breed. Seed Sci.49 (1): 79–89.
 
12.
Sobkowiak S., Zimnoch-Guzowska E., Zarzycka H. 2004b. Effect of various culture treatments on virulence and aggressiveness expression of Phytophthora infestans. 2004. Acta Agrobot. 57 (1–2): 131–143.
 
13.
Sujkowski L.S. 1983. Zmienność właściwości pasożytniczych grzyba Phytophthora infestans (Mont.) de Bary w stosunku do ziemniaka w cyklu rocznym. PhD thesis. Młochów, 63 pp.
 
14.
Sujkowski L.S. 1992. Zmienność wirulencji i agresywności u grzyba Phytophthora infestans(Mont.) de Bary w stosunku do ziemniaka. Habil. thesis, Potato Institute, Młochów, 56 pp.
 
15.
Świszczewska J., Osińska M., Piotrowski W. 1971. Pathogenicity of Phytophthora infestans Mont. de Bary races in dependence on the biotype, substrate and season. Biul. Inst. Ziemn. 8 (1): 21–27.
 
16.
Tooley P.W. 1988. Use of uncontrolled freezing for liquid nitrogen storage of Phytophthora species. Plant Dis. 72 (8): 680–682.
 
17.
Thurston H.D. 1957. The culture of Phytophthora infestans. Phytopathology 47 (3), p. 186.
 
18.
Webb K.M., Hill A.L., Laufman J., Hanson L.E., Panella L. 2011. Long-term preservation of a collection of Rhizoctonia solani using cryogenic storage. Ann. Appl. Biol. 158 (3): 297–304.
 
19.
Zarzycka H. 1995. Effect of culturing Phytophthora infestans on expression of its virulence and aggressiveness to potato. Phytopath. Pol. 9 (21): 81–88.
 
20.
Zarzycka H. 1996. Effect of various methods of maintenance of Phytophthora infestans on its pathogenicity to potato. Plant Breed. Seed Sci. 40 (1–2): 31–43.
 
21.
Zarzycka H., Sobkowiak S. 1997. Monitoring of changes in Polish population of Phytophthora infestans. Biul. Inst. Ziemn. 48 (1): 125–135.
 
22.
Zarzycka H. 2001. Isolation, maintenance and preservation of Phytophthora infestans isolates. p. 60–62. In: “The Methods of Evaluation and Selection Applied in Potato Research and Breeding” (H.J. Czembor, ed.). Monogr. Rozpr. Nauk. IHAR 10a, p. 131.
 
eISSN:1899-007X
ISSN:1427-4345