Phylogenic,molecular and decay analysis of Phialophora species causing soft rot of wood

Abstract

Soft rot decay of treated wood in ground contact and loss of service-life remains an important economic issue worldwide. This study determined the identity of unclassified Phialophora strains (204-1, 35-1, TS4M3, 25M3) isolated from wood in-service. Phylogenetic analysis clustered strains into a single clade homologous with Phialocephala dimorphospora. Sequencing of internal transcribed spacer (ITS) rDNA showed minor differences at single bases for strains 35-1 and 25M3. Compatibility occurred between TS4M3, 25M3 and 204-1 but not 35-1, Phialophora lagerbergii or Phialophora botulispora. Birch was more rapidly degraded than pine for all strains, with strain 35-1 least aggressive. Light microscopy showed Type I diffuse soft rot cavity formation reminiscent of P. dimorphospora. Growth rates at 15, 20 and 26°C were similar for TS4M3, 25M3 and 204-1 but always less for 35-1. Strains 35-1 and TS4M3 showed highest Cu-tolerance on CuSO₄ and micronized copper incorporated into malt extract agar plates. Results indicate the Phialophora spp. A strains represent P. dimorphospora as suggested by strong homology and decay morphology, although considerable intra-variation exists.

Keywords

Soft rot Phylogeny PCR Copper tolerance ASTM soil block test Decay testing

Get full access to this article

View all access options for this article.

References

Abrams

1948. Microbiological Deterioration of Organic Materials. Washington, DC: National Bureau of Standards, U.S. Dept. of Commerce (Miscellaneous publication 188).

Anagnost

S. E.

1998. Light microscopic diagnosis of wood decay. IAWA Journal. 19(2): 141–167. doi: 10.1163/22941932-90001517

Anagnost

S. E.

, Worrall

J. J.

and Wang

C. J. K.

1994. Diffuse cavity formation in soft rot of pine. Wood Science Technology. 28(3): 199–208. doi: 10.1007/BF00193328

AWPA Technical Sub-committee P6 . 2008. Standard Method of Testing Wood Preservatives by Laboratory Soil-Block Culture. Birmingham, AL: AWPA E10-08, American Wood Preservers Association.

Butcher

J. A.

and Nilsson

1982. Influence of variable lignin content amongst hardwoods of soft-rot susceptibility and performance of CCA preservatives. IRG/WP 1151.

Daniel

2014. Fungal and bacterial biodeterioration: white rots, brown rots, soft rots and bacteria. In: Schultz

T. P.

, Goodell

and Nicholas

(eds). Deterioration and Protection of Sustainable Biomaterials, Vol. 1158. Washington, DC. American Chemical Society, 23–58.

Daniel

and Nilsson

1988. Studies on preservative tolerant Phialophora species. International Biodeterioration. 24(4–5): 327–335. doi: 10.1016/0265-3036(88)90018-8

Daniel

and Nilsson

1989. Interactions between soft rot fungi and CCA preservatives in Betula verrucosa. Journal Institute Wood Science. 11: 162–171.

Daniel

and Nilsson

1998. Developments in the study of soft rot and bacterial decay. In: Bruce

and Palfreyman

J. W.

(eds). Forest Products Biotechnology, Ch 3, 37–62. Taylor & Francis, London. ISBN 0-203-48297-2

10.

Daniel

, Volc

and Nilsson

1992. Soft rot and multiple T-branching by the basidiomycete Oudemansiella mucida. Mycological Research. 96(1): 49–54. doi: 10.1016/S0953-7562(09)80995-7

11.

Dereeper

, Guignon

Blanc

Audic

Buffet

, Chevenet

, Dufayar

, Guindon

, Lefort

, Lescot

, Claverie

and Gascuel

2008. Phylogeny, fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Research. 36(Suppl. 2): W465–W469. doi: 10.1093/nar/gkn180

12.

Hale

M. D.

and Eaton

R. A.

1985a. The ultrastructure of soft rot fungi. II. Cavity-forming hyphae in wood cell walls. Mycologia. 77(4): 594–605. doi: 10.2307/3793358

13.

Hale

M. D.

and Eaton

R. A.

1985b. The ultrastructure of soft rot fungi. I. Fine hyphae in wood cell walls. Mycologia. 77, 447–463. doi: 10.2307/3793202

14.

Henningsson

and Nilsson

1971. Preliminära undersökningar över mikrobiella angrepp i impregnerat virke. Lahonturjunta. 2, 10–15 (in Swedish).

15.

Henningsson

and Nilsson

1976. Some aspects on microflora and the decomposition of preservative-treated wood in ground contact. Material und Organismen. 3: 307–318.

16.

Karunasekera

and Daniel

2013. Molecular identification and phylogenic analysis by sequencing the rDNA of copper-tolerant soft-rot Phialophora spp. International Biodeterioration Biodegradation. 82: 45–52. doi: 10.1016/j.ibiod.2013.01.019

17.

Larena

, Salazar

, Gonzalez

, Julian

M. C.

and Rubio

1999. Design of a primer for ribosomal DNA internal transcribed spacer with enhanced specificity for Ascomycetes. Journal Biotechnology. 75(2–3): 187–194. doi: 10.1016/S0168-1656(99)00154-6

18.

Leightley

L. E.

1979. Soft-rot decay in copper-chrome-arsenic treated eucalypt power transmission poles in Queensland, Australia – isolation and identification of soft-rot fungi. International Journal Wood Preservation. 1(4): 143–150.

19.

Leightley

L. E.

1980. Soft-rot wood decay in Eucalyptus power transmission poles in Queensland. Proceedings of Royal Society Queensland. 91: 91–20.

20.

Lindahl

B. D.

, de Boer

and Finlay

R. D.

2010. Disruption of root carbon transport into forest humus stimulates fungal opportunists at the expense of mycorrhizal fungi. The ISME Journal. 4(7): 872–881. doi: 10.1038/ismej.2010.19

21.

Molla

A. H.

, Fakhrul-Razi

, Abd-Aziz

, Hanafi

M. M.

and Alam

M. Z.

2001. In-vitro compatibility evaluation of fungal mixed culture for bioconversion of domestic wastewater sludge. World Journal Microbiological Biotechnology. 17(9): 849–856. doi: 10.1023/A:1013844306960

22.

Nilsson

and Henningsson

1978. Phialophora species occurring in preservative treated wood in ground contact. Material und Organismen. 13: 297–313.

23.

Nilsson

, Obst

J. R.

and Daniel

1988. The possible significance of the lignin content and lignin type on the performance of CCA-treated timber in ground contact. IRG/WP1357.

24.

Rambaut

1996. Se-Al. Sequence Alignment Editor. Version 1.0 alpha 1. Oxford: University of Oxford.

25.

Savory

1954. Breakdown of timber by Ascomycetes and fungi imperfecti. Annals of Applied Biology. 41(2): 336–347. doi: 10.1111/j.1744-7348.1954.tb01126.x

26.

White

T. M.

, Bruns

, Lee

and Taylor

1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J.J., White, T.J. (eds.), PCR protocol: a guide to methods and applications. Academic Press, San Diego, CA, 315–321.

27.

Zabel

R. A.

, Lombard

F. F.

, Wang

C. J. K.

and Terracina

1985. Fungi associated with decay in treated southern pine utility poles in the eastern United States. Wood and Fiber Science. 17: 75–91.

28.

Zainal

A. S.

1976. Micro-Morphological Studies of Soft rot Fungi in Wood. Ph.D. Thesis, University of London.