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Identification and analysis of Phytophthora cactorum genes up-regulated during cyst germination and strawberry infection

Current Genetics volume 57, Article number: 297 (2011) Cite this article

Abstract

The oomycete Phytophthora cactorum can cause economically important diseases on numerous host plants worldwide, such as crown rot on strawberry. To explore the molecular mechanisms underlying the pathogenicity of P. cactorum on strawberry, transcriptional analysis of P. cactorum during strawberry infection and cyst germination was performed by applying suppression subtractive hybridization (SSH) and effector-specific differential display (ESDD) techniques. Two SSH cDNA libraries were generated, enriched for P. cactorum genes expressed during infection or during cyst germination, respectively, and 137 unique differentially expressed genes were identified. To specifically select RxLR effector genes from P. cactorum, ESDD was performed using RxLR and EER motif-based degenerate primers. Eight RxLR effector candidate genes as well as 67 other genes were identified out of 124 selected fragments. The expression levels of 20 putatively up-regulated genes were further analyzed using real-time RT-PCR, showing that, indeed 19 of these 20 genes were up-regulated during at least one of the studied developmental stages or during strawberry crown invasion, relative to the mycelium. This study provides a first overview of P. cactorum genes that are up-regulated immediately prior to or during strawberry infection and also provides a novel method for selecting RxLR effector genes from the unsequenced genome of P. cactorum.

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Abbreviations

ESDD:

Effector specific differential display

Lib IN:

P. cactorum-strawberry interaction library

Lib GC:

P. cactorum germinating cyst library

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Acknowledgments

The authors would like to thank Abdelhameed Elameen for his technical assistance with differential display, Håvard Eikemo for providing P. cactorum isolate 10300 and Jahn Davik for providing strawberry plants. This work was supported by a grant from the Norwegian Research Council (No.179476/I30).

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Affiliations

  1. Plant Health and Plant Protection Division, Norwegian Institute for Agricultural and Environmental Research, Høgskoleveien 7, 1432, Ås, Norway

    Xiaoren Chen, Sonja Sletner Klemsdal & May Bente Brurberg

  2. School of the Horticulture and Plant Protection, Yangzhou University, Wenhui Eastern Road 12, Yangzhou, 225009, Jiangsu Province, China

    Xiaoren Chen

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  1. Xiaoren Chen
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  2. Sonja Sletner Klemsdal
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  3. May Bente Brurberg
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Corresponding author

Correspondence to May Bente Brurberg.

Additional information

The GenBank accession numbers for the sequences reported in this paper are, GW873048–GW873070 (Lib IN), GW874406–GW874527 (Lib GC), GW874528–GW874602 (ESDD).

Communicated by G. Braus.

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Chen, X., Klemsdal, S.S. & Brurberg, M.B. Identification and analysis of Phytophthora cactorum genes up-regulated during cyst germination and strawberry infection. Curr Genet 57, 297 (2011). https://doi.org/10.1007/s00294-011-0348-0

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  • DOI: https://doi.org/10.1007/s00294-011-0348-0

Keywords

  • Phytophthora cactorum
  • Strawberry
  • RxLR effectors
  • SSH
  • Differential display
  • Real-time PCR