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Identification of fungal genes involved in the preinfection events between ectomycorrhizal association (Pisolithus tinctorius and Pinus massoniana)

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Abstract

The preinfection stage of establishment of the mycorrhiza before physical contact between symbionts is crucial, as changes that occur throughout mycorrhiza formation are set in motion at this time. To improve the understanding of the molecular mechanisms involved in the fungus Pisolithus tinctorius during its interaction with its symbiotic partner Pinus massoniana, we set up a cellophane-separated culture system avoiding physical contact between the symbionts. A cDNA subtraction library representing the differentially expressed genes of P.tinctorius during the preinfection stage was constructed using suppression subtractive hybridization combined with mirror orientation selection (SSH/MOS). Differential screening of the SSH library allowed us to identify 133 unique cDNA clones. Sequences analysis of the expressed sequence tags (ESTs) showed that they represent 23 differentially expressed genes, of which 10 are involved in metabolism and energy, transcription and protein fate, membrane proteins and signaling, and cell rescue and defense, and 13 encoding hypothetical proteins with unknown function. Some fungal genes have not been previously identified in other ectomycorrhizal associations. Expression of the genes that code for ubiquitin-conjugating enzyme E2, 40S ribosomal protein, maternal g10 transcript, ADP ribosylation factor 6, and cytochrome P450 were evaluated by RT-qPCR, confirming the activation of these genes during the preinfection stage.

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Acknowledgments

This work was supported by the Natural Science Foundation of Zhejiang Province, China (Grant Nos. Y3090380, Y3110048, Y13C160011 and Y3110530) and the Project of Innovation team construction and Talents cultivation on Forest food research (Grant No. 2012bF20012).

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Authors and Affiliations

  1. Zhejiang Forestry Academy, Liuhe Road, 399, Hangzhou, 310023, Zhejiang Province, China

    Haibo Li, Huazheng Peng, Liling Wang, Hailong Wei, Nan Li & Qunying Jing

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  1. Haibo Li
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  2. Huazheng Peng
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  3. Liling Wang
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  4. Hailong Wei
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  5. Nan Li
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  6. Qunying Jing
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Correspondence to Haibo Li.

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Haibo Li and Huazheng Peng contributed equally to this work.

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Li, H., Peng, H., Wang, L. et al. Identification of fungal genes involved in the preinfection events between ectomycorrhizal association (Pisolithus tinctorius and Pinus massoniana). Mycol Progress 13, 123–130 (2014). https://doi.org/10.1007/s11557-013-0899-1

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