Abstract
The Pisolithus tinctorius symbiosis related protein expressed sequence tag (EST PtSRP) was previously identified in the first hours of the interaction between the fungus Pisolithus tinctorius and sweet chestnut Castanea sativa, and partially characterized as a fungal marker gene of ectomycorrhizal symbiosis formation. We used the 5’ rapid amplification of cDNA ends (RACE) to obtain the PtSRP mRNA 5’ region, and together with our previously reported 3’ mRNA region, the full mRNA sequence was assembled by use of bioinformatics tools and deposited to GenBank (Accession: GU733439). The full-length mRNA sequence (636 bp) revealed the locations of the 5’ and 3’ untranslated regions (UTRs) and contained the Kozak sequence (ccc aag ATG A) in the 5’ UTR. The in silico translated PtSRP open reading frame (ORF) codes for a 127 amino acid protein and contained four putative post-translational modification sites (two N-glycosylation and two phosphorylation). The protein secondary structure is postulated to be composed of one N-terminal hydrophobic transmembrane alpha helix and at least six hydrophilic beta-strands spread across the protein. Sub-cellular localization prediction suggests that the protein is involved in cellular secretory pathway, supported by the presence of a cleavage site motif close to the membrane anchor. The data presented herein indicate the role of PtSRP as a fungal membrane secreted protein involved in early stages of ectomycorrhizal formation, with application as a possible marker for nascent ectomy-corrhiza fungal development.
Highlights
The establishment of ectomycorrhiza involves controlled, intense gene expression in both partners that leads to drastic morphological and physiological changes, crucial to the development of mutualism and symbiotic harmony [1,2,3]
We present the full-length PtSRP fungal mRNA sequence, supported by sequencing of the 5’ region and our pre
The nearly perfect alignment of the 604 bp consensus fragment to the 636 bp contig supported the reliability of the contig; only two nucleotide differences (291 C/T and 306 A/C, Figure 2) were observed and they were in the putative open reading frame (ORF) region, with no changes to the amino acid
Summary
The establishment of ectomycorrhiza involves controlled, intense gene expression in both partners that leads to drastic morphological and physiological changes, crucial to the development of mutualism and symbiotic harmony [1,2,3]. The comparison of protein extracts from mycorrhizal and non-mycorrhizal mycelia in previous studies has shown differences that suggest specific gene activation during the symbiosis process [4,5,6]. These findings highlighted a new class of biomolecules thought to control the ectomycorrhiza symbiosis process: the ectomycorrhizins [7]. SRAPs (Symbiosis Related Acid Proteins) and hydrophobins are the most investigated and discussed classes of proteins These proteins were generally isolated from fully established mycorrhiza or those developing associations after several days of interaction [8,9]. We present the full-length PtSRP fungal mRNA sequence, supported by sequencing of the 5’ region and our pre-
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