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Molecular cloning of α-amanitin and characterization of its expression pattern in different parts and development stages of Amanita exitialis fruitbody

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Abstract

Amanita exitialis is a lethal mushroom in East Asia. α-Amanitin is one kind of the leading peptide toxins responsible for mushroom poisoning, which are mainly from the fatal amanitae. In the study, the full-length cDNA sequences of α-amanitin (α-AMA) and phallacidin (PHA) from A. exitialis were cloned and analyzed; in addition, the expression patterns of α-AMA in three parts as well as four developmental stages of the fruitbody were first characterized by quantitative real-time PCR. Results demonstrate that α-AMA and PHA were highly homologous to the toxin genes reported from A. bisporigera; the α-AMA could be expressed in all parts and stages of the fruitbody, from late elongation stage to late mature stage, α-AMA showed the highest expression level in the pileus, followed by the stipe and volva in sequence, while at the early elongation stage, the highest expression level was found in the stipe, followed by the pileus and volva. Further analysis revealed that the higher expression level of α-AMA was closely associated with the development stages, and the expression of α-AMA was accumulated in the more vigorous growth parts and stages, from which could be inferred that α-AMA might play an important role in the development of A. exitialis.

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References

  • Bresinsky A, Besl H (1990) A color atlas of poisonous fungi: a handbook for pharmacists, doctors and biologists. p.295. Wolfe, Wurzburg, Germany

  • Deng WQ, Li TH, Xi PG, Gan LX, Xiao ZD, Jiang ZD (2011) Peptide toxin components of Amanita exitialis basidiocarps. Mycologia 103:946–949

    Article  CAS  PubMed  Google Scholar 

  • Enjalbert F, Cassanas G, Andary C (1989) Variation in amounts of main phallotoxins in Amanita phalloides. Mycologia 81:266–271

    Article  CAS  Google Scholar 

  • Enjalbert F, Gallion C, Jehl F, Monteil H (1993) Toxin content, phallotoxin and amatoxin composition of Amanita phalloides tissues. Toxicon 31:803–807

    Article  CAS  PubMed  Google Scholar 

  • Enjalbert F, Gallion C, Guinchard C, Chaumont JP (1999) Distribution of the amatoxins and phallotoxins in Amanita phalloides. Influence of the tissues and the collection site. C R Acad Sci III Sci Vie Life Sci 322:855–862

    CAS  Google Scholar 

  • Hallen-Adams HE, Luo H, Scott-Craig JS, Walton JD (2007) Gene family encoding the major toxins of lethal Amanita mushrooms. Proc Natl Acad Sci U S A 104:19097–19101

    Article  Google Scholar 

  • Hu JS, Zhang P, Zeng J, Chen ZH (2012) Determination of amatoxins in different tissues and development stages of Amanita exitialis. J Sci Food Agric 92:2664–2667

    Article  CAS  PubMed  Google Scholar 

  • Kaya E, Yilmaz I, Sinirlioglu ZA, Karahan S, Bayram R, Yaykasli KO, Colakoglu S, Saritas A, Severoglu Z (2013) Amanitin and phallotoxin concentration in Amanita phalloides var. alba mushroom. Toxicon 76:225–233

    Article  CAS  PubMed  Google Scholar 

  • Li C, Oberlies NH (2005) The most widely recognized mushroom: chemistry of the genus Amanita. Life Sci 78:532–538

    Article  CAS  PubMed  Google Scholar 

  • Li P, Deng WQ, Li TH, Song B, Shen YH (2013) Illumina-based de novo transcriptome sequencing and analysis of Amanita exitialis basidiocarps. Gene 532:63–71

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(−Delta Delta C) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Luo H, Hallen-Adams HE, Walton JD (2009) Processing of the phalloidin proprotein by prolyl oligopeptidase from the mushroom Conocybe albipes. J Biol Chem 284:18070–18077

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Luo H, Hallen-Adams HE, Scott-Craig JS, Walton JD (2010) Colocalization of amanitin and a candidate toxin-processing prolyl oligopeptidase in Amanita Basidiocarps. Eukaryot Cell 9:1891–1900

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Luo H, Hallen-Adams HE, Scott-Craig JS, Walton JD (2012) Ribosomal biosynthesis of alpha-amanitin in Galerina marginata. Fungal Genet Biol 49:123–129

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:0034.1

    Article  Google Scholar 

  • Walton JD, Hallen-Adams HE, Luo H (2010) Ribosomal biosynthesis of the cyclic peptide toxins of Amanita mushrooms. Biopolymers 94:659–664

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wieland T (1986) Peptides of poisonous Amanita mushrooms. Springer, New York

    Book  Google Scholar 

  • Xue JH, Wu P, Chi YL, Xu LX, Wei XY (2011) Cyclopeptides from Amanita exitialis. Nat Prod Bioprospecting 1:52–56

    Article  CAS  Google Scholar 

  • Yang ZL (2005) Amanitaceae. Flora fungorum sinicorum. Vol. 27,ed.), Science, Beijing [in Chinese]

  • Yang ZL, Li TH (2001) Notes on three white Amanitae of section Phalloideae (Amanitaceae) from China. Mycotaxon 78:439–448

    Google Scholar 

  • Zhang P, Chen ZH, Hu JS, Wei BY, Zhang ZG, Hu WQ (2005) Production and characterization of amanitin toxins from a pure culture of Amanita exitialis. FEMS Microbiol Lett 252:223–228

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was funded by grants from the National Natural Science Foundation of China (Project Nos. 31101592).

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Correspondence to Wang-qiu Deng or Tai-hui Li.

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Validation of the 2-ΔΔCt method: Amplification of cDNA synthesized from different amounts of RNA. (A) and (B) were the cDNA of the pileus of the early mature stage and late elongation stage, respectively. All statistics were presented as means ± SE. (GIF 32 kb)

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Li, P., Deng, Wq. & Li, Th. Molecular cloning of α-amanitin and characterization of its expression pattern in different parts and development stages of Amanita exitialis fruitbody. Mycol Progress 13, 988 (2014). https://doi.org/10.1007/s11557-014-0988-9

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  • DOI: https://doi.org/10.1007/s11557-014-0988-9

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