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Background    Cynanchum komarovii  Al Iljinski is a desert plant that has been used as analgesic, anthelminthic and antidiarrheal, but also as a herbal medicine to treat cholecystitis in people. We have found that the protein extractions from  C. komarovii  seeds have strong antifungal activity. There is strong interest to develop protein medication and antifungal pesticides from  C. komarovii  for pharmacological or other uses.    Methodology/Principal Findings  An antifungal protein with sequence homology to thaumatin-like proteins (TLPs) was isolated from  C. komarovii  seeds and named CkTLP. The three-dimensional structure prediction of CkTLP indicated the protein has an acid cleft and a hydrophobic patch. The protein showed antifungal activity against fungal growth of  Verticillium dahliae ,  Fusarium oxysporum ,  Rhizoctonia solani ,  Botrytis cinerea  and  Valsa mali . The full-length cDNA was cloned by RT-PCR and RACE-PCR according to the partial protein sequences obtained by nanoESI-MS/MS. The real-time PCR showed the transcription level of  CkTLP  had a significant increase under the stress of abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), NaCl and drought, which indicates that CkTLP may play an important role in response to abiotic stresses. Histochemical staining showed GUS activity in almost the whole plant, especially in cotyledons, trichomes and vascular tissues of primary root and inflorescences. The CkTLP protein was located in the extracellular space/cell wall by CkTLP::GFP fusion protein in transgenic Arabidopsis. Furthermore, over-expression of CkTLP significantly enhanced the resistance of Arabidopsis against  V. dahliae .    Conclusions/Significance  The results suggest that the CkTLP is a good candidate protein or gene for contributing to the development of disease-resistant crops.

Purification and Characterization of a CkTLP Protein from Cynanchum komarovii Seeds that Confers Antifungal Activity