Fungicidal properties and insights on the mechanisms of the action of volatile oils from Amazonian Aniba trees

The Amazonian Aniba species are world-renowned for their essential oils (EOs). The molecules derived from EOs have been intensively investigated in regards to their potential for disease control in plants. The aim of this study was to investigate the antifungal properties of Aniba canelilla EO (ACEO) and Aniba parviflora EO (APEO) when used against eight phytopathogenic fungi. Gas chromatography-mass spectrometry (GC–MS) analysis of oils showed that 1-nitro-2-phenylethane (∼80%) and linalool (∼40%) are the major compounds in ACEO and APEO, respectively. The ACEO and APEO treatments displayed remarkable antifungal effects against Aspergillus flavus, Aspergillus niger, Fusarium oxysporum, Fusarium solani, Alternaria alternata, Colletotrichum gloeosporioides, Colletotrichum musae and Colletotrichum guaranicola, for which the IC50 values ranged from 0.05 to 0.28 μL mL−1 and 0.17 to 0.63 μL mL−1, respectively. Furthermore, the oil caused the inhibition of conidial germination by at least 83% for ACEO and 78% for APEO. The ACEO and APEO at 5 μL mL−1 induced leakage of nucleic acids and protein, suggesting that inhibition could be linked to the breakdown of membrane integrity of the conidia. In addition, the detection of fluorescent dye propidium iodide (PI) on F. solani conidia treated with ACEO and APEO indicates damage on the conidia cytoplasmic membrane. The findings of this study may be of biotechnological interest for the development of new plant protection products, with the advantage of being less harmful than the agrochemicals currently available. © 2019 Elsevier B.V

ClCPI, a cysteine protease inhibitor purified from Cassia leiandra seeds has antifungal activity against Candida tropicalis by inducing disruption of the cell surface

Infections caused by Candida tropicalis have increased significantly worldwide in parallel with resistance to antifungal drugs. To overcome resistance novel drugs have to be discovered. The objective of this work was to purify and characterize a cysteine protease inhibitor from the seeds of the Amazon rainforest tree Cassia leiandra and test its inhibitory effect against C. tropicalis growth. The inhibitor, named ClCPI, was purified after ion exchange and affinity chromatography followed by ultrafiltration. ClCPI is composed of a single polypeptide chain and is not a glycoprotein. The molecular mass determined by SDS-PAGE in the absence or presence of β-mercaptoethanol and ESI-MS were 16.63 kDa and 18.362 kDa, respectively. ClCPI was stable in the pH range of 7.0–9.0 and thermostable up to 60 °C for 20 min. ClCPI inhibited cysteine proteases, but not trypsin, chymotrypsin neither alpha-amylase. Inhibition of papain was uncompetitive with a Ki of 4.1 × 10 −7 M and IC 50 of 8.5 × 10 −7 M. ClCPI at 2.6 × 10 −6 M reduced 50% C. tropicalis growth. ClCPI induced damages and morphological alterations in C. tropicalis cell surface, which led to death. These results suggest that ClCPI have great potential for the development of an antifungal drug against C. tropicalis. © 201