# Low Nitrogen Concentration and Acidic pH Enhance the Antifungal Activity Against Botrytis cinerea of an Endophytic Alternaria sp. Isolated from Opuntia ficus-indica

**Authors:** Leonora Mendoza, Camilo Aguirre, Rodolfo Parada, Hristo Parada, Paulo Castro, Freddy Navarro, Araceli Vidal, Milena Cotoras

PMC · DOI: 10.3390/microorganisms14020376 · 2026-02-05

## TL;DR

A fungus from cacti shows stronger antifungal effects against Botrytis cinerea under acidic and low-nitrogen conditions, offering potential for sustainable agriculture.

## Contribution

The study identifies optimal culture conditions to enhance antifungal metabolite production by an endophytic Alternaria sp.

## Key findings

- Antifungal activity increased under acidic pH and low nitrogen conditions.
- Volatile compounds showed strongest inhibition at pH 4.5 and low ammonium tartrate.
- Ethyl acetate extracts inhibited Botrytis cinerea mycelial growth by 60% under optimized conditions.

## Abstract

Endophytic fungi from the Cactaceae family are an underexplored source of bioactive secondary metabolites with potential applications in sustainable agriculture. This study investigated an endophytic fungus obtained from healthy cladodes of Opuntia ficus-indica growing in the Chilean Andean Precordillera. The influence of culture conditions, specifically pH and nitrogen concentration, on the production of diffusible and volatile antifungal compounds against the phytopathogen Botrytis cinerea was evaluated using dual-culture (confrontation) and sandwiched Petri dish assays. Morphological characteristics and molecular analyses confirmed that the isolate belongs to the genus Alternaria. Antifungal activity increased significantly under acidic conditions and limited nitrogen availability. The strongest inhibition by volatile compounds occurred at pH 4.5 and the lowest concentration of ammonium tartrate. Furthermore, ethyl acetate extracts at 40 mg/L obtained from Alternaria sp. cultures grown at pH 4.5 with 2.3 g/L ammonium tartrate inhibited B. cinerea mycelial growth by 60%. The study provides a framework for improving the yield of antifungal metabolites produced by Alternaria, contributing to the development of biofungicides for gray mold control.

## Linked entities

- **Chemicals:** ammonium tartrate (PubChem CID 2724224), ethyl acetate (PubChem CID 8857)
- **Species:** Opuntia ficus-indica (taxon 371859), Botrytis cinerea (taxon 40559), Alternaria sp. (taxon 1715220)

## Full-text entities

- **Diseases:** Endophytic Fungus (MESH:D009181), gray (MESH:D055652), injury to (MESH:D014947), inflammatory (MESH:D007249)
- **Chemicals:** phenols (MESH:D010636), sodium hypochlorite (MESH:D012973), acetone (MESH:D000096), Silica (MESH:D012822), FeSO4 x 7H2O (-), ethyl acetate (MESH:C007650), glucose (MESH:D005947), flavonoids (MESH:D005419), chloramphenicol (MESH:D002701), sulfuric acid (MESH:C033158), tenuazonic acid (MESH:D013720), KCl (MESH:D011189), Tween  20 (MESH:D011136), anthocyanins (MESH:D000872), chloroform (MESH:D002725), bikaverin (MESH:C000269), harzianic acid (MESH:C088506), polymalic acid (MESH:C121060), ammonium tartrate (MESH:C029768), agar (MESH:D000362), carbon (MESH:D002244), ammonium (MESH:D064751), alternariol (MESH:C005197), salvic acid (MESH:C576147), K2HPO4 (MESH:C013216), N (MESH:D009584), gibberellic acid (MESH:C007842), patulin (MESH:D010365), quinones (MESH:D011809), silica gel (MESH:D058428), methanol (MESH:D000432), iprodione (MESH:C033148), polyketide (MESH:D061065), ethanol (MESH:D000431), polythene (MESH:D020959), kanamycin (MESH:D007612), alkaloids (MESH:D000470), vanillin (MESH:C100058), terpenoids (MESH:D013729), water (MESH:D014867)
- **Species:** Homo sapiens (human, species) [taxon 9606], Bacteria Latreille et al. 1825 (Bacteria stick insect, genus) [taxon 629395], Fusarium verticillioides (species) [taxon 117187], Botrytis cinerea (gray fruit mold, species) [taxon 40559], Solanum (genus) [taxon 4107], Fusarium fujikuroi (species) [taxon 5127], Cladosporium sphaerospermum (species) [taxon 92950], Conioselinum tenuissimum (species) [taxon 110318], Alternaria sp. (species) [taxon 1715220], Opuntia ficus-indica (Indian-fig, species) [taxon 371859], Aspergillus niger (species) [taxon 5061], Embothrium coccineum (Chilean firebush, species) [taxon 54937], Oscillospira sp. F (species) [taxon 227390], Fusarium graminearum (species) [taxon 5518], Saccharomyces cerevisiae (baker's yeast, species) [taxon 4932], Chenopodium quinoa (quinoa, species) [taxon 63459], Penicillium expansum (species) [taxon 27334], Opuntia humifusa (creeping prickly-pear, species) [taxon 446152], Stemphylium vesicarium (species) [taxon 119933], Fungi (kingdom) [taxon 4751], Aureobasidium pullulans (species) [taxon 5580], Leucostele chiloensis (species) [taxon 183649], Cladosporium cladosporioides (species) [taxon 29917], Vitis vinifera (wine grape, species) [taxon 29760], Aspergillus ochraceus (species) [taxon 40380], Trichoderma harzianum (species) [taxon 5544]

## Figures

7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12943664/full.md

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Source: https://tomesphere.com/paper/PMC12943664