*Article* **Antimalarials and Phytotoxins from** *Botryosphaeria dothidea* **Identified from a Seed of Diseased** *Torreya taxifolia*

**Mallika Kumarihamy 1,2,\*, Luiz H. Rosa 3, Natascha Techen 1, Daneel Ferreira 2, Edward M. Croom, Jr. 2, Stephen O. Duke 4,†, Babu L. Tekwani 1,‡, Shabana Khan 1,2 and N. P. Dhammika Nanayakkara 1,\***


**Abstract:** The metabolic pathways in the apicoplast organelle of *Plasmodium* parasites are similar to those in plastids in plant cells and are suitable targets for malaria drug discovery. Some phytotoxins released by plant pathogenic fungi have been known to target metabolic pathways of the plastid; thus, they may also serve as potential antimalarial drug leads. An EtOAc extract of the broth of the endophyte *Botryosphaeria dothidea* isolated from a seed collected from a *Torreya taxifolia* plant with disease symptoms, showed in vitro antimalarial and phytotoxic activities. Bioactivity-guided fractionation of the extract afforded a mixture of two known isomeric phytotoxins, FRT-A and flavipucine (or their enantiomers, sapinopyridione and (-)-flavipucine), and two new unstable *γ*lactam alkaloids dothilactaenes A and B. The isomeric mixture of phytotoxins displayed strong phytotoxicity against both a dicot and a monocot and moderate cytotoxicity against a panel of cell lines. Dothilactaene A showed no activity. Dothilactaene B was isolated from the active fraction, which showed moderate in vitro antiplasmodial activity with high selectivity index. In spite of this activity, its instability and various other biological activities shown by related compounds would preclude it from being a viable antimalarial lead.

**Keywords:** *Torreya taxifolia*; plant pathogenic fungi; *Botryosphaeria dothidea*; malaria; phytotoxin; *γ*-lactam alkaloids
