Supporting Information

1. Supporting Information Barceloneic acid C, a new polyketide from an endophytic fungus Phoma sp. JS752 and its antibacterial activities Xuekui Xia1,2†, Soonok Kim3†, Sunghee Bang1, Hyun-Jung Lee3, Changheng Liu2, Chanil Park4*, Sang Hee Shim1* 1School of Biotechnology, Yeungnam University, Gyeongsan 712-749, South Korea 2Key Laboratory for Applied Microbiology of Shandong Province, Biotechnology Center of Shandong Academy of Sciences, Jinan 250014, P. R. China 3National Institutue of Biological Resources, Incheon 404-708, South Korea 4Department of Marine Biology & Aquaculture, College of Marine Science, Gyeongsang National University, Gyeongnam 650-160, South Korea

2. Contents Figure S1. 1H-NMR spectrum of compound 1 (DMSO-d6, 300 MHz) Figure S2. 13C-NMR spectrum of compound 1 (DMSO-d6, 75 MHz) Figure S3. HSQC spectrum of compound 1 (DMSO-d6, 600 MHz) Figure S4. HMBC spectrum of compound 1 (DMSO-d6, 600 MHz) Figure S5. HREIMS of compound 1 S6. Materials and Methods S7. Possible biosynthetic pathway of compounds1-3 S8. HPLC chromatograms of compound 2 (initial state vs state after exposed to the same condition as used for its separation)

3. Figure S1. 1H-NMR spectrum of compound 1 (DMSO-d6, 300 MHz)

4. Figure S2. 13C-NMR spectrum of compound 1 (DMSO-d6, 75 MHz)

5. Figure S3. HSQC spectrum of 1 (DMSO-d6, 600 MHz)

6. Figure S4. HMBC spectrum of 1 (DMSO-d6, 600 MHz)

7. Figure S5. HREIMS of compound 1

8. S6. MATERIALS AND METHODS General Experimental Procedures. 1D and 2D NMR experiments were recorded on a VNS 300 and 600 MHz spectromete. Chemical shifts are expressed in (ppm), and referenced to the residual solvent signals. Mass spectra were recorded on a JEOL JMS600 spectrometer. Semi-preparative HPLC was performed on a Agilent system consisting of a vacuum degasser, quaternary pump, diode array detector (DAD) and a Luna 5u C18 (2) 100 Å column (250 x 10.00 mm, Phenomenex). Column chromatography (CC) was carried out on either silica gel (Merck KGaA, 70–230 mesh) or Sephadex TM LH-20 (GE Healthcare Sweden). TLC was performed on Merck KGaA precoated silica gel 60 F254 plates and spots were visualized under UV light (254 and 365 nm) or by spraying with 20% H2SO4 followed by heating.

9. Fungus material and Cultures. The fungal strain JR752 used in this study was isolated from isolated from reed plants (Phragmites communisTrinius) collected from a swamp at Seochun, South Korea. Fungal strains were identified by sequencing ITS regions with ITS1 and ITS4 primers. After homology search against NCBI nt DB with BlastN algorithm and phylogenetic analysis with ITS sequences from NCBI, JS752 were thought to belong Genus Phoma. After growing on PDA (potato dextrose agar) medium at 28 °C for 5 days, the fungus Phoma. sp. 752, was inoculated into Erlenmeyer flasks (500 mL) containing 200 mL of PDA medium. After incubation for 5 days at 28 °C on a rotary shaker at 150 rpm, 20 mL of culture liquid was transferred as the ‘‘seed’’ into 500 mL flasks, each preloaded with 80 g of rice in 120 mL of water, and grew for 28 days at 28 °C. with the relative humidity Antibacterial Assay. The antibacterial activities against five bacterial strains, Gram-positive Bacillus cereus, Bisteria monocytogenes and Staphylococcus Pseuditermedius, Gram-negative Escherichia coli, Salmonella typhimurium, were determined using a version of the 2-fold serial dilutions method. Compounds 1-3 were dissolved in DMSO to give a stock solution. Bacterial species were cultured overnight at 37 °C in LB broth and diluted to 106 cfu/mL when used. LB broth was used as a blank control, and DMSO was used as a negative control, while ampicillin was used as a positive control. The plates were incubated at 37 °C for 24 h. The results were determined by measuring the inhibition zone.

10. S7. Possible biosynthetic pathway of compounds1-3

11. S8. HPLC chromatograms of compound 2 (initial state vs state after exposed to the same condition as used for its separation) initial compound 2 after exposed to the same condition as used for its separation * Compound 2 was placed in the mixed solution of EtOAc/MeOH for 7 days and then evaporated in vacuo. That was again placed in the mixed solution of CH2Cl2/MeOH for 7 days, which was then evaporated in vacuo. The final treated compound was subjected to HPLC. As shown in the following figure, compound 2 was not converted, showing the same retention time as the initial.