The diversity of endophytic fungi of the sand coast plant Ipomoea pes-caprae in Taiwan

Study Extent

The study was conducted in northern and central Taiwan; The dataset covers natural vegetation at sand coasts of New Taipei City (Bali District and Shihmen District), Taoyuan (Guanyin District), Hsinchu County (Xinfeng Township) and Taichung City (Daan District and North District) and artificial plantations in the botanical gardens of Taipei City and Taichung City.

Sampling

One individual plant was collected per sampling (twice per year, representing the summer and the less hot seasons). Plants which were not conspicuously buried by sand during the collection time were removed with a trowel, individually placed in bags, returned to the laboratory and kept at 4°C until further processing for endophyte isolation within 48 hours after sampling. Altogether, 37 individuals of I. pes-caprae were collected from eight sites (27 from New Taipei, Taoyuan, Hsinchu and Taichung; 10 from Taipei and Taichung Botanical Gardens). Freshly collected healthy plants were cut into fragments (leaves, stems, roots) and surface-sterilized. Plant fragments were surface-sterilized under sterile conditions by agitation in 95% ethanol for 1 min, 6–12% sodium hypochlorite for 3 min (for stems) or 1.5 min (for roots and leaves), 95% ethanol for 0.5 min, and then rinsed in sterile water. All stems and roots were cut into six segments of approximately 1–2 cm, and each leaf into the petiole as well as three segments of ca. 0.6 cm diam. from the lamina. Three segments of stems and root and two segments of leaf lamina and the petiole were immediately placed onto malt extract agar (MEA) or corn extract agar (CMA) with 0.2% chloramphenicol. All isolates obtained from each plant sample were classified according to their morphological appearance into morphotypes. Representative isolates were identified to species as far as possible and deposited at the Bioresource Collection and Research Center, Hsinchu, Taiwan (BCRC). Dried cultures were deposited in the fungal specimen collection of the National Museum of Natural Science, Taichung, Taiwan (TNM).

Quality Control

Since surface-sterilization has to be adjusted whenever endophytes are isolated from a hitherto uninvestigated plant species, we optimized the methods for surface sterilization for the different plant parts of I. pes-caprae. The effectiveness was further controlled regularly by the imprint technique, i.d. by temporarily pressing the surface-sterilized plant fragments onto control media. Only if no growth occurred on these control media, but in the media with plant fragments, then the surface sterilization procedure was neither too weak nor too rigorous (Rodrigruez et al. 2008). DNA sequencing was performed by Mission Biotech (Nankang, Taipei) with the same primers used for the PCR. The forward and reverse DNA sequences were edited using CodonCode Aligner version 4.0.1 (CodonCode Corp., USA) and submitted to GenBank and the DNA Data Bank of Japan. The sequences were submitted to BLAST searches at GenBank (https://blast.ncbi.nlm.nih.gov/). Depending on the similarities of the used barcode and verifiability by morphology, the strains were identified to species, genus or higher taxonomic ranks. Only sequences cited in publications were considered for identification. Scientific names were crosschecked and updated with Index Fungorum (http://www.indexfungorum.org).

Method steps

1. Field collection, light microscopy, cultivation, DNA isolation, PCR, sequencing, sequence analyses

This dataset contains data from the Kingdom Fungi, Division Ascomycota, Basidiomycota and Zygomycota, corresponding to a total of 13 classes and 33 orders. It includes 177 different species of fungi for a total of 896 fungal strains.

1. Fungi

1. Taipei Botanical Garden in Taipei City (Latitude: 25.032205 Longitude: 121.509884) 2. Taichung Botanical Garden in Taichung City (Latitude: 24.156172 Longitude: 120.666314) 3. Wazihwei beach in New Taipei City (Latitude: 25.168699 Longitude: 121.414081) 4. Shimen beach in New Taipei City (Latitude: 25.292212 Longitude: 121.544561) 5. Daan beach in Taichung City (Latitude: 24.379000 Longitude: 120.583286) 6. Nanpu beach in Taichung City (Latitude: 24.343201 Longitude: 120.560261) 7. Sandy beach at Fongkeng fishing port in Hsinchu County (Latitude: 24.90521 Longitude: 120.96632) 8. Guanyin Beach in Taoyuan City (Latitude: 25.047 Longitude: 121.076)