FBIP: Mycorrhizal and root endophytic fungi associated with Orchids

Study Extent

South Africa (Eastern Cape, KwaZulu-Natal)

Sampling

1 g of orchid root material was collected from each orchid species, replicated 6 times, roots were surface sterilized and were etiher stored in RNA Later and frozen for molecular analysis or they were were sliced into smaller segments and placed onto suitable media to isolate endophytic fungi.

Quality Control

The coordinates of the location where samples were collected was recorded

Method steps

1. Fungal isolates: Sampled root sections were surface sterilized and plated onto Malt Extract Agar. After incubation sub-cultures were made to ensure pure cultures were maintained. DNA was extracted from the cultures and the ITS region was amplified using ITS1 and ITS 4 primers. After purification amplicons were sent for Sanger Sequencing and submitted to GenBank for identification. Root samples: Genomic DNA was extracted from root samples, followed by polymerase chain reaction (PCR) amplification of the ITS region using Miseq tagged primers. Amplicon were submitted to SAIAB for Illumina Next Generation Sequencing.Data was analysed using Mothur software.

The isolates and metagenomic sequences are mycorrhizal and other root endophytic fungi that are found within orchid roots

1. Fungi
   rank: phylum

South Africa (Eastern Cape, KwaZulu-Natal)