ASEAN Journal on Science and Technology for Development


Spiral nematodes Helicotylenchus sp. are known to infect a variety of plants, including coffee. This is in line with the studies and surveys of nematodes in Indonesia. However, to date, there has been no study reporting the nematode with molecular analysis. In this study, we surveyed the nematodes in Arabica coffee plantations in Bondowoso regency – Indonesia. These spiral nematodes were collected in DESS solution for morphological, morphometric and molecular analysis. The morphometric analysis was carried out under an Olympus BH-2 compound microscope with an attached calibrated Toupcam M20 camera. The molecular analysis was performed on the total DNA from a single nematode. The DNA was extracted using Nematodes Dissolving Solution (ISOHAIR, Nippon Gene Japan). PCR process was performed to amplify the small sub-unit area by using a pair of primers, namely the forward primer SSU F07 (5’ -AAA GAT TAA GCC ATG CAT G-3’) and reverse primer SSU R 81 (5’ -TGA TCC WKC YGC AGG TTC AC-3’). The results of the morphometric analysis showed that the nematodes had similar morphometry to Helicotylenchus dihystera. These results were confirmed through phylogenetic analysis which showed that the nematode sequence had a close phylogenetic distance to other H. dihystera sequences found in GenBank. In addition to the sequences documented in this study, both nematodes had homology up to 99.8% with other H. dihystera sequences. Based on the results of morphometric and molecular analysis, the spiral nematodes in this study were H. dihystera. Supported by morphometric and molecular data, this study has documented the first record of H. dihystera from Indonesia.

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