Morphological and molecular characteristics of Hemicriconemoides cocophillus from the origin of Robusta coffee plantation in Malang, East Java

Abstract

Abstract Hemicriconemoides cocophillus is one of the phytonematode associated with Robusta coffee plantation. Information regarding morphology, morphometry, and molecular characters of H. cocophillus has not been reported in Indonesia. This study is aimed to describe characters of H. cocophillus that was extracted from soil samples taken from Robusta coffee plantation in Malang, East Java. Nematode extraction was conducted using the flotation-centrifuge method. Morphology and morphometric characters were observed from the permanent nematode slides. Single nematode DNA extract was amplified at the D2D3 segment of 28S and ITS1-5.8S-ITS2 with universal primers. The amplicon was sequenced and analyzed for similarity and phylogenetic tree analysis. The female morphological key character of H. cocophillus are sheath looks clear and thin, stylet strong with anchor-shaped knobs, and tail tapering to a finely rounded tip. Female morphometrics key characters are n=5, L=446.5±22.5 μm, a=15.8±0.9, b=5.3±0.2, c=13.9±1.5, stylet length=51.6±2 µm, tail length=32.3±3.2 µm. Based on the D2D3 and ITS1-5.8S-ITS2 regions, H. cocophillus have similarity level of 99% and 98%, respectively with H. cocophillus from NCBI. Phylogenetic tree result using Maximum Likelihood at the D2D3 segment of 28S and ITS1-5.8S-ITS2 regions shows that H. cocophillus in this study was included in one clade with H. cocophillus from Mozambique.