Nucleopolyhedrovirus (NPV) is a potential biological control agent recommended to control corn cob borer larvae (Helicoverpa armigera Huber). NPV characters can be studied using various detection and ...identification methods. One technique to study the character of NPV is polymerase chain reaction (PCR) followed by analysis of the results of nucleotide and amino acid sequences. The molecular character of NPV in infected H. armigera larvae in corn plantations in Cibeureum Village, Dramaga District, Bogor Regency using DNA polymerase partial gene sequences needs to be done. The methods used to study the character of the NPV consisted of DNA isolation using a modified CTAB method, partial gene amplification of DNA polymerase using HearNPVF1 and HearNPVR1, and analysis of the level of homology of nucleotide and amino acid sequences compared to other countries and phylogeny. PCR amplification using specific primers was successfully carried out with the HearNPV DNA polymerase partial gene DNA band measuring about 1,200 bp. Phylogenetic analysis was also successfully carried out and showed that there was a high relationship between HearNPV isolates from Indonesia (Bogor, West Java) and NPV isolates that infect Helicoverpa from other countries such as: Spain, Australia, the Netherlands, India, Brazil, Russia, and China with nucleotide and amino acid homology values of 99%. HearNPV isolates from Bogor were in the same group as NPVs that attacked the Genus Helicoverpa from other countries, while NPVs from other genera were in separate groups based on phylogenetic analysis using Mega 7 software.
Phytoplasma as a phytopathogenic prokaryote with a wide host range is a pathogen that needs more attention in Indonesia. This pathogen is relatively difficult to detect and identify due to its ...complicated biological properties. This study involved detection of phytoplasmas by polymerase chain reaction (PCR) technique with P1/P7 primers from seven symptomatic plants, i.e. Bermuda grass white leaf, bamboo yellows, witches’ broom of peanut, soybean, yard long bean, and cactus, and sweet potato little leaf. The phytoplasma DNA of the 16S rRNA gene resulting from PCR amplification was examined by digestion reaction using three endonuclease enzymes AluI, RSaI, and MSeI to generate restriction fragment length polymorphism (RFLP) profile. The seven diseased plants were confirmed positive to be associated with phytoplasma as indicated by the PCR product of 1800 bp. Based on the RFLP profiles of the three enzymes, the phytoplasmas were divided into two groups, namely group I (Bermuda grass and bamboo) and group II (peanuts, soybeans, yard long beans, cactus, and sweet potatoes). Cactus phytoplasma is a sub-group (strain) because it has a slightly different fragment of MSeI RFLP profile.
Hyposidra talaca (Walker) is an important pest of tea plant. H. talaca can cause losses of between 40–100% in the dry season if proper control is not carried out. H. talaca has natural enemies such ...as predators, parasitoids, and pathogens. One of the entomopathogens is NPV. This study aims to obtain molecular characteristics through DNA polymerase sequences and determine the virulence level of NPV isolates from H. talaca. The same species from different locations can have genetic variability. Therefore, molecular characterization by polymerase chain reaction (PCR) on DNA polymerase sequences is one way to study the genetics of HytaNPV. NPV was isolated from infected H. talaca larvae collected from the field. The DNA isolates were used as templates for PCR for DNA polymerase gene amplification with an amplicon target of ±1,000 bp. A sequencing process followed the PCR provides nucleotide sequence. HytaNPV DNA polymerase sequencing results were aligned with GenBank’s BLAST data to provide information on the relationship of HytaNPV to NPVs isolated from other regions. Based on molecular character analysis using DNA polymerase gene sequence, HytaNPV Bogor has a homology level of 93.9% with HytaNPV isolated from India. HytaNPV Bogor has a genetic relationship with the NPV that infects Buzura suppressaria from China and Australia. HytaNPV Bogor is similar to the NPV that infects H. talaca from India. The bioassay of HytaNPV isolate against H. talaca showed the highest LT50 value of 1.92 days was found in concentration of 1.58 x 107 POBs/ml in second instar larvae.
Aphelenchoides besseyi merupakan nematoda penyebab penyakit pucuk putih yang terbawa benih padi. Gen AB FAR-1 diketahui sebagai gen penting yang mengendalikan patogenisitas A. besseyi. Penelitian ...dilakukan untuk mengetahui karakter gen AB FAR-1 yang diisolasi dari nematoda yang berasal dari benih padi. Ekstraksi nematoda dilakukan dengan metode corong Baerman dari benih 5 varietas padi “Ciherang“, “Inpari Sidenuk“, “Sintanur“, “Hibrida Prima“ dan “Pak Tiwi“. Ekstraksi DNA total nematoda menggunakan metode CTAB dilanjutkan dengan amplifikasi gen AB FAR-1 menggunakan primer spesifik FAR-F1/R1 dan analisis urutan nukleotidanya. Pita DNA spesifik gen AB FAR-1 berukuran 150 pb berhasil diamplifikasi dari semua sampel nematoda. Analisis sekuen menunjukkan bahwa gen AB FAR-1 tersebut memiliki homologi tertinggi (92.5 – 100%) dengan aksesi Genbank JQ686690.1, yaitu gen AB FAR-1 A. besseyi asal Cina. Walaupun memiliki homologi yang tinggi, terdapat beberapa perbedaan nukleotida pada sampel gen AB FAR-1 A. besseyi asal “Ciherang“, “Inpari Sidenuk“ dan “Hibrida Prima“. Analisis pohon filogenetika lebih lanjut mengelompokkan gen AB FAR-1 A. besseyi menjadi 2 grup, yaitu grup 1 terdiri atas gen AB FAR-1 A. besseyi asal Cina, “Sintanur“, “Hibrida Prima“ dan “Pak Tiwi“ dan grup 2 gen AB FAR-1 A. besseyi asal “Ciherang“, dan “Inpari Sidenuk“.
Rahayuwati S, Kusumah YM, Prawirosukarto S, Dadang, Santoso T. 2020. Genetic variability of Indonesian Oryctes rhinoceros nudivirus (OrNV) as genus of Alphanudivirus. Biodiversitas 21: 2047-2055. ...Oryctes rhinoceros nudivirus (OrNV) is known as a successful classical biological control agent. Recent reports have revealed differences in pathogenicity among OrNV isolates, with correlation suspected between virus virulence and genetic variability. The aim of the research is to study the genetic variability of OrNV isolates based on the DNA polymerase (DNA pol) gene and their phylogenetic relationship with Genus Alphanudivirus. Oryctes rhinoceros (Coleoptera: Scarabaeidae) as an OrNV host were collected from Sumatra, Belitung, Java, Kalimantan, and Sulawesi. Primers were designed to amplify complete gene of DNA pol. Sequencing, phylogenetic, and investigate genetic diversity scores were used to determine variability at nucleotide level. Five primers successfully amplified the entire DNA pol gene. Genetic variability of OrNV was high, from nine isolates found eight genetically different. Isolates could be divided into two groups of genetic diversity: high and low. High genetic diversity of OrNV supposed as an old population that occupied Sundaland and Wallacean paleogeography at first appearance in Miocene Era until early Pliocene 5 Mya. New population of OrNV had low genetic diversity were travel cross-island that supposed to correlate with oil palm plantation development and the genetic drift. The benefit information of the OrNV genetic variability is as preliminary guideline for virulence test. OrNV is a member of Nudiviridae family, Genus Alphanudivirus, and at recent moment they consist of eight species. Alphanudivirus infect Coleoptera, Diptera, Hemiptera and Orthoptera order that soil as immature niche may explain their closely related taxa.
Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) is one of the most important pests on cotton. Yield losses due to this pest can reach up to 70%. Pest control using the NPV has been proven to be ...effective in the field. However NPV isolates from different geographical locations could be showed variation in genetic and their biology. The purpose of this study was to compare morphology, biology, and molecular characteristics between three HearNPV isolates from cotton field in South Sulawesi, Central Java, and East Java. Scanning electron microscope (SEM) was used to look at morphological characters, whereas biological characters were observed by dipping shelled of young corn cob on each isolate suspension with a density of 2.5 x 103, 2.5 x 104, 2.5 x 105, 2.5 x 106, 2.5 x 107 POBs /ml and control then fed to the first instar larvae of H. armigera. Probit analysis was used to determine LC and LT. The molecular assay used polymerase chain reaction (PCR) with lef-8 gene. The results showed that the three isolates had the shape and size of polyhedra which was not significantly different with the mean diameter were 0.86 μm. The result showed that in 7 days post infection, HearNPV isolate from East Java was the most virulent isolates with the lowest LC50, which is 6.38 POBs/ml, but lowest LT50 value in 2.5 x 107 POBs/ml were showed both from East Java and South Sulawesi, which are 2.7 days post infection. Meanwhile, the DNA sequencing of the three isolates showed a high similarity (100% homology).
Spodoptera frugiperda is an invasive pest from the American continent that attacks corn (Zea mays) and rapidly invaded Africa and Asia. Two main factors that support migration and population ...distribution of this species are suitable habitats and human activities. To date, two genetic strains of S. frugiperda have been found in corn in Indonesia: the corn strain (CS) and the rice strain (RS). The most accurate gene markers to detect these strains are COI and Tpi, which are located in mitochondria and Z chromosome. This study aims to determine the existing strains of S. frugiperda and their distribution in various landscapes in Bogor Regency. The research was conducted from July 2020 to December 2021 in Bogor, West Java. Sampling of S. fungiperda was carried out from corn plants in Leuwisadeng, Pamijahan1, Pamijahan2, Kemang, Tenjolaya, Dramaga, Cigombong, Cijeruk, Tamansari, and Ciomas. Larval samples were collected and preserved using 96% ethanol, followed by DNA extraction, DNA amplification, electrophoresis, and DNA sequencing. Distribution data were analyzedusing QGIS and Google Earth Pro programs, and statistical analysis was performed using SPSS 22. Sequence data were edited using GeneStudio, aligned using ClustalW in BioEdit, and the phylogeny tree was reconstructed using the neighbor-joining method (bootstrap 1000x) using MEGA X. The obtained sequences were compared with sequences from the GenBank® database. The results showed the presence of two distinct strains of COI (COI-CSh4 and COI-RS) and one strain of Tpi (Tpi-C) in Bogor. The study found no relationship between thelandscape structure and genetic variation of S. frugiperda.
Cut flower of chrysanthemum has high aesthetic and economic values. Export volume of chrysanthemums from Indonesia are lower than other countries, due to the impact of insect pest. Insect pests, ...Aphis gossypii Glover and Macrosiphoniella sanborni Gillette are currently associated with cut flowers especially chrysanthemum. One control measures usually taken in quarantine is fumigation. Alternative fumigant, liquified phosphine formulations may potentially be applied for quarantine treatment. The objectives of this study were to determine the concentration and exposure time of liquified phosphine against these species and to evaluate the effect of fumigant on the physical quality of cut flowers. The experiment was conducted in four steps: (1) identification and mass rearing of A. gossypii and M. sanborni; (2) preliminary tests on adult and third instar nymphs; (3) determination of liquified phosphine with various concentration and exposure time against adult and third instar nymphs of A. gosspypii and M. sanborni and; (4) validation test of effective concentration of fumigant and exposure time and the effect on quality of cut flowers. The results of study showed concentration of fumigant causing 100% mortality A. gossypii and M. sanborni ere 500 ppm and 700 ppm, respectively with exposure time 12 hours. Concentrations of 700 ppm and exposure time up to 18 hours did not cause negative impact to the quality of chrysanthemum cut flowers.
Characterize molecular the Nucleopolyhedrovirus (NPV) Hyposidra talaca Wlk. of tea plantation at Gunung Mas Bogor. Hyposidra talaca is one of the most important pest in tea plantation, and generally ...attacks of leaves and shoots. This pest cause yield loss up to 40-100%. NPV can be pathogenic to the pest of H. talaca and can be developed as an alternative measure to control H. talaca in tea plantations and based management appears to be more ecofriendly and effective. However, information regarding characterisation molecular of HytaNPV is limited. The study conducted to characterize molecular the NPV of H. talaca by restriction nucleotide and amino acid, by using gens lef-8. Molecular identification used Polymerase Chain Reaction (PCR) consisted of DNA extraction, DNA amplication, and DNA electrophoresis. DNA amplication using gen lef-8 showed positif result with approximately 770 bp. Gen lef-8 can identified HytaNPV. DNA sequance showed that isolate HytaNPV Bogor had high homology of pathogonic NPV of genus Helicoverpa from Brazil, Australia, Spanyol and Netherland with homology nucleotide and amino acid reached 98% and 100%. Based on philogeny tree of HytaNPV was one group with pathogenic NPV of genus Helicoverpa.
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