Analisis Molekuler Helicoverpa armigera Nucleopolyhedrovirus (HearNPV) berdasarkan Gen DNA Polimerase

Abstract

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) atau ulat penggerek tongkol adalah hama penting pada komoditas jagung di Indonesia. Pengendalian H. armigera secara hayati menggunakan Nucleopolyhedrovirus (NPV) menjadi alternatif pengendalian yang sejalan dengan prinsip pengendalian hama terpadu (PHT). NPV tergolong ke dalam famili Baculoviridae yang memiliki inang spesifik. Helicoverpa armigera Nucleopolyhedrovirus (HearNPV) merupakan patogen serangga yang mampu mengendalikan ulat penggerek tongkol secara efektif. Penelitian ini bertujuan mengetahui hubungan kekerabatan HearNPV melalui analisis homologi dan filogenetik berdasarkan gen DNA polimerase (dnapol). Isolat NPV yang digunakan berasal dari koleksi Laboratorium Patologi Serangga yang diambil dari pertanaman jagung di Bogor, Indonesia. DNA dari gen DNA polimerase hasil ekstraksi kemudian diamplifikasi dengan teknik Polymerase Chain Reaction (PCR) menggunakan sepasang primer spesifik gen DNA polimerase SPTF dan SPTR yang didesain oleh Sapto (2022). Hasil amplifikasi digunakan untuk mengetahui informasi tentang hubungan kekerabatan HearNPV isolat Bogor dengan NPV yang telah terdaftar di situs GenBank. Hasil amplifikasi PCR menunjukkan satu pita DNA dari gen DNA polimerase HearNPV parsial berukuran sekitar ±1200 pb. Perunutan fragmen DNA HearNPV menghasilkan runutan nukleotida berukuran 1187 pb yang mengkodekan asam amino sebanyak 392. Analisis homologi menunjukkan bahwa isolat HearNPV asal Bogor memiliki hubungan kekerabatan yang paling dekat dengan NPV yang menginfeksi genus Helicoverpa dari negara Spanyol berdasarkan runutan nukleotida dan Australia, Spanyol, Rusia, Brazil, USA, Turki, serta Cina berdasarkan runutan asam amino. Berdasarkan analisis filogenetik, isolat HearNPV asal Bogor berada dalam kelompok yang sama dengan NPV yang menginfeksi genus Helicoverpa dari negara lain, yaitu Spanyol, Cina, India, Kenya, Turki, Brazil, Australia, Rusia, dan USA.

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) or corn earworm is one of the most important pests of corn in Indonesia. Biological control of H. armigera using Nucleopolyhedrovirus (NPV) is an alternative control that aligns with the principles of integrated pest management (IPM). NPV belongs to the Baculoviridae family which has a specific host. Helicoverpa armigera Nucleopolyhedrovirus (HearNPV) is an insect pathogen that can be used for controlling the earworm effectively. This study aims to determine the genetic relationship of HearNPV through homology and phylogeny analysis based on the DNA polymerase gene (dnapol). NPV isolates utilized in this study were sourced from the collection of the Insect Pathology Laboratory, gathered from corn plantations in Bogor, Indonesia. Total DNA from the extracted DNA polymerase gene was subsequently amplified by the Polymerase Chain Reaction (PCR) technique, utilizing a pair of DNA polymerase gene-specific primers SPTF and SPTR designed by Sapto (2022). The resulting amplification products were subjected to sequencing to elucidate the genetic relationship between HearNPV Bogor and NPV isolates already registered on GenBank. PCR amplification results showed a band of HearNPV partial DNA polymerase gene measuring approximately ±1200 bp. Sequencing of HearNPV DNA fragments produced a nucleotide sequence of 1187 bp which encodes 392 of amino acids. Homology analysis showed that HearNPV isolates from Bogor had a high genetic relationship with NPVs that infect the Helicoverpa genus from Spain based on nucleotide sequences and Australia, Spain, Russia, Brazil, the United States, Turkey, and China based on amino acid sequences, respectively. Based on the phylogenetic analysis, HearNPV isolates from Bogor are in the same group as NPVs that infect the Helicoverpa genus from other countries, such as Spain, China, India, Kenya, Turkey, Brazil, Australia, Rusia, and the United States.