Studi Kandidat Senyawa Alami Antijamur Dalam Menghambat Colletotrichum Sp. Patogen Penyebab Penyakit Antraknosa Pada Tanaman Cabai Secara In Silico, In Vitro Dan In Vivo

Winar, Devanska Titansa and Luqman Qurata Aini,, SP. M.Si., Ph.D and Muhammad Akhid Syib’li, ., SP., MP., Ph.D (2022) Studi Kandidat Senyawa Alami Antijamur Dalam Menghambat Colletotrichum Sp. Patogen Penyebab Penyakit Antraknosa Pada Tanaman Cabai Secara In Silico, In Vitro Dan In Vivo. Sarjana thesis, Universitas Brawijaya.

Abstract

Bioinformatika mulai banyak diterapkan dalam kegiatan pengembangan antijamur. Bioinformatika menggabungkan biologi molekuler dengan pendekatan teknik komputasi untuk mengolah data. Pada penelitian sebelumnya, senyawa alkaloid berberin terbukti secara In silico dapat menekan kinerja enzim kutinase. Enzim kutinase bekerja dengan memecah kutikula tanaman yang merupakan bagian penting dari proses penetrasi patogen Colletotrichum sp. ke dalam tanaman. Apabila kinerja kutinase dihambat, patogen akan lebih susah melakukan penetrasi dan infeksi akan berkurang. Tujuan penelitian ini adalah menemukan molekul senyawa alami yang bisa digunakan sebagai antijamur secara In silico, mengetahui nilai hasil docking dari kandidat senyawa alami dan mengobservasi pengaruh kandidat senyawa untuk menekan aktivitas enzim kutinase dan kejadian penyakit antraknosa. Penelitian ini dilakukan mulai bulan Oktober 2021 hingga Juni 2022 di Lab. Pengendalian Hayati 1, Dep. Hama dan Penyakit Tumbuhan, Fakultas Pertanian, Universitas Brawijaya Malang. Tahapan penelitian diawali dengan skrining molecular docking berbasis struktur menggunakan 1000 molekul senyawa yang terdaftar pada Protein Data Bank (PDB) terhadap protein kutinase Colletotrichum sp. Kemudian hasil docking dianalisis untuk pemilihan senyawa alami pengujian in vitro dan in vivo. Pengujian penghambatan enzim kutinase secara in vitro dilakukan menggunakan media Rhodamine Olive oil Agar (ROA) dengan lima perlakuan konsentrasi senyawa aktif (0%, 0,5%, 1%, 1,5%, 2%) dengan variabel pengamatan kualitatif yaitu zona pendaran kuning-oranye. Selanjutnya, uji intensitas serangan penyakit antraknosa pada cabai dilakukan menggunakan RAL dengan tiga ulangan dengan perlakuan perendaman cabai dengan larutan aquades, propineb dan larutan senyawa aktif dengan beberapa konsentrasi (2%, 4%, 6%, 8%, 10%). Data hasil pengamatan yang diperoleh dianalisis secara statistik menggunakan sidik ragam, apabila hasil menunjukkan berbeda nyata akan dilanjutkan dengan uji DMRT dengan taraf 5%. Hasil penelitian In silico didapatkan senyawa kurkumin dipilih menjadi kandidat untuk diuji lanjut dengan nilai binding energi -6,4kkal/mol, konstanta inhibisi 0,989 μM dan nilai Root Mean Square Deviation (RMSD) sejauh 2,461 Å. Pada uji In vitro dan In vivo menunjukkan penambahan konsentrasi kurkumin sudah mampu menghambat aktivitas dari enzim kutinase dan menekan kejadian penyakit antraknosa pada buah cabai.

English Abstract

Bioinformatics has been widely applied in antifungal drug discovery and development. Bioinformatics combines molecular biology with a computational method to analyze biological data. In previous, In silico reports, berberine alkaloids have been shown to inhibit the activity of the cutinase enzyme. Cutinase enzymes break down plant cuticles, which are necessary for the penetration of Colletotrichum sp. into the plant cuticle. Inhibition of cutinase activity makes pathogen penetration more difficult and infection of anthracnose may also decrease. This study aims to identify natural compound that can be used as antifungal based on in silico screening, determine the docking value of natural compound candidates and observe its potential to suppress cutinase activity and anthracnose disease. This research was conducted from October 2021 to June 2022 at the Lab. Biological Control, Dep. Pests and Plant Diseases, Faculty of Agriculture, Universitas Brawijaya Malang. The research begins with structural-based in silico screening using 1000 compounds listed on Protein Data Bank (PDB) against protein cutinase of Colletotrichum sp. The docking results were analyzed for the selection of natural compounds for in vitro and in vivo assays. In vitro cutinase inhibition assay was carried out using Rhodamine Olive oil Agar (ROA) media with five concentrations of selected compound extract (0%, 0.5%, 1%, 1.5%, 2%) with a yellow-orange fluorescence zone as the qualitative observed variable. The anthracnose disease intensity assay in chili was carried out using RAL with three replications by soaking chilies with aquades, propineb and a solution of the selected compound at several concentrations (2%, 4%, 6%, 8%, 10%). The data obtained were statistically analyzed using ANOVA and if there were significant differences between treatments then continued with the DMRT with a significant level of 5%. The results of the In silico study showed that curcumin was chosen as a candidate for further testing with a binding energy of -6.4 kcal/mol, an inhibition constant of 0.989 μM, and a Root Mean Square Deviation (RMSD) value of 2.461. The addition of curcumin concentration was able to inhibit the activity of the cutinase enzyme and suppress the incidence of anthracnose disease in chili peppers in both In vitro and In vivo assays.

Other obstract

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Item Type: Thesis (Sarjana)
Identification Number: 0522040190
Subjects: 600 Technology (Applied sciences) > 632 Plant injuries, diseases, pests > 632.6 Animal pests
Divisions: Fakultas Pertanian > Hama dan Penyakit Tanaman
Depositing User: Nur Cholis
Date Deposited: 16 Nov 2022 06:41
Last Modified: 16 Nov 2022 06:41
URI: http://repository.ub.ac.id/id/eprint/196358
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