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All Journal Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati
Jesmandt Situmorang
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

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Aplikasi Metode Sidikjari Protein (SDS-PAGE) untuk Identifikasi Isolat Bakteri Endogenik Indonesia (Bacillus thuringiensis Berliner) yang Patogenik terhadap Hama Kubis (Crocidolomia binotalis Zell) Christina L. Salaki; Langkah Sembiring; Jesmandt Situmorang; Niken Satut Nur Handayani
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 17, No 2 (2012): June 2012
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v17i2.135

Abstract

Isolat bakteri Bacillus thuringiensis endogenik Indonesia yang patogenik terhadap hama kubis (Crocidolomia binotalis Zell.) dikarakterisasi dan diidentifikasi secara kimiawi dengan metode sidikjari protein. Total protein selular terlarut 10 isolat terpilih (SLK2.3, SRNG4.2, TKO1, TK9, YPPA1, UG1A,BLPPN8.2, YWKA1, BAU3.2, LPST1) dan 2 strain acuan (B. thuringiensis serovar kurstaki ATCC 10792 dan B. thuringiensis serovar israelensis H14) diperoleh melalui pemecahan sel dengan sonikasi. Ekstrak sel disentrifugasi dengan kecepatan 13000 rpm selama 5 menit untuk memperoleh supernatant. Protein dipisahkan dengan metode elektroforesis (SDS-PAGE) untuk menghasilkan profil sidikjari protein yang didokumentasikan dalam bentuk file elektronik. Selanjutnya, sidikjari protein dianalisis secara kualitatif maupun secara kuantitatif menggunakan software MVSP (Multivariate Statistical Package) untuk menghasilkan dendrogram. Hasil penelitian menunjukkan bahwa profil protein yang dihasilkan oleh tiap-tiap isolat dan strain acuan memberikan pola yang bermakna sehingga dapat digunakan untuk mengkarakterisasi dan mengidentifikasi isolat secara tegas. Analisis terhadap dendrogram menunjukkan bahwa strain acuan B. thuringiensis serovar kurstaki HD1 dapat dibedakan secara tegas dan jelas dengan strain acuan B. thuringiensis serovar israelensis H14. Dengan demikian, dapat disimpulkan bahwa aplikasi profil sidikjari protein merupakan instrumen yang sangat ampuh untuk menyingkap keanekaragaman strain anggota B. thuringiensis yang disolasi dari habitat alami. Isolat yang diteliti diidentifikasi sebagai strain baru anggota B. thuringiensis serovar kurstaki karena menunjukkan kemiripan yang tinggi dengan strain acuan B. thuringiensis serovar kurstaki ATCC 10792 meskipun ke-10 isolat ini juga menunjukkan keanekaragaman genetik yang cukup tinggi.
Uji Patogenisitas Isolat Bakteri Indigenous (Bacillus thuringiensis) terhadap Serangga Hama Kubis (Crocidolomia binotalis Zell) Christina L. Salaki; Jesmandt Situmorang; Langkah Sembiring; Niken Handayani
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 14, No 3 (2009): October 2009
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v14i3.2582

Abstract

Pathogenicity of 34 indigenous B. thuringiensis isolates against C. binotalis were determined. The pathogenicity test was conducted by using leaf dipped method with various spore concentrations. Third instar larvae of C. binotalis were used as insect test. Mortality data of test larvae were used to determine the pathogenicity of the isolates in terms of 72 hours LC50 by using probit analysis. The results of experiments showed YPPA 1. was the most pathogenic isolate, producing 72 hours LC50 = 9.5 x 103 spore.ml-1 with LT50 (1.5 x 107 spore.ml-1) of 24.6 hours while the ACH 2.3 was found to be the least pathogenic isolate with 72 hours LC50 = 2.3 x 106 spore.ml-1 and LT50 (1.5 x 107 spoore.ml-1) of 40.7 hours. The shortest LT50 (1.5 x 107 spore.ml-1 was found to be 18.2 hours produced by TUS.1 with 72 hours LC50 = 3.9 x 105 spore.ml-1 whereas the longest LT50 (1.5 x 107 spore.ml-1) was found tobe 83.2 hours produced by the SLK 4.1 with 72 hours LC50 = 3.1 x 104 spore.ml-1. Therefore, it can be concluded that both YPPA.1 and TUS.1 isolates are potential candidate to be developed for biological control agent.
Analisis Keanekaragaman Isolat Bacillus thuringiensis yang Patogenik terhadap Serangga Hama Kubis (Crocidolomia binotalis) dengan Pendekatan Sistematika Numerik Christina L. Salaki; Jesmandt Situmorang; Langkah Sembiring; Niken Handayani
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 15, No 3 (2010): October 2010
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v15i3.2605

Abstract

Diversity of B. thuringiensis (Bt.) isolates pathogenic to C. binotalis was determined by using Numerical Systematic Method. Ten isolates were taken to represent 34 pathogenic isolates along with two reference strains namely B. thuringiensis serovar kurstaki and B. thuringiensis serovar israelensis. The test isolates were examined for 89 phenotypic characters by using convensional method for colonial and cell morphology (37 characters) as well as physiological characteristics (3 characters) but biochemical characterization (49 characters) was conducted by using commercial API-50 CHB procedures. All phenotypic characters existed in one of two mutually exclusive states and were either scored plus (1) of minus (0). The binary data were prepared in Programmer’s File Editor (PFE) software. The data then were analysed by using the Multi Variate statistical Package (MVSP) Plus-Version 3.1 using the Simple Matching Coefficient (SSM). Clustering was achieved using the UPGMA algorithm. The results were presented as dendrograms. It was obtained that the test isolates were clearly assigned to two distinct multimembered clusters defined by 79.6 similarity level (S-level) in the SSM, UPGMA analysis. The two distinct clusters represented by each of two widely known different group of Bt. strains, namely serovar israelensis and serovar kurstaki. The first cluster contained reference strain of B. thuringiensis serovar israelensis, and two of the isolates (Slk2.3, and YPPA1) and the second cluster contained another reference strain of B. thuringiensis serovar kurstaki, and 8 of the isolates. Therefore, it strongly suggested that the application of numerical-fenetic analysis could provide a tool to unravel the strain diversity belong to B. thuringiensis.
Analisis Filogenetik Burung Maleo (Macrocephalon maleo) Berdasarkan Sekuen Intron Satu Gen Rhodopsin (RDP1) Nukleus I Made Budiarsa; I Wayan Tunas Artama; Langkah Sembiring; Jesmandt Situmorang
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 15, No 2 (2010): June 2010
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v15i2.2693

Abstract

The phylogenetic relationships of the maleo (Macrocephalon maleo) were analyzed based on thefirst intron of rhodopsin nuclear gene sequence data obtained from 15 individuals, along withthose of 22 individuals taken from GenBank. The phylogenetic trees were reconstructed byNeighbor-Joining (NJ) method. Results indicated that 956 bp of RDP1 sequence, 414 (43.4%)sites were variable and 317 (33.2%) sites were phylogenetically-informative. The basecomposition for all species analyzed in this research were as follows: T 25.3%, C 26.3%, A18.5%, and G 29.9%. Analysis of RDP1 sequence produced trees that were remarkably wellresolved and had topologies at the marga level. The phylogenetic analysis showed that maleowas monophyly of Macrocephalon and closely related to Aepypodius, Talegalla, Leipoa andAlectura.
Co-Authors Christina Salaki I Wayan Tunas Artama Laksono Trisnantoro Langkah Sembiring Niken Handayani Niken Satut Nur Handayani