ANALISIS KEKERABATAN ISOLAT BAKTERI ENDOFIT RESISTEN AMPISILIN PADA AKAR TANAMAN OBAT

Jannah, Novi Dewi Khadikatul (2017) ANALISIS KEKERABATAN ISOLAT BAKTERI ENDOFIT RESISTEN AMPISILIN PADA AKAR TANAMAN OBAT. S1 thesis, Universitas Pendidikan Indonesia.

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Abstract

Penelitian mengenai analisis kekerabatan isolat bakteri endofit resisten ampisilin yang diisolasi dari akar tanaman obat Ageratum conyzoides dan Vetiveria zizanioides cultv. telah dilakukan. Penelitian bertujuan untuk mengetahui pola kekerabatan bakteri endofit yang resisten terhadap kandungan β-laktam pada antibiotik ampisilin. Metode yang digunakan dalam penapisan resistensi antibiotik adalah metode disk diffusion dengan konsentrasi ampisilin sebesar 10, 20 dan 30 µg/ml. DNA isolat yang resisten terhadap ampisilin kemudian diisolasi dan diukur kemurnian dan konsentrasinya menggunakan spektrofotometer. Analisis gen 16S rRNA dilakukan untuk mengidentifikasi bakteri secara molekuler menggunakan pasangan primer 63F dan 1387R. Hasil amplifikasi kemudian dilakukan elektroforesis gel menggunakan agarose 1% lalu dilihat dan diamati visualisasinya menggunakan UV transilluminator. Amplikon kemudian disikuensing dan dianalisis menggunakan program BLASTN yang dapat mengkomparasi sikuen nukleotida bakteri dengan sikuen nukleotida yang paling mirip pada database. Analisis data zona hambat yang terbentuk dilakukan dengan menggunakan uji Mann-Whitney U pada SPSS 16.0 for windows. Analisis filogenetik dilakukan dengan melakukan rekonstruksi pohon filogenetik menggunakan software MEGA7 dan metode maximum parsimony dengan bootstrap 500x. Setelah dilakukan pengujian terhadap sembilan isolat bakteri endofit, terdapat perbedaan tingkat resistensi yang signifikan. Isolat M dan H memiliki tingkat resistensi yang tinggi terhadap semua konsentrasi ampisilin. Hasil analisis bioinformatika menunjukkan bahwa isolat M memiliki kemiripan dengan Paenibacillus sp. dengan tingkat homologi sebesar 98% dan isolat H memiliki kemiripan dengan Pantoea sp. dengan tingkat homologi yang rendah, yaitu sebesar 79%. Genus Paenibacillus dan Pantoea diketahui memiliki resistensi terhadap ampisilin.,---The research about phylogenetic analysis on ampicillin-resistant endophytic bacteria isolated from Medicine Plants Root Ageratum conyzoides dan Vetiveria zizanioides cultv. has been done. This study aims to determine phylogenetic pattern of endophytic bacteria which have ampicillin-resistant to antibiotics that contain β-lactam. The methods were used to screening antibiotic resistance was disk diffusion with concentration of ampicillin 10, 20 and 30 µg/ml. DNA from isolates which of ampicillin-resistant was isolated and measured using spectrophotometer to determine the purity and the concentration of DNA. 16S rRNA gene analysis was used to identify bacteria molecularly using primers 63F and 1387R. Amplicon were electrophoresis using 1% agarose then the results were seen and observed using UV transilluminator. Amplicon were sequencing and analyzed using BLASTN program to compare bacterial nucleotide sequence with nucleotide sequences that are most similar to the database. Inhibition zone was analyzed with SPSS 16.0 for windows used Mann-Whitney U test. Phylogenetic analysis conducted by reconstructing phylogenetic trees using MEGA7 software with maximum parsimony bootstrap method 500x so that the phylogenetic pattern of endophytic bacteria tested is known. After testing the nine isolates of endophytic bacteria, there is a significant difference in the level of resistance. Isolates M and H have a high level of resistance against all concentrations of ampicillin. Bioinformatics analysis showed that the isolate M has high similarities to the Paenibacillus sp. with homology level of 98% and isolate H has poor similarities to the Pantoea sp. with percentage of homology is 79%. The genus Paenibacillus and Pantoea are known to have resistance to the ampicillin.

Item Type: Skripsi,Tesis,Disertasi (S1)
Additional Information: no.panggil : S BIO JAN a-2017; pembimbing : I Any Fitriani, II Peristiawati
Uncontrolled Keywords: Ampicillin, Endophytic Bacteria, Resistance, 16S rRNA, Ampisilin, Bakteri Endofit, Resistensi, 16S rRNA.
Subjects: B Philosophy. Psychology. Religion > BC Logic
Divisions: Fakultas Pendidikan Matematika dan Ilmu Pengetahuan Alam > Jurusan Pendidikan Biologi
Depositing User: Mr mhsinf 2017
Date Deposited: 29 Jan 2018 02:07
Last Modified: 29 Jan 2018 02:07
URI: http://repository.upi.edu/id/eprint/28828

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