Objective: The present study aimed to isolate and examine the characteristics of endophytic bacteria from kenikir (Cosmos caudatus Kunth.) leaves, to determine the endophytic bacteria capable of producing flavonoids, and to test their potency as anticancer and antimicrobial.Methods: The isolation of endophytic bacteria from C. caudatus Kunth. leaf was conducted by technical surface sterilization. Each of these isolates was produced from the insulation and then cultured on a liquid medium of 0.1% soluble starch, 0.5% peptone, and 0.15% yeast extract with a pH of 7 which have been incubated in room temperature for 5 days with agitation of 120 rpm and extracted with ethyl acetate solvent. The extract was then tested for flavonoid content using thin-layer chromatography method. The anticancer ability of flavonoids was tested by brine shrimp lethality test (BSLT) method, while that for antimicrobial, the test used paper disc method.Results: A total of 15 isolates of endophytic bacteria were successfully isolated from the kenikir leaves, but only 4 isolates produced flavonoids, among others PKM 1 (Serratia sp.), PKM 2 (Neisseria sp.), PKM 17 (Acinetobacter sp.), and PKM 19 (Yersinia sp.). BSLT test results showed that the flavonoids cause mortality in Artemia salina leach with LC50 as much as 16.736 in PKM 17, 17.267 in PKM 2, 18.672 in PKM 1, and 23.411 in PKM 19. The flavonoids also inhibited the growth of pathogens in human-based antimicrobial test results.Conclusion: Flavonoids produced by four endophytic bacterial isolates from kenikir leaves have great potential as anticancer and antimicrobial.
Abstrak. Thrixspermum merupakan anggrek yang memiliki ciri khas berbunga dalam waktu yang singkat. Akibatnya, dalam pengklasifikasian menggunakan morfologi cenderung sulit untuk dilakukan. Sehingga perlu adanya pengklasifikasian menggunakan cara alternatif misalnya dengan marka molekuler menggunakan DNA. Tujuan dari penelitian ini adalah mencari barkode DNA dari beberapa spesies Thrixspermum yang ada pada GenBank NCBI. Berdasarkan data dari NCBI, hanya terdapat 19 spesies dari genus Thrixspermum yang telah diteliti sekuennya. Dari total 19 spesies tersebut, setelah dilakukan pensejajaran sekuen menunjukkan bahwa pada lokus matK dan rbcL memiliki tingkat homologi yang tinggi, sementara pada lokus ITS menunjukkan lebih banyak variasi genetik. Terdapat dua spesies yakni T. annamense dan T. marguense yang berpotensi memiliki barcode pada lokus ITS. Selain itu, didapatkan pohon filogenetik dari 19 spesies Thrixspermum yang telah diketahui senkuennya pada NCBI.Abstract. Thrixspermum is an orchid that has the characteristic of flowering in a short time. As a result, classification using morphology tends to be difficult to perform. Therefore, an alternative method such as molecular markers using DNA is needed for classification purpose. This study was performed to identify DNA barcodes of several Thrixspermum species found in the NCBI database. Based on data from NCBI, there are only 19 species of the genus Thrixspermum with sequence have been explored. Of total 19 species, after sequence alignment showed that the matK and rbcL loci had high homology level, while at the ITS locus showed more genetic variant. There are two species namely T. annamense and T. marguense posses potential barcodes at ITS locus. In addition, a phylogenetic tree from 19 species of Thrixspermum was constructed based on the sequences deposited in NCBI.
Objective: The present study aimed to isolate and examine the characteristics of endophytic bacteria from kenikir (Cosmos caudatus Kunth.) leaves, to determine the endophytic bacteria capable of producing flavonoids, and to test their potency as anticancer and antimicrobial.Methods: The isolation of endophytic bacteria from C. caudatus Kunth. leaf was conducted by technical surface sterilization. Each of these isolates was produced from the insulation and then cultured on a liquid medium of 0.1% soluble starch, 0.5% peptone, and 0.15% yeast extract with a pH of 7 which have been incubated in room temperature for 5 days with agitation of 120 rpm and extracted with ethyl acetate solvent. The extract was then tested for flavonoid content using thin-layer chromatography method. The anticancer ability of flavonoids was tested by brine shrimp lethality test (BSLT) method, while that for antimicrobial, the test used paper disc method.Results: A total of 15 isolates of endophytic bacteria were successfully isolated from the kenikir leaves, but only 4 isolates produced flavonoids, among others PKM 1 (Serratia sp.), PKM 2 (Neisseria sp.), PKM 17 (Acinetobacter sp.), and PKM 19 (Yersinia sp.). BSLT test results showed that the flavonoids cause mortality in Artemia salina leach with LC50 as much as 16.736 in PKM 17, 17.267 in PKM 2, 18.672 in PKM 1, and 23.411 in PKM 19. The flavonoids also inhibited the growth of pathogens in human-based antimicrobial test results.Conclusion: Flavonoids produced by four endophytic bacterial isolates from kenikir leaves have great potential as anticancer and antimicrobial.
Anggrek adalah tumbuhan berbunga yang memiliki nilai komersial tinggi. Eksploitasi anggrek yang berlebihan menyebabkan keberadaannya di alam berkurang. DNA barcodingdapat menjadi alat yang efektif untuk deteksi anggrek langka seperti Paphiopedilum. Metode yang dilakukan dalam DNA barcodinganggrek Paphiopedilumadalah dengan studi in silicomenggunakan Genbank (NCBI). Sekuen DNA dikoleksi dari databasepada NCBI dengan cara menulis nama spesies dan gen yang dimaksud (matK, rbcL, dan ITS). Selanjutnya data dianalisis menggunakan CLUSTAL X untuk menentukan tingkat homologi antar sekuen melalui penjajaran sekuen, identifikasi sekuen yang berpotensi sebagai barcode, dan pembuatanpohon filogenetik. Analisis bioinformatik menunjukkan bahwa hasil alignmentdengan menggunakan lokus matKdan rbcLmemiliki tingkat homologi yang tinggi, meski demikian matKdapat menunjukkan karakter spesifik dari P. sangii yang dapat dijadikan sebagai barcode. Sedangkan ITSmemiliki tingkat variasi genetik yang lebih tinggi dan homologi yang rendah daripada matKdan rbcL. Barcode P. sangii dihasilkan dari lokus matK. Sedangkan pada lokus rbcL, semua spesies memiliki tingkat homologi yang tinggi, sehingga tidak mendapatkan sekuen yang berpotensi sebagai barcode. Variasi genetik yang tinggi dapat dipelajari pada lokus ITS, namun tidak ditemukan sekuen yang berpotensi sebagai barcode.
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