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CmBGI Gene Expression encoding β-glucosidase in melon (Cucumis melo L.) under stress condition Rachmawati, Yuanita; Aristya, Ganies Rizaa; Daryono, Budi Setiadi
BIOTROPIC The Journal of Tropical Biology Vol 1 No 2 (2017): Biotropic, Volume 1, Nomor 2, 2017
Publisher : Program Studi Biologi, Fakultas Sains dan Teknologi, Universitas Islam Negeri Sunan Ampel Surabaya

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Abstract

CmBGI is the enzymatic genes encoding β-glucosidase that involved in Abscisic Acid (ABA) metabolism of Cucumis melo L. β-glucosidase promotes the accumulation of glucose, fructose, and sucrose, and it might act as a regulator that mediates melon fruit ripening both climacteric and nonclimacteric. ABA mediates adaptive responses to abiotic and biotic stresses. Agricultural Balitbang in 1997 showed that there were approximately 158.600 ha of degraded land scattered in three zones of agroecosystems in Yogyakarta (DIY). One of them is Dlingo Bantul area which has a karst type critical land area. Karst provides stress to the certain plant growth. One way to conserve critical land is making this area for agriculture. Cultivar TACAPA and TA were superior melons that have been developed by Genetic Laboratory of Biology Faculty UGM. This preliminary research was conducted to examine molecular characterization of CmBGI gene expression in cultivar TACAPA and TA which are planted in normal condition medium and in critical land medium treatment. Total RNA was extracted from leaf tissue then Reversed Transcriptase (RT-PCR) to collect cDNA library. cDNA was amplified using specific primer. Spectrophotometry was conducted in λ260 nm and electrophoresis run in 1.5% agarose gel. Control of band chosen was Cm-Actin. CmBGI gene concentration of TACAPA and TA in normal condition medium are in succession 578.5 and 579.4 μg/ml then for critical land medium treatment 743.4 and 773.5 μg/ml. CmBGI band was showed both of TACAPA and TA as ± 1258 bp. Cm-actin was showed band of DNA as ± 445 bp. CmBGI gene concentration in critical land medium treatment which is given greater stress on melons are higher than normal condition. This suggests that the CmBGI gene is expressed more in cultivar TACAPA and TA melons when they are grown under stress condition.
KULTUR SEL BABY HAMSTER KIDNEY (BHK) MENGGUNAKAN MEDIA DULBECCO’S MODIFIED EAGLE MEDIUM (DMEM) Andiana, Mashita; Rachmawati, Yuanita; Andayani, Sri Susila
BIOTROPIC The Journal of Tropical Biology Vol 1 No 1 (2017): Biotropic, Volume 1, Nomor 1, 2017
Publisher : Program Studi Biologi, Fakultas Sains dan Teknologi, Universitas Islam Negeri Sunan Ampel Surabaya

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Abstract

Kultur sel merupakan suatu proses saat sel hidup ditempatkan ke dalam suatu media yang dapat membuat sel tersebut berkembang biak atau tumbuh secara in vitro, Kultur sel dapat berupa kultur sel primer maupun cell line, Metode dalam kultur sel terdiri atas kultur monolayer dan kultur suspensi. Pembuatan media kultur untuk pertumbuhan sel diusahakan memenuhi kriteria. Konstituen dasar dari media kultur yang paling banyak digunakan adalah BSS (Balanced Sald Solution). Untuk mendapatkan pertumbuhan sel yang optimal, media kultur ditambahkan serum. Serum yang biasa digunakan dalam kultur adalah serum anak sapi (calf serum), serum fetus sapi (foetal bovine serum), serum kuda dan serum manusia. Perhitungan sel menggunakan counting chamber. Metode yang digunakan adalah revival kemudian split sel, stor sel dan menghitung sel. Hasil revival  jumlah sel tertinggi didapat dari botol nomer 3 dengan jumlah sel 310 yaitu botol dengan sel BHK label tahun 2016, dan hasil split sel yang paling tinggi juga didapat dari botol kultur nomer 3 dengan jumlah sel 345 dan 311.
DETEKSI KONTAMINAN FRAGMEN DNA PENGKODE cyt b BABI PADA SAMPEL SOFTGELLCANDY TAK BERLABEL HALAL Rachmawati, Yuanita; Rokhim, Saiku; Munir, Misbakhul; Agustina, Eva
Indonesia Journal of Halal Vol 1 (1) 2018
Publisher : Pusat Kajian Halal Undip

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (205.92 KB) | DOI: 10.14710/halal.v1i1.3115

Abstract

Abstrak?Softgellcandy adalah permen bertekstur lunak yang diproses dengan penambahan komponen hidrokoloid seperti agar, gum, pektin, pati, kaegenan, gelatin dan lain-lain yang digunakan untuk modifikasi tekstur sehingga menghasilkan produk yang kenyal. Sayangnya distribusi Softgellcandy di pasaran seringkali terlepas dari pengawasan lembaga berwenang. Banyak ditemukan bermacam merk Softgellcandy yang tidak berBPOM maupun tidak berlabel Halal. Gelatin menjadi titik kritis kehalalan Softgellcandy. Penelitian ini menguji 15 sampel Softgellcandy tak berlabel halal yang dijual bebas di Surabaya dengan primer pengkode fragmen DNA cytochrome b Babi. Metode yang digunakan adalah konvensional PCR pada suhu 98oC-2 menit; 95oC-30 detik; 61oC-30 detik; 72oC-40 detik; dan 72oC-3 menit, selama 30 siklus. Visualisasi hasil PCR menggunakan elektroforesis 2% gel agarosa menunjukkan dari 15 sampel, 8 sampel terindikasi kontaminan DNA babi ditandai dengan pita DNA sebesar ?149bp. Pemerintah perlu melakukan monitoring lebih ketat terkait peredaran produk makanan tak berlabel halal yang dijual bebas di pasaran, mengingat Halal menjadi issue yang sangat sensitif di negara dengan mayoritas penduduk muslim terbesar di dunia ini. Karena halal adalah suatu keharusan.?Keywords: Softgellcandy, cyt b Babi, PCR
Phenotipical Characters of Melon (Cucumis melo L.) in Response to Karst Critical Land Rachmawati, Yuanita; Daryono, Budi Setiadi; Ariestya, Ganies Riza
Biotropic : The Journal of Tropical Biology Vol 2 No 1 (2018): Biotropic, Volume 2, Nomor 1, 2018
Publisher : Program Studi Biologi, Fakultas Sains dan Teknologi, Universitas Islam Negeri Sunan Ampel Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (712.581 KB) | DOI: 10.29080/biotropic.2018.2.1.1-10

Abstract

Yogyakarta Agroecosystem has 158,600 ha of critical land spread over three zones. Two areas are Karst Land, located on Agroecosystem II includes Gunungsewu Hills, Gunungkidul and III covers Dlingo Bantul Hills and Sentolo Hills Kulon Progo Regency.. Karst Land is certainly provides stress to plants. These research purposes are examining the phenotype character of superior melon Cultivar TACAPA compare to parents and offsprings phenotypes. The phenotype characters are based on plant height, leaf number, time of melon flowering, water content of plants, and fruit and seed productivity. This experiment was done by Split Plot Design with Completely Randomized Design (CRD) with 4 kinds of treatment (control plant media, Gunungsewu, Dlingo, and Sentolo), 7 experimental units cultivars: TACAPA, TA, TP, PT, AT, Action 434, PI 371795), and 4 replications. Research result reveals that most of the phenotypic characters including plant height, number of leaves, fruit weight, and number of seeds produced have relatively no significant effect between treatment and control, while the phenotypic first time flowering time and water content of the plant, have a noticeable difference.
Comparison of DNA Isolation Results with Simple Methods and Kits in Samples of Psidium guajava Leaves Rachmawati, Yuanita; Khoiriyah, Romyun Alvy
Biotropic : The Journal of Tropical Biology Vol 2 No 2 (2018): Biotropic, Volume 2, Nomor 2, 2018
Publisher : Program Studi Biologi, Fakultas Sains dan Teknologi, Universitas Islam Negeri Sunan Ampel Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (589.56 KB) | DOI: 10.29080/biotropic.2018.2.2.93-99

Abstract

DNA isolation is one of a series of methods that must be carried out on the basic techniques of Molecular Biology Analysis. Especially PCR-based molecular marking techniques. Many ways are done in DNA isolation. This study discusses the comparison of the results of DNA isolation using two methods. Simple DNA isolation methods and using Kit. Samples of Psidium guajava leaves used were taken from 15 different locations used. In general, DNA isolation methods include three steps, namely destruction, precipitation, and purification. Simple DNA isolation is done with detergents, alcohol groups, which are commonly available in the laboratory. Methods of DNA isolation with KIT are carried out according to the Promega Universal Wizard KIT protocol. The comparison results are seen from spectrophotometric absorption Å230 nm, Å260 nm, Å280 nm, Å320 nm, ratio Å260/Å230, ratio Å260/Å280 to see DNA purity, protein concentration before purification step, and DNA concentration produced. The results showed that there were no statistically significant differences in the results of DNA isolate spectrophotometry. However, the use of KIT with modified protocols is more recommended if researchers want to carry out DNA analysis more precisely and accurately.   Keywords: DNA isolation, spectrophotometry, DNA concentration and purity
KARAKTER GEN CMBG1 MELON (CUCUMIS MELO) PADA PENGARUH CEKAMAN TANAH KARST Aristya, Ganies Riza; Daryono, Budi Setiadi; Rachmawati, Yuanita
Sains & Matematika Vol 3, No 1 (2014): Oktober, Sains & Matematika
Publisher : Sains & Matematika

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Abstract

Lahan kritis berkapur, karst, memberikan cekaman abiotik pada tanaman karena hambatan hidrasi dan nutrisi. Asam absisat (ABA) adalah hormon yang diekspresikan tumbuhan pada kondisi cekaman abiotik. CmBG1 merupakan salah satu gen peregulasi hormon ABA pada melon yang akan terakumulasi saat tumbuhan mengalami cekaman. Tujuan penelitian ini adalah mendeskripsikan pengaruh lahan kritis karst terhadap ekspresi gen CmBG1 secara kualitatif dan kuantitatif melon hasil turunan kultivar TACAPA, yaitu kultivar PT dan AT yang ditanam pada medium tanah karst dari wilayah Agroekosistem II dan III Yogyakarta (Gunungsewu, Dlingo, maupun Sentolo). cDNA library diperoleh dari reverse transcription isolat RNA. cDNA diamplifi kasi dengan primer spesifi k, kemudian dispektrofotometri pada ?260 nm untuk mengetahui konsentrasi gen CmBG1. Ekspresi gen dianalisis dengan real time PCR dengan gen referensi Cm-actin. Uji kualitatif dilakukan dengan elektroforesis gel agarosa 1,5%. Hasil penelitian menunjukkan gen CmBG1 terdeteksi dengan ukuran ±1258 bp pada kultivar PT dan AT. Konsentrasi gen CmBG1 melalui spektrofotometri menunjukkan semua kultivar yang ditanam pada media kontrol memiliki konsentrasi yang lebih rendah bila dibandingkan media tanam dengan perlakuan lahan kritis baik Gunungsewu, Dlingo, maupun Sentolo. Hasil ini sama dengan uji ekspresi gen CmBG1 menggunakan analisis kuantitatif real time PCR. Karsts critical land gives abiotic stresses in plants because of hydration and nutrition disturbances. Abscisic acid (ABA) is a hormone that expressed in the plant in abiotic stress conditions. CmBG1 is one of the regulatory genes encoding hormone ABA in melon plants which is accumulated in stress condition. The purpose of this study was to describe the infl uence of karsts critical land on the genes expression of CmBG1 melon cultivars PT and AT qualitatively and quantitatively. The plants were grown in medium karst land of Agroecosystems II and III of Yogyakarta (Gunungsewu, Dlingo, and Sentolo). Total RNA was extracted from leaf tissue then Reversed Transcriptase (RT-PCR) to collect cDNA library. cDNA was amplifi ed using specifi c primer. Spectrophotometry was conducted in ?260 nm and electrophoresis run in 1.5% agarose gel. Control band and reference gene chosen in Real Time PCR was Cm-Actin. CmBGI band (± 1258 bp) was showed both on PT and AT. Cm-actin was showed band of DNA as ± 445 bp. CmBGI gene concentration in critical land medium treatment which is given greater stress on melons are higher than normal condition. This suggests that the CmBGI gene is expressed more in cultivar PT and AT melons when they are grown under stress condition. This result show similarly when using real time PCR.