Obtaining high-quality RNA is very important at an early stage of molecular biology research. To isolate RNA, high skill and caution are required in following laboratory procedures because RNA is easily degraded, especially samples from plant tissue culture. One of the parameters used to check the total RNA quality is RIN (RNA Integrity Number). The aim of this study was to obtain RNA extraction methods on oil palm leaves, callus and somatic embryos that were of good quality and high concentrations for transcriptomic analysis. RNA extraction was carried out using Plant RNA PureLink (Ambion), Genezol RNA Extraction (Geneaid) and RibospinTM Plant (Geneall) kit methods. The results showed that oil palm leaf, callus and somatic embryo RNA were successfully extracted using the RibospinTM (Geneall) kit. Based on the total RNA number of more than 4 μg and the RIN value of more than 7, the extracted RNA could be used in RNA sequencing for transcriptomic analysis. ABSTRAKMenghasilkan RNA berkualitas tinggi sangatlah penting pada tahap awal penelitian biologi molekuler. Untuk mengisolasi RNA diperlukan keterampilan dan kehati-hatian tinggi dalam mengikuti prosedur di laboratorium karena RNA lebih mudah terdegradasi, khususnya sampel hasil kultur jaringan tanaman. Salah satu parameter yang digunakan pada pengecekan kualitas RNA total adalah RIN (RNA Integrity Number). Penelitian bertujuan mendapatkan metode ekstraksi RNA pada daun, kalus dan embrio somatik kelapa sawit yang berkualitas baik dan memiliki konsentrasi tinggi untuk analisa transkriptomika. Ekstraksi RNA dilakukan menggunakan metode kit Plant RNA PureLink (Ambion), Genezol RNA Extraction (Geneaid) dan Ribospin TM Plant (Geneall). Hasil menunjukkan bahwa RNA daun, kalus dan embrio somatik kelapa sawit telah berhasil diekstraksi dengan menggunakan kit Ribospin TM (Geneall). RNA hasil ekstraksi tersebut dapat digunakan untuk sekuensing RNA dengan tujuan analisis transkriptomika, dilihat dari jumlah total RNA yang lebih dari 4 μg dan nilai RIN lebih dari 7.Kata Kunci: analisis RNA, embrio somatic, kalus, kelapa sawit, kualitas RNA a
Oil palm is a vegetable oil-producing plant (CPO) which provides the largest foreign exchange contribution compared to other crops and is widely used in food, medicine, cosmetic, and energy industries. Tissue culture technology is currently used to produce quality oil palm seeds. Oil palm shoots tend to grow and develop in clumps (groups) in vitro. Bipolar nature does not appear in all the shoots produced, so to produce plantlets it is necessary to do induction. This research aimed to obtain the right root induction media. A completely randomized design (CRD) with two factors was used with the first factor being the type of auxin (IAA, IBA, and NAA), and the second was the auxin concentration (0, 0.25, 0.5, and 0.75 ppm). In the eighth week after planting, the variables of root length, number of leaves, and shoot height were not significantly different except for the root number. The best root induction media for plantlet formation was the MS base medium with the addition of NAA type auxin at a concentration of 0.75 ppm. The plantlets formed a symbiosis with mycorrhiza which was applied at a dose of 4 g per polybag in the fourth month after planting.
One of the medicinal plants that is widely cultivated is the red ginger (Zingiber officinale Rosc. Var. Rubrum). The plant contains an active compound gingerol that is used as an ingredient for various treatments such as cough and flu. To meet the demand of medicinal and industrial raw materials, quality ginger seeds are needed. One alternative to producing seeds is to use tissue culture technology. This study aimed to obtain the best type and concentration of cytokinins in increasing the multiplication of red ginger shoots in vitro. This study used a factorial Completely Randomized Design (CRD) with 2 factors, namely the type of cytokinins (BAP, thidiazuron, zeatin, kinetin, and 2ip) and cytokinin concentrations (0, 0.1, and 1 ppm). The results showed that 1 ppm thidiazuron treatment produced the highest number of shoots and the highest shoot length in the first subculture. The responses in the second subculture showed that shoots from thidiazuron, 2ip, and BAP treatment media produced the highest number of shoots, roots, and leaves compared to kinetin and zeatin. Multiplication continued until the sixth subculture, and the best multiplication was found on shoots from 2ip treatment.
ABSTRAKKata kunci: Antibiotik, kontaminan jamur dan bakteri, kultur in vitro, metode sterilisasi, sagu ABSTRACT Natural sago (Metroxylon sagu Rottb.) forest can be found in large area in Maluku and Papua regions. There are wide genetic diversities of sago palm found in these areas. This palm grows along riverbanks and in swampy areas which are not suitable for other crops. Sago palm is propagated generatively by seed and vegetatively by suckers. With the purpose of establishing the in vitro culture method for a large-scale of mass clonally propagation of superior genotypes of sago palm, generating sterilized explants are very important. Young suckers (15-20 cm) obtained from areas of Papua Province were used as explants. The sterilization experiments were carrying out to support the tissue culture of sago palm. Sterilization was conducted using antibiotics in order to get rid of fungi and bacteria from inner part of explants tissues. The results showed that from all sterilization methods tested, the best result was treatment using alcohol and antibiotic as disinfectant agents.
ABSTRAK Pohon sagu (Metroxylon saguRottbKata kunci: Auksin, sitokinin, in vitro, sagu, inisiasi tunas ABSTRACT Sago palm (Metroxylon sagu Rottb.) has many advantages over other starch-producing crops especially for its higher yield, ability to grow along riverbanks and on swampy areas not suitable for other crops. With the purpose of establishing large-scale plantations, a large amount of uniform sago palm suckers are required. However, limited availability of uniform suckers has hindered the mass propagation and development of cultivated Sago palm. Alternatively, in vitro cultures were performed in order to obtain a large-scale of mass clonally propagation of superior genotypes of sago palm. The young suckers obtained from areas of Maluku Province were used as explants. In vitro culture was carried out through direct shooting. The explants were cultured on two kinds of media, which were MS and B5 media containing various growth hormones of auxins and cytokinins. The results showed that the treatment with BAP 2.0 ppm and NAA 2.0 ppm produced the highest number of shoots.
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