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
The availability of high-quality seeds is now a necessity. This is due to a government program to replace oil palm trees in smallholder plantations with high quality seeds. An efficient protocol to produce a large number of embryos is needed. To increase the number of embryogenic callus production, the callus proliferation experiment was carried out through suspension culture. This study aimed to examine the proliferation ability of embryogenic callus from three different oil palm clones, in several repeated subcultures. Liquid MS media added with 1 ppm 2.4-D and 0.1 ppm NAA were used. Embryogenic callus was weighed by 0.1 - 0.2 g, transferred into the liquid media, shaking at 60-80 rpm and 27 ºC for 8 weeks without light. Continues subcultures were repeated up to 7 times. The results showed that the growth rate of embryogenic callus increased in the third and fourth subcultures and then decreased in subsequent subcultures. It also revealed that the entire embryogenic callus from the first subculture up to seventh subculture still has the ability to regenerate into new plants. These results indicate that oil palm embryogenic callus can be proliferated by suspension culture with a limit up to the fourth subculture. Ketersediaan benih kelapa sawit berkualitas saat ini merupakan kebutuhan karena adanya program pemerintah untuk menggantikan tanaman sawit di kebun-kebun petani. Salah satu cara vegetatif yang dapat dilakukan adalah meningkatkan jumlah kalus embriogenik yang dihasilkan melalui pengembangan kultur suspensi. Penelitian ini bertujuan mengkaji kemampuan proliferasi kalus embriogenik dari tiga klon kelapa sawit, pada beberapa kali subkultur yang berulang. Media cair MS dengan penambahan 1 ppm 2,4-D dan 0,1 ppm NAA digunakan untuk memperbanyak 0,1–0,2 g kalus embriogenik, dikocok pada 60-80 rpm dan suhu 27 ºC tanpa cahaya selama 8 minggu. Subkultur berulang dilakukan hingga 7 kali. Hasil percobaan menunjukkan bahwa kemampuan proliferasi kalus dipengaruhi oleh genotip tanaman induk. Rata-rata kalus embriogenik dapat meningkat pada subkultur ke-3 dan ke-4 dan semakin menurun pada subkultur selanjutnya. Kalus embriogenik hasil proliferasi subkultur pertama hingga ke-7 dapat tumbuh menjadi calon tanaman baru. Hasil ini menunjukkan bahwa kalus embriogenik kelapa sawit dapat diperbanyak dengan kultur suspensi pada batas sampai subkultur ke-4.
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.
Abstract.Rubber from Hevea brasiliensis is the only commercial natural rubber in the world. Propagation of rubber trees usually done by grafting and seed germination. BPPT had been producing rubber tree by in vitro technique with embryo somatic methods. Validation of mother plant for in vitro propagation is important to compare between mother plant and propagated plants. The aim for this research was to validation of PB 260 clone that planted at Cikumpay Plantation by SSR marker. Sampling of 10 rubber leaves were done at Cikumpay Plantation based on GPS position from the area of PB 260 clone. Rubber leaves were isolated with CTAB modification method to obtained DNA. Four of SSR primers from rubber, i.e.: hmac 4, hmac 5, hmct 1, and hmct 5, were used as primers to amplification of rubber DNA. The result showed that no band that different from 10 rubber of PB 260 clone at Cikumpay Plantation. This research will continue to compare genomic validation between mother plant and propagated plants that had been produced from BPPT.
Potato is a food commodity that has the potential to support food diversification in Indonesia. There is an increasing demand for Atlantic potatoes as the raw material for processed potato products. The demand, which has not been met by the increased production, has been the cause of the ongoing potato import activities in Indonesia. The limitation of producing quality Atlantic potato seeds economically is one of the obstacles to increasing the production of Atlantic potatoes in Indonesia. The aim of this research was to study the effect of various table sugar concentrations as the carbon source and the type of the culture containers used for Atlantic potato shoot multiplication in vitro. The propagation was carried out in bioreactors and culture bottles with MS liquid medium + coconut water at a concentration of 150 mL/L medium, and 3 concentration levels of table sugar, namely 0; 7.5; and 15 g/L medium. The use of bioreactor significantly increased the height of the Atlantic potato plantlets. The use of bioreactor combined with table sugar addition decreased hyperhydricity level. The highest number of shoots, leaves, and roots were found at the table sugar concentration of 15 g/L medium in both containers. ABSTRAKKentang merupakan komoditas pangan yang berpotensi mendukung program diversifikasi pangan di Indonesia. Peningkatan permintaan terhadap kentang Atlantik sebagai bahan baku kentang olahan yang tak diimbangi dengan peningkatan produksi kentang Atlantik menjadi penyebab masih berlangsungnya impor kentang Atlantik di Indonesia. Keterbatasan menghasilkan benih kentang Atlantik berkualitas yang ekonomis merupakan salah satu hambatan dalam meningkatkan produksi kentang Atlantik di Indonesia. Penelitian ini bertujuan mempelajari pengaruh variasi konsentrasi sukrosa teknis sebagai sumber karbon dan penggunaan jenis wadah terhadap perbanyakkan tunas kentang Atlantik secara in vitro. Perbanyakkan tunas kentang Atlantik menggunakan media MS cair + 150 mL/L air kelapa dalam wadah bioreaktor dan botol kultur dengan 3 taraf konsentrasi sukrosa, yaitu 0; 7,5; dan 15 g/L media. Penggunaan bioreaktor secara signifikan meningkatkan tinggi planlet kentang Atlantik yang dihasilkan. Penggunaan bioreaktor yang dikombinasikan dengan penambahan sukrosa teknis menurunkan tingkat hiperhidrisitas. Tunas, daun, dan akar terbanyak dihasilkan oleh perlakuan sukrosa teknis 15 g/L media dalam kedua jenis wadah.
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