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Rizka Tamania SAPTARI
Indonesian Research Institute for Biotechnology and Bioindustry

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Propagasi in vitro tanaman kurma (Phoenix dactylifera L.) pada bioreaktor dengan perendaman sesaat Rizka Tamania SAPTARI; Masna Maya SINTA; Imron RIYADI; . PRIYONO; . SUMARYONO
E-Journal Menara Perkebunan Vol 88, No 2 (2020): Oktober,2020
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v88i2.394

Abstract

The cultivation of date palm in Indonesia has increased since the last decade. However, the superior date palm seedlings are still limited and most of them are imported from other countries. The mass supply of superior date palm seedlings can be provided by in vitro propagation in the bioreactor. Therefore, the research was conducted to develop a protocol of date palm in vitro propagation by using Temporary Immersion Bioreactor (TIB). The in vitro propagation was carried out through somatic embryogenesis technique using meristematic tissues isolated from offshoots of date palm female clone cv. Zambli as explants. The explants were sterilized and then cultured to produce embryogenic calli and somatic embryos. Afterwards, somatic embryos germination and plantlets formation were conducted in TIB with treatments of immersion period: 3, 10, and 30 minutes every 6 hours, with 8 replications, The results showed that the optimal somatic embryo germination in TIB was with the immersion period of 30 min every 6 h, resulting in the most formation of shoots and fresh biomass weight increment up to nearly threefold in 6 weeks. Thereafter, plantlets formation in TIB with immersion period of 10 min and 30 min every 6 h exhibited similar performances in producing more plantlets with higher total fresh weight and better vigor than those of 3 min every 6 h. However, there were more rooted plantlets in the TIB with immersion period of 10 min every 6 h. Based on the results, an in vitro propagation protocol via somatic embryogenesis in TIB has been successfully developed for mass propagation of date palm cv. Zambli, which produced plantlets with good vigor and rooting.
The in silico study of the COBRA gene family in sugarcane related to potential biomass content Riza Arief PUTRANTO; Galuh Wening PERMATASARI; Rizka Tamania SAPTARI
E-Journal Menara Perkebunan Vol 90, No 1 (2022): April, 2022
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v90i1.486

Abstract

AbstractSugarcane (Saccharum sp.) is potential as a biofuel and biomaterial source for its high cellulose content. Cellulose is the main constituent of the plant cell wall, as a linear chain arranged in a polysaccharide bundle, called cellulose microfibril. A gene named COBRA has been revealed to play role in the orientation of microfibril and cellulose crystallization. The COBRA gene in the Saccharum spp is under-explored. Therefore, the in silico study was conducted to explore the COBRA gene in Saccharum sp. By comparative genomics methods, the COBRA genes from Arabidopsis sp. (AtCOBLs) were compared to the Saccharum sp. (SoCOBLs). The conserved domain was then identified and the cluster system was constructed under a phylogenetic tree. Furthermore, each SoCOBLs protein was modelled to analyze its structure. According to the analysis, eleven of Saccharum sp. genomic scaffolds were successfully identified. Moreover, conserved domain identification resulted in nine SoCOBLs proteins. The phylogenetic tree showed two main clusters: I and II, differentiating those COBLs families based on the protein sequence, domain motif and amino acid properties. It leads to the variation of SoCOBLs protein structure as the results of the amino acid properties. Overall, the COBRA gene has been identified genomically in Saccharum sp. Yet, the function and tissue-specific expression are still unclear. It was predicted to act as the regulator of microfibril orientation and the cellulose synthesis process. Hence, further analyses by in vitro and in vivo are indispensable.[Keywords: cellulose, comparative genomic, Saccharum sp.]AbstrakTanaman tebu (Saccharum sp.) berpotensi sebagai sumber bahan bakar nabati dan biomaterial karena kandungan selulosanya yang tinggi. Selulosa merupakan komponen utama penyusun dinding sel tanaman, sebagai rantai lurus yang tersusun dalam gugusan polisakarida, yang disebut mikrofibril selulosa. Sebuah gen bernama COBRA telah diketahui berperan dalam menentukan arah mikrofibril dan kristalisasi selulosa. Gen COBRA pada spesies Saccharum spp. belum banyak dipelajari. Oleh karena itu, kajian in silico dilakukan untuk mempelajari gen COBRA pada Saccharum sp. Melalui metode perbandingan genomika, gen COBRA dari Arabidopsis sp. (AtCOBLs) dibandingkan dengan gen COBRA dari Saccharum sp. (SoCOBLs). Domain conserve pada gen kemudian diidentifikasi dan sistem klaster disusun dalam sebuah pohon filogeni. Setelah itu, dibuat model untuk menganalisis struktur dari protein SoCOBL. Dari hasil analisis, sebelas perancah genom Saccharum sp. berhasil diidentifikasi. Kemudian, identifikasi daerah lestari menghasilkan sembilan protein SoCOBL. Pohon filogeni menggambarkan dua klaster utama: I dan II, yang membedakan famili SoCOBLs tersebut berdasarkan sekuens protein, motif domain, dan karakteristik asam amino. Karakteristik asam amino menyebabkan variasi pada struktur protein-protein SoCOBL. Secara umum, gen COBRA telah teridentifikasi pada Saccharum sp., meskipun fungsi dan ekspresi spesifiknya pada jaringan masih belum diketahui. Diperkirakan gen tersebut berperan sebagai pengatur arah mikrofibril dan proses sintesis selulosa. Oleh karena itu, perlu adanya analisis lebih lanjut pada level in vitro dan in vivo.[Kata kunci: selulosa, genomika komparatif, Saccharum sp.] 
Modifikasi sistem kultur in vitro untuk meningkatkan vigor planlet stevia (Stevia rebaudiana Bert.) [Modification of in vitro culture system to increase the vigor of stevia (Stevia rebaudiana Bert.) plantlets] Rizka Tamania SAPTARI; . SUMARYONO
E-Journal Menara Perkebunan Vol 84, No 2 (2016): Desember 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (277.96 KB) | DOI: 10.22302/iribb.jur.mp.v84i2.211

Abstract

 Stevia (Stevia rebaudiana Bert.), a sweetener plant, has been mass propagated by tissue culture technique. Optimal conditions to increase vigor of stevia plantlets are needed to support the sustainability of in vitro plantlet stocks and increase plantlet survival rate during acclimatization. The aim of this research was to investigate the effect of different media, culture vessel sizes, and vessel closure types on the vigor of stevia plantlets. The plant material was derived from apical shoot cuttings of sterile stevia plantlets grown on WP medium without growth regulator. Several treatments used in this study were solid or double layer media; short or tall culture vessel; and polypropile screw cap or plastic film closures.  Growth of plantlets was determined after 3 weeks of culture. Temperature and light intensity inside the vessels were also observed. The results showed that the best treatment to increase the vigor of stevia plantlets was a double-layer medium in a tall culture vessel (diameter 7 cm and height 11 cm) with either screw cap or plastic film. It was exhibited by significantly bigger stem diameter, more and bigger leaves, longer roots, and higher biomass fresh weight than those of other treatments. Higher temperature was observed on tall culture vessel, whereas all treatments did not significantly affect light intensity inside the vessels.[Keywords: stevia, plantlet vigor, double-layer medium, culture vessel size, vessel closure]AbstrakStevia (Stevia rebaudiana Bert.), tanaman pemanis, telah diperbanyak melalui teknologi kultur jaringan. Kondisi kultur optimal untuk meningkatkan vigor planlet stevia masih diperlukan untuk mendukung keberlanjutan tanaman stock in vitro dan untuk meningkatkan daya hidup planlet ketika diaklimatisasi. Penelitian yang dilakukan bertujuan untuk menentukan pengaruh penggunaan jenis media, ukuran botol kultur, dan jenis penutup botol yang berbeda terhadap vigor planlet stevia. Material tanaman yang digunakan didapat dari potongan tunas apikal plantlet stevia steril yang ditumbuhkan pada media WP tanpa zat pengatur tumbuh. Perlakuan jenis media terdiri atas media padat dan media dua-lapis (double-layer), ukuran botol pendek dan tinggi, serta jenis tutup ulir berbahan polipropilen dan lembaran plastik transparan. Pengamatan pertumbuhan planlet dilakukan setelah 3 minggu, juga dilakukan pengamatan terhadap suhu dan intensitas cahaya di dalam botol kultur. Hasil penelitian memperlihatkan bahwa perlakuan terbaik untuk meningkatkan vigor planlet stevia adalah dengan menggunakan media dua-lapis dalam botol kultur (diameter 7 cm, dan tinggi 11 cm), baik dengan tutup ulir maupun plastik. Hal ini ditunjukkan dari diameter batang lebih besar, daun lebih banyak dan besar, akar lebih panjang, serta bobot segar biomassa lebih tinggi dibandingkan dengan perlakuan lainnya. Suhu lebih tinggi terukur pada perlakuan botol tinggi, sedangkan semua perlakuan tidak mempengaruhi secara nyata intensitas cahaya di dalam botol kultur.[Kata kunci: stevia, vigor, media dua-lapis, ukuran botol kultur, tutup botol]
Embriogenesis somatik dari pucuk tunas tanaman kurma (Phoenix dactylifera L.) Somatic embryogenesis from shoot tip of date palm (Phoenix dactylifera L.)) Rizka Tamania SAPTARI; . SUMARYONO
E-Journal Menara Perkebunan Vol 86, No 2 (2018): Oktober 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (345.41 KB) | DOI: 10.22302/iribb.jur.mp.v86i2.313

Abstract

 Date palm (Phoenix dactylifera L.) is the most important crop in the dry areas of the Middle East and North Africa. This palm has been introduced to many countries but has not been grown commercially in Indonesia. Date palm propaga-tion by seeds is easy but its progenies are varied and a half of them are male trees that will not produce fruits. Meanwhile, the propagation by offshoots is impractical and technically difficult. Tissue culture makes it possible to massproduce of genetically identicalsuperior date palms. This research aimed to develop somatic embryogenesis (SE) of date palm using shoot tipand young leaves of date palm seedling as explants. Steps on somatic embryogenesis are explant sterilization, callus initiation and proliferation, somatic embryos induction and maturation, and plantlets matura-tion and rooting. Calli emerged from shoot tip explants after  9 weeks of culture in a modified MS medium supplemented with 10 mg/L 2,4-D, 1 mg/L or  3 mg/L 2-iP, and 1.5 g/L active charcoal. The callus was able to bear somatic embryo in the modified MS medium without hormones. Somatic embryos then developed into plantlets, and roots of plantlets were effectively initiated in the medium supplemented with 0.5 mg/L NAA and 1 mg/L IBA.[Keywords:sterilization,  callogenesis, somatic embryo induction, plantlet rooting, clonal propagation]. Abstrak  Tanaman kurma (Phoenix dactyliferaL.) merupakan tanaman terpenting di wilayah kering Timur Tengah dan Afrika Utara. Palma ini telah menyebar ke banyak negara, namun belum ditanam secara komersial di Indonesia. Perbanyakan kurma dengan biji sangat mudah tetapi turunannya sangat beragam dan setengahnya merupakan tanaman jantan yang tidak berbuah. Perbanyakan dengan anakan (offshoots) secara komersial tidak praktis dan relatif sulit. Kultur jaringan memungkinkan untuk dihasilkan secara massal bibit tanaman kurma varietas unggul yang secara genetik seragam. Penelitian ini bertujuan untuk mengembangkan embriogenesis somatik menggunakan eksplan pucuk tunasdan daun muda dari bibit tanaman kurma. Pengembangan embriogenesis somatik terdiri dari tahap sterilisasi eksplan, inisiasi dan proliferasi kalus, induksi dan maturasi embrio somatik, serta pembesaran dan pembentukan akar planlet. Kalus terbentuk dari eksplan pucuk tunassetelah 9 minggu dikultur pada medium MS modifikasi yang ditambahkan 2,4-D 10 mg/L,  2-iP 1 mg/L atau 3 mg/L, dan arang aktif 1,5 g/L.Kalus berhasil diinduksi menghasilkanembrio somatik pada medium MS modifikasi tanpa penggunaan hormon. Embrio somatik kemudian berkembang hingga menjadi planlet, dan akar planlet secara efektif terinisiasipada medium yang ditambahkan NAA 0,5 mg/L dan IBA1 mg/L.  [Kata kunci :sterilisasi,  kalogenesis, induksi embrio somatik, pengakaran planlet, propagasi klonal].
Determination of the optimum initial callus weight for the efficient propagation of sugarcane in temporary immersion bioreactor Rizka Tamania SAPTARI; Imron RIYADI; Masna Maya SINTA; M Eko Riyo Bayu PRASETYO; Sylvia LINDAWATI; Sumaryono SUMARYONO
E-Journal Menara Perkebunan Vol 90, No 2 (2022): Oktober, 2022
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v90i2.505

Abstract

AbstrakBioreaktor perendaman sesaat (BPS) telah digunakan secara luas untuk propagasi skala massal berbagai tanaman penting, termasuk tanaman tebu. BPS menyediakan sistem kultur semi-otomatis dan kondisi optimal bagi pertumbuhan tanaman. Beberapa faktor menentukan pertumbuhan tanaman pada BPS, salah satunya densitas dari eksplan. Oleh karena itu, penelitian dilakukan untuk menentukan bobot awal yang optimal untuk kalus tebu yang dikulturkan pada BPS, serta mengevaluasi pengaruh perbedaan bobot awal kalus tersebut terhadap proliferasi dan regenerasi kalus tebu. Kalus tebu diinduksi dari daun muda yang masih menggulung dari empat varietas tebu unggul Indonesia. Bobot awal kalus yang dikultur ke dalam bejana TIB yaitu 0,05 g; 0,1 g; 0,2 g; 0,5 g; dan 1,0 g untuk setiap bejana. Kalus kemudian melalui tahap proliferasi pada BPS sebanyak tiga siklus, kemudian kalus diregenerasi pada BPS dengan perlakuan auksin dan sitokinin. Hasil penelitian menunjukkan bahwa 0,2 g merupakan bobot awal kalus yang efisien untuk proliferasi kalus tebu pada TIB, dimana eksponensial multiplikasi kalus tercapai pada bobot awal tersebut, yaitu untuk masing-masing varietas 130,3 kali (PSKA 942), 136,8 kali (PS 094), 21,3 (PS 881), dan 12,9 kali (PS 091) setelah 12 minggu. Densitas kalus pada TIB berkorelasi negatif dengan karakteristik fisikokimia medium. Hal ini menggambarkan variasi intensitas pertumbuhan dan metabolisme kalus dengan adanya perbedaan densitas pada BPS. Penggunaan BAP 0,2 mg L-1 bersama kinetin 0,2 mg L-1 paling sesuai untuk memacu regenerasi kalus tebu dengan menghasilkan jumlah tunas terbanyak dalam waktu relatif lebih cepat (1 – 2 minggu lebih cepat) dibandingkan perlakuan lainnya dan dengan tingkat kejadian pencoklatan yang rendah.[Kata kunci: kultur in vitro, kultur cair, proliferasi]AbstractTemporary immersion bioreactor (TIB) has been utilized for the mass-scale propagation of many important plants, including sugarcane. TIB facilitates a semiautomated culture system and provides optimal conditions for plant growth. Several factors determine plant growth in the TIB, such as explant density. Therefore, an experiment was carried out to determine the optimal initial weight of sugarcane calli and to evaluate its effect on the proliferation and regeneration in TIB. Sugarcane calli were induced from spindle leaves isolated from four Indonesian prime sugarcane varieties. The initial weights of the calli cultured in the TIB flasks were 0.05 g, 0.1 g, 0.2 g, 0.5 g and 1.0 g per flask. The calli were proliferated through three cycles in TIB, and subsequently regenerated in TIB with auxin and cytokinin treatments. The results of the experiments showed that 0.2 g was the most efficient initial weight for sugarcane callus proliferation in the TIB, resulting in an exponential multiplication rate of 130.3-fold (PSKA 942), 136.8-fold (PS 094), 21.3-fold (PS 881), and 12.9-fold (PS 091) within 12 weeks. In the TIB, callus density showed a negative correlation with the physicochemical properties of the medium, demonstrating various growth intensities or metabolic activities of calli at different densities in the TIB. The use of 0.2 mg L-1 BAP along with 0.2 mg L-1 kinetin was suitable for promoting the regeneration of sugarcane calli and producing the highest number of shoots in a relatively short amount of time (1 – 2 weeks faster) with low incidences of browning.[Keywords: in vitro culture, liquid culture, proliferation]