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Acclimatization and early growth of tissue culture-derived Stevia rebaudiana at low altitude area in Bogor, Indonesia (Aklimatisasi dan pertumbuhan awal Stevia rebaudiana asal kultur jaringan pada dataran rendah di Bogor, Indonesia) Masna Maya SINTA; Dian Mutiara AMANAH
E-Journal Menara Perkebunan Vol 87, No 1 (2019): April, 2019
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Aklimatisasi merupakan masa transisi sebelum kultur in vitro dapat ditanam di lingkungan ex vitro. Di daerah tropis, stevia seyogianya ditanam di dataran tinggi. Pengembangan klon stevia yang sesuai untuk dataran rendah di kawasan tropis sangat penting untuk memungkinkan penggunaan mekanisasi pada pertanaman stevia yang luas. Tujuan penelitian ini adalah menentukan pengaruh kondisi eksplan dan periode hardening terhadap daya hidup dan pertumbuhan pada tahap aklimatisasi dan pertumbuhan awal stevia klon BS 22 pada area terbuka di dataran rendah di wilayah tropis. Penelitian pertama dilangsungkan menggunakan umur tunas yang berbeda: 0, 1, 2, dan 3 minggu yang dikultur pada media padat sebagai sumber bahan eksplan. Penelitian kedua menggunakan satu buku stevia pada periode hardening dalam media cair selama 1, 4, 7 dan 10 hari. Aklimatisasi dilaksanakan dengan menanam eksplan dalam medium tumbuh campuran pada multi-tray dan diletakkan di dalam sungkup plastik tertutup selama 1 bulan. Tanaman yang berhasil hidup kemudian dipindah ke polibeg pada area terbuka dengan sinar matahari penuh. Pengamatan daya hidup dan pertumbuhan dilakukan pada akhir tahap aklimatisasi dan setelah 2 bulan di area terbuka. Hasil penelitian menunjukkan bahwa eksplan umur 1 minggu yang dikultur pada media padat mempunyai daya hidup tertinggi yakni 83%. Buku tunggal stevia yang dikultur pada medium cair pada tahap hardening selama 4 hari meningkatkan daya hidup menjadi 97% selama aklimatisasi 1 bulan. Setelah aklimatisasi, tinggi tanaman secara rata-rata adalah 2,6 cm dengan 10,6 helai daun. Tanaman yang dipindah ke area terbuka tumbuh pesat dengan tinggi tanaman mencapai 12 cm dengan 30 helai daun dan daya hidup 63% setelah 2 bulan. Hal ini menunjukkan bahwa stevia klon BS 22 mungkin sesuai untuk dataran rendah di daerah tropis. [Kata kunci: tanaman pemanis, eksplan tunas, tahap hardening, daya hidup, tropika] AbstractAcclimatization is a transition period before in vitro culture can be planted in ex vitro environment. In the tropical region, stevia is should be planted at high altitude areas. The development of stevia clones suitable for low land area in the tropics is important to make it possible to apply mechanization in a large scale stevia plantation. The purpose of this research was to determine the effect of explant conditions and hardening period on survival rate and growth during acclimatization stage and early growth of stevia clone B 22 in an open area at low altitude area in the tropics. The first experiment was conducted using different shoot ages: 0, 1, 2 and 3 weeks cultured on solid media as an explant material source. The second experiment was using single node of stevia in different hardening periods in liquid media for 1, 4, 7 and 10 days. Acclimatization was carried out by planting the explants on a mixture growing medium in multi-trays and placed inside a closed plastic tunnel for 1 month. The survival rate and growth parameters were observed at the end of acclimatization stage and after 2 months in the open area. The results show that 1-week explant age on solid media had the highest survival rate at 83%. Hardening single node of shoot in a liquid medium for 4 days increased the survival rate to 97% in 1 month acclimatization stage. After acclimatization, the plant height on average was 2.6 cm with 10.6 leaves. The survived plants planted in an open area grew rapidly to 12 cm in height with 30 leaves and survival rate 63% within 2 months. It indicates that stevia clone BS 22 may suitable for a low altitude area in the tropics. [Key words: sweetener plant, shoot explant, hardening period, survival rate, tropics]

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

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.

Front matter 2019 no 1 Masna Maya Sinta
E-Journal Menara Perkebunan Vol 87, No 1 (2019): April, 2019
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Back matter 2018 no 2 Masna Maya Sinta
E-Journal Menara Perkebunan Vol 86, No 2 (2018): Oktober 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Front matter 2018 no 2 Masna Maya Sinta
E-Journal Menara Perkebunan Vol 86, No 2 (2018): Oktober 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Back matter 2019 no 1 Masna Maya Sinta
E-Journal Menara Perkebunan Vol 87, No 1 (2019): April, 2019
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Induksi mutasi Stevia rebaudiana dengan perendaman kolkisin secara in vitro (Induced mutation of Stevia rebaudiana through colchicine soaking in vitro) Masna Maya SINTA; Ni Made Armini WIENDI; Syarifah Iis AISYAH
E-Journal Menara Perkebunan Vol 86, No 1 (2018): April, 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Stevia rebaudiana Bert. is a plant producing steviol glycosides that have 200-300 times sweeter than sucrose. These steviol glycosides are produced in the leaves and then spread to all parts of the plant including stems. The use of superior stevia planting material is important for stevia sugar industry. One of the stevia breeding programme is to increase genetic diversity through colchicine soaking to produce polyploid plants. Polyploid plants usually have higher vigor than diploid plants. The purpose of this research was to induce genetic diversity of stevia through colchicine soaking in vitro. Single nodes of sterile stevia clone BS were soaked in colchicine at the concentration of 0.01; 0.02; 0.04; 0.08 and 0.1% for 48 and 72 hours, and in sterile aquadest as a control. Plantlet subcultures were done until MV4 (mutant vegetative 4). Putative mutants were observed by plantlet vigor and stomata analyses on MV5. Vigor of plantlets was observed by counting the number of leaves, nodes, roots, fresh weight and dry weight of the plantlet. Stomata analysis was performed by calculating stomata density, stomata size and chloroplast number in stomata guard cells. Results showed that colchicine soaking treatment increased significantly fresh weight and dry weight of putative mutants. Colchicine soaking treatment increased chloroplast number on stomata guard cell and stomata size, but decreased stomata density. Stevia soaked in colchicine for 48 hours at concentration 0.01-0.04% produce putative mutants with high chromosome numbers. [Key words: poliploidy, stomata, chloroplast, mutant]AbstrakStevia rebaudiana Bert. merupakan tanaman penghasil glikosida steviol yang memiliki tingkat kemanisan 200-300 kali lebih tinggi dibandingkan sukrosa. Glikosida steviol ini diproduksi di daun yang kemudian disalurkan ke bagian tanaman lainnya termasuk batang. Penggunaan klon terbaik stevia merupakan salah satu kunci penting keberhasilan industri gula stevia. Salah satu program pemuliaan tanaman stevia adalah meningkatkan keragaman tanaman melalui mutasi dengan kolkisin sehingga menghasilkan tanaman poliploid. Tanaman poliploid umumnya memiliki vigor lebih baik dibandingkan tanaman diploid. Tujuan dari penelitian ini adalah untuk meningkatkan keragaman stevia melalui peren-daman kolkisin in vitro. Buku tunggal steril stevia klon BS direndam dalam kolkisin dengan konsentrasi 0,01; 0,02; 0,04; 0,08 dan 0,1% selama 48 dan 72 jam dengan perendaman dalam air steril sebagai kontrol. Sub kultur dilakukan hingga MV4 (mutan vegetatif 4). Pengamatan mutan putatif dilakukan meliputi analisis morfologi dan stomata pada MV5. Analisis morfologi dilakukan dengan mengamati jumlah daun, buku, akar, bobot basah serta bobot kering planlet. Analisis stomata dilakukan dengan menghitung kerapatan stomata, ukuran stomata serta jumlah kloroplas pada sel penjaga stomata. Hasil menunjukkan bahwa perendaman stevia pada kolkisin meningkatkan bobot basah serta bobot kering stevia in vitro. Perlakuan perendaman kolkisin meningkatkan jumlah kloroplas pada sel penjaga stomata serta ukuran stomata namun menurunkan kerapatan stomata. Perendaman stevia selama 48 jam pada konsentrasi kolkisin 0,01-0,04% menghasilkan mutan putatif dengan jumlah kromosom tertinggi.[Kata kunci: poliploidi, stomata, kloroplas, mutan]

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

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]

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