Anthurium adreanum Lindl. cv. Nitta is an ornamental plant for cut flower industries. In vitro propagation enables a large scale production of high quality seedlings of A. adreanum quickly. The aim of this experiment was to overcome some problems in the in vitro propagation of A. adreanum, such as contamination, lack of plantlets vigor and low rate of survival in the acclimatization. In vitro propagation of A. adreanum was conducted via organogenesis from leaf explant. Explant sterilization using desogerm, antioxidant, alcohol, rifampicin, and NaOCl was the best because they reduced browning and contamination until 0 %. Organogenesis of A. adreanum was successfully initiated using MS medium added with 2,4-D and BAP at 1 mg L -1 , which produced 74 shoots per explant. Shoots were best maturated in MS medium with a half strength of macro minerals added with 1 mg L -1 paclobutrazol. Survival rate increased by 7 and 14 days hardening treatment using double layer medium in the light culture room. Survival rate reached 89.3 % at 8 weeks after acclimatization.

Microenvironment inside the culture vessel such astemperature, light intensity, relative humidity, and aerationaffect growth and development of plantlets. This experimentwas conducted to determine the effect of different culturevessel closures on microenvironmental conditions inside thevessel and on growth of plantlets of oil palm. Shoots of oilpalm derived from somatic embryos were cultured on DFmedium for eight weeks in transparent culture bottlescovered with five different vessel closures e.i. screw cap withplastic wrap, screw cap, plastic wrap, aluminum foil, andautoclavable plastic. The culture vessels were placed in theculture room with light intensity 20 µmol/m 2 /sec for 12 hoursphotoperiod, at room temperature 26°C. Parametersobserved on plantlet growth were shoot height, biomass freshweight, leaf number, and leaf color grade, while onmicroenvironment were temperature and light intensity. Atthe end of experiment, the volume and fresh weight of theremaining medium were measured to determine evaporationrate of each treatment. Results show that the use of differentculture vessel closures affected the microenvironment insidethe vessel, the volume of the remaining medium, and thegrowth of the plantlets. The closure increased thetemperature by 1.6 – 2.6°C and decreased the light intensityby 1.7 – 8.7 µmol/m 2 /sec inside the culture vessels dependson the culture vessel closures. Culture vessels with aluminumfoil closure had the lowest temperature (28.9°C) and thelowest light intensity (10.8 µmol/m 2 /sec) gave the best resultin the growth of the plantlets. Better plantlets growth wasalso observed in the culture vessel with autoclavable plasticclosure that less expensive, therefore it can be used as analternative vessel closure for the growth of oil palm plantlets.AbstrakLingkungan mikro di dalam botol kultur seperti suhu,intensitas cahaya, kelembaban nisbi dan aerasi mem-pengaruhi pertumbuhan dan perkembangan planlet.Penelitian ini dilakukan untuk mengetahui pengaruhpenggunaan penutup botol kultur yang berbeda terhadapkondisi lingkungan mikro di dalam botol kultur danpertumbuhan planlet kelapa sawit. Planlet kelapa sawit asalembrio somatik dikulturkan dalam botol kultur bening berisimedium DF selama delapan minggu dan ditutup mengguna-kan lima jenis penutup botol yang berbeda yaitu tutup ulirdengan plastik wrap, tutup ulir, plastik wrap, aluminium foildan plastik tahan diautoklaf. Kultur diletakkan dalam ruangkultur, di bawah lampu TL dengan intensitas cahaya20 µmol/m 2 /detik dan suhu ruang 26 o C. Parameterpertumbuhan planlet yang diamati adalah tinggi planlet,bobot basah, jumlah daun dan kelas warna daun, sedangkanlingkungan mikro adalah suhu dan intensitas cahaya. Padaakhir eksperimen, volume dan bobot basah medium yangtersisa diukur untuk mengetahui tingkat penguapan padasetiap perlakuan. Hasil penelitian menunjukkan bahwapenggunaan penutup botol yang berbeda berpengaruhterhadap lingkungan mikro, volume medium tersisa dalambotol kultur dan pertumbuhan planlet. Penutup botolmeningkatkan suhu 1,6 – 2,6 o C dan menurunkan intensitascahaya 1,7 – 8,7 µmol/m 2 /detik di dalam botol tergantungpada jenis penutup botol yang digunakan. Botol kulturdengan penutup berbahan aluminium foil mempunyaiintensitas cahaya terendah (10,8 µmol/m 2 /detik) dan suhuterendah (28,9 o C) memberikan hasil terbaik pada pembesaranplanlet kelapa sawit. Pertumbuhan planlet yang baik jugaterdapat pada botol kultur dengan penutup plastik tahandiautoklaf yang lebih murah, sehingga penutup ini dapatdigunakan sebagai pilihan untuk pembesaran planlet kelapasawit.

No abstract

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]

Stevia rebaudiana Bertoni, a sweetener plant, has been cultivated in Indonesia since the early 1980s, but the yield is lower than in other countries. Five selected introduced stevia clones were planted at Megamendung, Bogor, Indonesia (6°39’ S, 106°56’ E, 800 m above sea level) on December 2014 until November 2015 to evaluate their growth and biomass yield. The growth and yield were observed from 6 to 12 months after planting. In addition, stevioside, rebaudioside A (reb A), and total steviol glycoside (TSG) contents of leaves were measured using high performance liquid chromatography (HPLC). At six months after planting, stem diameter was 8-11 mm, plant canopy diameter was 23-30 cm, plant height was 24-35 cm, and plant mortality was less than 5% except for clone BU at 30.7% due to Sclerotium attack. The plants flowered at different times, therefore had different harvest durations, from 4 weeks (clone BM) to 8 weeks (clone BP). The highest leaf yield was clone BM (6.04 ton ha-1 per year), followed by BX (4.91 ton ha-1 per year), and BP (4.38 ton ha-1 per year). From the five clones tested, clone BM was the best for leaf, TSG, and stevioside yields; whereas clone BS and BP were the best for reb A yield. Keywords: rebaudioside A, Stevia rebaudiana, stevioside, total steviol glycoside

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 leaves of sweetener plant Stevia rebaudiana contain secondary metabolites of steviol glycosides which are very sweet, with no calorie and zero glycemic index. Propagation of stevia by seeds is ineffective due to its low germination rate and diverse progenies. The tissue culture of stevia can be used to mass propagate rapidly and is commonly conducted by shoot multiplication. Up to now, the technology of somatic embryogenesis (SE) in stevia has not been successful yet. SE is developed to increase the production scale, rejuvenate clonal-propagated plants, and plant genetic transformation. The research objective was to develop protocols for the initiation, proliferation, and development of embryogenic calli of stevia as potential materials for SE. The explants used were young leaves, nodes, and internodes of axenic plantlets of stevia BX clone. The explants were cultured on MS solid media containing different concentrations of auxin and cytokinin for callus initiation. Callus emerged after 2-3 weeks of culture. The calli obtained were proliferated by subculturing several times as material stocks for indirect SE. MS solid media added with 1 µM 3,4-D and 16 mM CaCl2 gave the highest callus multiplication rate (4.7 times in 3 weeks). The selection of embryogenic calli was made continuously to obtain a pure line of embryogenic calli. Three types of calli attained were friable, fast-growing, yellowish calli, shiny nodular calli, and greenish nodular calli. Histological studies revealed that cells of the nodular calli had been differentiated to potentially formed somatic embryos.

Acclimatization of plantlets is a critical stage in themicropropagation of many plants. An experiment wasconducted to determine the effect of pre-condition periodand vessel closure on the growth and survival rate ofrubber (Hevea brasiliensis Muell.Arg.) plantlets derivedfrom in vitro microcutting during acclimatization.Plantlets were planted in plastic pots containing mixedgrowing media after being conditioned in ex vitroenvironment for 0, 3 and 6 days. Five closure vesseltreatments were closed pots placed in opened container,opened pots in closed glass container, closed pots inclosed glass container, opened pots in closed plasticcontainer, and closed pots in closed plastic container.Observation on leaf conditions, rooting frequency, andplant height were conducted at 1.5 months and on thepercentage of survive plantlets at 1.5 and 3 months afteracclimatization. The results showed that pre-conditionwas required to increase survival rate and growth of theplantlets. Pre-condition period of six days gave a highersurvival rate than 0 and 3 days which reached 100% and93% on opened pot in closed plastic container and closedpot in opened container, respectively after 1.5 monthsand was reduced to 80% after three months ofacclimatization. The highest formation of new leaves androots were also obtained on six days pre-conditionperiod. Plantlets with pre-condition for six days and wereplanted on closed pots in an opened container had thebest rooting frequency which was 90%. The resultshowed that the highest survival rate (80%) of rubberplantlets after three months was obtained when theplantlets were pre-conditioned in ex vitro conditions forsix days before acclimatization and planted on openedpots in a closed plastic container or closed pots in anopened container.AbstrakAklimatisasi planlet merupakan tahap kritis dalammikropopagasi tanaman. Penelitian dilakukan untuk me-nentukan pengaruh periode pra-kondisi dan penyung-kupan terhadap pertumbuhan dan daya hidup planletkaret (Hevea brasiliensis Muell.Arg.) asal stek mikro (invitro microcutting) selama aklimatisasi. Planlet ditanampada pot plastik berisi campuran media tanam setelahdikondisikan lebih dahulu pada lingkungan luar selama 0,3 dan 6 hari. Lima perlakuan penyungkupan adalahpenanaman planlet pada pot tertutup diletakkan dalamwadah terbuka, pot terbuka dalam wadah kaca tertutup,pot tertutup dalam wadah kaca tertutup, pot terbukadalam wadah plastik tertutup dan pot tertutup dalamwadah plastik tertutup. Pengamatan keadaan daun, pem-bentukan akar dan tinggi tanaman dilakukan pada 1,5bulan, sedangkan persentase planlet yang hidup diamatipada 1,5 dan 3 bulan setelah aklimatisasi. Hasil penelitianmenunjukkan bahwa periode pra-kondisi diperlukanuntuk meningkatkan daya hidup dan pertumbuhanplanlet. Pra-kondisi selama enam hari memberikan dayahidup planlet lebih tinggi dibandingkan dengan 0 dan 3hari yaitu 100% dan 93% pada perlakuan penanamanpada pot terbuka dalam wadah plastik tertutup dan pottertutup dalam wadah terbuka setelah 1,5 bulan danmenjadi 80% setelah tiga bulan. Penambahan daun barudan pembentukan akar tertinggi juga terdapat padaperlakuan pra-kondisi enam hari. Perlakuan pra-kondisienam hari dengan pot tertutup yang diletakkan dalamwadah terbuka memperlihatkan persentase pembentukanakar yang paling baik yakni 90%. Hasil peneltian me-nunjukkan bahwa daya hidup planlet karet tertinggi(80%) pada umur tiga bulan diperoleh apabila planletdipra-kondisi pada lingkungan ex vitro selama enam hari,kemudian ditanam pada pot terbuka dalam wadah plastiktertutup atau pot tertutup dalam wadah terbuka.