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OPTIMASI DAN EVALUASI METODE KRIOPRESERVASI PURWOCENG ROOSTIKA, IKA; DARWATI, IRENG; MEGIA, RITA
853-8212
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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ABSTRAKOptimasi dan evaluasi metode kriopreservasi perlu dilakukan dalammenentukan protokol standar untuk penyimpanan jangka panjang biakanpurwoceng. Penelitian ini bertujuan untuk mengetahui pengaruh kombinasiperlakuan pratumbuh, prakultur, dan formulasi media pemulih terhadapdaya tumbuh dan daya regenerasi tunas in vitro dan kalus embriogenikserta untuk mengevaluasi metode kriopreservasi melalui observasimorfologi, anatomi, dan sitologi. Penelitian dilakukan di LaboratoriumKultur Jaringan Kelompok Peneliti Biologi Sel dan Jaringan BB LitbangBiogen pada tahun 2008-2009. Teknik kriopreservasi yang digunakanadalah vitrifikasi (untuk apeks) dan enkapsulasi-vitrifikasi (untuk kalusembriogenik). Pada teknik vitrifikasi, tunas pucuk diberi perlakuanpratumbuh dengan sukrosa (3, 4, 5, dan 6%) selama 1 dan 2 minggu,perlakuan prakultur dilakukan pada media yang mengandung sukrosa 0,3M selama 1 dan 3 hari, perlakuan dehidrasi dengan PVS2 diberikan selama15 dan 30 menit, dan media pemulih yang diujikan adalah media dasar MSatau DKW dengan dan tanpa penambahan adenin sulfat 20 ppm. Padateknik enkapsulasi-vitrifikasi, kalus embriogenik dienkapsulasi terlebihdahulu dengan Na-alginat 3%, perlakuan dehidrasi dengan PVS2 diberikanselama 0, 30, dan 60 menit. Evaluasi metode teknik kriopreservasidilakukan melalui pengamatan morfologi secara visual, anatomi meristemdengan scanning electron microscope (SEM), pengujian viabilitas denganfluorescein diacetate (FDA), dan analisis ploidi secara flowcytometry.Hasil penelitian menunjukkan bahwa teknik enkapsulasi-vitrifikasi lebihbaik daripada teknik vitrifikasi untuk kriopreservasi purwoceng. Walaupunpersentase keberhasilan kriopreservasi rendah (10%), kalus embriogenikpurwoceng mampu berproliferasi dan beregenerasi menjadi ribuan embriosomatik dewasa. Evaluasi metode kriopreservasi dengan SEM dan FDAdapat diterapkan untuk memperkirakan keberhasilan teknik kriopreservasisecara dini sedangkan analisis flowcytometry dapat diterapkan untukmenguji stabilitas genetik bahan tanaman pasca-kriopreservasi.Kata kunci: Pimpinella pruatjan Molk., kriopreservasi, SEM, FDA,flowcytometryABSTRACTOptimization and evaluation of cryopreservation methods should beconducted to obtain standard protocol for long term conservation ofpruatjan. The objective of this study was to evaluate the effect ofcombined treatments of pregrowth, preculture, and recovery media to thesurvival and regeneration rate of in vitro shoots and embryogenic calli andto evaluate the cryopreservation methods by observing the morphological,anatomical, and cytological characters. The techniques of vitrification (forapex) and encapsulation-vitrification (for embryogenic calli) were appliedin this study. On vitrification technique, the apical shoots were pregrownon media containing of 3, 4, 5, and 6% sucrose for 1 and 2 weeks,precultured on media containing of 0,3 M sucrose for 1 and 3 days,dehydrated by PVS2 solution for 15 and 30 minutes, and planted onrecovery media (MS or DKW basal media supplemented with 20 ppmadenine sulphate). On encapsulation-vitrification technique, embryogeniccalli were encapsulated by 3% Na-alginate, dehydrated by PVS2 solutionfor 0, 30, and 60 minutes. The evaluation of cryopreservation methods wasdone through visual observation, SEM analysis, viability test, andflowcytometry determination. The result showed that encapsulation-vitrification was better than vitrification technique for cryopreservation ofpruatjan. The successful rate of this method was low (10%) but theembryogenic calli could proliferate and regenerate into thousands maturesomatic embryos. The evaluation by SEM and FDA can be applied asearly detection to estimate the successful of cryopreservation, whereasflowcytometryÂ analysisÂ mayÂ determineÂ theÂ geneticÂ stabilityÂ ofcryopreserved materials.Key words: Pimpinella pruatjan Molk., cryopreservation, SEM, FDA,flowcytometry

THE ECOLOGY AND DISTRIBUTION OF FREYCINETIA GAUD. (PANDANACEAE; FREYCINETOIDEAE) IN THE INDONESIAN NEW GUINEA SINAGA, NURHAIDAH IRIANY; MEGIA, RITA; HARTANA, ALEX; KEIM, ARY PRIHARDHYANTO
REINWARDTIA Vol 13, No 2 (2010): Vol. 13 No. 2
Publisher : Research Center for Biology

SINAGA, N. I., MEGIA, R., HARTANA, A., KEIM, A. P. Ecology and distribution of Freycinetia Gaud. (Pandanaceae; Freycinetoidea) in Indonesian New Guinea. Reinwardtia 13(2): 189-197. - The study mainly concerns with the species of Freycinetia that occurs in the Indonesian New Guinea, including the provinces of Papua and Papua Barat. The study indicates that almost all species of Freycinetia in the Indonesian New Guinea prefer high humidity and abundantly occur along rivers, except for the members of the group of species with imbricate leaves, which inhabit also secondary forests. Futhermore, the members of this group have never been found within the range of 1700 to 3000 m altitudes. This highest range of altitudes is specifically occupied by the members of the groups of species with semi imbricate and grass-like leaves. The costal forests are inhabited by the groups of species with semi and non imbricate leaves. Indonesian New Guinea shares many species with Papua New Guinea, except for the members of the group of species with semi imbricate leaves, which are more common in Indonesian New Guinea than in Papua New Guinea. On the contrary, the members of the group of species with grass-like leaves are more common in Papua New Guinea and becoming rare toward the Indonesian site and becoming absent in the Vogelklop (Birds head), except for F. polyclada which is commonly found in Sorong. Indonesian New Guinea possesses 34 species exclusively distributed in the area, while Papua New Guinea has 72 species. The two areas share 52 species. Only five species have extra New Guinean distributions, i.e. F. excelsa, F. funicularis, F. marginata, F. percostata, and F. scandens.

In Vitro Propagation of Bambusa balcooa as Alternative Material of Wood Nurhayani, Siti; Megia, Rita; Purnamaningsih, Ragapadmi
Biosaintifika: Journal of Biology & Biology Education Vol 10, No 1 (2018): April 2018
Publisher : Department of Biology, Faculty of Mathematics and Sciences, Semarang State University . Ro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v10i1.11079

A diversion of raw material from wood to bamboo is necessary. In vitro culture of bamboo can be used to provide a high number of seedling. The aim of this study was to increase the multiplication of a high quality Bambusa balcooa as a wood alternative material. Part of plants used was the sterile axillary shoot. The explants were planted on MS0 medium for 2 weeks and later on multiplication medium MS+0.3 mg/l BAP + 0.3mg/l TDZ. The shoots obtained were fragmented into clusters (3-5 shoots) used for the next multiplication stage using five different medium formulas: (1) MS0; MS containing: (2) 0.1 mg/l BAP, (3) 0.3 mg/l BAP, (4) 0.1 mg/l BAP + 0.1 mg/l TDZ and (5) 0.3 mg/l BAP + 0.1 mg/l TDZ. The results showed that MS medium containing 0.1 mg/l BAP + 0.1mg/l TDZ was the best medium for B. balcooa propagation. The shoots produced from aforementioned medium had a better quality compared to the other medium. Forty days after planting, the average number of shoots in this medium was 14.25. MS medium containing 0.3 mg/l BAP + 0.1 mg/l TDZ produced the highest number of shoot but in lower quality. Rooting medium containing 10 mg/l IBA + 5 mg/l NAA produced 9-16 root in 8 weeks. Vermicompost was more prevalent for the acclimatization of B. balcooa compared to compost. The use of B.balcooa resulted in in vitro propagation as a substitute alternative for wood is expected to save the environment from illegal logging. Â

Two New Records of Wild Bananas (Musa balbisiana and Musa itinerans) from Sulawesi Sulistyaningsih, Lulut Dwi; Megia, Rita; Widjaja, Elizabeth Anita
Makara Journal of Science Vol. 8, No. 1
Publisher : UI Scholars Hub

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The diversity of wild banana species in Sulawesi was investigated based on the morphological characteristics of herbarium specimens collected in Sulawesi and deposited in the Herbarium Bogoriense, Bogor, Indonesia (BO). New specimens were collected from Central, North, South, and Southeast Sulawesi, and digital type specimens were also used in this study. The aim of this study was to describe the diversity of wild banana species in Sulawesi as most Musaceae specimens stored at BO have not been identified. By examinating 110 sheets of herbarium specimens, five species of Musa, including two infraspecific taxa of M. acuminata housed in Sulawesi, were identified. Musa acuminata, M. celebica, and M. textilis were previously reported from Sulawesi. However, M. balbisiana and M. itinerans are two new records of wild banana species in Sulawesi. Identification keys, descriptions, distribution maps, and line-drawing illustrations of these two species are provided.

Two New Records of Wild Bananas (Musa balbisiana and Musa itinerans) from Sulawesi Sulistyaningsih, Lulut Dwi; Megia, Rita; Widjaja, Elizabeth Anita
Makara Journal of Science Vol. 18, No. 1
Publisher : UI Scholars Hub

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The diversity of wild banana species in Sulawesi was investigated based on the morphological characteristics of herbarium specimens collected in Sulawesi and deposited in the Herbarium Bogoriense, Bogor, Indonesia (BO). New specimens were collected from Central, North, South, and Southeast Sulawesi, and digital type specimens were also used in this study. The aim of this study was to describe the diversity of wild banana species in Sulawesi as most Musaceae specimens stored at BO have not been identified. By examinating 110 sheets of herbarium specimens, five species of Musa, including two infraspecific taxa of M. acuminata housed in Sulawesi, were identified. Musa acuminata, M. celebica, and M. textilis were previously reported from Sulawesi. However, M. balbisiana and M. itinerans are two new records of wild banana species in Sulawesi. Identification keys, descriptions, distribution maps, and line-drawing illustrations of these two species are provided.

DETEKSI INTEGRITAS GENOMIK PISANG HASIL IRADIASI IN VITRO BERDASARKAN PENANDA MIKROSATELIT Megia, Rita; Ratna, Nina Ratna
Makara Journal of Science Vol. 14, No. 2
Publisher : UI Scholars Hub

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Genomic Integrity Detection of In Vitro Irradiated Banana Using Microsatellite Marker. The research aims to detect genomic integrity of in vitro irradiated banana using microsatellite marker. These studies were done on banana cv. Pisang Mas irradiated by 15 Gy of gamma ray. The DNA was isolated from each accesion following Dixie. Amplification of DNA products were done by Perkin Elmer Gene Amp PCR 2400 using ten primers, and then electroforesis in agarose 1%. Finally a vertical polyacrylamide gel electroforesis was run and the products were visualized by silver staining. The result shown that among the primers tested, eight primers produced clear, discrete, and reproducible bands. Number of DNA band exhibited ranging from one to two, following the ploidy level of pisang Mas which is a diploid banana cultivar (AA). One band suggest homozygote allele while two bands showed heterozygote allele. Out of eight primers, six primers produced different allele among irradiated, in vitro, and in vivo control plant. Meanwhile, for the other two primers the allele were monomorph for all the accessions examined. Genomic modification was observed at all irradiated plants. The modification can happened at zygosity of certain allele that may change from heterozygote to homozygote or vice versa. While modification in allele size that underlying genomic instability could be caused by several genetic events such as deletion, insertion, and amplification of nucleotides.

Phylogenetical Study of Wild Banana Species (Musa L.) in Sulawesi Inferred from Internal Transcribed Spacer Region of Nuclear Ribosomal DNA Sequences Sulistyaningsih, Lulut Dwi; Megia, Rita; Widjaja, Elizabeth Anita
BIOTROPIA Vol. 21 No. 1 (2014): BIOTROPIA Vol. 21 No. 1 June 2014
Publisher : SEAMEO BIOTROP

Study to determine the phylogenetic relationship of wild banana species (Musa) in Sulawesi based on the ITS regions sequences has been done. A total of 28 samples including 16 ITS sequences from GeneBank were used. Species of Ensete and Musella were incorporated as an outgroup. Total DNA was extracted by the modified CTAB. ITS-5 and ITS-4 primers were used to amplify the ITS regions. Data sequences were edited by ChromasPro and were aligned by Muscle software.Some unique nucleotides were found in M. balbisiana, M. itinerans, and M. textilis sequences. Strict consensus tree revealed from Maximum Parsimony (MP) as well as Bayesian analysis showed Musa textilis was separated from other taxa in which M. textilis become as the basal sister to M. balbisiana. By using MP analysis, M. itinerans together with M. acuminata ssp. banksii, M. acuminata var. tomentosa, and M. celebica placed in the same clade, whereas Bayesian analysis showed M. itinerans was separated and placed in different clade.

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