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Cytological Analysis of Root Cultures of Artemisia cina Ermayanti, Tri Muji; Yanti, Oktavia; Hafiizh, Erwin Al
Annales Bogorienses Vol. 9 No. 2 (2004): Annales Bogorienses
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Artemisia cina is a medicinal plant species producing bioactive compounds which are potential a antitumor, antifungal and antibacterial. The aim of this study was to analyze the stability of chromosome number in root cultures of A. cina. Transformed root culture was established by infection of leaves of A. cina with Agrobacterium rhizogenes strains 07-20001, ATCC 15834, A4; and Agrobacterium tumefaciens strain R1000. Roots isolated from glasshouse plants, plantlets grown in solid and liquid MS medium were utilized for investigation of chromosome examination of untransformed roots. Chromosome examination was conducted by squashing method and chromosome numbers were calculated under microscope. The results showed that both untransformed and transformed root had in lability in the chromosome number, but had the modal number of chromosome x=8 with the diploid number of 2n =4x = 32. Roots isolated from glasshouse plants of A. cina had 53.7~ of cell with the diploid numbers of 2n = 3 and 46.3% of cells had chromosome numbers ranged from 2n = 12 to 2n = 64. Untransformed roots isolated from plantlets cultured in solid medw had only 36.1% or cells with chromo orne number of 20 = 32, and untransformed roots grown in liquid medium had 49.4% of cells with 2n =32. The chromosome numbers of A. cina transformed roots was affected by trains of Agrobacterium. Root transformed with the bacterium Strain 07-20001 showed the highest in normal chromosome number of 2n = 32 (62.4%) followed by roots transformed with strains ATCC 15834 (61.9%). R1000 (43.6%) and A4 (43.0%). The range of the chromosome number of untransformed roots was from 2n=17 to  2n=64 whilst that of transformed roots was from 2n=11 to 2n=66.
In vitro Growth Response of Taraxacum officinale Weber ex F.H. Wigg Regenerated from Different Type of Explants Ermayanti, Tri Muji; Martin, Andri Fadillah
Annales Bogorienses Vol. 15 No. 1 (2011): Annales Bogorienses
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Dandelion (Taraxacum officinale Weber ex F.H. Wigg) is a medicinal plant species, reported to have some active compounds useful as anti-cancer, anti-inflammatory, antioxidants, and diuretics. Study on in vitro secondary metabolic production and tissue culture of these plants has been reported. This research was aimed to investigate plant regeneration from leaf blade, petiole and root as explants grown in selected media. Three different type of explants were cultured in MS solid medium supplemented with BAP (0, 0.5, and 1.0 mg/l) combined with NAA (0, 0.5, 1.0, and 2.0 mg/l) to select the best medium for spontaneous regeneration. Explants were grown for 6 weeks, the viability of explants, and formation of callus, shoots as well as roots were recorded. The results showed BAP at 1 mg/l combined with at 0.5 mg/l of NAA was suitable for shoot formation, whereas NAA alone was suitable for root formation from leaf blade and petiole. Root was the best explant for shoot regeneration, callus was grown at the first two weeks, and multiple shoots were grown after 3 weeks. No roots were found from root explant. The best medium for multiple shoot regeneration was MS containing 1 mg/l BAP without or with addition of 0.5 mg/l NAA.
Comparison of the Reduction Effect of Sucrose and Table Sugar Concentration on Growth Characteristics of Red Ginger (Zingiber officinale Rocs.) Cultured in Liquid Medium Hapsari, Betalini Widhi; Ermayanti, Tri Muji; Rantau, Deritha E.; Rudiyanto, Rudiyanto
Annales Bogorienses Vol. 15 No. 1 (2011): Annales Bogorienses
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The aim of this research was to compare the reduction effect of sucrose or table sugar concentration on growth characteristics of red ginger cultured in MS liquid medium. Shoots of red ginger was cultured on MS liquid medium without addition of plant growth regulators, supplemented with 5, 10, and 20 g/l of sucrose or table sugar for 8 weeks. Resulted plantlets were acclimatized in a greenhouse to investigate their growth and survival rate. Numbers of stomata, chlorophyll concentration as well as cross section of leaves from plantlets grown in vitro were compared to those of transplants grown in the greenhouse. The results showed that the use of table sugar at concentration of 20 g/l gave the best growth of red ginger. Meanwhile, the reduction of table sugar from 20 to 10 g/l reduced growth and survival rate of in vitro shoots as well as that of transplants in the greenhouse. Only few shoots formed roots when they were grown on the medium containing 5 g/l of table sugar, and transplants failed to grow in the greenhouse. It found that the chlorophyll content of in vitro plantlets was lower than those of transplants grown in the glasshouse. However, the number of stomata of the in vitro plantlets was higher than that of transplants grown in the glasshouse. There was no anatomical abnormalities found on the cross section of leaves between in vitro plantlets and transplants grown in the greenhouse. The replacement of sucrose with table sugar may reduce the production cost of plantlets.
Detection of Potyvirus Using RT-PCR and ACP-ELISA of Dioscorea Species and In Vitro Shoot Multiplication of the Virus Free Plants Wulandari, Dyah Retno; Ermayanti, Tri Muji
Annales Bogorienses Vol. 15 No. 2 (2011): Annales Bogorienses
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Detection of Potyvirus using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Antigen Coated Plate-Enzyme Linked Immunoabsorbent Assay (ACP-ELISA) for Dioscorea alata, Dioscorea hispida, and Dioscorea esculenta was conducted in order to establish in vitro culture of virus-free of these species. Plants were collected from Yogyakarta, Lampung, Pasuruan, Jakarta and Bogor. Total RNA of plants grown in a greenhouse was then isolated according to Simple Direct Tube (SDT) method. Total RNA from symptomatic leaf of Yard Long Bean (Vigna unguiculata) infected with Bean Common Mosaic Potyvirus (BCMV) was used as the positive control treatment. RT-PCR assay with degenerate primers MJ1(F) and MJ2(R) was used to identify the Potyviruses infecting Dioscorea. ACP-ELISA with antibodies specific to group Potyvirus was carried out to detect Potyvirus from leaves samples. The Dioscorea virus-free species was then cultured on modified MS medium. Shoot tips or internodes were used as explants. The results showed that using both RT-PCR and ACP-ELISA, all species tested were free from virus. The growth response of explants on MS medium was varied depending on the plant species and the concentration of BAP.
Micropropagation of Tetramolopium klossii (Moore) Mattfeld, Endemic Subalpine and Alpine Plant Species to New Guinea Ermayanti, Tri Muji; Hafiizh, Erwin Al; Martin, Andri Fadillah; Mandessy, Ary; Hermono, Arief; Mukhsia, Andi
Annales Bogorienses Vol. 15 No. 2 (2011): Annales Bogorienses
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The protocol of micropropagation of Tetramolopium klossii (Moore) Mattfeld through adventitious shoot proliferation was established. This is the first report of T. klossii tissue culture. Seeds were surface-sterilized by dipping them in 10% of ethanol for 1 min followed by soaking in 1% of sodium hypochlorite for 10 min. The sterile seeds were germinated on MS medium solidified with 8 g/l of agar, supplemented with 20 g/l of sucrose. Two-month old seedlings formed some shoot clumps on MS solid medium without addition of any plant growth regulators. Small clumps containing 4-6 adventitious shoots were used for shoot proliferation on MS solid medium containing 0; 0.1; 0.5; 1.0 and 2.0 mg/l of BAP (benzylaminopurine) or kinetin. The results showed that medium MS without cytokinins (control treatment), and the addition of 0.1 or 0.5 mg/l of BAP was more effective for shoot proliferation compared to the medium with addition of BAP at higher concentration and the addition of kinetin. In further subcultures, multiple shoots grown on MS medium without cytokinins was more vigorous than that of shoots grown on other media, so that the shoots were ready for rooting. Medium containing 0.5 mg/l of BAP was the best for maintaining multiple shoot formation. Shoots formed roots on MS medium containing half strength of macro nutrients.
The Effect of Increase in NaCl Concentration on Growth and Proline Content of Purple Yam (Dioscorea alata L.) Grown In Vitro Martin, Andri Fadillah; Azizah, Farroh; Wulandari, Dyah Retno; Ermayanti, Tri Muji
Annales Bogorienses Vol. 16 No. 2 (2012): Annales Bogorienses
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Tuber of purple yam (Dioscorea alata L.) has been used as an alternative food in some areas in Indonesia. The tuber contains high carbohydrate, low glycemic index and gluten free, therefore, study on genetic improvement of this species is needed to increase the productivity and to find out new cultivars which can be cultivated in marginal lands. This research was aimed to investigate the effect of NaCl concentration on growth and proline content of purple yam grown in vitro. Shoot tips were cultured on MS (Murashige and Skoog) medium supplemented with NaCl at concentrations of 25; 50; 100; 200 and 250 mM. After six weeks in culture, height of shoots, number of nodes, number of leaves, as well as proline content were recorded. The results showed that shoots grown on MS medium supplemented with NaCl at 25 and 50 mM had better growth compared to control. The best medium for its growth was MS containing 50 mM of NaCl. Increase in NaCl level’s resulted in decrease of growth. The LD50 value was obtained at 183 mM of NaCl. The Highest proline concentration was achieved by shoots grown on the medium supplemented with 100 mM of NaCl. This result indicated that purple yam was tolerant to the increase of NaCl concentration up to 100 mM, on MS medium without addition of plant growth regulators.
Somatic Embryo Germination of Jatropha curcas L in Presence of Sucrose and Poly Ethylene Glycol (PEG) Rudiyanto, Rudiyanto; Efendi, Darda; Ermayanti, Tri Muji
Annales Bogorienses Vol. 18 No. 1 (2014): Annales Bogorienses
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Jatropha curcas L. is a potential source of a non-edible biofuel. Conventional propagation of J. curcas technique has some limitations. Somatic embryo can produce a large number of embryos and obtain a large number of plants all year round. Treatment of sucrose in combination with polyethylene glycol (PEG) was proven to enhance germination of somatic embryos in many plant species. The aim of the study was to investigate the effect of sucrose in combination of PEG on somatic embryo germination in J. curcas. Globular somatic embryos at 0.025-0.030 g fresh weight having 0.4-0.5 cm in diameter were grown on MS medium solidified with 3 g/l of Gelzan supplemented with sucrose at 20, 30, 40, and 50 g/l in combination with PEG at 0, 2.5, 5, 10, and 15%. Results showed that the best medium for germination of J. curcas somatic embryo cultures was MS medium supplemented with 20 and 30 g/l of sucrose in combination with 5% of PEG. The numbers of germinated embryos per clump had significant enhancement on those medium compared with the control (PEG free treatment) (2.65 to 5.65) and (2.55 to 5.50). In addition, those treatments resulted in the highest percentage of clumps forming germinated embryos (100%), with an average of normal germinated embryos at 94.163 and 96.065%. The addition of 40 and 50 g/l of sucrose in combination with 15% of PEG caused all embryos to fail at germinating.
In vitro Seed Germination and Shoot Multiplication of Seven Endemic Subalpine and Alpine Plant Species Grown on Mount Jaya, Papua, Indonesia Ermayanti, Tri Muji; Hafiizh, Erwin Al; Mandessy, Ary; Setyadi, Gesang; Mukhsia, Andi
Annales Bogorienses Vol. 18 No. 1 (2014): Annales Bogorienses
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Exploitation on plant population may put the endemic plants into an endangered state, hence, these plants will need to be conserved. In order to pursue conservation on endemic plants, we conducted in vitro seed germination and shoot multiplication of seven alpine and sub-alpine species endemic to Mount (Mt.) Jaya, in Papua, Indonesia, i.e. Tetramolopium klossii, Deschampsia klossii, Papuacalia cartenszensis, Epilobium hooglandii, Gaultheria novoguinensis, Rhododendron correoides, and Rhododendron culminicolum. These species are categorized as slow-growth plants found in higher altitude (over 3700 m above sea level) and low temperature of Mt. Jaya. Seeds were surface-sterilized using Na-hypochloride and germinated aseptically on Murashige and Skoog (MS) medium. Dytikinin benzyl adenine (BA) was used for shoot multiplication. Seedling cultures were maintained in a controlled environment with continuous low light intensity (800 lux) and at temperature 26-27oC. Results showed that most species had more than 80% of germination rate on MS medium after a week in culture. BA was required to enhance shoots multiplication. Woody Plant (WP) (Lloyd & McCown, 1981) medium gave better shoot multiplication for R. culminicolum.
In Vitro Induction of Tetraploid Pummelo ’Nambangan’ (Citrus maxima (Burm.) Merr.) by Colchicine Treatment Using Germinated Seed, Shoot Tip and Cotyledonary Node As Explants Wulandari, Dyah Retno; Purwito, Agus; Susanto, Slamet; Husni, Ali; Ermayanti, Tri Muji
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
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Tetraploid citrus are important for interploidal hybridization to create triploid seedless citrus. Colchicine is the most commonly used as antimitotic agent to induce polyploid plants. Tetraploid induction by colchicine in Pummelo ‘Nambangan’ was conducted in vitro using different types of explants. The aim of this research was to induce tetraploid pummelo ‘Nambangan’ by colchicine treatment using germinated seed, shoot tip and cotyledonary node as explants. Tetraploid shoot induction was conducted by soaking germinated seeds, shoot tips and cotyledonary nodes in 0.1 % colchicine for 1, 3 and 5 hours. Regenerant shoots were grown on MS medium and their growth was observed after four weeks in culture. Ploidy level was determined using flow cytometry analysis. Stomata density, length and width of stomatal guard cell were also recorded. The results showed that shoot elongation was inhibited by colchicine treatment. Soaking of shoot tip explants in 0.1 % colchicine for 1 hour resulted in 66.66 % of putative tetraploid shoots. Compared to diploid shoots, tetraploids had lower stomata density but bigger in guard cell size.
Response of Increasing NaCl Concentrations on Growth and Proline Content of Tacca leontopetaloides cultured in vitro Martin, Andri Fadillah; Hapsari, Betalini Widhi; Ermayanti, Tri Muji
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
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The effects of increasing NaCl concentrations on growth and proline content of Tacca leontopetaloides cultured in vitro were investigated. T. leontopetaloides were suspected to have high tolerance against salinity, thus the purpose of this research was to investigate the effect of increasing NaCl concentrations added on growth medium on growth and proline content of T. leontopetaloides grown in vitro. In vitro corms were cultured on MS medium supplemented with NaCl at concentrations of 0, 10, 25, 50, 75, 100, 150 and 200 mM, respectively. After six weeks in culture, shoots height, shoots number, leaves number, fresh weight, as well as their proline content were recorded. The results showed that fresh weight of shoots grown on MS medium supplemented with 10, 25 and 75 mM NaCl was higher compared to the control treatment. Fresh weight decreased when shoots were cultured on MS medium supplemented with NaCl at more than 100 mM. Proline content increased along with the increase of NaCl concentrations. Meanwhile, the height of shoots, number of shoots, and number of leaves decreased along with the increase of NaCl concentrations.
Co-Authors , Juliarni . Aryanti . Harsojo Adabiyah, Rifatul Agus Purwito Al Hafiizh, Erwin Ali Husni Andi Salamah Andry, Yuli Aryanti . Aryanti Aryanti Azizah, Farroh Azizah, Farroh Bambang Prajoga Bambang Prajoga Eko Wardoyo, Bambang Prajoga Eko Diah Ratnadewi Dwi Hastuti Dwi Kusuma Wahyuni Edy Setiti Wida Utami Edy SW Utami, Edy SW Efendi, Darda Hafiizh, Erwin Al Hafiizh, Erwin Al Hafiizh, Erwin Al Hamami Alfasani Dewanto Hamim Hamim Hapsari, Betalini Widhi Hapsari, Betalini Widhi Hapsari, Betalini Widhi Hermono, Arief Hermono, Arief Hery Purnobasuki Ida Ayu Putu Sri Widnyani Iman Rusmana JULIARNI JULIARNI Kartika Ika Priadi, Kartika Ika laela Sari, laela Laksono Trisnantoro Lelono, Arthur A. Lelono, Arthur A. Lizawati . Mandessy, Ary Mandessy, Ary Martin, Andri F Martin, Andri F Martin, Andri Fadillah Martin, Andri Fadillah Martin, Andri Fadillah Maulana, Evan Maulana, Evan Mukhsia, Andi Mukhsia, Andi Noorrohmah, Siti Noorrohmah, Siti Nugroho, Robertus Qonitatun Nisa, Qonitatun Rantau, Deritha E. Rantau, Deritha E. Rantau, Deritha Ellfy Rantau, Deritha Ellfy Rantau, Deritha Elly Rantau, Deritha Elly Rd. Selvy Handayani Rita Martaleta Dewi, Rita Martaleta Roedhy Poerwanto Rudiyanto Rudiyanto Rudiyanto Rudiyanto, Rudiyanto Setyadi, Gesang Setyadi, Gesang Siti Nur Hafida, Siti Nur Slamet Susanto Sobir Sobir Solly Aryza Sutedja, Lenny Wiguna Rahman Wijayanta, Ardian Nur Wijayanta, Ardian Nur Wulandari, Dyah Retno Wulandari, Dyah Retno Wulansari, Aida Wulansari, Aida Yanti, Oktavia Yanti, Oktavia Yefni Syafria