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EVALUASI PERTUMBUHAN STEVIA REBAUDIANA BERT. TETRAPLOID SECARA IN VITRO DAN DI LAPANG UNTUK PRODUKSI STEVIOSIDA DAN REBAUDIOSIDA-A Ermayanti, Tri Muji; Ratnadewi, Diah; Adabiyah, Rifatul
JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i2.3809

Abstract

ABSTRACTGenetic improvement through tetraploid induction of Stevia rebaudiana is important in order to increase the sweetener content, steviol glycoside. Tetraploid plants of several species after induction with colchicine and oryzalin have higher growth and secondary metabolite contents compared to the diploid plants. This study was aimed to evaluate growth as well as their stevioside and Reb-A content of S. rebaudiana tetraploid and diploid (control) plants cultured in vitro and grown in the field after acclimation process. This study used 3 tetraploid clones, namely B60.3H8, P1T22, P3T5, and 1 diploid clone as control. Shoot tips were cultured on MS medium without addition of plant growth regulators for 6 weeks, then they were acclimated in a greenhouse, followed by planting them in the field. Growth of shoot culture, plantlets in the greenhouse and plants in the field were observed. Analysis of stevioside and Reb-A was done by HPLC. The results showed that plantlets of diploid clone had higher in vitro growth and survival rate in the greenhouse than that of tetraploids. Tetraploid clone P1T22 had similar growth as diploid plants, but higher than the growth of tetraploid B60.3H8 and P3T5. Fresh and dry weights of B60.3H8 was similar with diploid plants, but higher than P1T22 and P3T5 tetraploid clones. The highest level of stevioside and Rebaudiosida-A was found in tetraploid B60.3H8 clone, the lowest was found in the diploid plants. The highest ratio of stevioside : Reb-A was found at B60.3H8 tetraploid clone.  Keywords: Stevia rebaudiana, in vitro, field, growth, Stevioside, Rebaudioside-A, tetraploid  
INDUKSI POLIPLOIDI TANAMAN KANGKUNG (IPOMOEA AQUATICA FORSSK.) KULTIVAR SALINA IN VITRO DENGAN ORYZALIN ratnadewi, diah; Rahmi, Putri; Witjaksono, Witjaksono
JURNAL BIOLOGI INDONESIA Vol 15, No 1 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i1.3760

Abstract

ABSTRACTWater spinach is a vegetable plant consumed by people all over the world. Its small morphology requires improvement to increase the size and productivity. An alternative way to do that is by increasing the ploidy of its chromosome. This paper describes in vitro induction of polyploidy of water spinach. Inoculum of in vitro seedling, in vitro shoots and nodal stem segments were immersed in MS solution containing oryzalin at concentrations of 0.00, 1.25, 2.50, 3.75 and 5.00 µM with immersion duration of 4, 8 and 24 hours. Oryzalin treatments reduced the growth variables of the in vitro shoots of all inocula tested, compared to control of no oryzalin treatment.  On immersion of 24 hours and high concentration of 5.00 µM, many inoculum failed to grow to the third passage of subculture. The best growth of in vitro shoot to the third passages occurred in the treatment combination of oryzalin 1.25 µM-8 h immersion and 2.50 µM-4 h immersion for seedling inoculum, treatment combination of 3.75 µM-4 h immersion for shoot tip inoculum and 1.25 µM-4 h immersion for nodal stem segment, compared to other treatments. Analysis of flow cytometry on 41 leaf samples from oryzalin treatment derived shoots showed 14.63% tetraploid, 36.59% mixoploid and 48.78% diploid. The efficiency of tetraploid formation reached  60%, provided only by the treatment of oryzalin 1.25 µM-8 h on seedling segments inoculum. Tetraploid shoots need to be proliferated, acclimatized, grown into planting materials and planted for agronomical analysis to provide evidence whether or not tetraploid water spinach is viable for commercial cultivation. 
Cinchona alkaloids are in extensive uses, not only for drugs but also for soft drink industries. They are harvested from the bark of trees Cinchona spp. after certain ages and therefore are available over a limited time. Cell culture is an alternative way to continuously produce such secondary metabolites in a much shorter time. Various substances were added in the normal growth media to promote quinoline alkaloids production by cell cultures of Cinchona ledgeriana. At the sixth week of culture, DIAH RATNADEWI; . SUMARYONO
HAYATI Journal of Biosciences Vol. 17 No. 4 (2010): December 2010
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.17.4.179

Abstract

Cinchona alkaloids are in extensive uses, not only for drugs but also for soft drink industries. They are harvested from the bark of trees Cinchona spp. after certain ages and therefore are available over a limited time. Cell culture is an alternative way to continuously produce such secondary metabolites in a much shorter time. Various substances were added in the normal growth media to promote quinoline alkaloids production by cell cultures of Cinchona ledgeriana. At the sixth week of culture, quinine and cinchonine contents were suppressed by paclobutrazol (PBZ), abscisic acid (ABA), or even by precursor tryptophan, while cinchonidine content was enhanced by 0.2 mg/l tryptophan to 43 fold of that produced by untreated cells (2.8% dry weight). At the seventh week of culture, the production of quinine and quinidine started to grow whereas the production of cinchonine and cinchonidine tended to decrease. An addition of 5 mg/l PBZ to culture media yielded the highest level of total quinine/quinidine after seven weeks, e.g. quinine 11 times more abundant and quinidine 23 fold higher compared to the untreated cells. Particularly the level of quinine which is the most demanded for medical and industrial purposes still need to be improved to approach to or even higher than that of extracted from the conventional source.
INDUKSI BIAK KALUS DAN BIAK SUSPENSI SEL Aquilaria malaccensis Lam. [Induction of Callus Culture and Cell Suspension Culture of Aquilaria malaccensis Lam.] Witjaksono, Witjaksono; Leksonowati, Aryani; Ratnadewi, Diah
BERITA BIOLOGI Vol 16, No 1 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2672.281 KB) | DOI: 10.14203/beritabiologi.v16i1.2687

Abstract

Aquilaria malaccensis Lam. is a plant species producing fragrant woody material that contains some resin. The compounds can be used as medicine and perfume. Sesquiterpenoid, one group of compounds has been found being synthesized and subsequently extracted from callus and cell suspension culture of Aquilaria species. The aim of this research was to find a method of producing friable calli and cell suspension cultures from leaves or internodes of A. malaccensis in vitro by using suitable plant growth regulators; cell suspension that will suitably serve as material to produce sesquiterpenoid afterwards. Calli were established in almost all treatments of auxin-cytokinin on both leaves and internod explants. The treatment of 10 mg/L IBA induced the highest percentage of callus coverage from leaves with a rather compact structure. The combined treatment of 1–2 mg/L 2.4-D and 0.2–0.3 mg/L BA induced friable callus formation in more than 80% of cultures with 27–32% callus coverage percentage.  The use of 2,4-D induced a better formation of cell suspension than Picloram, with maximum volume up to 7 mL. Cell suspension culture with fine and homogenous aggregate could be established in the medium supplemented with 0.5 –1 mg/L 2,4-D.
Induksi Poliploidi pada Tanaman Talas (Colocasia esculenta (L.) Schott) Kultivar Kaliurang dengan Perlakuan Kolkisin secara In Vitro Ermayanti, Tri Muji; Wijayanta, Ardian Nur; Ratnadewi, Diah
JURNAL BIOLOGI INDONESIA Vol 14, No 1 (2018): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (708.917 KB) | DOI: 10.14203/jbi.v14i1.3667

Abstract

ABSTRACTGenetic modification to increase productivity and other better growth characteristics of Kaliurang taro could be done by various methods; one of them is polyploid induction. Kaliurang taro has performed excellent traits, it is resistant to leaf blight disease and pests. Polyploid plants of Kaliurang taro are expected to have excellent char-acteristics and increased productivity. The objective to this study was to obtain an efficient method through in vitro induction of polyploidy using colchicine on Kaliurang taro. Aseptic plantlets of Kaliurang taro were soaked in colchicine solution at 0.05, 0.1 and 0.2% for 1, 2 and 3 days. Control was untreated plantlets. Each treatment consisted of 12 replicates. The colchicine-treated plantlets were then planted on MS medium containing 2 mg/L BAP, 1 mg/L thiamine and 2 mg/L adenine. Survival rate and vegetative growth of plantlets were observed every week for 8 weeks after planting. The results showed that increasing colchicine concentra-tion and the soaking period produced planlets with various survival rate. Flowcytometric analysis indicated that the treatment of soaking the plantlets in 0.05% colchicine for 1 day resulted in 14.3% of tetraploid plantlets. The most efficient of colchicine treatment was 0.2% for 3 days, resulting in 57.1% tetraploids, with the efficient value of 33.3%. There was chromosome multiplication from diploid to tetraploid which was confirmed through both flowcytometric analysis and chromosomes counting.Keywords: Taro (Colocasia esculenta L.), Kaliurang, in vitro, flowcytometer, chromosome multiplication, tetraploids
LOCALIZATION OF GFDD4-1 EXPRESSED PROTEIN IN Physcomitrella patens CELLS DIAH RATNADEWI
BIOTROPIA - The Southeast Asian Journal of Tropical Biology Vol. 13 No. 1 (2006)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (215.934 KB) | DOI: 10.11598/btb.2006.13.1.214

Abstract

The expression of a new dehydration-related gene of Physcomitrella patens, GFDD4-I, was traced for its localization in the plant cells. This revelation is useful to predict the possible roles of the protein in plant tolerance to environmental stress. This gene was fused to  gfp  marker gene and transfected into the plant protoplasts. Under a confocal laser microscope, it was detected that the GFDD4-1 protein associated with the OFF started to generate at the cell periphery and developed more intensively inwards to cytoplasm, forming vesicles and cystemal structures or network. The protein might be membrane protein which may involve directly in membrane maintenance or cellular protection against stress conditions. Key words : Protoplast transformation, protein localization, dehydration-related gene, GFP, Physcomitrella patens
INDUKSI POLIPLOIDI PADA TANAMAN TALAS (COLOCASIA ESCULENTA (L.) SCHOTT) KULTIVAR KALIURANG DENGAN PERLAKUAN KOLKISIN SECARA IN VITRO Ermayanti, Tri Muji; Wijayanta, Ardian Nur; Ratnadewi, Diah
JURNAL BIOLOGI INDONESIA Vol 14, No 1 (2018): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v14i1.3667

Abstract

ABSTRACTGenetic modification to increase productivity and other better growth characteristics of Kaliurang taro could be done by various methods; one of them is polyploid induction. Kaliurang taro has performed excellent traits, it is resistant to leaf blight disease and pests. Polyploid plants of Kaliurang taro are expected to have excellent char-acteristics and increased productivity. The objective to this study was to obtain an efficient method through in vitro induction of polyploidy using colchicine on Kaliurang taro. Aseptic plantlets of Kaliurang taro were soaked in colchicine solution at 0.05, 0.1 and 0.2% for 1, 2 and 3 days. Control was untreated plantlets. Each treatment consisted of 12 replicates. The colchicine-treated plantlets were then planted on MS medium containing 2 mg/L BAP, 1 mg/L thiamine and 2 mg/L adenine. Survival rate and vegetative growth of plantlets were observed every week for 8 weeks after planting. The results showed that increasing colchicine concentra-tion and the soaking period produced planlets with various survival rate. Flowcytometric analysis indicated that the treatment of soaking the plantlets in 0.05% colchicine for 1 day resulted in 14.3% of tetraploid plantlets. The most efficient of colchicine treatment was 0.2% for 3 days, resulting in 57.1% tetraploids, with the efficient value of 33.3%. There was chromosome multiplication from diploid to tetraploid which was confirmed through both flowcytometric analysis and chromosomes counting.Keywords: Taro (Colocasia esculenta L.), Kaliurang, in vitro, flowcytometer, chromosome multiplication, tetraploids
INDUKSI POLIPLOIDI TANAMAN KANGKUNG (IPOMOEA AQUATICA FORSSK.) KULTIVAR SALINA IN VITRO DENGAN ORYZALIN Rahmi, Putri; Witjaksono, Witjaksono; ratnadewi, diah
JURNAL BIOLOGI INDONESIA Vol 15, No 1 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i1.3760

Abstract

ABSTRACTWater spinach is a vegetable plant consumed by people all over the world. Its small morphology requires improvement to increase the size and productivity. An alternative way to do that is by increasing the ploidy of its chromosome. This paper describes in vitro induction of polyploidy of water spinach. Inoculum of in vitro seedling, in vitro shoots and nodal stem segments were immersed in MS solution containing oryzalin at concentrations of 0.00, 1.25, 2.50, 3.75 and 5.00 µM with immersion duration of 4, 8 and 24 hours. Oryzalin treatments reduced the growth variables of the in vitro shoots of all inocula tested, compared to control of no oryzalin treatment.  On immersion of 24 hours and high concentration of 5.00 µM, many inoculum failed to grow to the third passage of subculture. The best growth of in vitro shoot to the third passages occurred in the treatment combination of oryzalin 1.25 µM-8 h immersion and 2.50 µM-4 h immersion for seedling inoculum, treatment combination of 3.75 µM-4 h immersion for shoot tip inoculum and 1.25 µM-4 h immersion for nodal stem segment, compared to other treatments. Analysis of flow cytometry on 41 leaf samples from oryzalin treatment derived shoots showed 14.63% tetraploid, 36.59% mixoploid and 48.78% diploid. The efficiency of tetraploid formation reached  60%, provided only by the treatment of oryzalin 1.25 µM-8 h on seedling segments inoculum. Tetraploid shoots need to be proliferated, acclimatized, grown into planting materials and planted for agronomical analysis to provide evidence whether or not tetraploid water spinach is viable for commercial cultivation. 
EVALUASI PERTUMBUHAN STEVIA REBAUDIANA BERT. TETRAPLOID SECARA IN VITRO DAN DI LAPANG UNTUK PRODUKSI STEVIOSIDA DAN REBAUDIOSIDA-A Adabiyah, Rifatul; Ratnadewi, Diah; Ermayanti, Tri Muji
JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i2.3809

Abstract

ABSTRACTGenetic improvement through tetraploid induction of Stevia rebaudiana is important in order to increase the sweetener content, steviol glycoside. Tetraploid plants of several species after induction with colchicine and oryzalin have higher growth and secondary metabolite contents compared to the diploid plants. This study was aimed to evaluate growth as well as their stevioside and Reb-A content of S. rebaudiana tetraploid and diploid (control) plants cultured in vitro and grown in the field after acclimation process. This study used 3 tetraploid clones, namely B60.3H8, P1T22, P3T5, and 1 diploid clone as control. Shoot tips were cultured on MS medium without addition of plant growth regulators for 6 weeks, then they were acclimated in a greenhouse, followed by planting them in the field. Growth of shoot culture, plantlets in the greenhouse and plants in the field were observed. Analysis of stevioside and Reb-A was done by HPLC. The results showed that plantlets of diploid clone had higher in vitro growth and survival rate in the greenhouse than that of tetraploids. Tetraploid clone P1T22 had similar growth as diploid plants, but higher than the growth of tetraploid B60.3H8 and P3T5. Fresh and dry weights of B60.3H8 was similar with diploid plants, but higher than P1T22 and P3T5 tetraploid clones. The highest level of stevioside and Rebaudiosida-A was found in tetraploid B60.3H8 clone, the lowest was found in the diploid plants. The highest ratio of stevioside : Reb-A was found at B60.3H8 tetraploid clone.  Keywords: Stevia rebaudiana, in vitro, field, growth, Stevioside, Rebaudioside-A, tetraploid  
Pengaruh jumlah subkultur dan media sub-optimal terhadap pertumbuhan dan kemampuan regenerasi kalus tebu (Saccharum officinarum L.) (Effect of repeated subculture and suboptimum media on the growth of sugarcane calli (Saccharum officinarum L.)) Hayati MINARSIH; . Suharyo; Imron RIYADI; Diah RATNADEWI
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (657.274 KB) | DOI: 10.22302/iribb.jur.mp.v84i1.219

Abstract

Sugarcane (Saccharum officinarum L.) is an important crop for sugar production. One attempt to increase sugarcane productivity is through micropropagation and quality improvement of sugarcane seedlings in vitro. This research aimed to study the effect of repeated subcultures on callus capacity for regeneration and plant survival in acclimatization phase, as well as the influence of suboptimum media on the recovery capability of sugarcane callus to proliferate in vitro. Fourth subcultured sugarcane callus derived from young leaves were used as material in this research. Basic medium of Murashige and Skoog (MS) added with 3 mg/L 2,4-D, 10% coconut water, and 3% sucrose was used for callus initiation. For callus regeneration, the MS medium was supplemented with 2 mg/L BAP, 0.2 mg/L IAA, 10% coconut water, and 3% sucrose. Study on the effect of subculture numbers consisted of three stages, i.e. initiation, regeneration, and acclimatization, while the study on resting phase or the use of sub-optimal media included six treatment media and two pathways. Results showed that the fifth subcultures produced embryoid callus (91%), the highest non mucilaginous callus (97%), and the highest abnormality rate (6%). Results from the suboptimum media treatment, showed that B pathway (4 week resting phase) was better than the A pathway (8 week resting phase), based on fresh weight and callus abnormality percentage. A and B pathways indicated that the growth of callus can be recovered when it was grown back to the normal media and 1.5D-MS treatment of the resting phase showed the best growth and appearance.