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Effect of Cell Density and Benzyl Amino Purine on the Growth of Somatic Embryo of Citrus Mandarin Batu 55 (Citrus reticulata Blanco.) in Liquid Culture Februyani, Nawafila; Widoretno, Wahyu; Indriyani, Serafinah
The Journal of Experimental Life Science Vol. 6 No. 1 (2016)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (947.628 KB) | DOI: 10.21776/ub.jels.2016.006.01.01

Abstract

Citrus mandarin Batu 55 (Citrus reticulata Blanco.) is one of Indonesian fruits commodities that have high economic value and consumers demand. The propagation of citrus mandarin by plant tissue culture generally was carried out on solid medium. The liquid culture system could increase cell multiplication therefore it became alternative method of plant propagation through somatic embryogenesis. The effect of initial cell density and Benzyl Amino Purin (BAP) concentration in liquid media were investigated. The initial cells density and right concentration of BAP given in media can increase cell proliferation of somatic embryo in liquid culture. Globular somatic embryo were cultured on Murashige and Tucker media with initial cell density 4, 6, 8 and 10 mgL-1 and BAP 0, 0.25, 0.5, and 0.75 mgL-1. Growth evaluation of somatic embryo were obtained by weighing fresh and dry weight every 2 weeks for 8 weeks for initial cell density treatment and 6 weeks of BAP treatment. The result of the research showed that cell density affect the growth of somatic embryo of citrus mandarin. Somatic embryo with low cell density showed slower growth compared than high cell density. Peak growth occured in 6th cultured with cell density 10 mgL-1. In addition to cell density, the growth of somatic embryo in liquid culture was affected by BAP. The growth of somatic embryo on the media containing BAP showed better results than without BAP. The highest BAP concentration on media showed fresh and dry weight of somatic embryo increased. In this research, growth of somatic embryo is not optimal yet because fresh and dry weights of somatic embryo still increase with high concentration 0.75 mgL-1 of BAP.Keywords: Benzyl Amino Purine, cell density, citrus mandarin, liquid medium, somatic embryo.
Water and Chlorophyll Content and Leaf Anatomy of Patchouli Planlet (Pogostemon cablin Benth.) Resulted by Shoot-tip Culture Experience Hyperhydricity after Treatment of Modification Ammonium nitrate or Macro salt Concentration on MS medium (Murashige Skoog) Zen, Aisyah Rahmawati; Widoretno, Wahyu; Indriyani, Serafinah
The Journal of Experimental Life Science Vol. 6 No. 1 (2016)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1037.459 KB) | DOI: 10.21776/ub.jels.2016.006.01.10

Abstract

Hyperhydricity is a symptom of abnormal morphological and physiological function which inhibits the regeneration of plantlets. In general, the main symptom of hyperhydricity is a change in the condition of the plantlets which looks clear (Glassy) as a result of low levels of chlorophyll, the high water content in the plantlets, and the abnormal anatomical structure of the leaves. Hyperhydricity can be controlled by reducing cytokinin concentration, increasing gelling agent concentration, and reducing ammonium nitrate and macro salt concentration on medium. Objective of this research was to reduce hyperhydricity in shoot tip culture of patchouli by modification of ammonium nitrate and macro salt concentration on MS medium. The various treatment concentrations of ammonium nitrate were 0 mg.L-1 (0), 41.25 mg.L-1 (¼ concentration), 825 mg.L-1 (½ concentration), 1650 mg.L-1 (1 concentration) and macro salt MS with 0, ¼ MS, ½ MS, MS with 5 replications. Hyperhydricity on patchouli shoots could be lowered, as indicated by the decrease in water content from 96% to 90-91%, the increase in total chlorophyll content, and the increased number of palisade cells and stomata on the leaf treatment outcome. The concentration treatment of ammonium nitrate showed better results than the concentration of macros salt in increasing the total chlorophyll content, but it did not differ significantly in lowering water levels and increasing the number of palisade cells and stomata. ¼x concentration treatment of ammonium nitrate could increase chlorophyll content of 0.16 to 0.97 mg.g-1, but MS with 1x concentration showed the best result in the increase of number of palisade cells and stomata of the leaves. Keywords: Ammonium nitrate, Hyperhydricity, Macro Salt, Shoot-tip culture.
The Effect of Explant Types and Kinetin Concentration on In Vitro Callus Induction in Vetiveria zizanioides (L.) Nash Fauziah, Arbaul; Widoretno, Wahyu
The Journal of Experimental Life Science Vol. 7 No. 2 (2017)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1001.506 KB) | DOI: 10.21776/ub.jels.2017.007.02.04

Abstract

The aims of this research were to evaluate the effect of explant types and several kinetin concentrations on in vitro induction and growth of callus vetiver (Vetiveria zizanioides (L.) Nash). Crown and tiller of vetiver were cultured on Murashige and Skoog’s (MS) media supplemented with combination of 2,4-D 0.75 ppm and several kinetin concentrations (0, 0.3, 0.5, 0.75, and 1) ppm. The induction and growth of callus were influenced by type of explant and concentration of kinetin. Formation and growth of callus on tiller explant were faster than crown explant. Callus on tiller explant were formed one week after culture, while callus from crown explant were formed at four weeks after culture. Callus growth on tiller explant also was better than crown explant. Eight weeks after culture, callus fresh weight from tiller explant was 0.35 ± 0.09 g, while callus fresh weight from crown explant was only 0.16 ± 0.08 g. The addition of kinetin in the medium combined with 2,4-D was able to increase callus growth and the optimum concentration of kinetin used was 0.5 ppm. The addition of kinetin more than 0.5 ppm in the medium decreased the callus fresh weight.Keywords: callus, crown, in vitro culture, tiller, Vetiveria zizanioides.
Effect of Bulbils Position on Leaf Branches to Plant Growth Responses and Corms Quality of Amorphophallus muelleri Blume Mastuti, Retno; Harijati, Nunung; Arumingtyas, Estri Laras; Widoretno, Wahyu
The Journal of Experimental Life Science Vol. 8 No. 1 (2018)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1140.975 KB) | DOI: 10.21776/ub.jels.2018.008.01.01

Abstract

Bulbil (corm leaf) is one material source of vegetative propagation in Amorphophallus muelleri Blume. Based on the position in branches there are two types of bulbil, middle and edge bulbils, which are different in shape and size. It has been reported that bulbil size affected not only the growth response of seedlings and plants but also the quality of the produced corms. Therefore, the objectives of this experiment were to study the effect of bulbils origin on A. muelleri Blume (Porang) growth and the quality of harvested corms. The quality of corms was assessed based on glucomannan and (CaOx) content. Bulbils from the middle and edge of branches were grown in polybag (f 5 cm) containing compost:fertilizer (1:1) mixed media under 40% shade. Each bulbil origin was repeated 8 times. The results showed that growth responses of plant from middle bulbil were significantly better than that from edge bulbil. However, at the harvesting time, the weight and diameter of corms derived from both types of bulbils were not significantly different. The glucomannan and calcium oxalate (CaOx) content of harvested corms grown from middle bulbil tends to be higher than that from edge bulbil.Keywords: branching, calcium oxalate (CaOx), germination, glucomannan, Porang.
Effect of Colchicine on In Vitro Growth and Ploidicity of Crown Vetiver Plant (Vetiveria zizanioides L. Nash) Sinta, Anisa Firmandanur; Widoretno, Wahyu
The Journal of Experimental Life Science Vol. 10 No. 1 (2020)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (780.439 KB) | DOI: 10.21776/ub.jels.2019.010.01.02

Abstract

The goal of this research was to evaluate the effect of colchicine on the formation and growth of shoot from explant crowns by in vitro, as well as to obtain polyploidy vetiver plant (Vetiveriazizanioides L. Nash). Induction of polyploidy vetiver plants carried out by culturing explant crowns on MS media supplemented by 2 mg.L-1 BAP and colchicine (0, 30, 60, 90, 120 mg.L-1) for three weeks. Explant and formed shoot regenerated on MS media containing 1 mg.L-1 NAA. The generated plantlets acclimatized on the growing media (cocopeat: husk charcoal: compost = 1: 1: 2). Plant ploidy level of the plants regenerated from treated explant estimated by chromosome counting in root tips. The effect of colchicine on media was able to induce polyploidy in vetiver plants. Five mixoploids were obtained from explant treated colchicine. The vetiver mixoploid plants obtained were 20% and 62.5% from colchicine 60 mg.L-1 and 90 mg.L-1, respectively. The vetiver mixoploid plants consist of diploid (2n=2x=20) and triploid cells (2n=3x=30) or diploid (2n=2x=20) and tetraploid (2n=4x=40). The addition of colchicine in concentrations above 30 mg.L-1 caused inhibition of shoot formation and growth, even a concentration of 120 mg.L-1 caused explant death. However, 30 mg.L-1 colchicine could increase the number of formed shoots, while only 60 mg.L-1 and 90 mg.L-1 could be induced the mixoploidy in the vetiver plant. The results showed that colchicine treatment could increase ploidicity in vetiver plants in vitro, but caused inhibition of shoot formation and growth. Keywords : Chromosome, colchicine, in vitro, polyploid, Vetiveria zizanioides L. Nash.
Effect of Benzyl Adenine Concentration on Callus Induction of Geranium Plants (Pelargonium graveolens L'Her) from Petiole and Leaf Explants Huda, Moch Faizul; Indriyani, Serafinah; Widoretno, Wahyu
The Journal of Experimental Life Science Vol. 10 No. 1 (2020)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (880.869 KB) | DOI: 10.21776/ub.jels.2019.010.01.04

Abstract

Geranium plant (Pelargonium graveolens L'Her) is one of the geranium oil producing plants that has many benefits. Callus culture is a technique that can be used to plant multiplication and increase production of secondary metabolites. This study aims to determine the effect of the concentration of Benzyl Adenine on the formation of geranium callus from petiole and leaf explants. Callus induction was carried out by culturing petiole and leaf explants on MS medium + 0.1 mg.L-1 NAA + Benzyl Adenine (0; 0.5; 1; 1.5 and 2 mg.L-1). Callus morphological parameters, percentage of callus formation, and time of first callus formation were observed. The formation of geranium callus influenced by the explant type and the concentration of Benzyl Adenine. In the 2nd week, the geranium callus was initiated, light green colored with a compact callus texture. At 4th week, the percentage of callus formation containing NAA 0.1 mg.L-1 of petiole and leaf explants was 20% and 8%, whereas the percentage of callus formation on medium containing 0.1 mg.L-1 NAA combined with 0.5-2 mg.L-1 Benzyl Adenine of petiole and leaf explants was 52-80% and 24-52%. The best percentage of callus formation was found on the culture medium containing 1 mg.L-1 Benzyl Adenine, equaled 80% of petiole explants, and 52% of leaf explants. Keywords: BA, Callus, Leaf, Petiole, Pelargonium graveolens L'Her.  
The Effect of Gamma Irradiation on the Growth and Multiplication of the In Vitro Shoot of Patchouli (Pogostemon cablin Benth.) Banyo, Yunia Efrice Banyo Efrice; Indriyani, Serafinah; Widoretno, Wahyu
The Journal of Experimental Life Science Vol. 10 No. 2 (2020)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2020.010.02.11

Abstract

The objective of this research was to evaluate the effect of gamma irradiation on shoot growth and multiplication of Patchouli (Pogostemon cablin Benth.) Two weeks-old in vitro shoots were irradiated gamma-ray, at doses of 0, 15, 30, 45, 60, and 75 Gy. The control shoot was not irradiated. The irradiated shoots were cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg.L-1 NAA and 0.3 mg.L-1 BA and incubated in a growth room for eight weeks at a temperature of 25±2°C. The results showed that the gamma irradiation inhibited the growth and multiplication of shoots. Explants irradiated with high-dose gamma-ray (45-75 Gy) had not formed shoot in four weeks of culture, while 58.3-83.3% of the explants without irradiation or irradiated at low doses 15 and 30 Gy formed shoots. The higher irradiation doses increased percentage of browning explants and reduced the percentage of forming shoots. Within the eight weeks of culture, explant without irradiation was able to form shoots at the percentage of 100% with 24 shoots per explant, while explants irradiated at 15-45 Gy were able to grow form shoots at the percentage of 77.7-95.5%. The high doses-irradiated explants (60 and 75 Gy) were only able to form shoots less than 13-20%, with 2-3 shoots per explant. Keywords: Gamma rays (Gy), in vitro shoot, Pogostemon cablin Benth.
The Effect of Ethyl Methane Sulfonate (EMS) on The In Vitro Shoot Regeneration of Vetiver (Vetiveria zizanioides [L.] Nash) Widoretno, Wahyu; Indriyani, Serafinah
The Journal of Experimental Life Science Vol. 10 No. 3 (2020)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2020.010.03.01

Abstract

Production of vetiver in Indonesia is low, and its oil quality is not in line with market demand due to the low quality of plant raw materials. In vitro mutagenesis using ethyl methanesulfonate (EMS) has the potential to produce superior vetiver variants. This study aimed to determine the effect of EMS on the growth and shoot formation of in vitro vetiver plants. In vitro mutagenesis was performed by culturing in vitro shoots on MS medium containing EMS with various concentrations (0.02; 0.04; 0.06; 0.08; 0.1%) for 4 weeks. The results showed that the addition of 0.02-0.1% EMS to the medium affected on in vitro shoot growth and the ability to form new shoots. EMS increased the percentage of dead explants and decreased the average number of shoots, the number of shoots formed per explant, and the shoot fresh weight. The higher the EMS concentration in the medium, the smaller the percentage of survived explants and the lower ability to form new shoots with the average number of shoots formed less. The percentage of survived explants on the medium without the addition of EMS and on the medium with the addition of EMS with low concentrations of 0.02 and 0.04% reached 100% with an average number of shoots formed ranging from 8.5-10 shoots/explant. Meanwhile, the percentage of survived explants on medium with the addition of EMS with a high concentration of 0.08-0.1% was only 12.5-25% with an average number of shoots formed was less than 2 shoots/explant.Keywords: EMS, in vitro shoots, Vetiveria zizanioides.
The Sensitivity of Leersia hexandra Sw. to Gamma-Ray Irradiation Sukmasari, Pritha Kartika; Widoretno, Wahyu; Siswanto, Dian
The Journal of Experimental Life Science Vol. 11 No. 1 (2021)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2021.011.01.02

Abstract

Gamma-ray irradiation as a physical mutagen has high penetrating power. Therefore, it is most often used to increase genetic variability or produce new mutant plants. This research was conducted to obtain the lethal dose of gamma-rays in Leersia hexandra plants. The used plant part was a single node stolon which had a length of 10 cm with the node in the middle of the stolon. The irradiation doses given were 0, 25, 50, 75, 100 Gy. Stolons were inserted into plastic clips and irradiated using a Cobalt-60 gamma irradiation source at the Center for Irradiation and Radioisotope Applications (PAIR), National Nuclear Energy Agency (BATAN) Jakarta. Gamma-irradiation has a significant effect on inhibiting shoot growth. The growth of irradiated Leersia hexandra with the best number of plants and the highest shoots was obtained at a dose of 25 Gy and decreased with increasing irradiation dose. The lethal dose (LD50) was determined eight days after irradiation using CurveExpert 1.4 software. Leersia hexandra plants that can sprout and regenerate followed the linear equation y = 1.02 - 7.5x with LD50 at 68.85 Gy and LD20 at 29.36 Gy.Keywords: gamma irradiation, genetic variation, Leersia hexandra, lethal dose, mutant plant.