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Genetic Variation Analysis of EMS-Induced Chili Pepper (Capsicum frutescens L.) Mutants Using SSR Markers Dwinianti, Edia Fitri; Mastuti, Retno; Arumingtyas, Estri Laras
Journal of Tropical Life Science Vol 9, No 3 (2019)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (273.472 KB)

Mutation induction with chemical mutagen ethyl methane sulfonate (EMS) capable of producing genetic variation in plants. The purpose of this research was to evaluate genetic variation of EMS-induced chili pepper mutants using SSR molecular marker. Various concentration of EMS (0.01%; 0.02%; 0.04%) were applied to chili pepper seeds of Genotypes 2, 7, and 11. The genomic DNA was isolated from mutants and control plants then used for PCR analysis using three SSR primers namely CA26, CA52 and CA96. The three SSR primers were able to detect genetic variation in the three genotypes of EMS-induced chili pepper mutants and control plants. A total of 18 alleles have been identified in all three SSR loci with PIC values ranging from 0.792 to 0.893. The three genotypes of chili pepper plants had different sensitivity to EMS mutation induction. Genotype 11 plants showed higher sensitivity to EMS treatment than genotypes 2 and 7 based on the amount of genomic changes. Dendrogram constructed based on genetic similarity values distributed the mutants and control plants into three main groups with similarity coefficients ranging from 0.34 to 0.90. Similar genotype either control plant or mutants, especially Genotypes 2 and 7 were group into similar group. However, for the Genotype 11 plants, large genome changes cause mutant plants G11K1, G11K2, G11K3 had a low genetic similarity to their control plants, so the mutants were separated in a different group from the control plant. Those indicate that EMS mutation induction capable of increasing genetic variation in chili pepper plants based on SSR molecular marker.

MICROSATELLITE MARKER FOR GENETIC VARIATION ANALYSIS IN LOCAL CHILI PEPPER (CAPSICUM FRUTESCENS L.) INDUCED BY ETHYL METHANE SULFONATE (EMS) Juliandari, Ria Reinnata; Mastuti, Retno; Arumningtyas, Estri Laras; Arumningtyas, Estri Laras
Journal of Tropical Life Science Vol 9, No 2 (2019)
Publisher : Journal of Tropical Life Science

Mutation using Ethyl Methane Sulfonate (EMS) is a simple and quick method to produce genetic variation in chili pepper. In this study, a total of 3 genotypes of local chili pepper (Capsicum frutescens L.), i.e. Genotype 2 (G2), Genotype 7 (G7), and Genotype 11 (G11) were induced by EMS with concentrations of 0% (K0), 0.01% (K1), 0.02% (K2), and 0.04% (K3). Genetic variation analysis in mutant was performed based on 3 microsatellite markers CA 19, CA 27, CA 62. Those molecular markers successfully detected the genetic variation in chili pepper mutant based on the number and size of microsatellite alleles variation. The 3 genotypes of chili pepper mutant produced a total of 15 alleles with the average Polymorphism Information Content (PIC) value of 0.82. Compared to the control plant, genetic variations in genome level were observed in local chili pepper. Furthermore, the treatment of EMS with concentration of 0.04% produced the most notable genetic variation in 3 genotypes of local chili pepper.

Genetic Diversity Study Among Six Genera of Amaranth Family Found in Malang Based on RAPD Marker Fatinah, Arik Arubil; Arumingtyas, Estri Laras; Mastuti, Retno
Journal of Tropical Life Science Vol 2, No 3 (2012)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar

Genera of amaranth family tend to have phenotypic variation partly caused by environmental factor. Phenotypic variation was the result of interaction between genetic and environmental factors. One of molecular markers that is widely used for detecting genetic variation is RAPD. RAPD is used for polymorphism detections and is now possible for identifiying a large number of loci and ascribes unambiguous taxonomic and genetic relationships among different taxa. Members of amaranth family found in Indonesia are Amaranthus, Celosia, Aerva, Alternanthera, Achyranthes, Gomphrena, Salsola, and Iresine. Six genera of which (Amaranthus, Celosia, Aerva, Alternanthera, Achyranthes, and Gomphrena) were observed in this study. DNA was extracted from fresh young leaves using Doyle and Doyles method with modification in the extraction buffer used. RAPD analyses were carried out with 20 decamer primers from Kit A of Operon Technology. DNA was amplified using master cycler gradient Eppendorf with 35 cycles. RAPD products were separated on 1,5 % agarose gels and detected by staining with ethidium bromide. There were 374 bands generated in 18 random primers. The number of monomorphic bands, polymorphic bands, and the percentage of polymorphism were 21 bands, 353 bands, and 94,38 % respectively. The high number and percentage of polymorphic bands revealed genomic DNA variation. This variation is in accordance with phenotypic variation detected in this experiment. Therefore, it can be concluded that, based on DNA polymorphism detected by RAPD, Amaranth family can be classified into two sub families namely Amaranthoideae and Gomphrenoideae.

The Dynamic of Calcium Oxalate (CaOx) in Porang Corms (Amorphophallus muelleri Blume) at Different Harvest Time Chairiyah, Nurul; Harijati, Nunung; Mastuti, Retno
Journal of Tropical Life Science Vol 11, No 1 (2021)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.11.01.05

The research aims to observe the influence of harvesting time on the change of calcium oxalate (CaOx) content and crystal density in Porang corms. The corms were harvested at different times, i.e., (1) two weeks before the plants shed (R0-1), (2) when the plants shed (R0), and (3) two weeks after the plants shed (R0+1). CaOx was obtained using the modified extracting method. Microscopic observations were obtained from the slices of the edge and center part of porang corms. Parameter observed including CaOx content, corm weight, shape, and density of CaOx crystal. CaOx content and crystal density in corms were analyzed using One way ANOVA. If the results are significant, it will be followed by Tukey Test Î± 0.05. In the meantime, the relation between CaOx content and corm weight was analyzed using Correlation Test Bivariate. The results showed that CaOx content was relatively higher in porang corms, i.e., 15.98 Â± 0.60g/100g. On the other hand, the increasing of CaOx content might improve corm weight. The total density of druse, styloid, and prism crystal was pretty high in corms obtained when the plants shed compared to another harvest time, i.e., 1,494 Â± 286; 31,280 Â± 17,406 and 6,256 Â± 1,533 crystals/cm2 . Raphide crystal density, by contrast, increased in corms obtained after the plants shed, i.e.,1,656 Â± 368 crystals/cm2 . Total CaOx crystal density in the edge parts of corms harvested when the plants shed was proportionately higher than in the other harvest times, i.e., 12,292 Â± 4,687.89 crystals/cm2 . In contrast, CaOx crystal densities in the center parts of corms were not much different at three harvesting times. The density of druse and prism crystals was somewhat higher in the center part of corms than in the edge parts. In opposition to, the density of raphide and styloid crystals was fairly higher in the edge part of corms than it was in the center parts. However, only raphide crystal density found in the edge and center part of corms was significantly affected by harvest time from all these results.

Genetic Diversity Study Among Six Genera of Amaranth Family Found in Malang Based on RAPD Marker Arik Arubil Fatinah; Estri Laras Arumingtyas; Retno Mastuti
Journal of Tropical Life Science Vol. 2 No. 3 (2012)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar

Genera of amaranth family tend to have phenotypic variation partly caused by environmental factor. Phenotypic variation was the result of interaction between genetic and environmental factors. One of molecular markers that is widely used for detecting genetic variation is RAPD. RAPD is used for polymorphism detections and is now possible for identifiying a large number of loci and ascribes unambiguous taxonomic and genetic relationships among different taxa. Members of amaranth family found in Indonesia are Amaranthus, Celosia, Aerva, Alternanthera, Achyranthes, Gomphrena, Salsola, and Iresine. Six genera of which (Amaranthus, Celosia, Aerva, Alternanthera, Achyranthes, and Gomphrena) were observed in this study. DNA was extracted from fresh young leaves using Doyle and Doyles method with modification in the extraction buffer used. RAPD analyses were carried out with 20 decamer primers from Kit A of Operon Technology. DNA was amplified using master cycler gradient Eppendorf with 35 cycles. RAPD products were separated on 1,5 % agarose gels and detected by staining with ethidium bromide. There were 374 bands generated in 18 random primers. The number of monomorphic bands, polymorphic bands, and the percentage of polymorphism were 21 bands, 353 bands, and 94,38 % respectively. The high number and percentage of polymorphic bands revealed genomic DNA variation. This variation is in accordance with phenotypic variation detected in this experiment. Therefore, it can be concluded that, based on DNA polymorphism detected by RAPD, Amaranth family can be classified into two sub families namely Amaranthoideae and Gomphrenoideae.

Microsatellite Marker for Genetic Variation Analysis in Local Chili Pepper (Capsicum frutescens L.) Induced by Ethyl Methane Sulfonate (EMS) Ria Reinnata Juliandari; Retno Mastuti; Estri Laras Arumningtyas
Journal of Tropical Life Science Vol. 9 No. 2 (2019)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.09.02.08

Mutation using Ethyl Methane Sulfonate (EMS) is a simple and quick method to produce genetic variation in chili pepper. In this study, a total of 3 genotypes of local chili pepper (Capsicum frutescens L.), i.e. Genotype 2 (G2), Genotype 7 (G7), and Genotype 11 (G11) were induced by EMS with concentrations of 0% (K0), 0.01% (K1), 0.02% (K2), and 0.04% (K3). Genetic variation analysis in mutant was performed based on 3 microsatellite markers CA 19, CA 27, CA 62. Those molecular markers successfully detected the genetic variation in chili pepper mutant based on the number and size of microsatellite alleles variation. The 3 genotypes of chili pepper mutant produced a total of 15 alleles with the average Polymorphism Information Content (PIC) value of 0.82. Compared to the control plant, genetic variations in genome level were observed in local chili pepper. Furthermore, the treatment of EMS with concentration of 0.04% produced the most notable genetic variation in 3 genotypes of local chili pepper.Â

Assessing Exogenous L-Glutamine and PEG Influence on Soybean Adventitious Roots Culture: Lateral Root Morphology and Amino Acid Profiles: L-Glutamine and PEG Influence on Adventitious Roots Culture Ardhian, Dhike; Mastuti, Retno; Nahampun, Hartinio Natalia
Journal of Tropical Life Science Vol. 15 No. 1 (2025)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.15.01.01

Soybean (Glycine max L. Merr.) plays an important role in global agriculture and provides a primary source of plant nutrition, notably amino acids. However, tropical climates often result in lower amino acid levels than subtropical climates. This study explored the effects of L-glutamine and PEG on enhancing the amino acid content in in vitro soybean adventitious root. In this study, in vitro adventitious roots from the Detap1 soybean variety were treated in a liquid medium with various concentrations of L-glutamine (0, 1, 2, 4, 8, and 16 g.L-1) or PEG (0, 10, 20, 40, 80, and 160 g.L-1) for 8 days. The root biomass was quantified, and the amino acid profile was analyzed using HPLC. L-glutamine or PEG treatments significantly reduced root biomass by inhibiting lateral root initiation. Meanwhile, 20 amino acids were detected in the control and L-glutamine or PEG treatments. The application of a 16 g.L-1 L-glutamine significantly increased amino acid content. PEG treatments also had similar results in root growth inhibition and increased amino acid accumulation. Multivariate analysis showed significant separation between the control and treatment groups, with glutamine playing a crucial role in the group separation. This study demonstrates the potential of in vitro technology to improve amino acid production through adventitious root culture, providing insights into stress responses and metabolic regulations

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