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All Journal Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences Makara Journal of Science
Fatinah, Arik Arubil
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Agriculture, Biological Sciences & Forestry Environmental Science

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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

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Abstract

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.
Employing SecA Recombinant Protein to Generate Polyclonal Antibodies for the Rapid Identification of Phytoplasma Fatinah, Arik Arubil; Rifa'i, Muhaimin; Arumingtyas, Estri Laras; Widyarti, Sri
Makara Journal of Science Vol. 29, No. 3
Publisher : UI Scholars Hub

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Phytoplasma, a plant pathogen that threatens plant health, can induce diseases such as yellow wilt, virescence, phyllody, and witches’ broom. Although early detection methods using polyclonal and monoclonal antibodies have been developed, synthetic peptides offer advantages such as higher titers, predictable antigenic characteristics and superior sensitivity and specificity. Epitope mapping plays a key role in designing peptide antigens and includes gene synthesis, transformation, expression assessment, large-scale production, and purification. A study using New Zealand male White rabbits was conducted to generate antibodies against phytoplasma. The SecA (395–470) sequence, obtained from the preprotein translocase meets the criteria of being non-homologous, nonallergic, and antigenic. The three-dimensional structure of SecA (395–470) has a sequential similarity of 77–97% with certain phytoplasma types, and the protein purity exceeds 90%. The production of polyclonal antibodies was successful, achieving titers greater than 1:512,000. The SWISS model was used to predict the 3D structure of SecA in phytoplasma species, revealing structural homology with other phytoplasma species. The recombinant protein antigen SecA was able to induce high-titer antibody formation (>1:512.000). The developed lateral flow immunoassay (LFA) detects phytoplasma in plants using purified and conjugated antibodies. The peptide design, derived from Aster yellows witches-broom (SrI), effectively detects phytoplasma from various groups, especially Cactus witches’ broom phytoplasma (SrII-C) and Peanut witches’ broom phytoplasma (SrII-A). The SrI group phytoplasma was chosen as a reference sequence due to its` ability to infect plants across the broadest host range. LFIA was tested on samples from both phytoplasma-infected and healthy plants using nested PCR. The SecA sequence was successfully produced and used as an immunogen candidate against phytoplasmas.
Co-Authors Estri Laras Arumingtyas Retno Mastuti Rifa'i, Muhaimin Sri Widyarti