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Dyah Retno Wulandari
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI)

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Detection of Potyvirus Using RT-PCR and ACP-ELISA of Dioscorea Species and In Vitro Shoot Multiplication of the Virus Free Plants Dyah Retno Wulandari; Tri Muji Ermayanti
Annales Bogorienses Vol. 15 No. 2 (2011): Annales Bogorienses
Publisher : National Research and Innovation Agency (BRIN)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2011.v15.n2.1-8

Abstract

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.
The Effect of Increase in NaCl Concentration on Growth and Proline Content of Purple Yam (Dioscorea alata L.) Grown In Vitro Andri Fadillah Martin; Farroh Azizah; Dyah Retno Wulandari; Tri Muji Ermayanti
Annales Bogorienses Vol. 16 No. 2 (2012): Annales Bogorienses
Publisher : National Research and Innovation Agency (BRIN)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2012.v16.n2.13-18

Abstract

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.
In Vitro Induction of Tetraploid Pummelo ’Nambangan’ (Citrus maxima (Burm.) Merr.) by Colchicine Treatment Using Germinated Seed, Shoot Tip and Cotyledonary Node As Explants Dyah Retno Wulandari; Agus Purwito; Slamet Susanto; Ali Husni; Tri Muji Ermayanti
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
Publisher : National Research and Innovation Agency (BRIN)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2015.v19.n1.29-36

Abstract

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.
Co-Authors Agus Purwito Ali Husni Andri Fadillah Martin Farroh Azizah Slamet Susanto Tri Muji Ermayanti