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All Journal ANNALES BOGORIENSES
Tri Muji Ermayanti
Research Centre for Biotechnology, Indonesian Institute of Sciences (LIPI), Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Medicine & Pharmacology

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Micropropagation of Tetramolopium klossii (Moore) Mattfeld, endemic subalpine and alpine plant species to New Guinea Tri Muji Ermayanti; Erwin Al Hafiizh; Andri Fadillah Martin; Ary Mandessy; Arief Hermono; Andi Mukhsia
ANNALES BOGORIENSES Vol 15, No 2 (2011): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (415.898 KB) | DOI: 10.14203/ann.bogor.2011.v15.n2.9-15

Abstract

The protocol of micropropagation of Tetramolopium klossii (Moore) Mattfeld through adventitious shoot proliferation was established. This is the first report of T. klossii tissue culture. Seeds were surface-sterilized by dipping them in 10% of ethanol for 1 min followed by soaking in 1% of sodium hypochlorite for 10 min. The sterile seeds were germinated on MS medium solidified with 8 g/l of agar, supplemented with 20 g/l of sucrose. Two-month old seedlings formed some shoot clumps on MS solid medium without addition of any plant growth regulators. Small clumps containing 4-6 adventitious shoots were used for shoot proliferation on MS solid medium containing 0; 0.1; 0.5; 1.0 and 2.0 mg/L of BAP (benzylaminopurine) or kinetin. The results showed that medium MS without cytokinins (control treatment), and the addition of 0.1 or 0.5 mg/L of BAP was more effective for shoot proliferation compared to the medium with addition of BAP at higher concentration and the addition of kinetin. In further subcultures, multiple shoots grown on MS medium without cytokinins was  more vigorous  than that of shoots grown on other media, so that the shoots were ready for rooting. Medium containing 0.5 mg/L of BAP was the best for maintaining multiple shoot formation. Shoots formed roots on MS medium containing half strength of macro nutrients.   Keywords: Tetramolopium klossii, micropropagation, Murashige and Skoog medium, shoot multiplication
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 : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (273.445 KB) | 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, D. hispida and D. 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.   Keywords: Dioscorea spp., Potyvirus, RT-PCR, ACP-ELISA, in vitro shoot multiplication
Co-Authors Andi Mukhsia Andri Fadillah Martin Arief Hermono Ary Mandessy Dyah Retno Wulandari Erwin Al Hafiizh