Annales Bogorienses

Number 2, 2011

1-8
Detection of Potyvirus using RT-PCR and ACP-ELISA of Dioscorea species and in vitro shoot multiplication of the virus free plants

Authors: Dyah Retno Wulandari, Tri Muji Ermayanti

Abstract

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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

9-15
Micropropagation of Tetramolopium klossii (Moore) Mattfeld, endemic subalpine and alpine plant species to New Guinea

Authors: Tri Muji Ermayanti, Erwin Al Hafiizh, Andri Fadillah Martin, Ary Mandessy, Arief Hermono, Andi Mukhsia

Abstract

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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

17-23
The Investigation of Polycyclic Aromatic Hydrocarbon and Oil Degrading Bacteria Isolated from The Marina Port Ancol, Jakarta Bay

Authors: Puspita Lisdiyanti, Yopi Yopi, Tutik Murniasih

Abstract

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Number of views: 324

Polycyclic Aromatic Hydrocarbons (PAHs) as well as crude oil are widespread environmental pollutants. The contamination of air, soil, freshwater (surface water and groundwater), and marine environments by PAHs as well as crude oil has been reported. Of concern to public health is the fact that many PAHs or their metabolites are mutagenic, carcinogenic, or both. North Java coastal area such as Jakarta Bay is the polluted marine area in Indonesia as a result from anthropogenic wastes and the oil spill. Although evaporation and photo-oxidation play an important role in oil detoxification, ultimate and complete degradation is accomplished mainly by marine micro flora, and being dominant in this function. Certain bacteria are well-known could consume and degrade the PAHs as well as crude oil. Therefore investigating the potential PAH and oil degrading marine bacteria is important. In this study, we collected sample from oil polluted area in Marina Port Ancol, Jakarta Bay and isolated four PAH substrates and Arabian crude oil degrading marine bacteria using enrichment method and direct isolation method. As result, 223 strains could degrade PAHs, among these strains, 94 strains could degrade phenanthrene, 23 strains degrade fluoranthene, 92 strains could degrade dibenzotiophen, 14 strains could degrade phenotiazin and 106 isolates degrade crude oil.

25-30
Alkane Degradation and Detection of Mono-xygenase Gene from Alcanivorax sp. from Jakarta Bay

Authors: Ahmad Thontowi, Yopi Yopi

Abstract

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Number of views: 377

Alkanes is a major component of crude oil that can be hydrolized by enzyme alkane monooxygenase from bacteria. Nine oil-degrading bacteria were analyzed their capability to degrade alkanes (pristane and paraffin). The result of growth test on paraffin and pristane were showed that 9 isolates could be devided into two groups. First group (BL09, BL31 and BL45) could degrade both paraffin and pristane, and second group (BL01, BL06, BL44, BL057, BL058 and BL071) preferred to degrade paraffin than pristane. Three isolates (BL09, BL31 and BL45) have activity to decrease paraffin and pristane until less 50% remain. Based on homology analysis of 16S rRNA gene sequences showed that isolates No. BL09, BL31 and BL45 were identified as Alcanivorax sp. and the partial sequences of the alkB gene from those three isolates are showing 66-68% of identity compare with some mono-oxygenase gen from database of genbank.

31-36
Ecological Study of Rare-Actinomycetes in Soils and Leaf-Litters

Authors: Puspita Lisdiyanti, Shanti Ratnakomala, Roni Ridwan, Yantyati Widyastuti, Misa Otoguro, Ando Katsuhiko

Abstract

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Number of views: 325

A total of 268 strains of Actinomycetes were isolated from soil and leaf-litter samples collected in Jambi (Sumatera) and Cibinong (West Java) using three selective isolation methods. All the isolates were identified by morphological characteristic and by analysis of 16S rDNA sequence. On the basis of their morphology and 16S rDNA sequence, 164 isolates were belonged to the Streptomyces Group and 104 isolates were belonged to the Rare-Actinomycetes (Non-Streptomyces) Group. Furthermore, 40% of isolates in Streptomyces Group and 62% of isolates in Non-Streptomyces Group are supposed to be new taxa. It is indicated the richness of Actinomycetes in these two area. The genus Streptomyces is the most abundant in soil samples, occupying 60- 75% of all isolates; while the genus Actinoplanes is mainly found in leaf-litter samples (70%). The use of selective isolation media is important to elucidate the microbial diversity.