Annales Bogorienses

1-8

Authors: Dyah Retno Wulandari, Tri Muji Ermayanti

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

17-23

Authors: Puspita Lisdiyanti, Yopi Yopi, Tutik Murniasih

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.

31-36

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

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.

1-6

Authors: Muhaimin Rifa'i

Naturally occurring CD4+CD25+ regulatory T (Treg) cells, a component of the innate immune response which play a key role in the maintenance of self-tolerance, have become the focus of numerous studies over the last decade. These cells have the potential to be exploited to treat autoimmune disease. These cells inhibit the immune response in an antigen-nonspecific manner by interacting with other T cells. These T cell populations actively control the properties of other immune cells by suppressing their functional activity to prevent autoimmunity but also influence the immune response to allergens as well as against tumor cells and pathogens. In this experiment we showed that induced regulatory T cells have a protective effect on mice model of rheumatoid arthritis (RA). RA mice which were injected intraperitoneally with Andrographis paniculata substrate or injected with induced regulatory T cells showed the effects of recovery. We further showed that the generation of leukocyte including B cells can be promoted by the administration of A. paniculata substrate. Tissue damage from free radicals that arise due to imperfect metabolism can be prevented by such treatment in RA model mice. Recovery effects occurred in RA model mice involves the increasing number of CD4 +CD25+ regulatory T cells.

15-20

Authors: Betalini Widhi Hapsari, Tri Muji Ermayanti, Deritha E. Rantau, Rudiyanto Rudiyanto

The aim of this research was to compare the reduction effect of sucrose or table sugar concentration on growth characteristics of red ginger cultured in MS liquid medium. Shoots of red ginger was cultured on MS liquid medium without addition of plant growth regulators, supplemented with 5, 10, and 20 g/l of sucrose or table sugar for 8 weeks. Resulted plantlets were acclimatized in a greenhouse to investigate their growth and survival rate. Numbers of stomata, chlorophyll concentration as well as cross section of leaves from plantlets grown in vitro were compared to those of transplants grown in the greenhouse. The results showed that the use of table sugar at concentration of 20 g/l gave the best growth of red ginger. Meanwhile, the reduction of table sugar from 20 to 10 g/l reduced growth and survival rate of in vitro shoots as well as that of transplants in the greenhouse. Only few shoots formed roots when they were grown on the medium containing 5 g/l of table sugar, and transplants failed to grow in the greenhouse. It found that the chlorophyll content of in vitro plantlets was lower than those of transplants grown in the glasshouse. However, the number of stomata of the in vitro plantlets was higher than that of transplants grown in the glasshouse. There was no anatomical abnormalities found on the cross section of leaves between in vitro plantlets and transplants grown in the greenhouse. The replacement of sucrose with table sugar may reduce the production cost of plantlets.

27-32

Authors: Yuli Sulistyowati, Agus Rachmat, Fatimah Zahra, Syamsidah Rahmawati, Satya Nugroho

Rice stem borer (Scirpophaga sp.) is considered as one of the primary pests causing yield decrease in Indonesia. Genetic engineering is considered as one of the most effective way to improve the resistance of rice plants to yellow stem borer since no resistance gene has been found in rice and its wild relatives. A double T-DNA binary vector p2TDNAcryIAb wich carry two independent T-DNA was used to transform Indonesian elite rice cv Ciherang using Agrobacterium-mediated transformation to generate transgenic rice that free from selectable marker genes. One T-DNA containing a selectable marker hygromycin phosphotransferase (hpt) gene and the other carrying the cry1Ab gene. As many as 37 putative transgenic plants from 14 embryos were generated. PCR analysis demonstrated that 22 out of 37 putative transgenic rice contain cry1Ab gene. All Ciherang rice carrying the cry1Ab gene expressing Cry1Ab protein as shown by the immunostrip assay. Further work will be carried out to determine the transgenes copy number, to select marker free transgenic rice, and to evaluate the resistance of transgenic rice against neonates larvae of yellow stem borer.