Sukarti Moeljopawiro

Genetic Mapping Refines DFNB3 to 17p11.2, Suggests Multiple Alleles of DFNB3, and Supports Homology to the Mouse Model shaker-2

The nonsyndromic congenital recessive deafness gene, DFNB3, first identified in Bengkala, Bali, was mapped to a approximately 12-cM interval on chromosome 17. New short tandem repeats (STRs) and additional DNA samples were used to identify recombinants that constrain the DFNB3 interval to less, similar6 cM on 17p11.2. Affected individuals from Bengkala and affected members of a family with hereditary deafness who were from Bila, a village neighboring Bengkala, were homozygous for the same alleles for six adjacent STRs in the DFNB3 region and were heterozygous for other distal markers, thus limiting DFNB3 to an approximately 3-cM interval. Nonsyndromic deafness segregating in two unrelated consanguineous Indian families, M21 and I-1924, were also linked to the DFNB3 region. Haplotype analysis indicates that the DFNB3 mutations in the three pedigrees most likely arose independently and suggests that DFNB3 makes a significant contribution to hereditary deafness worldwide. On the basis of conserved synteny, mouse deafness mutations shaker-2 (sh2) and sh2J are proposed as models of DFNB3. Genetic mapping has refined sh2 to a 0.6-cM interval of chromosome 11. Three homologous genes map within the sh2 and DFNB3 intervals, suggesting that sh2 is the homologue of DFNB3.

Congenital non-syndromal autosomal recessive deafness in Bengkala, an isolated Balinese village.

Bengkala is an Indonesian village located on the north shore of Bali that has existed for over 700 years. Currently, 2.2% of the 2185 people in this village have profound congenital deafness. In response to the high incidence of deafness, the people of Bengkala have developed a village specific sign language which is used by many of the hearing and deaf people. Deafness in Bengkala is congenital, sensorineural, non-syndromal, and caused by a fully penetrant autosomal recessive mutation at the DFNB3 locus. The frequency of the DFNB3 mutation is estimated to be 9.4% among hearing people who have a 17.2% chance of being heterozygous for DFNB3.

Overexpression of PaFT gene in the wild orchid Phalaenopsis amabilis (L.) Blume

To shorten vegetative stage and induce transition from vegetative to reproductive stage in orchids, we overexpressed Phalaenopsis amabilis Flowering LocusT (PaFT) gene under the control of Ubiquitin promoter into protocorm of Indonesian Wild Orchid Phalaenopsis amabilis (L.) Blume. The dynamic expression of vegetative gene Phalaenopsis Homeobox1 (POH1) and flowering time gene PaFT has been analyzed. Accumulation of mRNA was detected in shoot and leaves of both transgenic and non transgenic plants by using Reverse transcriptase-PCR (RT-PCR) with specific gene primers for POH1 and PaFT in 24 months old plants. To analyze the POH1 and PaFT genes, three pairs of degenerate primers PaFT degF1R1, F2R2 and F3R3 that amplified 531 bp PaFT cDNA were used. We detected 700 bp PaFTcDNA from leaves and shoots of transgenic plants, but not in NT plants. POH1 mRNA was detected in plants. PaFT protein consists of Phospatidyl Ethanolamine-Binding Protein (PEBP) in interval base 73-483 and CETS family protein at base 7-519, which are important motif for transmembrane protein. We inserted Ubipro::PaFT/pGAS101 into P. amabilis protocorm using Agrobacterium. Analysis of transgenic plants showed that PaFTmRNA was accumulated in leaves of 12 months after sowing, although it is not detected in non transgeic plants. Compare to the wild type (NT plants), ectopic expression of PaFT shows alter phenotype as follows: 31% normal, 19% with short-wavy leaves, 5% form rosette leaves and 45% produced multishoots. Analysis of protein profiles of trasgenic plants showed that a putative PaFT protein (MW 19,7 kDa) was produced in 1eaves and shoots.This means that at 12 months, POH1 gene expression gradually decreased/negatively regulated, the expression of PaFT gene was activated, although there is no flower initiation yet. Some environmental factors might play a role to induce inflorescens. This experiment is in progress.

Antioxidant and antibacterial activities of agarwood (Aquilaria malaccensis Lamk.) leaves

Several factors involve in the synthesis of secondary metabolites in plants, including leaves maturity. The objectives of this study were to compare the antioxidant and antibacterial activities of agarwood Aquilaria malaccensis Lamk. leaves at different stages of maturity as well as to identify the group of compounds responsible to the corresponding activities. Young and old leaves of agarwood were used. Leaves were powdery dried and extracted using three different polarity solvents including chloroform, methanol and water using Soxhlet apparatus. The potential extract was fractionated using Vacuum Liquid Chromatography (VLC). Antioxidant activity was analysed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) while antibacterial activity was evaluated using paper disc diffusion assay. Thin Layer Chromatography (TLC) followed by spraying with specific reagents was applied to identify groups of bioactive compounds in the potential fraction. The results showed that the methanol extract of the old leaves was the mos...

Effect of Biofertilizer Addition on Nitrous Oxide Emission

Application of nitrogen fixing biofertilizer, such as Azotobacter, has a potential for reducing nitrous oxide (N2O) emission. The aim of this study was to examine the effect of nitrogen fixing biofertilizer addition to common practices of urea and fresh cattle manure usages for maize (Zea mays L.) growing on N2O emission. The field experiment was conducted at Gunung Kidul, Yogyakarta, Indonesia. The treatments were addition of fresh cattle manure (M), fresh cattle manure added with nitrogen fixing biofertilizer (MB), urea (U), urea added with nitrogen fixing biofertilizer (UB), and control (no N fertilizer added). Nitrogen contents of the added urea and fresh cattle manure were adjusted to be equal. Urea and fresh cattle manure were given three times throughout the experiment period, i.e. 12, 30, and 48 days after planting (DAP). Urea was given at a rate of 44, 29, and 15 kg.ha -1 , respectively while fresh cattle manure was given at a rate of 6000, 4000, and 2000 kg.ha -1 , respectively. The emitted N2O was collected using a closed-chamber method at 24, 42, 60, and 72 DAP and were determined using Gas Chromatograph. Soil properties including available N (NH4 + -N and NO3 - -N) and organic C contents were also analyzed. On the harvesting time, the harvest index and the grain yield were determined. Biofertilizer addition influence decomposition process of cattle manure and urea that led to mineralization and nitrification of residual organic matter and hence to cause soil NH4 + N in the order concentration of M treatment > MB > U > UB > C, and soil NO3 - -N of MB treatment > M > U > UB > C. Reduction of NO3 - N was resulted in the highest N2O emission of M > U > MB > UB > C (P UB > U > M> C treatments. Available mineral N and soil organic C contents strongly affected N2O emission (P < 0.01). The results suggested that biofertilizer addition to common agricultural practices reduce N2O emission and simultaneusly increased grain yield, and harvest index of maize.