Judul: Modifikasi kandungan lignin kayu sengon (Paraserianthes falcataria (L.) Nielsen) melalui rekayasa Gen 4- coumarate CoA Ligase (4CL)
Abstrak: Separation of lignin from cellulose requires high input of chemicals and energy resulted in high cost production and high ecological risks. The presence of lignin in plant cells is a factor limiting the efficiency of processing lignocellulosic materials of wood-based industries including pulp and paper industry either chemically or biologically (biopulping) process using white rot fungi. Lignin content and composition affect the efficiency of pulping process. Low-lignin wood or wood containing more reactive lignin which means syringyl content is higher than guaiacyl are more easily separated from cellulose. Therefore, it would be advantage to improve the efficiency process of pulp and paper industry. Sengon (P. falcataria) as a fast growth species is useful as a raw material for pulp because its characteristic which possess good fiber dimensions, high quality of physical and mechanical properties of its paper sheets. This study was aimed at (1) isolating the cDNA encoding 4-coumarate CoA ligase (4CL) from sengon, (2) constructing a binary vector of 4CL gene, with antisense orientation sengon transformation, (3) producing transgenic sengon plants which have characteristics low-lignin or higher syringyl ratio by introducing binary vector construct containing 4CL gene fragment with antisense orientation. Modification of the content or composition of lignin through recombinant DNA technology (genetic engineering) has been carried out in several stages: (1) quantitative analysis and histochemical lignin of sengon (P. falcataria), (2) embryogenesis shoots induction of sengon (3) isolation and cloning of cDNA fragments of genes encoding 4-coumarate: Coenzyme A ligase from sengon, and (4) genetic transformation on sengon by using gene fragment of antisense 4-coumarate CoA ligase (4CL) via A. tumefaciens. Lignin content determination of sengon collected from several areas in Indonesia indicated that lignin content of sengon ranged low to moderate category (16.58-35.59%). Lignin histochemical assay of transverse section of stems by using phloroglucinol-HCl staining showed that the initiation of lignin deposition was noted in 2 weeks old seedling. Quantitative and qualitative assessment through histochemical assay showed that lignin content in trees was varied depending on heights and location. Shoots induction from cotyledon nodes was selected as the most appropriate regeneration system for genetic transformation to improve sengon wood quality for pulp and paper industry. The cDNA fragment of 342 bp in size was obtained using degenerate primer designed using CODEHOP technique. Blast analysis result showed that deduction amino acid sequences of one out of two RT-PCR products nucleotide was highly homologous with the 4CL conserved region from Rubbus ideaus, Oryza sativa, Populus tomentosa, Populus balsamifera, Betulla platyphilla, Nicotiana tabacum, and Arabidopsis thaliana with identity ranging from 78-90%. The 4CL fragment has been successfully constructed and cloned for antisense orientation in pCAMBIA 2301. Recombinant expression vector of antisense 4CL has been successfully introduced into sengon via Agrobacterium tumefaciens. Gene integration test by PCR method showed that 19 out of 112 transformed sengon seedlings which resistant to kanamycine were transgenic seedlings. Lignin content determination of transgenic seedlings stem including histochemical assay, Klason lignin content and FTIR analysis to determine the S/G ratio was indicated that two transgenic sengon seedlings namely 4CLAS-4 and 4CLAS-1 possessed lower lignin content (15:53%) and
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Judul: Cloning Method of Pest Resistance Gene from Sengon (Paraserianthes falcataria).
Abstrak: Sengon is one of fast growing trees species, which is widely cultivated in plantations forest in Indonesia due to its multipurpose nature, and easy to grow. The plantation, however, faces serious problem of stem borer (Xystrocera festiva) attack, which was reported could destroyed sengon plantation. Meanwhile, several studies have reported the role of trypsin inhibitors (TI) and alpha-amylase inhibitor (AAI) in resistance in trees to insects. This research aimed at cloning a possible pest resistance gene from sengon. Sengon genomic DNA was isolated using CTAB buffer combined with binding columnn of GenElute plant Genomic DNA Miniprep Kit. Designed primer pairs did not generate amplicon. PCR products were obtained from published degenerate primers as two amplicons, with predicted size of 350 bp and 550 bp for AAI gene, and 600 bp and 800 bp for TI gene. Cloning with pGEM-T Easy vector resulted in several white colonies. Sequencing from selective colonies with long and short amplicon of AAI and TI sequences with 693 nucleotide, 278 nucleotide and 832 nucleotide, 278 nucleotide, respectively. BLASTN and BLASTX the sequences did not confirm that the inserts were AAI and TI genes. Further research is still needed to identify pest resistance gene in sengon tree.
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Judul: Distemper Pada Anjing
Abstrak: Distemper merupakan penyakit infeksius pada bimbingan anjing yang menduduki tempet teratas dan penyakit ini tersebar di seluruh bagian dunia dimana terdapat anjing-anjing yang dipelihara, terutama pada anjing-anjing muda di kota-kota besar, pada kelompok-kelompok anjing atau pada situasi lain dimana banyak terdapat kontak antara anjing. Penyakit ini disebabkan oleh virus yang termasuk dalam famili Paramyxoviridae dan genus Morbilivirus. Infeksi virus ini sering disertai oleh infeksi sekunder oleh kuman Staphylococcus, streptococcus, Brucella bronchiseptica dan kuman Salmonella, inilah yang kadang-kadang sebagai penyebab kematian.
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