Judul: Soybean transformation with proteinase inhibitor 11 gene through agrobacterium and particle bombardment
Abstrak: The objectives of these research were (1) to obtain the best protocols of soybean transformation using Agrobacterhm-mediated bransformation and Particle Bombardment, (2) to transfer and regenerate soybean explants transformed with the proteinase inhibitor I1 gene, and to obtain soybean plants that contained the proteinme inhibitor II gene and resistant to pod barer. Research was included three experiments i.e 1) Soybean transformation using gus gen, 2) Soybean trmsfarmation using proteinase inhibitor (pin) II gene, and 3) Evaluation of soybean transformant plants. Two soybean cdtivars (Wilis and Tidar) were used in this experiments. Both cdtivars were chosen because it shown good response on early in vitro plant regeneration study and very popular to the farmers, especially cv. Wilis. On the first activity, soybean explants were inoculated with A. tumefaciens strain EHA 105 which is contained plasmid pCambia 1301 with p s gene in T-DNA region. The treatments were included an optical density (0.5; 1 and 1.5), inoculation time (60 and 90 minutes), co-cultivation time (3 and 5 days) and type of explants (young embryo and cotyledon). Result indicated that the best protocol for inoculation was using young cotyledon as explants with 1 x 10' celYrnl of Agrobacterium for 90 minutes inoculation and 5 days co-cdtivation. While on the particle bombardment method, young embryos and cotyledons were bombarded with pRQ6 plasmid. Treatment were included Helium gas pressure (1 1 00 and 1 300 psi), shooting distance (5 and 7 cm) and number of bombardment (once and twice). The best treatment was using young cotyledon as explant, bombarded once or twice with gold particles at 1 100 psi of the Helium gas pressure and 5 crn of shooting distance. On the second activity, 1539 young cotyledon explants from Wilis and 984 explants from Tidar were inoculated with A. tumefaciens which is contained proteinase inhibitor II gene on plasmids pGApin LI construct. Result from plant regeneration and selection of tsansformant explants on media with 200 mgA Kariamycine (spesific for nptII gene selection) indicated that c.v Wilis were better than Tidar, because its produced more plantlets/plants 8 plants (coded: A W I -AW8), while Tidar produced only one plant (coded: AT& On the Particle Bombardment experiments, 1274 young cotyledon explants of Wilis and 1 763 explants of Tidar were bombarded using gold particle coated with pTWa plasmids (contained pinn and bar genes). Result of plant regeneration and selection of transformant explants on media with 3 mg~l Basta (spesific for bar gene selection) indicated that cv. Tidar produced three plants (TPI-TP3) while cv.Wi1is produced only 2 plants (WPI and WP2). On the third activity, transformant plants were evaluated through molecular and bioassay analysis. Molecular analysis of nine soybean plants obtained from Agrobacterium-mediated transformation (8 Wilis and 1 Tidar) using Polymerase Chain Reaction (PCR) technique indicated that only event ATI (Tidar) was positive to pin I1 gene test, while 8 plants from event AW (Wilis) were negative. PCR test of five soybean plants obtained from bombardments experiments (2 Wilis and 3 Tidar ) indicated that no plants were positive to pin 11 gene, except WP2 (Wilis). Seeds fiom these positive
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Judul: Transformasi Genetik Tembakau dengan Gen BtCspB Penyandi Cold Shock Protein Melalui Perantara Agrobacterium tumefaciens
Abstrak: Cold shock protein (Csp) is essential for organisms to survive in the abiotic stress condition. It protects RNA from misfolding caused by various abiotic stresses. BtCsp gene had been isolated from Bacillus thuringiensis, fused to promoter of ubiquitin in the T-DNA of pCambia 1300int, and introduced into Agrobacterium tumefaciens LBA4404. We suppose that plant overexpressing this gene would be tolerant to abiotic stress including cold heat and drought stress. This research had an objective to transform genetically Nicotiana tabacum cv Samsun by BtCsp gene under the control of pUbiquitin promoter and NosT terminator mediated by A. tumefaciens. Leaf disks were inoculated by the method of co-cultivation with A. tumefaciens LBA4404 harboring pCambia1300int-BtCspB. We compared the effect of shoaking time between 20 and 30 minutes on the transformation efficiency. The transformation calli were selected on the medium containing 20 mg/l and 50 mg/l hygromcin and regenerated on MS medium containing 1 mg/l BAP and 0,1 mg/L IAA. Based on resistant calli to 50 mg/l hygromicin, the efficiency of transformation between 20 and 30 minutes was selatively equal i.e 65% and 57.5% respectively. PCR analaysis showed that BtCspB gene under the control of Ubquitin promoter is integrated into N. tabacum genome of To generation. Genetic analysis showed that hpt gene was inherited to the progeny T1 generation following Mendalian law. Based on chi-square analysis, the ratio of resistant and sensitive plant To hygromycin in T1 generation was 3:1, indicating that only one copy of hpt gene was integrated into the genome of transgenic plant in To generation.
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Judul: Hamiltonian formulation for linear waves motion in two layers fluid
Abstrak: The surface wave could be considered as a wave that separates two fluids, namely water and air. Based on this assumption, it is introduced the interfacial wave, a wave between two layers of fluid with different density. The formulation of interfacial waves motion begins with deriving the base equation of irrotational ideal fluid. Furthermore, according to irrotational fluid assumption, the base equation can be stated in velocity potential. In this derivation, the fluids domain is assumed to be restricted by rigid lid boundary conditions, both at the upper and lower limit. Therefore, the interfacial waves motion can be explained in a hamiltonian formulation. In the hamiltonian formulation, total energy is defined as the sum of kinetic and potential energy. The hamiltonian system is obtained from reduction of kinetic energy by using the Dirichlet Neumann Operator. The resulted kinetic energy equation is nonlinear. Therefore, this form is linearized by first part of the Taylor expansion. This linearization gives a dispersion relation of linear wave. Based on this dispersion relation, the phase speed of the linear wave depends on the density ratio of the two layers fluid.
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