Judul: Various Types Of Pstv Cp Gene Constructs To Protect Transgenic Nicotiana Benthamiana From Pstv Inoculation And Transformation Of Pstv Cp Gene Into Peanut
Abstrak: Disertasi ini dirnulai dari BAB I. Pendahuluan Umum dan BAB II. Tinjauan Pustaka. Selanjutnya ditulis hasil-hasil serangkaian percobaan yang mengarah kepada pembentukan tanaman Nicotiana henthamiana dan kacang tanah tahan PStV. terutama dengan memanfaatkan teknik transformasi gen dengan bantuan Agrobacterium tumefaciens. Gen ketahanan terhadap PStV dalam penelitian ini diambil dari gen coat protein PStV itu sendiri, terdiri atas empat tipe konstruksi yaitu pBINRCPL pBINRCP2. pBINRCP3, dan pBINRCP4. Untuk mengetahui dengan cepat efektivitas keempat gen ini dalam memberikan sifat ketahanan tanaman terhadap PStV digunakan tanaman model. yaitu N benthamiana. Setelah diketahui konstruksi gen yang paling efekti1~ konstruksi ini akan diintroduksikan ke tanaman kacang tanah. Dengan demikian dalarn penelitian ini digunakan dua tanaman yaitu N henthamiana sebagai tanaman model dan kacang tanah (Arachis hypogaea L) sebagai tanaman utarna yang diinginkan memperoleh sifat ketahanan terhadap PStV. Untuk keberhasilan regenerasi tanaman transgenik diperlukan adanya empat syarat yaitu: (i) tersedianya gen yang akan diintroduksikan; (ii) metode introduksi gen tersebut ke tanaman target: (iii) metode regenerasi tanaman yang efektif secara in virro: dan (iv) kemampuan untuk mengekspresikan gen yang telah terintegrasi dalam genom tanaman. Untuk memperoleh tanarnan transgenik N. benthamianu juga diperlukan keempat syarat di atas. Syarat pertama yaitu tersedianya gen yang akan diintroduksikan. Gen-gen yang digunakan dalam penelitian ini adalah gen cp PStV dan dua gen marker [gen /3-glucoronidase (gus) dan gen green fluorescent protein (g/p)]. Gen cp dan gen gus sudah dikonstruksi oleh peneliti lain sedangkan gen gfp dikonstruksi dalam penelitian ini (dilakukan di QABC. Queensland. Australia). Konstruksi dan kloning gen gfjJ dilakukan untuk memperoleh gen marker yang Iebih baik dibandingkan gen marker lain (seperti gen gus dan gen luc). Dengan tersedianya konstruksi gen marker ini. pengembangan metode transforrnasi tanaman dapat dilakukan tanpa menggunakan esei yang sifatnya destruktif jadi jaringan/eksplan tidak rnati seperti yang terjadi pada esei GUS. Dari penelitian yang ditulis pada BAB III ini dihasilkan dua plasmid yaitu pSAQl (singkatan dari Sholeh Avivi QABC) dan pSAQ2. Gen gfp dalam plasmid basil rekonstruksi dapat diekpresikan dengan baik pada tanaman tembakau W38. Plasmid hasil penelitian ini digunakan untuk membandingkan efektifitas transfer gen pada BAB VI. I, Peanut stripe virus (PStV) is one of the most important virus diseases in peanut. It causes severe reduction in yield and seed quality of peanut. Hence it is a significant constraint to peanut production in Indonesia. In this study, we aim to generate genetically-modified commercial peanut cultivars that are resistant to PStV infection by expressing viral coat protein sequence in transgenic plants. To achieve this objective, several experiments have been conducted, such as: (1) Construction of non-destructive selectable marker gene, green fluorescens protein (g/p), for expression in plants; (2) Evaluation of various types of PStV cp transgene(s) to protect transgenic Nicotiana benthamiana from PStV; (3) The development of procedures for obtaining efficient somatic embryos in peanut; (4) Agrobacterium-mediated transformation of peanut somatic embryos; and ( 5) Regeneration of transgenic peanut carrying PStV cp transgene(s). The PStV cp transgene was isolated from coat protein gene sequence of PStV isolate from Malang (Indonesia), and was constructed in four different types: pBINRCP 1 (full length translatable), pBINRCP2 (full length translatable, modified DAG motif), pBINRCP3
Keyword: Peanut Stripe Virus(PStV), Nicotiana benthamiana

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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