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S. Suharsono
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Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

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Molecular cloning of gene fragment encoding 4-coumarate: Coenzyme A ligase of Sengon (Paraserianthes falcataria) Sri N. Hartati; Enny Sudarmonowati; S. Suharsono; Kurnia Sofyan
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1105.206 KB) | DOI: 10.22146/ijbiotech.7816

Abstract

4-coumarate:Coenzyme A ligase (4CL) plays an important role in lignin biosynthetic pathway thatcatalyzed the activation of coumaric acid, caffeic acid or ferulic acid to be a syringil monomer. Ligninbiosynthesis control through 4CL down regulating would support lower lignin wood production. Theobjective of this study was to clone conserved region cDNA of gene encoding 4CL. Gene fragment isolation wasconducted by means of reverse transcriptase polymerase chain reaction (RT-PCR) using degenerateheterologous primer. The RT-PCR products were purified, sequenced and analyzed to select the highlyhomologous fragment to 4CL. BLASTanalysis result showed that deduction of amino acid sequences from oneof two RT-PCR product nucleotide was highly homologous with the 4CL conserved region from Rubbus ideaus,Oryza sativa, Populus tomentosa, Populus balsamifera, Betulla platyphilla, Nicotiana tabacum, and Arabidopsisthaliana with identity ranging from 78-90%.Key words: 4-coumarate: Coenzyme A ligase, lignin, sengon
Comparative Analysis of Rice Transformation Using Agrobacterium tumefaciens and Rhyzobium leguminosarum Syamsidah Rahmawati; Osmat Azzam Jefferson; Didy Sopandie; S. Suharsono; Inez Hortense Slamet-Loedin
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (287.394 KB) | DOI: 10.22146/ijbiotech.7821

Abstract

This study was aimed to study the effectiveness of Rhizobium transformation system compared to the most widely used Agrobacterium mediated transformation system on three rice cultivars, Ciherang (Indica), Nipponbare (Japonica), and Rojolele (Javanica). Six day old calli induced from immature embryos were inoculated with Rhizobium leguminosarum bv trifolii ANU845 and Agrobacterium tumefaciens LBA288 that harbored with vector pCAMBIA 5106. This plasmid contained a minimum set of transfer machinery genes and had a gusplus and an hptII gene driven by 35S CaMV promoter in the T-DNA. The results showed that the transformation frequencies (number of PCR positive plants per number of calli inoculated) ranging from 0 to 12.05 % depend on the genotype and transfer agent used. The highest transformation frequency (12.05%) was obtained in Ciherang transformed with R. leguminosarum. Most of the transgenic rice obtainedby Rhizobium transformation were normal in morphology and fertile similar to those obtained by Agrobacterium transformation. Integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis in T0 and T1 generations.Key words : Rhizobium leguminosarum, immature embryos, Agrobacterium tumefaciens
Co-Authors Didy Sopandie Enny Sudarmonowati Inez Hortense Slamet-Loedin Kurnia Sofyan Osmat Azzam Jefferson Sri N. Hartati Syamsidah Rahmawati