Indonesian Journal of Biotechnology
Vol 22, No 2 (2017)

Agrobacterium tumefaciens-mediated transformation of Jatropha curcas L. with a polyhydroxyalkanoate gene (phaC)

Chesara Novatiano (Biotechnology Study Program, School of Life Sciences and Technology, Insধtut Teknologi Bandung, Jalan Ganesha No. 10, Bandung 40133, Indonesia)
Adi Pancoro (School of Life Sciences and Technology, Insধtut Teknologi Bandung, Jalan Ganesha No. 10, Bandung 40133, Indonesia)
Erly Marwani (School of Life Sciences and Technology, Insধtut Teknologi Bandung, Jalan Ganesha No. 10, Bandung 40133, Indonesia)



Article Info

Publish Date
13 Feb 2018

Abstract

Polyhydroxybutyrate is a component of bioplastics that is synthesized under the control of enzymes encoded by pha multigenes. The genes are naturally present in Ralstonia eutropha. However, the production of bioplastics in bacteria is inefficient because the bacterial biomass is relatively small compared with plants or fungi. As such, engineering techniques have been developed that enable pha genes to be inserted into plant biomass, and then be expressed in the biomass of the plant to produce polyhydroxybutyrate. The objectives of this study were to transform the tissue of Jatropha curcas using the phaC gene (a pha gene), to regenerate the transformed plant, and to confirm the presence of the inserted genes with PCR. The genetic transformation of J. curcas was mediated by Agrobacterium tumefaciens strain GV3101 containing pARTC by dipping the cotyledon tissue of J. curcas in a suspension of the bacterium for 30 min, followed by cocultivation for 3 d on Murashige and Skoog (MS) medium. The tissue was then placed on a selection medium, i.e. MS medium containing 13.3 µM BAP and 0.05 µM IBA with the addition of 20 mg/L kanamycin. The results showed that 12.35% of the tissue survived and regenerated into a shoot after 1–2 months. Molecular analysis of the transformed tissue was performed using phaC and nptII primers, in order to detect the presence of the phaC and nptII genes. Specific bands were detected at 659 bp and 700 bp, corresponding to the nptII primer and phaC primer, respectively.

Copyrights © 2017






Journal Info

Abbrev

ijbiotech

Publisher

Subject

Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

Description

The Indonesian Journal of Biotechnology (IJBiotech) is an open access, peer-reviewed, multidisciplinary journal dedicated to the publication of novel research in all aspects of biotechnology, with particular attention paid to the exploration and development of natural products derived from ...