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Title: Overexpression of 3-hydroxy-3-methylglutaryl coenzyme A reductase in Parthenium argentatum (guayule)

item Dong, Niu
item Ponciano, Grisel
item McMahan, Colleen
item Coffelt, Terry
item LAUREN, JOHNSON - Yulex Corporation
item ROBERT, CREELMAN - Mendel Biotechnology
item Whalen, Maureen
item KATRINA, CORNISH - The Ohio State University

Submitted to: Industrial Crops and Products
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/19/2012
Publication Date: 2/14/2013
Citation: Dong, N., Ponciano, G.P., McMahan, C.M., Coffelt, T.A., Lauren, J., Robert, C., Whalen, M.C., Katrina, C. 2013. Overexpression of 3-hydroxy-3-methylglutaryl coenzyme A reductase in Parthenium argentatum (guayule). Industrial Crops and Products. 46(2013):15-24. DOI: 10.1016/j.indcrop.2012.12.044.

Interpretive Summary: Guayule is under development as a new industrial crop in the US as a source natural rubber, organic resins, and bioenergy. Genetic modification of guayule for increased rubber yields could significantly contribute to economic sustainability of this new crop. Here we report results from a single gene modification of guayule designed to increased rubber and/or resin production. In the laboratory, increased rubber was found for a modified guayule line; that was not duplicated in the field, probably due to the overwhelming influence of environment in the harsh semi-arid cropland used for cultivation. However, the genetically improved lines had improved survival rates following pollarding (cut at the stem just above the base) harvest. Guayule is often left to re-grow in the field after the initial harvest, then re-harvested. Use of the improved lines might improve productivity in this scenario.

Technical Abstract: Natural rubber biosynthesized via the isoprenoid pathway by domestic plant sources, such as guayule (Parthenium argentatum) may be more economically sustainable with improved yields, through breeding or targeted metabolic engineering of the isoprenoid pathway. The enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is considered a key regulatory enzyme of isoprenoid carbon flux in mammals, in microbial systems, and possibly in plants. The objective of our study was to modify isoprenoid production in guayule through overexpression of the HMGR gene. A transformation plasmid pND4-HMGR(tAN) was constructed with a modified binary vector and the HMGR gene from A. nidulans containing only the 465 amino acid catalytic domain driven by a constitutive promoter. Five independent transgenic lines were obtained via leaf disc Agrobacterium-mediated transformation. In the laboratory, the NR content of two-month old in vitro plantlets showed a 65% increase in rubber over the vector control for one line (HMGR6), and lower resin production for another (HMGR2). In field evaluations, the genetically modified HMGR6 line was differentiated from control lines in size, biomass, and plant morphology descriptors, but not in rubber or resin content. Remarkably, the survival rate of all HMGR-modified plants following pollarding harvest, was better than controls, with the highest survival rate for line HMGR6. In conclusion, we report the first genetic modification of guayule to overexpress the isoprenoid pathway enzyme HMGR. Survival during regrowth was significantly improved for HMGR overexpressing plants, suggesting enhanced carbon flux to important secondary isoprenoid metabolites, such as growth phytohormones.