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Manipulating expression of tonoplast transporters.

Methods in molecular biology (Clifton, N.J.) (2012-08-17)
Zhigang Li, Man Zhou, Qian Hu, Shane Reighard, Shuangrong Yuan, Ning Yuan, Bekir San, Dayong Li, Haiyan Jia, Hong Luo
ABSTRACT

Plant vacuoles have multifaceted roles including turgor maintenance, cytosolic pH and ionic homeostasis, plant protection against environmental stress, detoxification, pigmentation, and cellular signaling. These roles are achieved through the coordinated activities of many proteins in the tonoplast (vacuolar membrane), of which the proton pumps and ion transporters have been modified for improved abiotic stress tolerance in transgenic plants. Here we describe a method to manipulate vacuolar H(+)-pyrophosphatase in turfgrass and evaluate the impact of the modified tonoplast on the phenotype, biochemistry, and physiology of the transgenics. Creeping bentgrass (Agrostis stolonifera L.) plants overexpressing an Arabidopsis vacuolar H(+)-pyrophosphatase AVP1 exhibited improved growth and enhanced salt tolerance, likely associated with increased photosynthesis, relative water content, proline production, and Na(+) uptake. These transgenic plants also had decreased solute leakage in the leaf tissues and increased concentrations of Na(+), K(+), Cl(-), and total phosphorus in the root tissues. Similar strategies can be employed to manipulate other tonoplast transporters and in other plant species to produce transgenic plants with improved performance under various abiotic stresses.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Pyrophosphatase, Inorganic from Escherichia coli, recombinant, expressed in E. coli, lyophilized powder, ≥90%, ≥800 units/mg protein
Sigma-Aldrich
Pyrophosphatase, Inorganic from baker′s yeast (S. cerevisiae), powder, ≥500 units/mg protein (E1%/280)