The Trafficking of wheat prolamins in rice endosperm

IN Discussion Topics, ONCYTE AVID

Wheat is the most important crop in Europe due in part to high relative crop yields over a range of environments. “(Wheats) success as a food crop is also due to the unique properties which form the gluten fraction… It (is) necessary to understand the pathways and mechanisms which determine the synthesis, processing, trafficking and deposition of storage components in the developing grain.” (1) 

To study the pattern and pathway of wheat high molecular weight (HMW) subunit 1Dx5 deposition within rice endosperm, tissue sections were examined by light and electron microscopy.  An array of antibodies were used to determine deposition pattern, localization within storage protein bodies and temporal relevance to distribution levels of HMW subunit 1Dx5. Transgenic rice seeds expressing wheat HMW subunit 1Dx5 were harvested at 7, 10, 14, 17, 20 and 25 days after flowering.  Seeds were frozen, cryosectioned and tissue printed with the anti-R2 HWM antibody on Oncyte® Film Slides.  Expression patterns for antibodies to purified rice glutelin subunits, prolamins, and polyclonal anti-R2 HMW were detected at the different stages of development.  Immunohistochemistry results from samples probed with purified rice glutelin subunits, prolamins and polyclonal anti-R2 HMW antibodies indicate that HMW glutenin subunits are concentrated on the outer layers of the rice endosperm cells.

ONCYTE® Nitrocellulose Film Slides

Tissue printing onto membranes such as nitrocellulose is a technique employed to study the localization of proteins, nucleic acids, and soluble metabolites from tissue slices. 

Porous nitrocellulose (PNC) film slides are the premier high binding microarray substrate technology. 

  • 3D structure offers increased surface area for binding potential
  • high signal to noise ratio
  • multiplex capability
  • compatible with colorimetric, chemiluminescent, fluorescent and near-IR detection strategies