Moderate heat stress reduces the pH component of the transthylakoid proton motive force in light-adapted, intact tobacco leaves.

We measured the DeltaPsi and DeltapH components of the transthylakoid proton motive force (pmf) in light-adapted, intact tobacco leaves in response to moderate heat. The DeltaPsi causes an electrochromic shift (ECS) in carotenoid absorbance spectra. The light-dark difference spectrum has a peak at 518 nm and the two components of the pmf were separated by following the ECS for 25 s after turning the light off. The ECS signal was deconvoluted by subtracting the effects of zeaxanthin formation (peak at 505 nm) and the qE-related absorbance changes (peak at 535 nm) from a signal measured at 520 nm. Heat reduced DeltapH while DeltaPsi slightly increased. Elevated temperature accelerated ECS decay kinetics likely reflecting heat-induced increases in proton conductance and ion movement. Energy-dependent quenching (qE) was reduced by heat. However, the reduction of qE was less than expected given the loss of DeltapH. Zeaxanthin did not increase with heat in light-adapted leaves but it was higher than would be predicted given the reduced DeltapH found at high temperature. The results indicate that moderate heat stress can have very large effects on thylakoid reactions.

Analysis of the maize polyubiquitin-1 promoter heat shock elements and generation of promoter variants with modified expression characteristics.

The maize polyubiquitin-1 (Ubi-1) promoter is one of a few select promoters used to express foreign genes in monocots, such that recombinant proteins can be produced at commercially viable levels. Modifying the activity, specificity and responsiveness of such promoters provides a means to achieve desired levels and patterns of expression of genes encoding target products. Ubi-1 is constitutively expressed but is further induced by heat shock. The promoter contains two overlapping sequences with similarity to defined heat shock elements and we show that these sequences are also present upstream of the Ubi-1 homologue isolated from teosinte. Both the maize and teosinte promoters can mediate a heat shock response in transgenic maize. We have dissected the overlapping maize Ubi-1 promoter heat shock elements and demonstrate that the 3' element is required to mediate a heat shock response. The Ubi-1 promoter is particularly active in tissues consisting of rapidly dividing cells, and within the seed it is strongly biased towards driving expression in the embryo. However, replacement of the heat shock elements with a trimer of a basic domain/leucine zipper factor binding site of a pea lectin promoter shifts the balance in seed expression towards the endosperm. The Ubi-1 variants described here differ in their overall activity in the seed, but they all show potential for driving high levels of heterologous gene expression in maize.