Development of a quantitative analysis protocol of heat shock protein 70 in heat stressed sugar beet (Beta vulgaris L.) plants.

摘要

Quantitative real-time PCR (RT-PCR) has already been used to study the expression profile of several plant gene families (Yokoyama and Nishitani, 2001; Mladek et al., 2003; Charrier et al., 2002). This study demonstrates the potential of RT-PCR to measure the expression of heat shock protein (HSP) genes in sugar beet. Young plants underwent heat stress for 5.5 h followed by RT-PCR of hsp70 mRNA using the housekeeping gene tubulin (tub) as a reference gene and the fluorogenic dye SYBR Green I. Expression of hsp70 mRNA peaked at 1 h of heat stress and decreased at 5.5 h. This method proves very sensitive, quantitating the target hsp70 transcript in 6.25 ng total RNA..