Concerning the colour, the fungus B. cinerea can attack the grape berry and introduce the oxidative enzyme laccase into the berry and hence into grape juice. Laccase targets phenolics such as the red colour compounds in red wine and oxidizes them into brown-coloured compounds. Furthermore, the association of B. cinerea with other, less visible, fungi frequently leads to the development of organoleptic defects in grapes and sometimes in wines (La Guerche et al., 2006). The strategy most widely adopted by winegrowers to reduce the impact of grey selleckchem mould is the systematic application of chemical fungicides, based on a preset calendar that takes into account the phenological growth

stages of the grapevine. This reduction policy will have an impact on Botrytis resistance to fungicides (Leroux, 2004) and on the environment. Indeed, the contamination of agricultural soils with

inorganic (Cu-based) and organic pesticides (including their residues) presents a major environmental and toxicological click here concern (Komárek et al., 2010). Although there are alternative methods to synthetic fungicides, such as the application of antagonistic microorganisms and the application of natural antimicrobial substances, it is essential to monitor the disease development and particularly the concentration of fungal spores. Indeed, monitoring disease development will allow better disease management, and will reduce cost and improve grape quality. Spores can be identified and quantified by light microscopy (Aylor, 1998; Hunter et al., 1999). However, this is not straightforward. Indeed, it is a time-consuming technique that needs expertise for the accurate identification of spores. Antibody immunoassays have been used for the early detection of B. cinerea (Kennedy et al., 2000). However, taking into account the low sensitivity and the limited dynamic range of the method, it is not well adapted for quantification, although it can be used to confirm the nature of the agent (Suarez et al., 2005). Molecular techniques for the identification of spores have

already been published (West et al., 2008), most of which are based on detection by standard PCR methods (Zhou et al., 2000; Calderon et al., 2002; Chew et al., 2006). However, under these conditions, quantification is not Fossariinae precise. One way to assess for the presence of specific spores more accurately and to avoid some of the problems that accompany the other methodologies is real-time quantitative PCR (qPCR). Numerous quantitative assays utilizing real-time PCR have been developed to specifically detect microbial targets in many types of samples, including, but not limited to, moulds (Alaei et al., 2009; Carisse et al., 2009; Luo et al., 2010). Advantages of utilizing qPCR for spore enumeration over classic culture-based methods include its enhanced specificity and reduced processing time, leading to quicker results. Cadle-Davidson (2008) reported a qPCR method based on Taqman chemistry for monitoring B.