Development of a rapid and highly sensitive direct-PCR assay to detect a single conidium of <i>Botrytis cinerea</i> Pers.:Fr <i>in vitro</i> and quiescent forms <i>in planta</i>

Authors

  • K. Gindro
  • R. Pezet
  • O. Viret
  • H. Richter

DOI:

https://doi.org/10.5073/vitis.2005.44.139-142

Keywords:

Botrytis cinerea, “Direct-PCR”, Vitis vinifera

Abstract

“Direct-PCR” amplifications of Botrytis cinerea-specific genomic sequences, without any DNA purification step or time consuming sample preparation, were developed. A single copy sequence of 0.7 Kb in the Botrytis cinerea genome was amplified in reactions containing no more than 1 x 105 to 1 single conidium. As a demonstrative application, this assay was applied to detect B. cinerea in different parts of immature grape berries (at ‘pea size’), when previously inoculated with conidia at flowering. Using this method we showed the presence of quiescent Botrytis in the receptacle area only. Cloning and sequencing of the fragment confirmed the single sequence gene of B. cinerea. These results demonstrate that the method is easy to apply and of sufficiently high sensitivity to detect the presence of B. cinerea in immature grape berries. Its use for studies on the development of grey mould and improved control of the disease in vineyards is discussed.


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Published

2015-04-16

Issue

Vol. 44 No. 3 (2005): Vitis

Section

Article

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