Identification of genes expressed during the compatible interaction of grapevine with Plasmopara viticola through suppression subtractive hybridization (SSH)

Abstract

Grapevine (Vitis vinifera) is the most widely cultivated and economically important fruit crop, but is susceptible to a large number of diseases. Downy mildew, caused by the obligate biotrophic oomycete pathogen Plasmopara viticola, is a common disease present in all regions where vines are cultivated. We used suppression subtractive hybridization (SSH) to generate two cDNA libraries enriched for transcripts induced and repressed, respectively, in the susceptible grapevine cultivar Chasselas 24 h after inoculation with P. viticola. Differential screening on glass slide microarrays yielded over 800 putative genes that were upregulated in response to P. viticola infection and over 200 that were down-regulated. One hundred and ninety four of these, were sequenced, identified and functionally categorised. Transcript abundance of twelve genes over a 48 h time course was examined by reverse transcriptase quantitative real-time PCR (RT-qPCR). Ten of these genes were induced/enhanced by P. viticola challenge, confirming the results of the SSH. The vast majority of the genes identified are related to defence. Interestingly, many genes involved in photosynthesis were down-regulated.

Characterization of Erysiphe necator-Responsive Genes in Chinese Wild Vitis quinquangularis

“…The largest gene functional category, defense (8.3%), detected in this study was similar to the previous study which categorized disease/defense, accounting for 24% in grapevine infected by P. viticola [8]. Moreover, defense-related genes were observed to overlap between this study and the previous study [8] in the susceptible grapevine cultivar Chasselas in response to P. viticola , including pathogenesis-related protein, chitinase, lipid transfer protein (LTP), and thaumatin-like protein (TLP) (Table 1). This indicated that there might be common genes in grapevines in response to fungal and oomycete diseases, and putative grapevine defense genes induced in susceptible plants were also induced in resistant plants.…”

“…Riesling with P. viticola [22], in compatible grapevine cv. Chasselas with P. viticola [8] and in rice with Magnaporthe grisea [23]. Taking these results together, the induction of PR-10 may play an important role in plant defense against pathogens.…”

“…Plant LTPs increase cutin layer thickness to protect leaf cells from fungus damage, retard growth of fungal pathogens and lead to cell death [14,24]. Legay et al [8] found an interesting feature that the LTP structure showed similarity with elicitin: a small lipid-binding protein that is secreted by the phytopathogenic oomycetes Phytophthora and Pythium , which triggers the hypersensitive response (HR) and SAR in tobacco.…”

“…This technique has been successfully used to isolate functional genes that are responsive to pathogen infection in plants [8–10]. In this study, to better understand the molecular basis of the high resistance to PM by Chinese wild V. quinquangularis , we constructed an SSH cDNA library to determine the differential expression of PM-responsive genes.…”

Downy mildew resistance induced by Trichoderma harzianum T39 in susceptible grapevines partially mimics transcriptional changes of resistant genotypes

“…Transcriptional changes associated with P . viticola infection of susceptible grapevines have been related to a weak defence response [4] and to the establishment of a compatible interaction [5,7,9,10]. The response of resistant genotypes has been characterized by strong and rapid transcriptional reprogramming of processes related to defence, signal transduction, and secondary metabolism, which are either not induced or induced to a lesser extent in susceptible grapevines [4,8,11-14].…”