Nitric oxide plays an important role in the pathogenesis of Pseudoidium neolycopersici, the causative agent of tomato powdery mildew. S-nitrosoglutathione reductase, the key enzyme of S-nitrosothiol homeostasis, was investigated during plant development and following infection in three genotypes of Solanum spp. differing in their resistance to P. neolycopersici. Levels and localization of reactive nitrogen species (RNS) including NO, S-nitrosoglutathione (GSNO) and peroxynitrite were studied together with protein nitration and the activity of nitrate reductase (NR). GSNOR expression profiles and enzyme activities were modulated during plant development and important differences among Solanum spp. genotypes were observed, accompanied by modulation of NO, GSNO, peroxynitrite and nitrated proteins levels. GSNOR was down-regulated in infected plants, with exception of resistant S. habrochaites early after inoculation. Modulations of GSNOR activities in response to pathogen infection were found also on the systemic level in leaves above and below the inoculation site. Infection strongly increased NR activity and gene expression in resistant S. habrochaites in contrast to susceptible S. lycopersicum. Obtained data confirm the key role of GSNOR and modulations of RNS during plant development under normal conditions and point to their involvement in molecular mechanisms of tomato responses to biotrophic pathogens on local and systemic levels.

• Klíčová slova
• Nitric oxide, Powdery mildew, Pseudoidium neolycopersici, Reactive nitrogen species, S-nitrosoglutathione reductase, Solanum spp., Tomato,
• MeSH
• aldehydoxidoreduktasy metabolismus   MeSH
• Ascomycota patogenita   MeSH
• genotyp MeSH
• nemoci rostlin * mikrobiologie   MeSH
• reaktivní formy dusíku metabolismus   MeSH
• Solanum lycopersicum enzymologie   mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldehydoxidoreduktasy MeSH
• formaldehyde dehydrogenase, glutathione-independent MeSH Prohlížeč
• reaktivní formy dusíku MeSH

Resistant Lactuca spp. genotypes can efficiently modulate levels of S-nitrosothiols as reactive nitrogen species derived from nitric oxide in their defence mechanism against invading biotrophic pathogens including lettuce downy mildew. S-Nitrosylation belongs to principal signalling pathways of nitric oxide in plant development and stress responses. Protein S-nitrosylation is regulated by S-nitrosoglutathione reductase (GSNOR) as a key catabolic enzyme of S-nitrosoglutathione (GSNO), the major intracellular S-nitrosothiol. GSNOR expression, level and activity were studied in leaves of selected genotypes of lettuce (Lactuca sativa) and wild Lactuca spp. during interactions with biotrophic mildews, Bremia lactucae (lettuce downy mildew), Golovinomyces cichoracearum (lettuce powdery mildew) and non-pathogen Pseudoidium neolycopersici (tomato powdery mildew) during 168 h post inoculation (hpi). GSNOR expression was increased in all genotypes both in the early phase at 6 hpi and later phase at 72 hpi, with a high increase observed in L. sativa UCDM2 responses to all three pathogens. GSNOR protein also showed two-phase increase, with highest changes in L. virosa-B. lactucae and L. sativa cv. UCDM2-G. cichoracearum pathosystems, whereas P. neolycopersici induced GSNOR protein at 72 hpi in all genotypes. Similarly, a general pattern of modulated GSNOR activities in response to biotrophic mildews involves a two-phase increase at 6 and 72 hpi. Lettuce downy mildew infection caused GSNOR activity slightly increased only in resistant L. saligna and L. virosa genotypes; however, all genotypes showed increased GSNOR activity both at 6 and 72 hpi by lettuce powdery mildew. We observed GSNOR-mediated decrease of S-nitrosothiols as a general feature of Lactuca spp. response to mildew infection, which was also confirmed by immunohistochemical detection of GSNOR and GSNO in infected plant tissues. Our results demonstrate that GSNOR is differentially modulated in interactions of susceptible and resistant Lactuca spp. genotypes with fungal mildews and uncover the role of S-nitrosylation in molecular mechanisms of plant responses to biotrophic pathogens.

• Klíčová slova
• Bremia lactucae, Golovinomyces cichoracearum, Lettuce downy mildew, Lettuce powdery mildew, Nitric oxide, Pseudoidium neolycopersici,
• MeSH
• aldehydoxidoreduktasy metabolismus   MeSH
• konfokální mikroskopie MeSH
• nemoci rostlin mikrobiologie   MeSH
• odolnost vůči nemocem fyziologie   MeSH
• oomycety patogenita   MeSH
• polymerázová řetězová reakce MeSH
• regulace genové exprese u rostlin MeSH
• S-nitrosothioly metabolismus   MeSH
• salát (hlávkový) enzymologie   fyziologie   MeSH
• western blotting MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldehydoxidoreduktasy MeSH
• formaldehyde dehydrogenase, glutathione-independent MeSH Prohlížeč
• S-nitrosothioly MeSH

The level of resistance induced in different tomato genotypes after β-CRY treatment correlated with the upregulation of defence genes, but not sterol binding and involved ethylene and jasmonic acid signalling. Elicitins, a family of small proteins secreted by Phytophthora and Pythium spp., are the most well-known microbe-associated molecular patterns of oomycetes, a lineage of fungus-like organisms that include many economically significant crop pathogens. The responses of tomato plants to elicitin INF1 produced by Phytophthora infestans have been studied extensively. Here, we present studies on the responses of three tomato genotypes to β-cryptogein (β-CRY), a potent elicitin secreted by Phytophthora cryptogea that induces hypersensitive response (HR) cell death in tobacco plants and confers greater resistance to oomycete infection than acidic elicitins like INF1. We also studied β-CRY mutants impaired in sterol binding (Val84Phe) and interaction with the binding site on tobacco plasma membrane (Leu41Phe), because sterol binding was suggested to be important in INF1-induced resistance. Treatment with β-CRY or the Val84Phe mutant induced resistance to powdery mildew caused by the pathogen Pseudoidium neolycopersici, but not the HR cell death observed in tobacco and potato plants. The level of resistance induced in different tomato genotypes correlated with the upregulation of defence genes including defensins, β-1,3-glucanases, heveins, chitinases, osmotins, and PR1 proteins. Treatment with the Leu41Phe mutant did not induce this upregulation, suggesting similar elicitin recognition in tomato and tobacco. However, here β-CRY activated ethylene and jasmonic acid signalling, but not salicylic acid signalling, demonstrating that elicitins activate different downstream signalling processes in different plant species. This could potentially be exploited to enhance the resistance of Phytophthora-susceptible crops.

• Klíčová slova
• Defence-related genes, Elicitins, Pseudoidium neolycopersici, Resistance, Signalling, Sterol binding,
• MeSH
• cyklopentany metabolismus   MeSH
• ethyleny metabolismus   MeSH
• fungální proteiny metabolismus   MeSH
• interakce hostitele a patogenu MeSH
• kyselina salicylová metabolismus   MeSH
• listy rostlin metabolismus   mikrobiologie   MeSH
• nemoci rostlin mikrobiologie   MeSH
• oxylipiny metabolismus   MeSH
• peroxid vodíku metabolismus   MeSH
• Phytophthora MeSH
• Pythium MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulátory růstu rostlin metabolismus   MeSH
• signální transdukce * MeSH
• Solanum lycopersicum metabolismus   mikrobiologie   fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cryptogein protein, Phytophthora cryptogea MeSH Prohlížeč
• cyklopentany MeSH
• ethylene MeSH Prohlížeč
• ethyleny MeSH
• fungální proteiny MeSH
• jasmonic acid MeSH Prohlížeč
• kyselina salicylová MeSH
• oxylipiny MeSH
• peroxid vodíku MeSH
• reaktivní formy kyslíku MeSH
• regulátory růstu rostlin MeSH