Mycorrhiza induced resistance in potato plantlets challenged by Phytophthora infestans

Biological control of soil-borne pathogens by arbuscular mycorrhizal (AM) fungi has been repeatedly demonstrated. However, their role in the control of above-ground hemibiotrophic pathogens is less conclusive. Here, we investigated in vitro the impact of an AM fungus on Phytophthora infestans in potato plants. The leaf infection index was decreased in mycorrhizal potato plants. Real-Time Quantitative PCR revealed the induction of two pathogenesis related genes (PR1 and PR2) in the leaves of mycorrhizal plants shortly after infection with P. infestans. These results suggested a systemic resistance in mycorrhizal plants, related to the priming of the two PR genes in potato.     

Phosphite compounds reduce disease severity in potato seed tubers and foliage

Phosphites (Phi) are alkali metal salts of phosphorous acid, with the ability to protect plants against different pathogens. In this research, the effect of Phi applied to potato plants on severity of three important potato diseases in Argentina was assessed. Seed tubers and foliage of potato cvs Shepody and Kennebec were treated with Phi to assess effects on resistance against Phytophthora infestans, Fusarium solani and Rhizoctonia solani. Protection resulting from Phi treatment in seed tubers was high against P. infestans, intermediate against F. solani, and low against R. solani. In addition, seed tubers treated with calcium or potassium phosphites (CaPhi and KPhi, respectively) at 1% of commercial product emerged earlier than untreated ones. When Phi were foliarly applied two or four times at different doses, high levels of protection against P. infestans were achieved in both cultivars. Higher protection was observed in Kennebec when CaPhi was applied, while in Shepody this was true for KPhi. Expression of β-1,3-glucanases was induced at different times after treatment but no correlation between β-1,3-glucanases expression and foliar protection level was found. On the other hand, Phi positive protection effects did not produce negative effects in plant growth. Leaves from CaPhi-treated plants showed a darker green colour than leaves from control plants; also an increase in Rubisco protein and a delay in crop senescence was observed.     

Involvement of Iron and Ferritin in the Potato–Phytophthora infestans Interaction

This work shows that the infection of potato (Solanum tuberosum) detached leaves by the late blight pathogen Phytophthora infestans, was drastically reduced by adding deferoxamine, an exogenous iron chelator. Reactive oxygen species in leaves inoculated with P. infestans were also reduced after adding deferoxamine. A leaf ferritin cDNA fragment was obtained by PCR and used as probe for screening a tuber cDNA library. A cDNA (named StF1) encoding the iron-storing potato ferritin was cloned. StF1 is 915 bp in length and has an open reading frame of 230 amino acids that contains the information for the mature 28 kDa subunit of potato ferritin. StF1 was used as probe in northern blot hybridizations to analyze expression of the ferritin gene. In leaves, ferritin mRNA accumulated in response to pathogen attack. In tubers, ferritin mRNA increased upon treatment with the elicitor eicosapentaenoic acid. These results suggest that iron plays a role in the potato-P. infestans interaction.     

四种化合物诱导马铃薯抗晚疫病的效果及其相关防御基因表达分析

为探究水杨酸、壳聚糖、麦角甾醇和纤维二糖4种化合物诱导马铃薯抗晚疫病的效果及其作用机理,采用整株喷雾法、挑战接种法、菌丝生长速率法分析4种化合物对马铃薯晚疫病的预防作用及对其病原菌致病疫霉Phytophthora infestans的离体抑制效果,利用实时荧光定量PCR技术测定4种化合物诱导后马铃薯病程相关蛋白和防御酶基因的调控表达情况。结果显示,水杨酸、壳聚糖、纤维二糖和麦角甾醇分别处理马铃薯植株24 h后接种致病疫霉,在多个浓度下对晚疫病均有不同程度的诱导抗性,其中在1 000 μmol/L浓度（壳聚糖浓度为1 000 mg/L）时诱导抗性最好,较对照显著提高,诱抗效果分别为54.73%、61.99%、59.73%和48.59%。壳聚糖对马铃薯晚疫病的诱抗效果高于其它3种化合物,且只有壳聚糖能显著抑制致病疫霉的菌丝生长和孢子囊形成,说明壳聚糖除了能诱导作物产生抗病性外,还具有直接抑菌作用。4种化合物诱导后马铃薯病程相关蛋白基因PR1、过氧化物酶基因POD和多酚氧化酶基因PPO的表达量在施用早期均显著升高。表明这些化合物能诱导马铃薯对晚疫病产生抗性,可能与马铃薯体内过氧化物酶、多酚氧化酶的活性和抗病信号传导途径关键基因PR1的表达有关。     

An Aspartic Protease With Antimicrobial Activity is Induced after Infection and Wounding in Intercellular Fluids of Potato Tubers

Aspartic proteases (APs) one of the main proteinase classes, have different physiological functions in animals, fungi and viruses. In plants, knowledge of the biological roles of APs is less well developed. An AP has been purified from potato tuber and leaves (Guevara et al., 1999, 2001). In this paper, the changes in the level of AP in response to infection by Phytophthora infestans (P. infestans) and wounding were studied in intercellular washing fluids (IWFs) from tuber disks of two potato cultivars differing in their susceptibility to P. infestans. A differential induction was observed between both cultivars: in the resistant cultivar, induction was higher and faster in infected tissues than in wounded ones. In the susceptible cultivar, a lower and later accumulation was observed than in the resistant cultivar. In addition, AP had a direct inhibitory effect on the germination of cysts of P. infestans and conidia of Fusarium solani. The pattern of accumulation and in vitro activity of AP suggest that this enzyme may have a role in the defense response of potato.     

Interactions Between the Potato Cyst Nematode Globodera rostochiensis and Diseases Caused by Rhizoctonia solani AG3 in Potatoes Under Field Conditions

Findings from 2 years of field experiments investigating the relationship between Globodera rostochiensis and Rhizoctonia solani on unique field sites are reported. In 2000, a field experiment was positioned on land that had previously been used for experimental work investigating integrated potato cyst nematode (PCN) management methods. This study had produced an ‘untypical’ mosaic of PCN population densities ranging from 5 to 221 eggs g⁻¹ soil. In 2001, the field experiment was conducted on a different field site and overlaid on a focus of G. rostochiensis population densities ranging from 11 to 108 eggs g⁻¹ soil. In each experiment, potatoes (cv. Désirée) were grown in plots with similar population densities of G. rostochiensis that were either uninoculated or inoculated with R. solani. A series of potato plant harvests were undertaken to investigate the effects of nematode infestation on the incidence and severity of R. solani diseases and the associated development of plants. In both experiments, a clear relationship was found between the density of G. rostochiensis juveniles present in potato roots and the incidence of stolons infected by R. solani, 6 weeks after planting. For the first time this interaction has been determined under field conditions. The results of the study suggest that the interaction between nematode and fungus is indirect and possible mechanisms are discussed.     

<Emphasis Type="Italic">N</Emphasis>,<Emphasis Type="Italic">N</Emphasis>-dimethylsphingosine,an inhibitor of sphingosine kinase,induces phytoalexin production and hypersensitive cell death of Solanaceae plants without generation of reactive oxygen species

Plant recognition of elicitors derived from pathogens induces various resistant reactions, including production of reactive oxygen species, hypersensitive cell death and accumulation of phytoalexins. Previously, we isolated a ceramide elicitor from Phytophthora infestans, which activates O₂ ⁻ production of potato suspension-cultured cells. In this study, we employed nine ceramide-related chemicals to test their elicitor activity. Although, none of the tested chemicals induced O₂ ⁻ production, N,N-dimethylsphingosine (DMS) induced accumulation of phytoalexin in potato tubers. In potato, tobacco and Nicotiana benthamiana, DMS also induced rapid cell death. DMS-treated potato cells stained with 4′,6-diamidino-2-phenylindole (DAPI) showed chromatin condensation, and isolated DNA from DMS-treated cells had ladder pattern, confirming that DMS-induced plant cell death is a hypersensitive reaction-like programmed cell death. Involvement of ceramide signaling in induction of plant defense reactions is discussed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effects of three esca-associated fungi on Vitis vinifera L.: II. Characterization of biomolecules in xylem sap and leaves of healthy and diseased vines

Secondary metabolites and host defense compounds were shown to occur in xylem sap, and leaves of Vitis vinifera cv. Italia and cv. Matilde naturally infected by the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme). Samples of xylem sap and leaves were collected from healthy vines and from vines showing severe symptoms of brown wood-streaking caused by Pch and Tmi, or from vines with symptoms of both brown wood-streaking and white rot caused by Fme. Xylem sap collection was carried out during the early spring of 2003 and 2004, corresponding to the phenological phases: (A) cotton bud; (B) green tip; (C) leaves out; (D) stretched out leaves; and (E) visible clusters. In the present work we have studied the accumulation of biomolecules (pentaketides and α-glucans), host defense compounds (benzaldehydes, benzoic acid and cinnamic acid derivatives, flavonols, flavanols, flavan-3-ol derivatives and stilbenes) at different stages of grapevine development. Accumulation and changes in total phenolics and recurring phenolics, and of three phytotoxic secondary metabolites (scytalone, isosclerone and pullulan) were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that phenolic concentrations are strongly related to the cv.     

Effects of three esca-associated fungi on Vitis vinifera L.: I. Characterization of secondary metabolites in culture media and host responses to the pathogens in calli

Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) produced scytalone, isosclerone and pullulans in liquid cultures, as well as in calli. Secondary metabolites and host defense compounds were shown to occur in calli of Vitis vinifera cv. Italia and cv. Matilde infected by the esca-associated fungi Pch, Tmi and Fomitiporia mediterranea (Fme). Calli of both cvs. were grown as dual cultures with Pch, Tmi and Fme. The fungi grew well in the presence of calli of both cvs., but callus growth was reduced. Accumulation and changes of total phenolics and recurring phenolics, and of two phytotoxic pentaketides and pullulans were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that concentrations of phenolics are strongly related to the cv. The paper discusses the possible relationship between melanin biosynthesis in Pch and Tmi, which utilize pentaketide metabolites as intermediates and their pathogenicity related to phytotoxity of scytalone and isosclerone.     

Effects of three esca-associated fungi on Vitis vinifera L.: III. Enzymes produced by the pathogens and their role in fungus-to-plant or in fungus-to-fungus interactions

When the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme) were grown in liquid stationary cultures, it was seen that they were able to live in media containing resveratrol (RES) or tannic acid (TA) as the sole carbon source and that the fungi were able to convert both compounds. Particular attention is paid here to detecting RES and TA conversion. Pch, Tmi and Fme were partially inhibited by RES or TA. Pch, Tmi and Fme produced extracellular tannase, laccase and peroxidase enzymes in liquid or agarized cultures, whether glucose was present or not. When colonies of Pch, Tmi and Fme were confronted, they showed spatially and temporally heterogeneous patterns of laccase and peroxidase activity. The results indicate the non-synergistic, competitive association of Pch and Tmi and the inhibition of Fme growth. Muconic acid, a well-known intermediate in a large number of lignin and phenol oxidative processes, can partly or completely inhibit the lignolytic agent Fme, but is tolerated by Pch and Tmi. An explanation for wood pigmentation patterns by Pch, Tmi and Fme is given.