Abstract: Elicitins and lipid-transfer proteins are small cysteine-rich lipid-binding proteins secreted by oomycetes and plant cells, respectively, that share some structural and functional properties. In spite of intensive work on their structure and diversity at the protein and genetic levels, the precise biological roles of lipid-transfer proteins remains unclear, although the most recent data suggest a role in somatic embryogenesis, in the formation of protective surface layers and in defence against pathogens. By contrast, elicitins are known elicitors of plant defence, and recent work demonstrating that elicitins and lipid-transfer proteins share the same biological receptors gives a new perspective to understand the role played by lipid binding proteins,mainly the early recognition of intruders in plants.
Abstract: Palmivorein, a new member of the elicitin family, was purified from the culture filtrate of Phytophthora palmivora isolated from the rubber tree, Hevea brasiliensis. The elicitin was obtained by ammonium sulfate precipitation and further purified using ion-exchange and gel filtration. The molecular weight, isoelectric point, amino acid composition and N-terminal sequences of this molecule are reported and compared to other known elicitins. Palmivorein, as determined by SDS-PAGE, is a small protein of Mr ca. 10,000. It is classified as an a-elicitin according to its acidic pI and the valine residue at position 13. Like other elicitins, the P. palmivora elicitin causes tissue necrosis on tested tobacco leaves. It also causes severe wilting and necrosis of Hevea tissue, and leaves of the susceptible rubber clone (with respect to P. palmivora) are much more sensitive to this elicitin than those that are resistant.
Abstract: he rapid and effective activation of disease resistance responses is essential for plant defense against pathogen attack. These responses are initiated when
pathogen-derived molecules (elicitors) are recognized by the host. A cDNA encoding elicitor, the major secreted extracellular glycoprotein of Phytophthora colocasiae,a pathogen of taro (Colocasia esculenta) plants, was isolated, sequenced and characterized. The expression of the corresponding elicitor gene during the disease cycle of P. colocasiae was analyzed. Elicitor was shown to be expressed in mycelium grown in culture media, whereas it was not expressed in sporangiospores and zoospores. In planta, during infection of taro, particularly during the biotrophic stage, expression of elicitor was down-regulated compared to in vitro. The highest levels of expression of elicitor were observed in in vitro grown mycelium and in late stages of infection when profuse sporulation and leaf necrosis occur. The elicitation of the suspension-cultured taro cells was effective in the induction of the enzyme activity of L-phenylalanine-ammonia lyase, peroxidase and lipoxygenase as well as the expression of defense-related endochitinase gene. All these biological activities were exerted within a low concentration range. The glycoprotein represents a powerful tool to investigate further the signals and their transduction pathways involved in induced disease resistance. It may also be useful to engineer broad disease protection in taro plant against Phytophthora leaf blight.
Abstract: Recent advances in cloning avirulence genes from a rust fungus and three oomycete species have provided the novel insight that these eukaryotic plant pathogens deliver small proteins into the host cell cytoplasmwhere they are recognized by resistance proteins. Anne Rehmany et al. have recently identified a potential host-targeting signal in oomycete avirulence proteins from Hyaloperonospora parasitica, Phytophthora sojae and Phytophthora infestans that might be involved in transporting proteins into the host cell. This signal is surprisingly similar to the host targeting signal used by the malaria pathogen Plasmodium fulciparum to target virulence proteins to the mammalian host cell.
Abstract: We previously isolated, by differential display and 5 RACE (rapid amplification of cDNA ends), cDNAs corresponding to genes activated following cryptogein treatment of tobacco cell suspensions, among them tcI 7 (tcI for tobacco cryptogein Induced), a gene encoding a L L-subunit of proteasome. Here, we
report that tcI 7 was up-regulated in tobacco plants treated with
elicitins (cryptogein and parasiticein) that have been shown to induce a systemic acquired resistance (SAR). Moreover,subsequent inoculation of tobacco with the pathogen Phytophthora parasitica var. nicotianae (Ppn) was shown to induce an additional activation of tcI 7 in tobacco plants pretreated with cryptogein. We also showed an up-regulation of tcI 7 by salicylic acid (SA). Moreover, accumulation of tcI 7 transcripts after treatment with cryptogein or with SA only occurred in NahG 9- tobacco plants that do not express the salicylate hydroxylase and thus are able to accumulate SA and develop a SAR. Suppressed accumulation of tcI 7 transcripts in NahG 8+ tobacco plants after cryptogein or SA treatment correlated with the loss of SAR. H2O2 was also shown to up-regulate tcI 7 in tobacco plants.Using gene walking by PCR we cloned and sequenced the 5 flanking region of tcI 7 containing hypothetical regulatory sequences, especially myb and NF-U UB boxes, that could be responsible for the regulation of tcI 7 by salicylic acid and H2O2 respectively.
Abstract: Expression profiling using cDNA-AFLP is commonly used to display the transcriptome of a specific tissue or developmental stage. Here, cDNA-AFLP was used to identify transcripts in a segregating F1 population of Phytophthora infestans, the oomycete pathogen that causes late blight. To find transcripts derived from putative avirulence (Avr) genes germinated cyst cDNA from F1 progeny with defined avirulence phenotypes was pooled and used in a bulked segregant analysis (BSA). Over 30,000 transcript derived fragments(TDFs) were screened resulting in 99 Avr-associated TDFs as well as TDFs with opposite pattern. With 142 TDF sequences homology searches and database mining was carried out. cDNA-AFLP analysis on individual F1 progeny revealed 100% co-segregation of four TDFs with particular AVR phenotypes and this was confirmed by RT-PCR. Two match the same P. infestans EST with unknown sequence and this is a likely candidate for Avr4. The other two are associated with the Avr3b-Avr10-Avr11 locus. This combinedcDNA-AFLP/BSA strategy is an efficient approach to identify Avr-associated transcriptome markers that can complement positional cloning.
Abstract: Plant–pathogen interactions are still classically described using concepts that make a distinction between qualitative and quantitative aspects linked to these concepts. This article first describes these aspects, using the terminology associated with them. It then presents some recent experimental observations that demonstrate that such concepts share either common or closely related
mechanisms at the cellular and molecular levels. The emergence of a more global vision and understanding of the interactions between plants and their parasites is discussed.
Abstract: We examined the role of salicylic acid (SA) accumulation in the tobacco hypersensitive response (HR) and systemic acquired resistance(SAR) induced by PB90, a 90 kDa protein elicitor that is secreted by Phytophthora boehmeriae. The elicitor induced HR of a consistent shape and size on tobacco plants expressing the bacterial gene nahG. Salicylate hydroxylase is encodec by nahG and inactivates SA by converting it into catechol. The mutant nahG does not accumulate SA. In contrast, infiltration of a wild type tobacco (cv. Xanthi nc) leaves with the elicitor caused the increase of SA levels. The same SA levels were observed in leaves not treated by the elicitor. HR appears to be mediated by a SA-independent signaling pathway. PB90 treatment resulted in enhanced resistance of wild-type plants to infection by black
shank fungus, P. nicotianae, and TMV, but not in nahG plants. Moreover, the elicitor-induced expression of an SAR marker gene encoding PR-1a was suppressed in nahG plants. These results indicate that SA mediates SAR but not HR in tobacco treated with PB90. During plant elicitor interactions, HR and SAR may be regulated by distinct signal pathways, or SA may function as an intermediate signal upstream of SAR but downstream of HR, and HR may not be a direct defense mechanism against pathogen infection.
Abstract: The heterotrimeric G-protein pathway is a ubiquitous eukaryotic signalling module that is known to regulate growth and differentiation in many plant pathogens. We previously identified Pigpa1, a gene encoding a G-protein a subunit from the potato late blight pathogen Phytophthora infestans. P. infestans belongs to the class oomycetes, a group of organisms in which signal transduction processes have not yet been studied at the molecular level. To elucidate the function of Pigpa1, PiGPA1-deficient mutants were obtained by homology-dependent gene silencing. The Pigpa1-silenced mutants produced zoospores that turned six to eight times more frequently, causing them to swim only short distances compared with wild type. Attraction to the surface, a phenomenon known as negative geotaxis, was impaired in the mutant zoospores, as well as autoaggregation and chemotaxis towards glutamic and aspartic acid. Zoospore production was reduced by 20–45% in different Pigpa1-silenced mutants. Transformants expressing constitutively active forms of PiGPA1, containing amino acid substitutions (R177H and Q203L), showed no obvious phenotypic differencesfrom the wild-type strain. Infection efficiencies on potato leaves ranged from 3% to 14% in the Pigpa1 -silenced mutants, compared with 77% in wild type showing that virulence is severely impaired. The results prove that PiGPA1 is crucial for zoospore motility and for pathogenicity in an importan oomycete plant pathogen.
Abstract: Oomycetes are phylogenetic relatives of brown algae that cause many destructive diseases of plants, as well as several animal and human diseases. Because of oomycetes distinct physiology, most fungicides are ineffective against them. With the aid of genetic and genomic tools, oomycete genes encoding secreted proteins that control the outcome of infection are being identified. Ongoing genomics efforts promise to identify further genes and create the possibility of new control measures.
Abstract