Functional analysis of RXLR effectors from the New Zealand kauri dieback pathogen Phytophthora agathidicida.

New Zealand kauri is an ancient, iconic, gymnosperm tree species that is under threat from a lethal dieback disease caused by the oomycete Phytophthora agathidicida. To gain insight into this pathogen, we determined whether proteinaceous effectors of P. agathidicida interact with the immune system of a model angiosperm, Nicotiana, as previously shown for Phytophthora pathogens of angiosperms. From the P. agathidicida genome, we defined and analysed a set of RXLR effectors, a class of proteins that typically have important roles in suppressing or activating the plant immune system. RXLRs were screened for their ability to activate or suppress the Nicotiana plant immune system using Agrobacterium tumefaciens transient transformation assays. Nine P. agathidicida RXLRs triggered cell death or suppressed plant immunity in Nicotiana, of which three were expressed in kauri. For the most highly expressed, P. agathidicida (Pa) RXLR24, candidate cognate immune receptors associated with cell death were identified in Nicotiana benthamiana using RNA silencing-based approaches. Our results show that RXLRs of a pathogen of gymnosperms can interact with the immune system of an angiosperm species. This study provides an important foundation for studying the molecular basis of plant-pathogen interactions in gymnosperm forest trees, including kauri.

Gymnosperm glandular trichomes: expanded dimensions of the conifer terpenoid defense system.

Glandular trichomes (GTs) are defensive structures that produce and accumulate specialized metabolites and protect plants against herbivores, pathogens, and abiotic stress. GTs have been extensively studied in angiosperms for their roles in defense and biosynthesis of high-value metabolites. In contrast, trichomes of gymnosperms have been described in fossilized samples, but have not been studied in living plants. Here, we describe the characterization of GTs on young stems of a hybrid white spruce. Metabolite and histological analysis of spruce GTs support a glandular function with accumulation of a diverse array of mono-, sesqui- and diterpenes including diterpene methylesters. Methylated diterpenes have previously been associated with insect resistance in white spruce. Headspeace analysis of spruce GTs showed a profile of volatiles dominated by monoterpenes and a highly diverse array of sesquiterpenes. Spruce GTs appear early during shoot growth, prior to the development of a lignified bark and prior to accumulation of terpenes in needles. Spruce GTs may provide an early, terpene-based chemical defense system at a developmental stage when young shoots are particularly vulnerable to foliage and shoot feeding insects, and before the resin duct system characteristic of conifers has fully developed.

Cross-reactivity of pollen allergens: impact on allergen immunotherapy.

OBJECTIVE: To provide guidelines for the rational formulation of allergen immunotherapy extracts based on knowledge of pollen allergen and epitope cross-reactivity. DATA SOURCES: A PubMed search was performed for articles published from 1966 to 2007 using the keywords pollen, allergen, and cross-reactivity. Older literature was found through cross-referencing of older articles and older reviews on pollen cross-reactivity. STUDY SELECTION: Articles that dealt with crude pollen extracts and characterized allergens that addressed cross-reactivity were selected for inclusion in this review. RESULTS: In addition to unique allergens, several families of botanic proteins have similarities that allow them to act as pan-allergens. Although frequently these are minor allergens, in some circumstances they may also be major allergens. Recent studies have investigated nonspecific lipid transfer proteins, calcium-binding proteins, pathogenesis-related protein families, and profilins. Calcium-binding proteins and nonspecific lipid transfer proteins are responsible for pollen-fruit interactions and pollen cross-reactivity. Clarification of pollen allergen enzymatic activity helps explain the ubiquitous nature of these proteins. CONCLUSION: Characterization of specific pollen allergens and their protein families has provided insight into cross-reactivity. Clarification of these relationships allows for consolidation or substitution in formulation of inhalant extracts.

[Application of the serological method for evaluation of relations between gymnosperms and dicotyledonous plants]. / Ispol'zovanie serologicheskogo metoda dlia otsenki otnoshenii golosemennykh i dvudol'nykh.

An attempt has been undertaken to evaluate interrelations of gymnospermous and dicotyledonous plants on the basis of immunochemical studies of seed proteins. For this purpose, 12 antisera were raised to proteins of taxa representing four gymnosperm classes: Ginkgoopsida, Cycadopsida, Coniferopsida, and Gnetopsida. Seed proteins of eight dicotyledonous subclasses (after Takhtadzhyan, 1987) were used. The representatives of all dicotyledonous subclasses gave immunochemical reactions with those of all gymnospermous classes. The data obtained suggest the presence of sufficiently close immunochemical relations between gymnosperms and dicotyledons. Samples were found among the representatives of subclasses Dilleniidae, Hamamelididae, and Rosidae, which gave satisfactory reactions with eight to ten antisera to proteins of dicotyledonous seeds. Analysis of the data we obtained suggests that gymnospermous and dicotyledonous plants took their origin from a common pragymnospermous ancestor and later evolved independently or that dicotyledons separated from gymnosperms at an early stage of their evolution before divergence of the latter into several phyletic lineages.

Cupressaceae pollinosis: identification, purification and cloning of relevant allergens.

Allergy to Cupressaceae pollen is a worldwide pollinosis caused by several species. Pollen extracts prepared from allergenic species belonging to this family are characterised by low protein and high carbohydrate content. The allergenic components represented in the pollen extracts from different species of the Cupressaceae family show high levels of cross-reactivity when probed with human IgE from allergic subjects and share a number of common epitopes also identified by polyclonal rabbit antisera and monoclonal antibodies. A close relationship has also been described with the Taxodiaceae and Podocarpaceae families. Although both proteic and carbohydrate epitopes appear to be involved in IgE recognition and allergenic cross-reactivity, a large portion of the IgE reactivity of Cupressaceae-allergic patients seems to be associated with sugar moieties present on the relevant allergenic molecules. From this point of view, Cupressaceae/Taxodiaceae allergens constitute a particularly useful model to study IgE cross-reactivity, as they have been shown to display different levels of homology. Moreover, the availability of the purified allergens, together with their recombinant counterparts, may shed light on the actual role played by carbohydrate in allergic sensitisation, IgE recognition and allergenic cross-reactivity.

Seasonal rhinitis in a cat sensitized to Japanese cedar (Cryptomeria japonica) pollen.

A cat showing seasonal allergic symptoms of rhinitis was examined for reactivities to Japanese cedar (Cryptomeria japonica, CJ) pollen allergen by intradermal skin test (IDST), Prausnitz-Kustner (P-K) test, and lymphocyte blastogenic response. In IDST for 26 common allergens. the cat showed a positive reaction to CJ pollen allergen. P-K test using CJ pollen allergen also showed a positive reaction, indicating the presence of serum IgE specific to CJ pollen. In the lymphocyte blastogenic response, the stimulation index in the presence of CJ pollen allergen was 2.4. These data suggested that the seasonal rhinitis observed in the cat was caused by the sensitization to CJ pollen allergen.

Clinical and immunologic evaluation of Cedrus deodara pollen: a new allergen from India.

BACKGROUND: Allergy to pollen from gymnosperms is well documented in the West. However, many allergenic species are native to the Himalayan region of India, and Cedrus deodara (Pinaceae) was selected for allergologic investigation. The objective was to define the allergologic and immunochemical aspects of C. deodara pollen. METHODS: Pollen antigen from C. deodara (CD) was prepared and characterized by biochemical and biologic assays. Specific IgE binding was determined by means of ELISA and immunoblotting. RESULTS: CD pollen antigen caused marked skin sensitivity in 7.5% of an atopic population. A significantly elevated level of CD-specific IgE antibodies was observed in 65.8% of the skin-positive patients. Immunoblotting showed protein fractions of 37, 44, 58, and 78 kDa with 100% binding with the patients' sera suspected to be due to carbohydrate moieties. CONCLUSIONS: Patients from the Himalayan region, where CD occurs naturally, were sensitized more than patients from distant places. The immunochemical characterization revealed multiple protein fractions from low to very high molecular mass (14-126 kDa) mostly in the acidic pI range. CD pollen has been recognized as a new allergen from India for the first time. The role of pollen as a causative agent of respiratory allergic disorders is very well established, as is evident from the recent increase of reports from across the world (1-4). India is blessed with the richest flora on the earth, from alpine tundra to Rajasthan desert. Consequently, it provides considerable variation in the quality and quantity of airborne pollen in different ecogeographic regions of the country (5-8). Although studies on the allergenic properties of airborne pollen from various species have been carried out by several workers in India (9-12), information on allergy to aerial pollen from Himalayan tree species has been completely