Yeheb (<Emphasis Type="Italic">Cordeauxia</Emphasis><Emphasis Type="Italic">edulis</Emphasis>) extract deters feeding and oviposition of <Emphasis Type="Italic">Plutella xylostella</Emphasis> and attracts its natural enemy

The effects of yeheb (Cordeauxia edulis Hemsl.) leaf extract on feeding and oviposition by diamondback moth (DBM) (Plutella xylostella L.) and the behavior of DBM parasitoid, Cotesia vestalis (Haliday), were studied. Volatile organic compounds (VOCs) from the headspace of intact and DBM-damaged broccoli plants sprayed with yeheb extracts (YE) were also analyzed. Larval feeding and growth, and oviposition by adult DBM were strongly inhibited by the extract. Cotesia vestalis were attracted to volatile blends from intact or DBM-damaged broccoli plants sprayed with YE over intact plants sprayed with water or methanol. Analyses of VOCs in the headspace of broccoli plants revealed that both intact and DBM-damaged plants sprayed with YE showed remarkable differences in sesquiterpene compounds compared to intact control treatments. These combined negative effects of YE on DBM fitness together with positive effects on the parasitoid show that yeheb is a potential source of compounds for use in integrated pest management to control damage caused by DBM.     

Characterization of volatile organic compounds emitted by kiwifruit plants infected with <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">actinidiae</Emphasis> and their effects on host defences

Key message

Specific VOC emissions from infected plants allow their recognition and elicit defence responses in neighboring plants, which are, however, insufficient to induce resistance.

Abstract

A wide range of volatile organic compounds (VOCs) is released during plant–pathogen interactions both by the pathogens and the hosts. Some of these VOCs are specific for the different diseases and are known to play a role in the pathogenicity or in plant defence responses. Besides, disease-specific VOCs may serve as markers for diagnostic protocols, which allow a non-destructive and rapid screening of bulk samples of plant material. This work aimed to verify the feasibility of a VOC-based diagnosis and to investigate the possible biological role of VOCs released during the plant–pathogen interactions. The volatile emissions from Pseudomonas syringae pv. actinidiae in axenic cultures and from inoculated in vitro kiwifruit plants were characterized by gas chromatography–mass spectrometry (GC–MS) and proton transfer reaction–time-of-flight-mass spectrometry (PTR–ToF-MS). By GC–MS, several putative biochemical markers, such as 1-undecene, were identified. PTR–ToF-MS resulted highly effective in screening the plant material for latent infections. To develop a more user-friendly, portable and less expensive diagnosis system, two different electronic nose models were tested for the early diagnosis of P. syringae pv. actinidiae in asymptomatic plant material. Our experiments demonstrated the feasibility of the electronic nose-based screening of infected plant material. Concerning the biological role of the VOCs released during the plant–pathogen interactions, the exposure of healthy plants to VOCs from infected ones influences the plant growth and induces the stimulation of protective responses. However, after the infection, P. syringae pv. actinidiae is able to selectively inactivate the induced plant defences.

     

Close-Range Host Searching Behavior of the Stemborer Parasitoids <Emphasis Type="Italic">Cotesia sesamiae</Emphasis> and <Emphasis Type="Italic">Dentichasmias busseolae</Emphasis>: Influence of a Non-Host Plant <Emphasis Type="Italic">Melinis minutiflora</Emphasis>

Studies were conducted on the host searching behavior of the larval parasitoid Cotesia sesamiae (Cameron) (Hymenoptera: Braconidae) and the pupal parasitoid Dentichasmias busseolae Heinrich (Hymenoptera: Ichneumonidae), both of which attack lepidopteran (Crambidae, Noctuidae) cereal stemborers. The behavior of D. busseolae was observed in a diversified habitat that consisted of stemborer host plants (maize, Zea mays L. and sorghum, Sorghum bicolor (L). Moench (Poaceae)) and a non-host plant (molasses grass, Melinis minutiflora Beauv. (Poaceae)), while C. sesamiae was observed separately on host plants and molasses grass. In previous olfactometer studies, C. sesamiae was attracted to molasses grass volatiles while hboxD. busseolae was repelled. The aim of the present study was to investigate the influence of molasses grass on close-range foraging behavior of the parasitoids in an arena that included infested and uninfested host plants. Dentichasmias busseolae strongly discriminated between host and non-host plants, with female wasps spending most of the time on infested host plants and least time on molasses grass. Likewise, C. sesamiae spent more time on uninfested and infested host plants than it did on molasses grass in single choice bioassays. While on infested plants, the wasps spent more time foraging on the stem, the site of damage, than on other areas of the plant. Overall, the results indicate that presence of the non-host plant does not hinder close range foraging activities of either parasitoid.     

Plant growth-promoting rhizobacteria modulate root-system architecture in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> through volatile organic compound emission

Extensive communication occurs between plants and microorganisms during different stages of plant development in which signaling molecules from the two partners play an important role. Volatile organic compounds (VOCs) emission by certain plant-growth promoting rhizobacteria (PGPR) has been found to be involved in plant growth. However, little is known about the role of bacterial VOCs in plant developmental processes. In this work, we investigated the effects of inoculation with twelve bacterial strains isolated from the rhizosphere of lemon plants (Citrus aurantifolia) on growth and development of Arabidopsis thaliana seedlings. Several bacterial strains showed a plant growth promoting effect stimulating biomass production, which was related to differential modulation of root-system architecture. The isolates L263, L266, and L272a stimulated primary root growth and lateral root development, while L254, L265a and L265b did not significantly alter primary root growth but strongly promoted lateral root formation. VOC emission analysis by SPME-GC-MS identified aldehydes, ketones and alcohols as the most abundant compounds common to most rhizobacteria. Other VOCs, including 1-octen-3-ol and butyrolactone were strain specific. Characterization of L254, L266 and L272a bacterial isolates by 16S rDNA analysis revealed the identity of these strains as Bacillus cereus, Bacillus simplex and Bacillus sp, respectively. Taken together, our data suggest that rhizospheric bacterial strains can modulate both plant growth promotion and root-system architecture by differential VOC emission.     

The floral volatile,methyl benzoate,from snapdragon (<Emphasis Type="Italic">Antirrhinum majus</Emphasis>) triggers phytotoxic effects in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Previously it has been shown that the floral scent of snapdragon flowers consists of a relatively simple mixture of volatile organic compounds (VOCs). These compounds are thought to be involved in the attraction of pollinators; however, little is known about their effect on other organisms, such as neighboring plants. Here, we report that VOCs from snapdragon flowers inhibit Arabidopsis root growth. Out of the three major snapdragon floral volatiles, myrcene, (E)-β-ocimene, and methyl benzoate (MB), MB was found to be primarily responsible for the inhibition of root growth. Ten micromoles MB reduced root length by 72.6%. We employed a microarray approach to identify the MB target genes in Arabidopsis that were responsible for the root growth inhibition phenotype in response to MB. These analyses showed that MB treatment affected 1.33% of global gene expression, including cytokinin, auxin and other plant-hormone-related genes, and genes related to seed germination processes in Arabidopsis. Accordingly, the root growth of cytokinin (cre1) and auxin (axr1) response mutants was less affected than that of the wild type by the volatile compound: roots of the treated mutants were reduced by 45.1 and 56.2%, respectively, relative to untreated control mutants. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Host Plant Perception and Selection in the Sibling Species <Emphasis Type="Italic">Macrolophus melanotoma</Emphasis> and <Emphasis Type="Italic">Macrolophus pygmaeus</Emphasis> (Hemiptera: Miridae)

The electroantennogram responses (EAGs) of Macrolophus melanotoma and Macrolophus pygmaeus (Hemiptera: Miridae) exposed to volatile compounds (VOCs) of host and non-host plants were compared. The VOCs were identified by gas chromatography. Hosts and non-hosts eliciting similar EAGs were tested in olfactory assays against plants without a significant EAGs for the two Macrolophus species. No characteristic VOC profile was found for hosts and non-hosts. Terpenes predominated in many hosts and carboxylic acids in non-hosts, but no specific VOCs were characteristic of host plants. Significant EAGs (maximum deflection values in mV) were recorded in plants with very different VOC profiles, both hosts and non-hosts. The EAGs were higher for M. melanotoma than for M. pygmaeus, and were higher for males than for females. In M. melanotoma the EAGs were greater with hosts than with non-hosts, but they were similar in M. pygmaeus. The EAGs were correlated with the concentrations of sesquiterpenes and alcohols in both species. In olfactory assays, M. melanotoma and M. pygmaeus preferred their respective hosts, but they did not discriminate between non-host with and without significant EAGs. According to the results, Macrolophus species are expected to rely on ubiquitous VOCs for the identification of their hosts. The variation in the EAGs between M. melanotoma and M. pygmaeus is attributed to the variation in the proportions of olfactory receptor neurones with different sensitivity to VOCs (e.g. sesquiterpenes). Host plant selection is discussed in the light of the perception of VOCs and the processing of information by the central nervous system.     

A key volatile infochemical that elicits a strong olfactory response of the predatory mite <Emphasis Type="Italic">Neoseiulus californicus</Emphasis>, an important natural enemy of the two-spotted spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Herbivore-induced plant volatiles (HIPVs) emitted from lima bean leaves infested with the two-spotted spider mites Tetranychus urticae strongly attract the predatory mites Neoseiulus californicus. Among these HIPVs, methyl salicylate and linalool can attract the predators. Three green-leaf volatiles (GLVs) of (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate and (E)-2-hexenal, found in the odor blends from T. urticae-infested leaves and physically damaged leaves, can also attract the predators. To search for a strong predator attractant, the olfactory responses of N. californicus to each synthetic compound or their combinations were investigated in a Y-tube olfactometer. When presented a choice between a mixture of the five compounds (i.e. the two HIPVs and the three GLVs) and T. urticae-infested leaves, N. californicus did not discriminate between these odor sources. The same trend was observed when either a mixture of the two HIPVs or methyl salicylate vs. T. urticae-infested leaves were compared. In contrast, the predators preferred T. urticae-infested leaves to linalool, each of the three GLVs, or a mixture of the three GLVs. These results indicated that methyl salicylate is a strong predator attractant, and its potential attractiveness almost equaled that of the blend of HIPVs from T. urticae-infested leaves.     

Phenolic compounds induced by <Emphasis Type="Italic">Bemisia tabaci</Emphasis> and <Emphasis Type="Italic">Trialeurodes vaporariorum</Emphasis> in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L. and their relationship with the salicylic acid signaling pathway

Changes in the levels of secondary compounds can trigger plant defenses. To identify phenolic compounds induced by Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) in tobacco (Nicotiana tobacco L.), the content changes of 11 phenolic compounds in plants infested by B. tabaci MEAM1 or Trialeurodes vaporariorum were compared. The chlorogenic acid, catechin, caffeic acid, p-coumaric acid, rutin, and ferulic acid contents in B. tabaci MEAM1-infested tobacco plants increased significantly, having temporal and spatial effects, compared with uninfested control and T. vaporariorum infested plants. The contents were 4.10, 2.84, 2.25, 3.81, 1.46, and 1.91 times higher, respectively, than those in the control. However, a T. vaporariorum nymphal infestation just caused smaller chlorogenic acid, catechin, caffeic acid, and rutin contents increase, which were 2.33, 2.13, 1.59, and 3.19 times higher, respectively, than those in the control. In B. tabaci MEAM1 third-instar nymph-infested plants, chlorogenic acid, catechin, caffeic acid, and rutin increased more significantly in systemic than in local leaves. Salicylate-deficient plants inhibited the induction of the content of 10 phenolic compounds, but not caffeic acid, after a B. tabaci MEAM1 nymphal infestation. Thus, the elevated levels of phenolic compounds induced by B. tabaci MEAM1 were correlated with the salicylic acid signaling pathway and induced the responses of defense-related phenolic compounds.     

Cyclolavandulyl butyrate: an attractant for a mealybug parasitoid, <Emphasis Type="Italic">Anagyrus</Emphasis><Emphasis Type="Italic">sawadai</Emphasis> (Hymenoptera: Encyrtidae)

In this study, we discovered and isolated an attractant for a mealybug-parasitic wasp Anagyrus sawadai from an esterification product prepared with commercialized lavandulol (2-isopropenyl-5-methyl-4-hexen-1-ol) and butyryl chloride. Using gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy, we determined the structure of the compound to be (2,4,4-trimethyl-2-cyclohexenyl)methyl butyrate (cyclolavandulyl butyrate). This is a novel compound as far as we know, although the alcohol moiety has been known as a cyclization product from lavandulol. Cyclolavandulyl butyrate has two enantiomers, and the (−)-isomer, which is suggested to have S configuration, showed higher attractiveness. A potential use for the A. sawadai attractant for mealybug management in agricultural fields is discussed.     

Effects of <Emphasis Type="Italic">Paecilomyces fumosoroseus</Emphasis> and <Emphasis Type="Italic">Encarsia formosa</Emphasis> on the control of the greenhouse whitefly: preliminary assessment of a compatability study

The effect of separate and combined activity of Paecilomyces fumosoroseus Wize (Brown and Smith) Trinidadian strain T11 and the parasitoid, Encarsia formosa Gahan, was assessed on populations of the greenhouse whitefly, Trialeurodes vaporariorum (Westwood), infesting Phaseolus vulgaris L. (French bean) and Pelargonium x domesticum (regal geranium) plants in replicate experiments. When infested bean and geranium plants were exposed to E. formosa for 2 days, and 4 days later sprayed with P. fumosoroseus blastospores, whitefly percent mortality was 99.5% and 75.5%, 94.6% and 59.4% for experiments 1 and 2, respectively. Treatment of infested bean plants with either E. formosa or P. fumosoroseus resulted in 87.8% and 78.7%, 73.1% and 97.0% whitefly mortality for experiments 1 and 2, respectively, while similar treatment of infested geranium plants resulted in 9.2% and 52.8%, 34.3% and 64.5% whitefly mortality for experiments 1 and 2, respectively. Our results support the use of E. formosa and P. fumosoroseus in combination in Experiment 1 for the treatment of whitefly infested P. vulgaris plants since a significant difference in mortality is observed than when either E. formosa or P. fumosoroseus is applied alone. However, in experiment 2, the combination treatment on P. vulgaris was no more effective than spraying P. fumosoroseus alone. On P. x domesticum plants, only P. fumosoroseus alone is needed for efficient control of the whitefly compared to the combination treatment. The relative timing of parasitoid oviposition and fungal infection are critical in determining the outcome of the interaction and are plant host dependent.     

Tissue culture of <Emphasis Type="Italic">Sinningia speciosa</Emphasis> and analysis of the <Emphasis Type="Italic">in vitro</Emphasis>-generated <Emphasis Type="Italic">tricussate whorled phyllotaxis</Emphasis> (<Emphasis Type="Italic">twp</Emphasis>) variant

Two protocols were developed for the efficient regeneration of Sinningia speciosa from leaf explants via two developmental pathways. The first method involved formation of callus and buds, followed by subsequent root growth, in Murashige and Skoog medium (MS) containing 2.0 mg l⁻¹ 6-benzylaminopurine (BA) and 0.2 mg l⁻¹ α-naphthalene acetic acid (NAA), with a regeneration efficiency of 99.0%. The second method involved producing callus and roots, followed by subsequent formation of buds, in MS medium supplemented with 1.0–5.0 mg l⁻¹ NAA, and resulted in a regeneration efficiency of 90.4%. Our experiments indicate that the root-first pathway resulted in a lower plant regeneration efficiency. Through five continual generations using the buds-first method, a total of 215 regenerated plants were obtained in the last generation, and eight exhibited a phenotype we named tricussate whorled phyllotaxis (twp). Six of the regenerated twp variant plants maintained their tricussate whorled phyllotaxis phenotype, showing no other abnormalities, while one reverted to a wild type before flowering and another formed two rounds of sepals. Physiological analysis revealed that the twp plants responded differently than wild type to exogenous NAA and 2,3,5-triiodobenzoic acid (TIBA), while high-performance liquid chromatography (HPLC) analysis showed that the levels of endogenous indole-3-acetic acid (IAA) and gibberellin (GA) were lower in twp than wild-type plants. These results suggest that the formation of the twp mutant may be related to phytohormones and that the twp variant could be an important material for investigating the molecular mechanism of plant phyllotaxis patterning.     

An Endophytic <Emphasis Type="Italic">Phomopsis</Emphasis> sp. Possessing Bioactivity and Fuel Potential with its Volatile Organic Compounds

An unusual Phomopsis sp. was isolated as endophyte of Odontoglossum sp. (Orchidaceae), associated with a cloud forest in Northern Ecuador. This fungus produces a unique mixture of volatile organic compounds (VOCs) including sabinene (a monoterpene with a peppery odor) only previously known from higher plants. In addition, some of the other more abundant VOCs recorded by GC/MS in this organism were 1-butanol, 3-methyl; benzeneethanol; 1-propanol, 2-methyl and 2-propanone. The gases of Phomopsis sp. possess antifungal properties and an artificial mixture of the VOCs mimicked the antibiotic effects of this organism with the greatest bioactivity against a wide range of plant pathogenic test fungi including: Pythium, Phytophthora, Sclerotinia, Rhizoctonia, Fusarium, Botrytis, Verticillium, and Colletotrichum. The IC₅₀ values for the artificial gas mixture of Phomopsis sp. varied between 8 and 25.65 μl/mL. Proton transfer reaction-mass spectrometry monitored the concentration of VOCs emitted by Phomopsis sp. and yielded a total VOC concentration of ca. 18 ppmv in the head space at the seventh day of incubation at 23°C on PDA. As with many VOC-producing endophytes, this Phomopsis sp. did survive and grow in the presence of the inhibitory gases of Muscodor albus. A discussion is presented on the possible involvement of VOC production by the fungus and its role in the biology/ecology of the fungus/plant/environmental relationship.     

Terpenoids dominate the bouquet of volatile organic compounds produced by <Emphasis Type="Italic">Passiflora edulis</Emphasis> in response to herbivory by <Emphasis Type="Italic">Heliconius erato phyllis</Emphasis> (Lepidoptera: Nymphalidae)

In response to injury, plants produce volatile organic compounds (VOCs) that usually differ depending on the type of damage they have suffered (e.g., mechanical damage, herbivory, and oviposition). The objectives of this study were to identify and compare the bouquet of volatiles emitted by passion vine plants (Passiflora edulis) after injury caused by mechanical damage (MD), herbivory (HB), and oviposition (OV) by the lepidopteran, Heliconius erato phyllis. Following injury, extracts of plant emissions were collected from each treatment every 24 h for three days and were analyzed by GC and GC/MS. Results show that plants emitted 12 volatiles before and after damage, namely terpenoids, ketones, and aldehydes. Although no significant differences were detected between the three treatments individually, if the entire bouquet of volatiles is analyzed, samples collected at 24 h were different from samples collected at 48 and 72 h. However, terpenoid emission increased significantly in HB plants after 24 h. HB plants emitted approximately 6300, 50, 46, 11, 6, and 3.6 times more (3E,7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), (E)-β-ocimene, (Z)-β-farnesene, (E)-β-caryophyllene, and farnesane, respectively, compared to control plants. OV plants displayed a peak of emission of (E)-β-ocimene after 72 h, which distinguished them from HB plants. MD plants showed a general increase of VOCs versus undamaged control plants. Furthermore, it has been suggested that (E)-β-ocimene may be sequestered by larvae of H. erato phyllis as a component of the odoriferous bouquet of the abdominal scent glands present in adult males, which play a role in sexual communication.     

Volatiles of bacterial antagonists inhibit mycelial growth of the plant pathogen <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Bacterial antagonists are bacteria that negatively affect the growth of other organisms. Many antagonists inhibit the growth of fungi by various mechanisms, e.g., secretion of lytic enzymes, siderophores and antibiotics. Such inhibition of fungal growth may indirectly support plant growth. Here, we demonstrate that small organic volatile compounds (VOCs) emitted from bacterial antagonists negatively influence the mycelial growth of the soil-borne phytopathogenic fungus Rhizoctonia solani Kühn. Strong inhibitions (99–80%) under the test conditions were observed with Stenotrophomonas maltophilia R3089, Serratia plymuthica HRO-C48, Stenotrophomonas rhizophila P69, Serratia odorifera 4Rx13, Pseudomonas trivialis 3Re2-7, S. plymuthica 3Re4-18 and Bacillus subtilis B2g. Pseudomonas fluorescens L13-6-12 and Burkholderia cepacia 1S18 achieved 30% growth reduction. The VOC profiles of these antagonists, obtained through headspace collection and analysis on GC-MS, show different compositions and complexities ranging from 1 to almost 30 compounds. Most volatiles are species-specific, but overlapping volatile patterns were found for Serratia spp. and Pseudomonas spp. Many of the bacterial VOCs could not be identified for lack of match with mass-spectra of volatiles in the databases.     

Host plant volatiles serve to increase the response of male European grape berry moths, <Emphasis Type="Italic">Eupoecilia ambiguella,</Emphasis> to their sex pheromone

The European grape berry moth is an important pest in vineyards. Males respond to the female-produced sex pheromone released from a piezo nebulizer in a dose-dependent manner in a wind tunnel: <50% arrive at the source at 5–50 pg/min (underdosed), 80% arrive at 100 pg/min to 10 ng/min (optimal) and <20% arrive at 100 ng/min (overdosed). Males responding to overdosed pheromone show in flight arrestment at 80 cm from the source. Host plant chemostimuli for Eupoecilia ambiguella increase the responses of males to underdosed and overdosed pheromone. (Z)-3-hexen-1-ol, (+)-terpinen-4-ol, (E)-β-caryophyllene and methyl salicylate released with the underdosed pheromone cause a significant increase in male E. ambiguella flying to the source. Time–event analysis indicates a positive correlation between faster activation and probability of source contact by the responding males. The four host plant compounds added to the overdosed pheromone permitted males to take off faster and with a higher probability of flying to the source. This suggests that perception of host plant products with the sex pheromone facilitates male E. ambiguella to locate females on host plants, lending credence to the hypothesis that plant products can signal rendezvous sites suitable for mating. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Accumulation of salicylic acid,jasmonic acid and phytoalexins in rice, <Emphasis Type="Italic">Oryza sativa</Emphasis>, infested by the white-backed planthopper, <Emphasis Type="Italic">Sogatella furcifera</Emphasis> (Hemiptera: Delphacidae)

In order to clarify the mechanism of induced resistance to blast disease in rice, Oryza sativa, that had been previously infested by the white-backed planthopper, Sogatella furcifera Horváth, we first investigated the accumulation of salicylic acid (SA) and jasmonic acid (JA) in rice plants infested by the planthopper. The results confirmed that infestation of S. furcifera strongly stimulates the production of SA and JA in rice. These results indicate that both salicylate- and jasmonate-mediated pathways (SA and JA pathways), which are involved in the general defense system in plants, were activated in rice infested by S. furcifera. Further results confirmed that S. furcifera infestation induces accumulation of a major rice diterpenoid phytoalexin, momilactone A, and a flavonoid phytoalexin, sakuranetin, which are well known as antimicrobial chemicals, particularly in blast disease caused by the blast fungus, Magnaporthe oryzae B. Couch. All these results strongly suggest the following hypothetical mechanism of induced-resistance to M. oryzae in rice infested by S. furcifera. First, S. furcifera releases some elicitor-active compounds, which might be produced in the salivary glands, into the rice plant during feeding. Next, the defense signal systems, SA- and JA-mediated pathways, are activated by the elicitor. Finally, phytoalexins are induced in rice as antimicrobial compounds mainly through activation of the JA-mediated pathway.     

Isolation and characterization of <Emphasis Type="Italic">NgRLK1</Emphasis>, a receptor-like kinase of <Emphasis Type="Italic">Nicotiana glutinosa</Emphasis> that interacts with the elicitin of <Emphasis Type="Italic">Phytophthora capsici</Emphasis>

Elicitins, extracellular proteins from Phytophthora fungi, elicit a hypersensitivity response (HR), including systemic acquired resistance, in some plants. The elicitin capsicein (~10 kDa) was purified by FPLC from culture filtrates of P. capsici. Purified native and recombinant capsicein induced a hypersensitive response in leaves of the non-host plants Nicotiana glutinosa and Brassica rapa subsp. pekinensis. To search for candidate capsicein-interacting proteins from N. glutinosa, a yeast two-hybrid assay was used. We identified a protein interactor that is homologous to a serine/threonine kinase of the plant receptor-like kinase (RLK) group and designated it NgRLK1. The ORF of NgRLK1 encodes a polypeptide of 832 amino acids (93,490 Da). A conserved domain analysis revealed that NgRLK1 has structural features typical of a plant RLK. NgRLK1 was autophosphorylated, with higher activity in the presence of Mn²⁺ than Mg²⁺.     

Expression of the <Emphasis Type="Italic">Escherichia coli</Emphasis> heat-labile enterotoxin B subunit in transgenic watercress (<Emphasis Type="Italic">Nasturtium officinale</Emphasis> L<Emphasis Type="Italic">.</Emphasis>)

A gene encoding the B subunit of the enterotoxigenic Escherichia coli heat-labile enterotoxin (LTB) was adapted to the optimized plant coding sequence, and fused to the endoplasmic reticulum retention signal SEKDEL in order to enhance its expression level and protein assembly in plants. The synthetic LTB (sLTB) gene was placed into a plant expression vector under the control of the CaMV 35S promoter, and subsequently introduced into the watercress (Nasturtium officinale L.) plant by the Agrobacterium-mediated transformation method. The integration of the sLTB gene into the genomic DNA of transgenic plants was confirmed by genomic DNA PCR amplification. The assembly of plant-produced LTB protein was detected by western blot analysis. The highest amount of LTB protein produced in transgenic watercress leaf tissue was approximately 1.3% of the total soluble plant protein. G_M1-ganglioside enzyme-linked immunosorbent assay indicated that plant-synthesized LTB protein bound specifically to G_M1-ganglioside, which is the receptor for biologically active LTB on the cell surface, suggesting that the plant-synthesized LTB subunits formed biologically active pentamers.