Utilization of chemical inducers of resistance and Cryptococcus flavescens OH 182.9 to reduce Fusarium head blight under greenhouse conditions

Four chemicals [salicylic acid (SA), sodium salt of salicylic acid (NaSA), isonicotinic acid (INA), and DL-β-amino-n-butyric acid (BABA)] and the yeast antagonist Cryptococcus flavescens (=C. nodaensis nomen nudum) OH 182.9 were evaluated separately or together for the ability to reduce Fusarium head blight (FHB) of wheat in the greenhouse. When sprayed onto wheat heads at 3 days prior to pathogen challenge with Gibberella zeae, NaSA and INA at 10 mM significantly reduced FHB severity compared to the non-treated disease control. Applied at concentrations of 1 and 5 mM at 3 days before pathogen challenge, NaSA or INA in combination with OH 182.9 did not significantly reduce FHB severity compared to either treatment alone, though the lowest disease severity values frequently were associated with the combination treatments. When sprayed onto wheat heads just beginning to emerge from boot at 10 days prior to pathogen inoculation, NaSA, INA, and BABA at 1 mM significantly reduced FHB severity indicating that induced systemic resistance was at least partially responsible for the reduction of FHB disease. Induced FHB resistance was achieved by treating wheat with INA at concentrations as low as 0.1 mM. In only one instance was 100-kernel weight affected by any chemical or combination of chemicals with OH 182.9 treatment. Data from our studies in the greenhouse suggest that chemical inducers can induce resistance in wheat against FHB, and that further efforts are warranted to explore the potential of improved control of FHB disease by incorporating chemical inducers with the FHB biocontrol agent OH 182.9.     

Inducible expression of bacterio-opsin in transgenic tobacco and tomato plants

The development of new strategies to enhance resistance of plants to pathogens is instrumental in preventing agricultural losses. Lesion mimic, the spontaneous formation of lesions resembling hypersensitive response lesions in the absence of a pathogen, is a dramatic phenotype occasionally induced upon expression of certain transgenes in plants. These transgenes simulate the presence of a pathogen and, therefore, activate the plant anti-pathogen defense mechanisms and induce a state of systemic resistance. Lesion mimic genes have been successfully used to enhance the resistance of a number of different plants to pathogen attack. However, constitutive expression of these genes in plants is associated with the spontaneous formation of lesions on leaves and stems, reduced growth, and lower yield. We tested the possibility of using a wound-inducible promoter to control the expression of bacterio-opsin (bO), a transgene that confers a lesion mimic phenotype in tobacco and tomato plants when constitutively expressed. We found that plants with inducible expression of bO did not develop spontaneous lesions. Nevertheless, under controlled laboratory conditions, they were found to be resistant to infection by pathogens. The activation of defense mechanisms by the bO gene was not constitutive, and occurred in response to wounding or pathogen infection. Furthermore, wounding of transgenic tobacco plants resulted in the induction of systemic resistance to pathogen attack within 48 h. Our findings provide a promising initial assessment for the use of wound-inducible promoters as a new strategy to enhance pathogen resistance in transgenic crops by means of lesion mimic genes.     

<Emphasis Type="Italic">Piriformospora indica</Emphasis> and <Emphasis Type="Italic">Sebacina vermifera</Emphasis> increase growth performance at the expense of herbivore resistance in <Emphasis Type="Italic">Nicotiana attenuata</Emphasis>

A Sebacinales species was recovered from a clone library made from a pooled rhizosphere sample of Nicotiana attenuata plants from 14 native populations. Axenic cultures of the related species, Piriformospora indica and Sebacina vermifera, were used to examine their effects on plant performance. Inoculation of N. attenuata seeds with either fungus species stimulated seed germination and increased growth and stalk elongation. S. vermifera inoculated plants flowered earlier, produced more flowers and matured more seed capsules than did non-inoculated plants. Jasmonate treatment during rosette-stage growth, which slows growth and elicits herbivore resistance traits, erased differences in vegetative, but not reproductive performance resulting from S. vermifera inoculation. Total nitrogen and phosphorous contents did not differ between inoculated and control plants, suggesting that the performance benefits of fungal inoculation did not result from improvements in nutritional status. Since the expression of trypsin proteinase inhibitors (TPI), defensive proteins which confer resistance to attack from Manduca sexta larvae, incur significant growth and fitness costs for the plant, we examined the effect of S. vermifera inoculation on herbivore resistance and TPI activity. After 10 days of feeding on S. vermifera-inoculated plants, larval mass was 46% higher and TPI activity was 48% lower than that on non-inoculated plants. These results suggest that Sebacina spp. may interfere with defense signaling and allow plants to increase growth rates at the expense of herbivore resistance mediated by TPIs.     

Community‐level changes in Banksia woodland following plant pathogen invasion in the Southwest Australian Floristic Region

Question: Does the introduced pathogen Phytophthora cinnamomi change Banksia woodland α‐ or β‐diversity and what are the implications for species re‐colonization? Location: High rainfall zone of the Southwest Australian Floristic Region (SWAFR). Methods: We measured pathogen‐induced floristic change along a disease chronosequence, and re‐sampled historic quadrats in Banksia attenuata woodlands of the SWAFR. The chronosequence represents three disease stages: (1) healthy vegetation with no disease expression; (2) the active disease front; and (3) diseased vegetation infected for at least 15 years. Comparative data were obtained by resampling diseased plots that were historically disease‐free when established in 1990. Results: β‐diversity differed substantially for both chronosequence and historic data, while α‐diversity was maintained, as measured by plot species richness and Simpson's reciprocal index. Species of known pathogen susceptibility were significantly reduced in cover–abundance, including the structurally dominant species; Banksia attenuata, B. ilicifolia and Allocasuarina fraseriana. Although these species remained present on diseased sites, there were overall reductions in canopy closure, leaf litter and basal area. These declines were coupled with an increase of species with unknown susceptibility, suggesting potential resistance and capacity to take advantage of altered site conditions. Conclusions: This study highlights the ability of an introduced plant pathogen to alter community floristics and associated stand variables. Species cover–abundances are unlikely to recover due to a reduced seed source, altered site conditions and pathogen persistence at the landscape level. However, maintenance of α‐diversity suggests continued biological significance of Phytophthora‐affected sites and the formation of novel ecosystems, themselves worthy of conservation.     

Defining Plant Resistance to Phytophthora cinnamomi: A Standardized Approach to Assessment

Phytophthora cinnamomi is a soil‐borne plant pathogen that causes devastating disease in agricultural and natural systems worldwide. While a small number of species survive infection by the pathogen without producing disease symptoms, the nature of resistance, especially under controlled conditions, remains poorly understood. At present, there are no standardized criteria by which resistance or susceptibility to P. cinnamomi can be assessed, and we have used five parameters consisting of plant fresh weight, root growth, lesion length, relative chlorophyll content of leaves and pathogen colonization of roots to analyse responses to the pathogen. The parameters were tested using two plant species, Zea mays and Lupinus angustifolius, through a time course study of the interactions and resistance and susceptibility defined 7 days after inoculation. A scoring system was devised to enable differentiation of these responses. In the resistant interaction with Z. mays, there was no significant difference in fresh weight, root length and relative chlorophyll content in inoculated compared with control plants. Both lesion size and pathogen colonization of root tissues were limited to the site of inoculation. Following inoculation L. angustifolius showed a significant reduction in plant fresh weight and relative leaf chlorophyll content, cessation of root growth and increased lesion lengths and pathogen colonization. We propose that this technique provides a standardized method for plant–P. cinnamomi interactions that could be widely used to differentiate resistant from susceptible species.     

<Emphasis Type="Italic">Colletotrichum gloeosporioides</Emphasis> can Overgrow <Emphasis Type="Italic">Colletotrichum kahawae</Emphasis> on Green Coffee Berries First Inoculated with <Emphasis Type="Italic">C. kahawae</Emphasis>

Colletotrichum gloeosporioides is a weak pathogen of coffee that infects ripe berries at dark red stage causing necrotic lesions, but only penetrates up to the second superficial layers of the pericarp at the rose and pink stages. C. kahawae, the causal agent of coffee berry disease (CBD) and responsible for 70–80% of crop loss, infects berries at any stage of development. When green berries are first inoculated with C. kahawae and then at 2, 72 or 96 h later with C. gloeosporioides, the necrotic lesions were significantly larger than in the controls, and were much more evident when the berries were incubated at the optimum growth temperature of 28 °C for C. gloeosporioides. Isolations from the lesions induced by the first inoculations with C. kahawae followed by inoculation with C. gloeosporioides revealed that all or most of the time the recovered isolates of the latter. Thus, C. gloeosporioides can overwhelm C. kahawae under conditions of higher environmental temperature and humidity and may enhance the CBD infection process under field conditions.     

Role of cyanide production by Pseudomonas fluorescens CHA0 in the suppression of root-knot nematode, Meloidogyne javanica in tomato

Summary Pseudomonas fluorescens strain CHA0 produces hydrogen cyanide (HCN), a secondary metabolite that accounts largely for the biocontrol ability of this strain. In this study, we examined the role of HCN production by CHA0 as an antagonistic factor that contributes to biocontrol of Meloidogyne javanica, the root-knot nematode, in situ. Culture filtrate of CHA0, resulting from 1/10-strength nutrient broth yeast extract medium amended with glycine, inhibited egg hatch and caused mortality of M. javanica juveniles in vitro. The bacterium cultured under high oxygen-tension conditions exhibited better inhibitory effects towards nematodes, compared to its cultivation under excess oxygen situation. Growth medium amended with 0.50 or 1.0 mM FeEDDHA further improved hatch inhibition and nematicidal activity of the strain CHA0. Strain CHA77, an HCN-negative mutant, failed to exert such toxic effects, and in this strain, antinematode activity was not influenced by culture conditions. Exogenous cyanide also inhibited egg hatch and caused mortality of M. javanica juveniles in vitro. Strains CHA0 or CHA77 applied in unsterilized sandy-loam soil as drench, caused marked suppression of root-knot disease development incited by M. javanica in tomato seedlings. However, efficacy of CHA77 was noticeably lower compared to its wild type counterpart CHA0. An increased bioavailability of iron following EDTA application in soil substantially improved nematode biocontrol potential of CHA0 but not that of CHA77. Soil infestation with M. javanica eggs resulted in significantly lower nematode population densities and root-knot disease compared to the juveniles used as root-knot disease-inducing agents. Strain CHA0 significantly suppressed nematode populations and inhibited galling in tomato roots grown in soil inoculated with eggs or juveniles and treated with or without EDTA. Strain CHA0 exhibited greater biocontrol potential in soil inoculated with eggs and treated with EDTA. To demonstrate that HCN synthesis by the strain CHA0 acts as the inducing agent of systemic resistance in tomato, efficacy of the strain CHA0 was compared with CHA77 in a split root trial. The split-root experiment, guaranteeing a spatial separation of the inducing agent and the challenging pathogen, showed that HCN production by CHA0 is not crucial in the induction of systemic resistance in tomato against M. javanica, because the HCN-negative-mutant CHA77 induced the same level of resistance as the wild type but exogenous cyanide in the form of KCN failed to trigger the resistance reaction. In the root section where both nematode and the bacterium were present, strain CHA0 reduced nematode penetration to a greater extent than CHA77, suggesting that for effective control of M. javanica, a direct contact between HCN-producing CHA0 and the nematode is essential.     

Induction of systemic acquired resistance in Zea mays also enhances the plant’s attractiveness to parasitoids

Plants under attack by caterpillars emit volatile compounds that attract the herbivore’s natural enemies. In maize, the caterpillar-induced production of volatiles involves the phytohormone jasmonic acid (JA). In contrast, pathogen attack usually up-regulates the salicylic acid (SA)-pathway and results in systemic acquired resistance (SAR) against plant diseases. Activation of the SA-pathway has often been found to repress JA-dependent direct defenses, but little is known about the effects of SAR induction on indirect defenses such as volatile emission and parasitoid attraction. We examined if induction of SAR in maize, by chemical elicitation with the SA-mimic benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), attenuates the emission of volatiles induced by Spodoptera littoralis or exogenously applied JA. In addition, we determined how these treatments affected the attractiveness of the plants to the parasitoid Microplitis rufiventris in a six-arm-olfactometer. BTH treatment alone resulted in significant systemic resistance of maize seedlings against the pathogen Setosphaeria turcica, but had no detectable effect on volatile emissions. Induction of SAR significantly reduced the emission rates of two compounds (indole and (E)-β-caryophyllene) in JA-treated plants, whereas no such negative cross-talk was found in caterpillar-damaged plants. Surprisingly, however, BTH treatment prior to caterpillar-feeding made the plants far more attractive to the parasitoid than plants that were only damaged by the herbivore. Control experiments showed that this response was due to plant-mediated effects rather than attractiveness of BTH itself. We conclude that in the studied system, plant protection by SAR activation is compatible with and can even enhance indirect defense against herbivores.     

Trunk injection of fosetyl‐aluminium controls the root disease caused by Phytophthora cinnamomi on Quercus ilex woodlands

In Spain, Quercus open woodlands are animal ranching systems of organic production seriously threatened by the exotic pathogen Phytophthora cinnamomi. The root disease it causes kills thousands of oaks annually. Effective disease management needs to integrate different techniques, and the use of a resistance inducer such as fosetyl‐Al can play a key role, because the use of potassium phosphite is prohibited in Spain. In a woodland where the pathogen recently arrived, 60 holm oaks in three different defoliation classes (asymptomatic, slight and moderate defoliation) were selected for trunk injection with pressurised capsules containing 4% of commercial fosetyl‐Al or water (controls). Holm oaks were checked periodically for defoliation and presence of the pathogen in roots and rhizosphere soil. Three years after treatments, defoliation was significantly lower in oaks treated with fosetyl‐Al, which even increased canopy cover, in comparison with control oaks, independent of the initial defoliation class considered. Chlamydospore density in rhizosphere soil, as well as the presence of the pathogen into the roots, was not significantly influenced by fosetyl‐Al treatments, although a trend to a lower presence of P. cinnamomi in roots was observed in treated oaks at every soil inoculum density detected. This study has shown that fosetyl‐Al, a phosphonate registered as a fungicide in the European Union, provides protection to holm oaks against P. cinnamomi, even exhibiting a therapeutic effect on pre‐existing infections. Consequently, this effective measure should be considered as part of the integrated approach to control this highly destructive pathogen in holm oak woodlands.     

Containment and spot eradication of a highly destructive,invasive plant pathogen (<Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis>) in natural ecosystems

The invasive plant pathogen Phytophthora cinnamomi (Stramenopila, Oomycota) has been introduced into 15 of the 25 global biodiversity hotspots, threatening susceptible rare flora and degrading plant communities with severe consequences for fauna. We developed protocols to contain or eradicate P. cinnamomi from spot infestations in threatened ecosystems based on two assumptions: in the absence of living hosts, P. cinnamomi is a weakly competitive saprotroph; and in the ecosystems we treated, the transmission of the pathogen occurs mainly by root-to-root contact. At two P. cinnamomi-infested sites differing in climate and vegetation types, we applied increasingly robust treatments including vegetation (host) destruction, fungicides, fumigation and physical root barriers. P. cinnamomi was not recovered at three assessments of treated plots 6–9 months after treatments. Given the high rates of recovery of P. cinnamomi from untreated infested soil and the sampling frequency, the probability of failing to detect P. cinnamomi in treated soil was <0.0003. The methods described have application in containing large infestations, eradicating small infestations and protecting remnant populations of threatened species.     

对增强辣椒疫霉菌抗性的研究

病原物诱导型启动子能精确控制抗病基因在侵染位点的表达,是抗病基因工程的有效工具。prp1-1是来自马铃薯谷胱甘肽巯基转移酶基因启动子的一个273bp的片段,能够快速准确地启动被侵染位点抗病基因的表达;Rs-AFP2是具有对致病性丝状真菌的广谱抗性。该研究构建prp1-1调控Rs-AFP2基因表达的载体,经农杆菌介导转化法导入辣椒。逆转录PCR检测发现,转基因辣椒只在受到疫霉菌孢子侵染时,才由prp1-1启动Rs-AFP2基因的转录。用疫霉菌孢子灌根接种转基因辣椒T1代植株,35株T1代辣椒中有29株表现出明显的疫霉菌抗性。另将23株T1代辣椒种于人工气候箱,发现其形态和发育特征与相同条件下的非转基因植株无明显区别。研究表明,prp1-1调控Rs-AFP2的诱导表达达到了增强辣椒疫霉菌抗性的目的,而且避免了负面效应的发生。     

Relationship of Pseudomonas syringae pv. tabaci races to the rhizosphere of Wisconsin-grown tobacco

Isolates of Pseudomonas syringae pv. tabaci, including 21 strains of the wildfire pathogen and 2 strains of the angular leafspot pathogen, were isolated from 143 rhizosphere and soil samples collected from 11 tobacco fields in Wisconsin. These pathogens were isolated by inoculating rhizosphere and soil washings into tobacco leaves and isolating the bacteria from wildfire or angular leafspot lesions that developed on the leaves. The wildfire isolates were from the rhizospheres of tobacco and Panicum capillare and from soil. While the majority of these were from wildfire-diseased fields, one isolate was from a field without disease symptoms; both angular leafspot isolates were from fields without angular leafspot symptoms. The majority of wildfire isolates were race 1, but three were race 0, and one was a new race. In three fields multiple races of wildfire were found. Both angular leafspot isolates were race 1. Two wildfire and one angular leafspot isolates were from fields where the cultivars were resistant to the races isolated.     

Expression of the Hypersensitive Response-assisting Protein in <Emphasis Type="Italic">Arabidopsis</Emphasis> Results in Harpin-dependent Hypersensitive Cell Death in Response to <Emphasis Type="Italic">Erwinia carotovora</Emphasis>

Active defense mechanisms of plants against pathogens often include a rapid plant cell death known as the hypersensitive cell death (HCD). Hypersensitive response-assisting protein (HRAP) isolated from sweet pepper intensifies the harpin_Pss-mediated HCD. Here we demonstrate that constitutive expression of the hrap gene in Arabidopsis results in an enhanced disease resistance towards soft rot pathogen, E. carotovora subsp. carotovora. This resistance was due to the induction of HCD since different HCD markers viz. Athsr3, Athsr4, ion leakage, H₂O₂ and protein kinase were induced. One of the elicitor harpin proteins, HrpN, from Erwinia carotovora subsp. carotovora was able to induce a stronger HCD in hrap-Arabidopsis than non-transgenic controls. To elucidate the role of HrpN, we used E. carotovora subsp. carotovora defective in HrpN production. The hrpN⁻ mutant did not induce disease resistance or HCD markers in hrap-Arabidopsis. These results imply that the disease resistance of hrap-Arabidopsis against a virulent pathogen is harpin dependent.     

The <Emphasis Type="Italic">ABI2</Emphasis>-dependent abscisic acid signalling controls HrpN-induced drought tolerance in <Emphasis Type="Italic">Arabidopsis</Emphasis>

HrpN, a protein produced by the plant pathogenic bacterium Erwinia amylovora, has been shown to stimulate plant growth and resistance to pathogens and insects. Here we report that HrpN activates abscisic acid (ABA) signalling to induce drought tolerance (DT) in Arabidopsis thaliana L. plants grown with water stress. Spraying wild-type plants with HrpN-promoted stomatal closure decreased leaf transpiration rate, increased moisture and proline levels in leaves, and alleviated extents of damage to cell membranes and plant drought symptoms caused by water deficiency. In plants treated with HrpN, ABA levels increased; expression of several ABA-signalling regulatory genes and the important effector gene rd29B was induced or enhanced. Induced expression of rd29B, promotion of stomatal closure, and reduction in drought severity were observed in the abi1-1 mutant, which has a defect in the phosphatase ABI1, after HrpN was applied. In contrast, HrpN failed to induce these responses in the abi2-1 mutant, which is impaired in the phosphatase ABI2. Inhibiting wild-type plants to synthesize ABA eliminated the role of HrpN in promoting stomatal closure and reducing drought severity. Moreover, resistance to Pseudomonas syringae developed in abi2-1 as in wild-type plants following treatment with HrpN. Thus, an ABI2-dependent ABA signalling pathway is responsible for the induction of DT but does not affect pathogen defence under the circumstances of this study.Hong-Ping Dong and Haiqin Yu contributed equally to this study and are regarded as joint first authors.     

Interactions Between Plant Growth Promoting Fungi and Arbuscular Mycorrhizal Fungus Glomus Mosseae and Induction of Systemic Resistance to Anthracnose Disease in Cucumber

Cucumber plants were treated with plant growth promoting fungi (PGPF), Phoma sp. (isolates GS8-2 and GS8-3) and Penicillium simplicissimum (isolate GP17-2) with or without the arbuscular mycorrhizal fungus (AMF) Glomus mosseae. Induction of systemic resistance in cucumber against the anthracnose disease caused by Colletotrichum orbiculare was tested to evaluate the nature of the interaction between the PGPF and AMF. Root colonizing ability of each fungal species as influenced by their interaction was also evaluated. Plant roots were pre-inoculated with each PGPF isolate and/or G. mosseae for four weeks and leaves were then challenge inoculated with the pathogen C. orbiculare. Plants treated with each PGPF isolate showed considerable protection against the disease, but the treatment of G. mosseae had no significant effect on disease development. However, combined inoculation of Phoma GS8-2 or GS8-3 with G. mosseae reduced the level of disease protection induced by single inoculation of each Phoma isolate. In contrast, the high levels of protection induced by the P. simplicissimum GP17-2 were not altered by combining it with G. mosseae. Root colonization of both Phoma sp. isolates was also suppressed by the presence of the G. mosseae, but such an effect was not found on the population development of P. simplicissimum. The percent cucumber root length colonized by G. mosseae was not affected by any of the PGPF isolates tested.     

Geographic range, impact, and parasitism of lepidopteran species associated with the invasive weed Lantana camara in South Africa

Six isolates of Trichoderma were screened for antagonism to Armillaria in tea stem sections buried in the soil. The inability of Armillaria to invade Trichoderma-colonized stem sections and the reduction of its viability in the plant materials following invasion of these by Trichoderma were used as indicators of antagonism. Four isolates of the species Trichoderma harzianum significantly (P<0.001) reduced the incidence of the pathogen in the plant materials. Isolate T4 completely eliminated the pathogen from plant materials in sterile soil and also antagonized two different isolates of the pathogen in nonsterile soil. Application of this T. harzianum isolate to the soil as a wheat bran culture significantly (P<0.001) reduced viability of Armillaria in woody blocks of inoculum. Soil amendment with coffee pulp also reduced the inoculum viability but did not affect the incidence of Trichoderma in the blocks of inoculum. We conclude that the direct application of wheat bran-formulated T. harzianum into soil surrounding woody Armillaria inoculum sources can suppress the pathogen. Further, no organic amendment is needed to enhance development of the antagonist in the soil as a pre-requisite to suppressing the pathogen.     

Current and projected global distribution of Phytophthora cinnamomi,one of the world's worst plant pathogens

Globally, Phytophthora cinnamomi is listed as one of the 100 worst invasive alien species and active management is required to reduce impact and prevent spread in both horticulture and natural ecosystems. Conversely, there are regions thought to be suitable for the pathogen where no disease is observed. We developed a climex model for the global distribution of P. cinnamomi based on the pathogen's response to temperature and moisture and by incorporating extensive empirical evidence on the presence and absence of the pathogen. The climex model captured areas of climatic suitability where P. cinnamomi occurs that is congruent with all available records. The model was validated by the collection of soil samples from asymptomatic vegetation in areas projected to be suitable by the model for which there were few records. DNA was extracted, and the presence or absence of P. cinnamomi was determined by high‐throughput sequencing (HTS). While not detected using traditional isolation methods, HTS detected P. cinnamomi at higher elevations in eastern Australia and central Tasmania as projected by the climex model. Further support for the climex model was obtained using the large data set from south‐west Australia where the proportion of positive records in an area is related to the Ecoclimatic Index value for the same area. We provide for the first time a comprehensive global map of the current P. cinnamomi distribution, an improved climex model of the distribution, and a projection to 2080 of the distribution with predicted climate change. This information provides the basis for more detailed regional‐scale modelling and supports risk assessment for governments to plan management of this important soil‐borne plant pathogen.     

Phosphite stimulated histological responses of Eucalyptus marginata to infection by Phytophthora cinnamomi

Phosphite is used to protect plants from the soil borne pathogen, Phytophthora cinnamomi. Although phosphite stimulates resistance to P. cinnamomi, this is the first histological study of its effect on Eucalyptus marginata, an economically important forest tree in Western Australia. Clonal lines of E. marginata, considered resistant and susceptible to P. cinnamomi, were underbark inoculated with P. cinnamomi. 4 days later, they were treated with 0, 2.5, 5 or 10 g L⁻¹ phosphite. Transverse hand sections were stained for suberin and lignin, and histological responses to infection were examined. Defence responses were stimulated at all phosphite concentrations in both clonal lines, and the genotypic difference in lesion length was eliminated within 8 days of treatment. In the resistant line, suberin production was stimulated while in the susceptible line both lignin and suberin were stimulated. By 2 days after treatment, phosphite stimulated a faster rate of suberin production in the resistant line than the susceptible line, but by 4 days after treatment, there was no difference in the increase between the lines. Damage caused by P. cinnamomi was found to extend furthest in the cortex and outer phloem in transverse sections in both genotypes. In the presence of P. cinnamomi, phosphite stimulated mitosis as part of the defence response, with meristematic activity involved in the compartmentalisation of damaged tissue (formation of periderm) and closure of healthy tissue (callus). Phytotoxicity had a detrimental effect in healthy tissues and this was more apparent in the resistant line, where it did not provide the best protection from lesion extension and plant mortality, suggesting phytotoxicity could disrupt defence responses. Phosphite increases the capacity of susceptible and resistant E. marginata clonal lines to wall-off and contain P. cinnamomi colonisation through lignin and suberin deposition, and increased meristematic activity.     

Priming,signaling, and protein production associated with induced resistance by <Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> KPS46

Bacillus amyloliquefaciens KPS46 is a rhizobacterium that induces systemic protection in soybean (Glycine max L.) against several diseases and enhances plant growth. In this study, treatment of soybean seed with KPS46 provided protection to leaves from bacterial pustule, caused by Xanthomonas axonopodis pv. glycines (Xag). KPS46 treatment also increased phenolic content and β-1,3-glucanase and peroxidase activity levels in leaves over non-treated plants. Differential expression of these traits was more rapid and pronounced when KPS46 treated plants were infected with Xag, this pattern indicating priming. Also associated with induced resistance by KPS46 was increased production of salicylic acid (SA) and jasmonic acid (JA) in soybean leaves, suggesting both SA- and JA-dependent signaling pathways are systemically triggered by KPS46 seed treatment. When KPS46 was applied to Arabidopsis roots, however, resistance against Pseudomonas syringae pv. tomato (Pst) was induced only in host genotypes with intact jasmonate, ethylene, and auxin sensitivity. Thus, induced resistance against Pst by KPS46 was SA independent and JA/ethylene dependent. Proteins induced in soybean leaves by KPS46 seed treatment and by the seed treatment in combination with pathogen inoculation were determined by proteomic analysis. Among 20 proteins upregulated in KPS46-treated plants, compared with non-treated plants, only three were defense related. In plants that received both KPS46 treatment and inoculation with Xag, nine of the 20 upregulated protein, as compared with proteins produced Xag inoculated plants having no KPS46 treatment, were defense related. This pattern of increased induction of defense-related proteins following pathogen infection of KPS46 treated plants supported priming by KPS46. Aside from proteins with defense-related function, most of the proteins induced by KPS46 were involved in metabolism and energy conversion, reflecting the strong direct positive effect that KPS46 has on soybean growth.     

Induction of Resistance in Melon to Didymella bryoniae and Sclerotinia sclerotiorum by Seed Treatments with Acibenzolar-S-methyl and Methyl Jasmonate but not with Salicylic Acid

The plant resistance activator acibenzolar‐S‐methyl (BTH), the signalling molecules salicylic acid (SA) and methyl jasmonate (MeJA) were tested by seed treatment for their ability to protect melon seedlings from gummy stem blight and white mould disease caused by the soil‐borne fungal pathogens Didymella bryoniae and Sclerotinia sclerotiorum, respectively. Didymella bryoniae infection on melon seedlings was completely suppressed by MeJA treatment. Necrotic lesions akin hypersensitive response occurred on all inoculated seedlings and prevented pathogen diffusion into healthy tissues. Didymella bryoniae infection was restricted following BTH seed treatment as well, although the percentage of necrotic lesions in comparison with the water soaked lesions was significantly lower than that from MeJA‐induced seedlings. BTH protected melon seedlings against S. sclerotiorum by the occurrence of a high percentage of necrotic lesions. A lower level of resistance was also achieved by MeJA seed treatment. The augmented level of resistance of tissues from BTH and MeJA‐treated seeds was associated with rapid increases in the activity of the pathogenesis‐related proteins chitinase and peroxidase. MeJA also determined a rapid and transient accumulation of lipoxygenase. Moreover, BTH and MeJA treatments determined the differential induction of particular de novo synthesized isoenzymes of these proteins. Results indicate that BTH and MeJA applied to melon seeds may activate on seedlings diverse metabolic pathways leading to the enhancement of resistance against distinct pathogens.