Host Plant Response Research Articles

Identification of defence-related genes expressed in coffee and citrus during infection by Xylella fastidiosa

One of the phytosanitary problems of coffee cultivation in Brazil is Coffee Leaf Scorch (CLS) disease, caused by the phytopathogenic bacterium Xylella fastidiosa. Due to the economic importance of coffee to Brazil and the losses caused by X. fastidiosa, a cDNA library (RX1) was constructed using infected coffee stems. This library is one of the 37 coffee EST libraries constructed using different organs and tissues and biological conditions (Coffee Genome Project-CafEST). The objective of this study was to identify genes potentially involved in defence processes in response to X. fastidiosa infection by in silico analysis of the transcripts from the RX1 library as well as compare the coffee ESTs to citrus Xylella-infected ESTs. Clustering analysis of the RX1 library grouped a total of 7,501 sequences into 3,248 contigs, 19 of which were not found in the other 36 libraries. Additionally, 119 contigs were considered differentially expressed in comparison with the other libraries and according to statistical criteria. The global analysis of these contigs showed several genes involved in dehydration and photosynthesis. A total of 2,235 singlets were also obtained in the RX1 library and several of these genes are classically involved in defence processes. The comparison to a Xylella-infected citrus EST library revealed several genes similarly modulated in both species, indicating common defence mechanisms in both host plants in response to X. fastidiosa. The results obtained showed that water deprivation and response to osmotic and oxidative stress were expressed in a similar way in both coffee and citrus libraries. This is the first study to propose a common mechanism shared by citrus and coffee plants in response to the same pathogen.

Aphid-host plant interactions: does aphid honeydew exactly reflect the host plant amino acid composition?

Plants provide aphids with unbalanced and low concentrations of amino acids. Likely, intracellular symbionts improve the aphid nutrition by participating to the synthesis of essential amino acids. To compare the aphid amino acid uptakes from the host plant and the aphids amino acid excretion into the honeydew, host plant exudates (phloem + xylem) from infested and uninfested Vicia faba L. plants were compared to the honeydew produced by two aphid species (Acyrthosiphon pisum Harris and Megoura viciae Buckton) feeding on V. faba. Our results show that an aphid infestation modifies the amino acid composition of the infested broad bean plant since the global concentration of amino acids significantly increased in the host plant in response to aphid infestations. Specifically, the concentrations of the two amino acids glutamine and asparagine were strongly enhanced. The amino acid profiles from honeydews were similar for the two aphid species, but the concentrations found in the honeydews were generally lower than those measured in the exudates of infested plants (aphids uptakes). This work also highlights that aphids take large amounts of amino acids from the host plant, especially glutamine and asparagine, which are converted into glutamic and aspartic acids but also into other essential amino acids. The amino acid profiles differed between the host plant exudates and the aphid excretion product. Finally, this study highlights that the pea aphid, a “specialist” for the V. faba host plant, induced more important modifications into the host plant amino acid composition than the “generalist” aphid M. viciae.

Effect of Raw Cow Milk and Gliocladium virens in Pearl Millet against Downy Mildew Disease Caused by Sclerospora graminicola

Pearl millet [Pennisetum glaucum (L.) R. Br.] downy mildew (DM) is caused by the fungus Sclerospora graminicola (SACC.) SCHRŐT. is the most widespread and destructive disease of pearl millet affecting yield and quality in all the millet cultivating tracts of India. Since pearl millet is a crop of low economic value grown by resource-poor farmers, conventional technological interventions are not cost feasible. Integration of indigenous knowledge with biocontrol agents appeared as a logical strategy in the present case. Studies were, therefore, undertaken to manage DM in rainfed crop of pearl millet using raw cow milk together with Gliocladium virens. Seed and soil treatments resulted in the lowest disease incidence. Biochemical constituents (metabolites and oxidative enzymes) were analysed to determine possible mode of action of Raw Cow Milk (RCM) and Gliocladium virens. A considerable increase in sugars, phenols and ortho-dihydroxy phenols (OD) in healthy and DM infected leaves of treated pearl millet plants was recorded when compared to untreated controls. A marked increase in all the photosynthetic pigments in both healthy and diseased treated plants was observed. The induction of resistance was accompanied by increased activities of defense related enzymes. It is assumed that the combination of RCM and G. virens is capable of stimulating different systemic responses in host plant.

Growth promotion of pineapple 'vitória' by humic acids and burkholderia spp. during acclimatization

In vitro propagation of pineapple produces uniform and disease-free plantlets, but requires a long period of acclimatization before transplanting to the field. Quicker adaptation to the ex vitro environment and growth acceleration of pineapple plantlets are prerequisites for the production of a greater amount of vigorous, well-rooted planting material. The combination of humic acids and endophytic bacteria could be a useful technological approach to reduce the critical period of acclimatization. The aim of this study was to evaluate the initial performance of tissue-cultured pineapple variety Vitória in response to application of humic acids isolated from vermicompost and plant growth-promoting bacteria (Burkholderia spp.) during greenhouse acclimatization. The basal leaf axils were treated with humic acids while roots were immersed in bacterial medium. Humic acids and bacteria application improved shoot growth (14 and 102 %, respectively), compared with the control; the effect of the combined treatment was most pronounced (147 %). Likewise, humic acids increased root growth by 50 %, bacteria by 81 % and the combined treatment by 105 %. Inoculation was found to significantly increase the accumulation of N (115 %), P (112 %) and K (69 %) in pineapple leaves. Pineapple growth was influenced by inoculation with Burkholderia spp., and further improved in combination with humic acids, resulting in higher shoot and root biomass as well as nutrient contents (N 132 %, P 131 %, K 80 %) than in uninoculated plantlets. The stability and increased consistency of the host plant response to bacterization in the presence of humic substances indicate a promising biotechnological tool to improve growth and adaptation of pineapple plantlets to the ex vitro environment.

Phytophagous insect fauna tracks host plant responses to exotic grass invasion

The high dependence of herbivorous insects on their host plants implies that plant invaders can affect these insects directly, by not providing a suitable habitat, or indirectly, by altering host plant availability. In this study, we sampled Asteraceae flower heads in cerrado remnants with varying levels of exotic grass invasion to evaluate whether invasive grasses have a direct effect on herbivore richness independent of the current disturbance level and host plant richness. By classifying herbivores according to the degree of host plant specialization, we also investigated whether invasive grasses reduce the uniqueness of the herbivorous assemblages. Herbivorous insect richness showed a unimodal relationship with invasive grass cover that was significantly explained only by way of the variation in host plant richness. The same result was found for polyphagous and oligophagous insects, but monophages showed a significant negative response to the intensity of the grass invasion that was independent of host plant richness. Our findings lend support to the hypothesis that the aggregate effect of invasive plants on herbivores tends to mirror the effects of invasive plants on host plants. In addition, exotic plants affect specialist insects differently from generalist insects; thus exotic plants affect not only the size but also the structural profile of herbivorous insect assemblages.

Histological responses of host and non-host plants to Hyaloperonospora parasitica

Differences in Hyaloperonospora parasitica development and plant tissue responses were compared for 10 cruciferous hosts (including both resistant and susceptible genotypes), 3 leguminous and 1 graminaceous non-host species. Cotyledons, or true leaves in the case of Triticum aestivum and Pisum sativum, were studied at 2, 8, 24 h and 3, 5, 7 days post inoculation (dpi). The high levels of zoosporangial germination observed on all species tested, as well as on glass slides, suggested that inhibition of germination did not play a significant role in distinguishing host versus non-host resistance. During the early stages of infection, at spore germination and host penetration, there was no evidence of a clear-cut difference between Brassica host species which displayed a hypersensitive, partially resistant or susceptible reaction compared with non-host species. Haustoria formation was the key infection phase for the establishment of biotrophy. Across all tested species, haustoria were initiated inside the epidermal cells. However, there were significant differences in the frequency and timing of haustorial formation and the final size of haustoria among the tested species at early infection stage. Fully developed haustoria were never observed in Raphanus raphanistrum, Triticum aestivum, Lupinus angustifolius nor Trifolium subterraneum. Instead, the haustorium development appears to abort in the penetrated epidermal cells of these species. Although haustoria were formed in the epidermal and mesophyll cells of Sinapsis alba and Pisum sativum, subsequent hyphal growth and/or continued haustoria formation were rare or few, respectively. Hypersensitive reaction was the key resistance response observed among the host and non-host resistant species tested. It is noteworthy that, in the initial stages of pathogenesis, there was no differentiating point that separated the non-host species from those that were hosts.

Do the costs and benefits of fungal endophyte symbiosis vary with light availability?

• Here, we examined whether fungal endophytes modulated host plant responses to light availability. First, we conducted a literature review to evaluate whether natural frequencies of endophyte symbiosis in grasses from shaded habitats were higher than frequencies in grasses occupying more diverse light environments. Then, in a glasshouse experiment, we assessed how four levels of light and the presence of endophyte symbioses affected the growth of six grass species. • In our literature survey, endophytes were more commonly present in grasses restricted to shaded habitats than in grasses from diverse light environments. • In the glasshouse, endophyte symbioses did not mediate plant growth in response to light availability. However, in the host grass, Agrostis perennans, symbiotic plants produced 53% more inflorescences than nonsymbiotic plants at the highest level of shade. In addition, under high shade, symbiotic Poa autumnalis invested more in specific leaf area than symbiont-free plants. Finally, shade increased the density of the endophyte in leaf tissues across all six grass species. • Our results highlight the potential for symbiosis to alter the plasticity of host physiological traits, demonstrate a novel benefit of endophyte symbiosis under shade stress for one host species, and show a positive association between shade-restricted grass species and fungal endophytes.

Calcium signaling during the plant-plant interaction of parasitic Cuscuta reflexa with its hosts

The plant parasite Cuscuta reflexa induces various responses in compatible and incompatible host plants. The visual reactions of both types of host plants including obvious morphological changes require the recognition of Cuscuta ssp. A consequently initiated signaling cascade is triggered which leads to a tolerance of the infection or, in the case of some incompatible host plants, to resistance. Calcium (Ca2+) release is the major second messenger during signal transduction. Therefore, we have studied Ca2+ spiking in tomato and tobacco during infection with C. reflexa. In our recently published study1 Ca2+ signals were monitored as bioluminescence in aequorin-expressing tomato plants after the onset of C. reflexa infestation. Signals at the attachment sites were observed from 30 to 48 h after infection. In an assay with leaf disks of aequorin-expressing tomato which were treated with different C. reflexa plant extracts it turned out that the substance that induced Ca2+ release in the host plant was closely linked to the parasite´s haustoria.

Cerato-platanin elicits transcription of defence-related genes earlier than Ceratocystis platani on Platanus acerifolia

The ascomycete Ceratocystis platani (Walter) Engelbrecht & Harrington is the causal agent of canker stain, the most dangerous disease affecting the plane tree. Cerato-platanin (CP) is a protein elicitor from C. platani that acts as a pathogen-associated molecular pattern (PAMP), eliciting defence responses in both host and non-host plants. Using plane leaves as an experimental model, we compared the capacity of CP and C. platani to cause alterations in the transcription level of five Platanus acerifolia (Ait.) Willd. genes, three strictly related to a defence response, and two non-defence related. The results suggest that CP elicits the transcription of defence-related genes earlier than does C. platani. The expression of the cp gene in C. platani inoculated on plane leaves and grown in an axenic culture was also studied. For the first time CP has been shown to be released by C. platani when inoculated on P. acerifolia. The cp gene was expressed sooner when C. platani was inoculated on the leaves, than when it was grown in axenic culture, suggesting that the host-tree has a positive effect on the expression of this elicitor protein.

Metabolic profiling reveals local and systemic responses of host plants to nematode parasitism

The plant parasitic beet cyst nematode Heterodera schachtii induces syncytial feeding structures in Arabidopsis roots. The feeding structures form strong sink tissues that have been suggested to be metabolically highly active. In the present study, metabolic profiling and gene targeted expression analyses were performed in order to study the local and systemic effects of nematode infection on the plant host. The results showed increased levels of many amino acids and phosphorylated metabolites in syncytia, as well as high accumulation of specific sugars such as 1-kestose that do not accumulate naturally in Arabidopsis roots. A correlation-based network analysis revealed highly activated and coordinated metabolism in syncytia compared to non-infected control roots. An integrated analysis of the central primary metabolism showed a clear coherence of metabolite and transcript levels, indicating transcriptional regulation of specific pathways. Furthermore, systemic effects of nematode infection were demonstrated by correlation-based network analysis as well as independent component analysis. 1-kestose, raffinose, α,α-trehalose and three non-identified analytes showed clear systemic accumulation, indicating future potential for diagnostic and detailed metabolic analyses. Our studies open the door towards understanding the complex remodelling of plant metabolism in favour of the parasitizing nematode.

Adaptive Traits of Riptortus pedestris Nymphs (Heteroptera: Alydidae) for Locating Host Plants

Abstract Riptortus pedestris (F.) (Heteroptera: Alydidae) females oviposit not only on host plants but also on nonhosts, which may impose high costs on nymphs in terms of locomotion energy and time searching for host plants. Therefore, we hypothesized that second-instar nymphs have developed adaptive traits that help them access host plants, because the first feeding stage occurs during the second instar in this bug. We compared responses to the aggregation pheromone, relative leg lengths, and locomotion performance using a servosphere locomotion compensator as tests of physiological, morphometric, and behavioral traits, respectively, among the instars. We also investigated the effects of delayed feeding in the second instar on subsequent survival and development. Our results indicated that second-instar nymphs might have responded more sensitively to synthetic aggregation pheromone than other instars. Morphological measurements showed that second-instar nymphs had the longest relative leg length compared with other instars. The experiment using the servosphere revealed that second-instar nymphs had higher locomotion performance than did older nymphs, which may allow second-instar nymphs to walk a distance comparable to older nymphs, although their body size is much smaller. However, we did find that more than a 48-h delay in feeding after the first-instar molt decreased subsequent survival rates and that a later first-feeding led to a longer developmental period during the second instar. We concluded that R. pedestris nymphs have evolved various adaptive traits to enhance the probability of accessing host plants in response to the costly oviposition habit of adult females that lay eggs on nonhosts.

Chitosan in plant protection.

Chitin and chitosan are naturally-occurring compounds that have potential in agriculture with regard to controlling plant diseases. These molecules were shown to display toxicity and inhibit fungal growth and development. They were reported to be active against viruses, bacteria and other pests. Fragments from chitin and chitosan are known to have eliciting activities leading to a variety of defense responses in host plants in response to microbial infections, including the accumulation of phytoalexins, pathogen-related (PR) proteins and proteinase inhibitors, lignin synthesis, and callose formation. Based on these and other proprieties that help strengthen host plant defenses, interest has been growing in using them in agricultural systems to reduce the negative impact of diseases on yield and quality of crops. This review recapitulates the properties and uses of chitin, chitosan, and their derivatives, and will focus on their applications and mechanisms of action during plant-pathogen interactions.

Plant–eriophyoid mite interactions: cellular biochemistry and metabolic responses induced in mite-injured plants. Part I

This review is a comprehensive study of recent advances related to cytological, biochemical and physiological changes induced in plants in response to eriophyoid mite attack. It has been shown that responses of host plants to eriophyoids are variable. Most of the variability is due to individual eriophyoid mite-plant interactions. Usually, the direction and intensity of changes in eriophyoid-infested plant organs depend on mite genotype, density, or the feeding period, and are strongly differentiated relative to host plant species, cultivar, age and location. Although the mechanisms of changes elicited by eriophyoid mites within plants are not fully understood, in many cases the qualitative and quantitative biochemical status of mite-infested plants are known to affect the performance of consecutive herbivorous arthropods. In future, elucidation of the pathways from eriophyoid mite damage to plant gene activation will be necessary to clarify plant responses and to explain variation in plant tissue damage at the feeding and adjacent sites.

Mycorrhizal root colonisation and the subsequent host plant response of soil less grown tomato plants in the presence and absence of the mycorrhizal stimulant, Mycotech

A hydroponic trial was carried out to assess the effect of inoculating tomato plants with AM fungi, in the presence and absence of a mycorrrhizal stimulant (Mycotech). Four treatments were applied: (1) AM inoculant, (2) mycorrhizal stimulant, (3) AM + stimulant and (4) stimulant alone. Arbuscular mycorrhizal treated plants were 14% root colonized, whilst AM + stimulant treated plants, were 25% colonized. No colonization was observed in non-AM and non-AM + Stimulant treated plants. However, AM + stimulant treated plants did not show any significant improvement over AM treated plants, in either the plant nutrient concentrations, except Cu, or the growth of the plants, despite a root colonization advantage. In general, there was also no improvement in colonized plants over non-colonized plants, possibly due to the abundant supply of nutrients. However, stimulant treated plants performed better than the other plants in terms of their P, K, Ca, Mg and Mn nutrition. The causes of this increase remain unknown. This study has shown that the combined use of mycorrhizal stimulant and AM have a definite advantage for improving root colonization levels, but more research with regard to nutrient supply in the system, is required to ensure further benefits to the plants. Key words: Arbuscular mycorrhizal, dry matter, hydroponics, inoculant, macronutrients, micronutrients, tomatoes

In Vitro Physiological Responses of Fusarium oxysporum f. sp. niveum to Exogenously Applied Syringic Acid

Plant-microbe interactions are often accompanied by allelochemicals, such as syringic acid, released from the host plant. To explore the role of phenolic acids released from crop host plants in response to pathogen invasion, we examined the allelopathic effect of an artificially applied syringic acid on Fusarium oxysporum f. sp. niveum. We demonstrated that the growth and the conidial germination rate of F. oxysporum f. sp. niveum were stimulated at lower concentrations of syringic acid, though inhibited by higher dosage compared with control. The yield of fungus mycotoxin was increased from 60.9% to 561.5%. We conclude that syringic acid can be considered as a allelochemical inducer, stimulating the relative virulence factors of invading pathogens.

Stress magnitude matters: different intensities of pulsed water stress produce non‐monotonic resistance responses of host plants to insect herbivores

Abstract 1. Water stress may increase or reduce the suitability of plants for herbivores. The recently proposed ‘pulsed stress hypothesis’ suggests consideration of stress phenology (pulsed vs. continuous stress) to explain these conflicting effects of plant water stress on herbivore performance.2. This hypothesis was tested for the effect of differing stress intensity on performance and preference of insect herbivores belonging to different feeding guilds, namely leaf‐chewing insects (Spodoptera littoralis caterpillars) and phloem‐feeding insects (Aphis pomi aphids), on apple plants (Malus domestica). The plants were non‐stressed or exposed to a low or high intensity of pulsed water stress.3. Plant responses to the different stress levels were generally monotonic. Growth, stomatal conductance (gs), leaf water, and old‐leaf nitrogen concentration decreased, whereas young‐leaf nitrogen concentration and leaf mass per area (LMA) increased with increasing stress intensity. The stable isotope composition of foliar carbon (δ13C) responded non‐monotonically to the drought treatments. The δ13C values were highest in low‐stress plants, intermediate in high‐stress plants, and lowest in non‐stressed plants.4. The preference and performance responses of the caterpillars were also non‐monotonic. Non‐stressed plants were intermediately, low‐stress plants least, and high‐stress plants most attractive or suitable. Aphid population growth was highest on non‐stressed plants and lowest on low‐stress plants.5. The results highlight the importance of water stress intensity for the outcome of interactions between herbivores and drought‐affected plants. They show that pulsed water stress may enhance or reduce insect herbivore performance and plant resistance, depending on stress intensity.

Gall midge attack intensity and host-plant response in a Neotropical coastal ecosystem

The gall inducer Clusiamyia nitida Maia, 1996 (Diptera, Cecidomyiidae) often infests the shrub Clusia lanceolata (Camb.) (Clusiaceae) in the Neotropical vegetation of restinga of Rio de Janeiro State, Brazil. Leaves of Clusia lanceolata host up to 20 spheroid galls and show variation in their shape. We aimed to evaluate the effect of gall's intensity on leaves of Clusia lanceolata, and the extension of gall's impact on adjacent non-galled leaves. We analyzed the effect of the number of galls on leaf area, biomass, specific area and leaf appearance from 509 leaves of 14 individual plants. The results showed that differences of individual plants, pairs of leaves, and gall presence were responsible for more then 90% of variation on infested leaves. Variation on parasitic intensity level created differences in leaf response. Under moderate gall attack characterized by scattered galls on a leaf, the increase of the number of galls caused an increase of leaf biomass and area, and a decrease of specific area. The specific area was smaller also under high attack intensity, characterized by coalescent galls on a leaf. In those cases of extremely high parasitic intensity, galled leaves became deformed and the surface area was severely reduced. Leaf deformation due to gall attack led to early leaf abscission, indicated by the 90% of deformed leaves found in the youngest leaf pair of the branch. There was insufficient evidence that the impact of galls on leaf morpho-physiological parameters extended beyond the attacked leaves, because ungalled leaves did not change significantly when their opposite leaf had been galled.

Effect of awaaLmutation on lipopolysaccharide composition, oxidative stress survival, and virulence inErwinia amylovora

Erwinia amylovora, the causal agent of fire blight, is an enterobacterial pathogen of Rosaceous plants including apple and pear. We have been studying the response of E. amylovora to oxidative stress because, during infection, the bacterium elicits an oxidative burst response in host plants. During the screening of a transposon mutant library for hydrogen peroxide sensitivity, we identified a mutant carrying an insertion in waaL, a gene involved in lipopolysaccharide biosynthesis, that was more sensitive to hydrogen peroxide than the parental wild-type strain. We also confirmed that a waaL mutant of Pseudomonas aeruginosa exhibited an increased sensitivity to hydrogen peroxide compared with the wild-type strain. The E. amylovora waaL mutant was also reduced in virulence, showed a decrease in twitching motility, and was more sensitive to polymyxin B than the wild type. Each of these phenotypes was complemented by the cloned waaL gene. Our results highlight the importance of the lipopolysaccharide layer to virulence in E. amylovora and the unexpected finding of an additional function of lipopolysaccharide in protection from oxidative stress in E. amylovora and P. aeruginosa.

Host plant response and phenotypic plasticity of a galling weevil (Collabismus clitellae: Curculionidae)

The response of a host plant to gall-inducing insects varies both among and within plants, so that different levels of resources are available to the insects. The weevil Collabismus clitellae Boheman induces galls on the shoots of Solanum lycocarpum St Hil. in south-east Brazil. Galls are found on a range of parts within an individual plant and are more abundant on smaller plants. In the present study, the host plant response as a possible influence on the performance of C. clitellae both between and within plants was tested. Gall abortion increased with plant height. Within plants, gall size was positively related to shoot diameter and number of chambers within the gall. The increase in gall larval density (number of individuals per gall volume unit) resulted in smaller adults and reduced developmental rates, probably because of resource limitation within the gall. The number of eggs laid by females increased with shoot diameter. Females laid more eggs on thicker shoots, where there are fewer chances to form galls with high larval density. However, this relationship was weak and a large variation was found for adult sizes. The availability of high quality sites is limited to smaller plants and thicker shoots located on the basal region of the plant. The phenotypic plasticity of this insect species in adult size and development time allows individuals growing on low quality sites to reach maturity, thus enhancing exploitation of the host plant.

Systemic Disease Protection Elicited by Plant Growth Promoting Rhizobacteria Strains: Relationship Between Metabolic Responses, Systemic Disease Protection, and Biotic Elicitors

A study of plant defensive systemic responses induced by three plant growth promoting rhizobacteria (PGPR) on Arabidopsis thaliana Col 0 against Pseudomonas syringae pv. tomato DC3000 at the biochemical and transcriptional levels is reported in this paper. All three strains decreased disease severity when applied to A. thaliana prior to pathogen inoculation. At the biochemical level, each of the three strains induced ethylene (ET) when incubated with 1-amino-cyclopropane-1-carboxylic acid, and salicylic acid (SA) production in the plant. Plants treated with each of the three strains were also reduced in salicylic acid production after pathogen challenge compared to untreated controls. This effect was more marked in plants treated with Chryseobacterium balustinum AUR9, the strain most effective in decreasing disease severity. The expression level of PR1, a transcriptional marker of the SA-dependent pathway in C. balustinum AUR9-treated plants, is fourfold that of controls while the expression of PDF1.2, a transcriptional marker for the SA-independent pathway, is not induced. C. balustinum cell wall lipopolysaccharides, being putative bacterial elicitor molecules, are able to reproduce this systemic induction effect at low doses. From these observations, we hypothesize that certain PGPR strains are capable of stimulating different systemic responses in host plants. With C. balustinum AUR9, the SA-dependent pathway is stimulated first, as indicated by increases in SA levels and PR1 expression, followed by induction of the SA-independent pathway, as indicated by the increases in ET concentrations. The effects of both pathways combined with respect to disease suppression appear to be additive.