The inhibitory effects of rose powdery mildew infection on the oviposition behaviour and performance of beet armyworms

In this study, we investigated whether the oviposition behaviour and performance of the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), on the rose cultivar Rosa chinensis Jacq. (Rosaceae) were affected when the plants were infected by rose powdery mildew, Podosphaera pannosa (Wallr.: Fr.) de Bary (Erysiphales). The bioassays revealed that the moths significantly avoided ovipositing on mildew‐infected rose leaves when compared to healthy leaves. Pupal weights, emergence rates, and fecundity decreased when the caterpillars were fed mildewed rose leaves. Further laboratory bioassays aimed to elucidate the effects of two volatile headspace extracts (separately collected from healthy and mildewed rose plants) on the oviposition behaviour and performance of the moths. The moths clearly preferred to oviposit on healthy rose leaves that were not sprayed with additional volatiles rather than on healthy leaves sprayed with the volatile extracts from mildewed plants. The mean number of eggs laid on the former leaves was more than six times higher than that laid on the latter leaves. Olfactory bioassays demonstrated that ovipositing moths were significantly more attracted to volatiles emitted by healthy rose leaves than to those emitted by mildew‐infected leaves. Similar results were obtained when comparisons were made between the volatile extracts collected from healthy and mildewed rose plants. Thus, volatiles from mildew‐infected roses have a strong inhibitory effect against the moths. These results indicated that rose volatiles play a role in the oviposition behaviour of the moths, and that the volatiles induced by powdery mildew might be used for insect control.     

The production of toxins by Botrytis fabae in relation to growth of lesions on field bean leaves at different humidities

Filtrate extracts from liquid cultures of B. fabae and extracts from spreading chocolate spot lesions contained at least two heat-stable, light-labile phytotoxic compounds. Lesions similar in appearance to those of chocolate spot developed after injection of fractions containing these compounds into healthy bean leaves. Of 15 plant species injected with an extract from lesions, Vicia faba appeared to be the most susceptible to damage. Evidence is presented to support the hypothesis that under conditions of low humidity the concentrations of toxic compounds in an infected leaf become high enough to kill healthy cells surrounding infected tissue. The dead tissue then dries out preventing further fungal growth and lesion spread. In saturated air, however, the toxic compounds diffuse throughout the lamina and become too dilute to kill uninfected tissue. Tissue does not become desiccated and the fungus continues to spread.     

GC-MS analysis of headspace and liquid extracts for metabolomic differentiation of citrus Huanglongbing and zinc deficiency in leaves of 'Valencia' sweet orange from commercial groves

Introduction – Citrus Huanglongbing (HLB) is considered the most destructive citrus disease worldwide. Symptoms‐based detection of HLB is difficult due to similarities with zinc deficiency. Objective – To find metabolic differences between leaves from HLB‐infected, zinc‐deficient, and healthy ‘Valencia’ orange trees by using GC‐MS based metabolomics. Methodology – Analysis based on GC‐MS methods for untargeted metabolite analysis of citrus leaves was developed and optimized. Sample extracts from healthy, zinc deficient, or HLB‐infected sweet orange leaves were submitted to headspace solid phase micro‐extraction (SPME) and derivatization treatments prior to GC‐MS analysis. Results – Principal components analysis achieved correct classification of all the derivatized liquid extracts. Analysis of variance revealed 6 possible biomarkers for HLB, of which 5 were identified as proline, β‐elemene, (‐)trans‐ caryophyllene, and α‐humulene. Significant (P < 0.05) differences in oxo‐butanedioic acid, arabitol, and neo‐inositol were exclusively detected in samples from plants with zinc deficiency. Levels of isocaryophyllen, α‐selinene, β‐selinene, and fructose were significantly (P < 0.05) different in healthy leaves only. Conclusion – Results suggest the potential of using identified HLB biomarkers for rapid differentiation of HLB from zinc deficiency. Copyright © 2010 John Wiley & Sons, Ltd.     

The effect of a foliar disease (rust) on the development of Gastrophysa viridula (Coleoptera: Chrysomelidae)

Abstract.

• 1 Gastrophysa viridula Degeer (Coleoptera: Chrysomelidae) and the pathogenic rust fungus Uromyces rumicis (Schum.) Wint. both occur on leaves of Rumex crispus L. and R.obtusifolius L. Individual stages of beetle development, and egg laying, were compared on healthy and infected leaves of each plant species in the laboratory. Oviposition choice was investigated in the field and laboratory.
• 2 Beetles reared on infected leaves of each species had greater larval mortality and slower development than those reared on healthy leaves. Although larvae feeding on infected leaves consumed up to 2.5 times more dry weight than those reared on healthy leaves, they had a lower relative growth rate and pupated at a lower weight. These changes were consistent with the reduced nutritive quality of rust-infected Rumex leaves.
• 3 Fecundity of beetles reared on infected leaves of both species was considerably reduced. Eggs laid by beetles feeding on infected R.crispus leaves also had a reduced viability.
• 4 The beetle developed consistently poorer on healthy R.crispus than on healthy R.obtusifolius throughout its life-cycle. Differences in larval performance were greater between host species than between infected and healthy leaves.
• 5 Oviposition was similar on infected and healthy R.crispus in both the laboratory and field. However, adults consumed less, and laid fewer eggs on infected than on healthy R.obtusifolius. The pattern of egg laying on different aged leaves was affected by rust infection: a greater proportion of eggs was laid on the older, infected leaves, than on the equivalent aged leaves on the healthy plants. Few larvae survived from eggs laid on rusted leaves in the field.

     

Physiology of Rice Tungro Virus Disease: Involvement of Abscisic Acid-Like Substance in Susceptible Host-Virus Interactions

Stunting was severe in susceptible rice (Oryza saliva L.) cultivar ‘Taichung Native 1’ infected with tungro virus (RTV) compared to less-susceptible cultivar ‘IR 20’. The senescence of detached leaves of RTV-infected susceptible cultivar incubated in water in dark was accelerated compared to the healthy leaves as measured by the loss of total chlorophyll content. The transpiration rate of RTV-infected leaves of the susceptible cultivar was much lower than the healthy and RTV-infected leaves of the less-susceptible cultivar. Partially purified extracts obtained from RTV-infected leaves effectively inhibited GA-induced α-amylase synthesis in barley endosperms, and rice seedling growth, and they accelerated senescence of detached rice leaves. In all the three bibassays the ABA-like activity was significantly greater in the extracts from the RTV-infected susceptible cultivar than in extracts from the less-susceptible cultivar.     

Physiology of Rice Tungro Virus Disease: Increased Cytokinin Activity in Tungro-Infected Rice Cultivars

Tungro virus infection decreased the chlorophyll content in the susceptible rice (Oryza sativa) cultivar ‘Taichung Native 1’, while it increased the pigment in the less-susceptible cultivar ‘IR 20’ during the later stages of infection. The virus was recovered from the infected susceptibile cultivar as early as 3 days after inoculation. Relatively low amounts of virus were recovered from the less-susceptible cultivar between 12 and 30 days after inoculation. The senescence of detached leaves of virus-infected less-susceptible cultivar incubated in water in darkness was considerably delayed compared to healthy leaves. The cytokinin activity in the extracts from inoculated less-susceptible cultivar was 69% higher than in the extracts from the healthy plants.     

AN ANALYSIS OF THE GROWTH RESPONSE OF YOUNG TOMATO PLANTS TO INFECTION BY VERTICILLIUM ALBO-ATRUM: II. THE PRODUCTION OF GROWTH SUBSTANCES

Bioassays on ether-soluble acid extracts from healthy and Verticillium -infected tomato plants, showed the presence of substances inhibiting growth of wheat coleoptiles in both healthy and infected leaves and stems, but the amounts were greater in the infected.
Assays of infected stems and leaves showed increases in growth-promoting activity expressed as indole-3-acetic acid equivalents (IAAe), up to 200% of those for healthy controls.
Similar assays of cultures of V. albo-atrum showed growth-promoting activity. No acid substance capable of inhibiting the growth of wheat tissue was detected in the culture filtrate. IAA was identified by colour test with Ehrlich's reagent on chromatograms from extracts of both infected stems and fungal culture filtrates.
The vertical distribution of IAAe was determined in healthy and infected plants at the eight-leaf stage by assaying individual leaves and four stem segments separately. In healthy plants the IAAe content was greatest in the young leaves (6–8) but no gradient was observed as between leaves 1–5. In infected leaves increases over the controls were found in leaves, 1, 3 and 6 and a decrease in leaf 8.
In healthy stems IAAe was highest in the distal segment and infected stems showed higher values at all four levels, with the relative increase greatest in the distal region.
It is suggested that the major part of the Verticillium syndrome including petiolar epinasty, tylosis, pith hyperplasia and the formation of adventitious roots is the result of an accumulation of growth substances in infected tissue.     

Effects of cultural conditions on the mycelial growth of healthy and virus-infected cultivated mushroom, Agaricus bisporus

There was a significant inverse correlation (P= 0=001) between concentrations of mushroom viruses 1 and 2 in sporophores and amounts of mycelial growth on malt agar of isolates taken from them. Increasing virus concentrations decreased linear growth of one mushroom strain from 76 mm (healthy) to 35 mm (mildly infected) and 7 mm (severely infected) when incubated at 25°C for 3 wk. Mycelial growth rates of isolates from healthy and from virus-infected mushrooms were compared on eleven agar media. All media clearly differentiated between healthy and severely infected isolates, but fewer separated healthy from mildly infected isolates. Those that did contained maltose, sucrose or starch as carbon source. Media containing peptone usually gave better differentiation than those with other sources of nitrogen, but the best differentiation was obtained with malt agar. Growing healthy and infected isolates on a range of media affected their subsequent growth on malt agar, the growth of some isolates apparently being changed permanently after 2 months on some of the different media. Whereas none of the infected isolates grew less rapidly after this treatment, the growth of some of the mildly infected isolates improved to such an extent that they could no longer be distinguished from healthy isolates. After heat-treatment (1–6 wk at 33°C), mycelial growth rates of infected isolates were increased, but viruses 1 and 2 were not always eliminated unless the heat-treatment was begun immediately after subculture. Mycelial growth rate and colony characters are not infallible criteria of the presence or absence of virus, a feature of particular significance when checking the health of mushroom spawn.     

Phenolic compounds in apple leaves after infection with apple scab

Leaves of the scab-susceptible apple (Malus domestica) cultivar Golden Delicious were harvested from May to August 2008 and 2009. Some leaves were healthy and some infected with fungus Venturia inaequalis. The phenolic compounds were analysed in healthy leaves, infected leaves and in the scab spot tissue. In comparison to healthy leaves, the infected leaves showed higher contents of hydroxycinnamic acid, flavanols and phloridzin, while lower contents on procyanidins, quercetins and phloretin. The total amount of phenolic compounds in the infected tissue was 10 to 20 % higher than in the healthy leaves. Accumulation of phenolic compounds is a post-infection response, and probably their further transformation is a prerequisite for plant resistance.     

Enzyme Changes Associated with Host-Parasite Interactions between Pearl Millet and Downy Mildew Fungus

During the pathogenesis of pearl millet by the downy mildew (Sclerospora graminicola [Sacc.] Schroet.), the activities of peroxidase (PO) and indoleacetic acid oxidase (IAAO) and their isozyme pattern determined by isoelectric focusing on polyacrylamide gels, were observed in extracts of leaves and ears at different stages of development. The PO activity in extracts from infected plants of a susceptible cultivar was found to be higher than in healthy plants. The higher activity was most probably due to the acceleration of host senescence by the pathogen. Quantitative differences in the isozyme patterns of PO and IAAO were found. In inoculated plants of the resistant cultivar, no symptoms developed under the conditions used for infection of the susceptible cultivar and the changes in enzyme activities after inoculation were not significant. The results indicated that different proteins are synthesized in the two cultivars.     

Transmission characteristics and some other properties of bean yellow vein-banding virus, and its association with pea enation

Bean yellow vein-banding virus (BYVBV) has been found occasionally in mixed infection with pea enation mosaic virus (PEMV) in spring-sown field beans (Vicia faba minor) in southern England. Glasshouse tests confirmed that, like PEMV, BYVBV is transmissible by manual inoculation and by aphids in the persistent manner. However, BYVBV can be transmitted by aphids only from plants that are also infected with a helper virus, usually PEMV. Thus after separation from PEMV by passage through Phaseolus vulgaris it was no longer aphid-transmissible. It became aphid-transmissible again only after re-mixing in plants with PEMV or with a substitute helper, bean leaf roll virus (BLRV). It was not transmitted by aphids that fed sequentially on plants singly infected with PEMV and BYVBV. Thus the interaction between BYVBV and PEMV (or BLRV) that enables BYVBV to be transmitted by aphids seems to occur only in doubly infected plants. However, it was not transmitted by aphids from plants doubly infected with BYVBV and broad bean wilt virus (BBWV). BYVBV and PEMV were transmitted more readily by Acyrthosiphon pisum than by Myzus persicae; neither virus was transmitted by Aphis fabae. Phenol extracts of BYVBV-infected leaves were more infective than phosphate buffer or bentonite-clarified extracts and were sometimes infective when diluted to 1/1000. The infectivity of BYVBV in phosphate buffer extracts of leaves singly infected with BYVBV, unlike that in extracts of leaves doubly infected with BYVBV and PEMV (or BLRV), was destroyed by treatment with organic solvents. BYVBV infected 11 of 28 plant species that were inoculated with phenol extracts; seven of the infected species were legumes. No transmission of BYVBV was detected through seed harvested from infected field bean plants. Isometric particles c. 30 nm in diameter were seen in extracts of plants doubly infected with BYVBV and PEMV but not in extracts of plants infected with BYVBV alone. Leaves of plants infected with BYVBV, alone or with PEMV, contained membrane-bound structures c. 50–90 nm in diameter associated with the tonoplast in cell vacuoles. These structures were not found in healthy leaves. BYVBV has several properties in common with other known aphid-borne viruses that are helper-dependent and transmitted in a persistent manner. Possibly, as suggested for some of them, aphid transmission of BYVBV depends on the coating of its nucleic acid with helper virus coat protein.     

Transmission by Laodelphax striatellus Fallen of Rice black‐streaked dwarf virus from Frozen Infected Rice Leaves to Healthy Plants of Rice and Maize

Rice black‐streaked dwarf virus (RBSDV) is transmitted naturally to important crops such as rice, maize, barley and wheat in a persistent manner by the planthoppers, Laodelphax striatellus, Unkanodes sapporona and Unkanodes albifascia. Insect vector transmission tests are the basis for identifying viral incidence, evaluating the resistance of varieties and selecting resistance sources for rice and maize breeding. A simple, rapid and reliable method is described by which virus‐free small brown planthoppers (L. striatellus) acquired RBSDV from frozen infected rice leaves and transmitted it to healthy rice and maize plants. After feeding on frozen infected rice leaves, the planthoppers were tested by RT‐PCR for the presence of virus after 10, 15, and 22 days, respectively. The percentages of RBSDV‐containing insects were 0, 25 and 71.43% of L. striatellus fed on frozen infected rice leaves compared to 0, 28.25 and 71.43% of L. striatellus fed on fresh infected rice leaves, respectively. In transmission tests, three of eight rice seedlings (37.5%) and four of eight maize seedlings (50%) were inoculated by the planthoppers that had fed previously on frozen leaves and had allowed a 22 days latent period and showed typical disease symptoms. As a positive control, four of eight rice seedlings (50%) and four of six maize seedlings (66.67%) became infected. All rice and maize plants expressing disease symptoms were identified as virus‐positive by RT‐PCR. These results indicated that the planthoppers acquired RBSDV from frozen infected leaves and transmitted the virus to healthy plants.     

Demonstration of Levan and Alginate in Bean Plants (Phaseolus vulgaris) Infected by Pseudomonas syringae pv. phaseolicola

A simple method for the extraction of extracellular polysaccharides (EPS) from plant tissue was developed. The polysaccharides of bacterial and plant origin present in the crude leaf extracts were separated by column chromatography on DEAE-fractogel, and the bacterial polymers were identified by IR spectroscopy. In extracts from infected leaves as well as in exudates (ooze) from leaf axils, alginate (an acetylated mannuronan) and levan (β-2,6-fructofuranan) were detected as the major components amounting up to 80% of the crude extracts. A race-1 isolate of P. phaseolicola synthesized both levan and alginate in about equal amounts in planta, whereas a race-2 strain produced EPS composed almost solely of alginate. Extraction of healthy leaves yielded low amounts of complex polysaccharides. These consisted mainly of galactose, arabinose, and galacturonic acid. Neither fructose nor mannuronic acid were detected. Kinetic studies indicated that the main production of bacterial EPS in planta was correlated with the appearance of the water-soaked symptom in leaves. However, before water-soaking became apparent, alginate was detected in infected leaves (1, day after inoculation). The high amount of extractable material (ca. 50 mg levan plus alginate per g of dry weight of diseased tissue) suggests that the bacterial EPS is responsible for the typical water-soaked appearance of lesions after bacterial infection. Since alginate was predominantly synthesized by the more virulent race-2 isolate, this component of bacterial EPS was suspected to be a decisive factor of virulence of P. phaseolicola. A possible function of alginate during pathogenesis is discussed.     

Reproduction of sugar beet mosaic and tobacco mosaic viruses in anthocyanized beet plants

During our studies on the interaction of anthocyanins and plant virus diseases, reproduction of sugar beet mosaic (SBMV) and tobacco mosaic viruses (TMV) was investigated. Experiments were carried out in leaves of sugar beet,Beta vulgaris cv. Dobrovicka N and its spontaneous anthocyanized mutant. SBMV induces a systemic infection while TMV is responsible for primary local symptoms in sugar beet leaves only. Our quantitative analyses onAmaranthus caudatus L. andChenopodium quinoa Wilid. showed a significant decrease in concentration of SBMV in juice extracted from anthocyanized beet plants as compared with extracts from normal green infected plants. Significant differences were also obtained when SBMV — containing juice was tested in mixtures with healthy extracts from anthocyanized and normal green plants. Also the intensity of TMV symptoms in beet leaves was considerably decreased in leaves of antho-eyanized plants.     

Dark and light green tissues of tobacco leaves systemically infected with tobacco mosaic virus

There are significant changes in the structure of the upper tobacco (Nicotiana tabacum L.) leaves systemically infected with tobacco mosaic virus (TMV) especially in the light green tissue (LGT). Dark green areas (DGI) had intermediate status between healthy tissue and LGT. DGI contained significantly less infectious TMV and viral antigen than the LGT. The DGI, LGT and healthy tissues did not differ in the permeability of cell membranes and in the set of acidic pathogenesis-related (PR) proteins but the total content of PR-proteins in the healthy plants was higher than in the infected ones with the DGI being intermediate between healthy tissue and LGT. The crude leaf extracts from DGI and LGT showed less total ribonuclease activity and ribonuclease isozymes in comparison with control.     

Alterations in peroxidase activity and peroxidase isozymes in virus-infected plants

Peroxidase activity and isozyme patterns were investigated in two leguminous species infected with viruses which produced either local necrotic or systemic chlorotic symptoms. Highest peroxidase activity was recorded when the hosts reacted to infection with necrotic local lesions. No new virus-specific isozymes were found as a result of infection, but some isozymes, apparently associated with senescence, appeared earlier in extracts from leaves showing necrosis than in extracts from healthy leaves, or from infected leaves showing only very mild chlorosis. Increase in peroxidase activity was accompanied by alteration in isozyme pattern.     

Studies on the Hypertrophic Disease of Cherry (Genus Prunus), So-called “Witch’s Broom” Caused by Taphrina wiesneri

Metabolites of Taphrina wiesneri (Rath.) Mix. were examined. Brassicasterol, stearic acid, and p-hydroxyphenylacetic acid were isolated in crystalline form. p-Hydroxybenzoic acid and vanillic acid were identified by paper chromatography and UV measurement. Palmitic acid was identified by gas-chromatography. The fungus produced usually these compounds on any one of four kinds of medium used. p-Hydroxyphenylacetic acid promoted germination of rape seeds at the concentration of 20 ppm in water and showed inhibition at 250 ppm.

Phenolic acids and their related compounds in Japanese flowering cherry leaves infected by Taphrina wiesneri were examined. In the acidic and neutral extracts of infected cherry leaves (I), eighteen compounds positive to diazotized sulfanilic acid and two fluorescent compounds were detected by paper chromatography. Of these compounds, coumarin, 3, 4-dihydrocoumarin, melilotic acid, o- and p-coumaric acids, p-hydroxybenzoic melilotic acid, ferulic acid and caffeic acid were identified. Melilotic acid and coumarin were obtained in crystalline form. The amount of melilotic acid in I was higher than that in healthy leaves independent of sample source, although increased with the growth of cherry leaves.     

The sesquiterpene α-copaene is induced in tomato leaves infected by Botrytis cinerea

Abstract

Explorative experiments were done to investigate the possibility that tomato plants infected by Botrytis cinerea have a different emission of volatile organic compounds (VOC) than healthy plants. This was done by headspace analysis of volatiles emitted by detached leaves of infected and healthy plants. Principal component analysis (PCA) of GC-FID chromatograms revealed clearly separated clusters between infected and control leaves and identification of an interesting compound. In further analysis by GC-MS, the significantly distinctive component (p≤0.05) was identified as the sesquiterpene α-copaene. In previous work on herbivore damage, α-copaene was not distinctive, which may suggest that α-copaene may be specifically associated to fungal infections in tomato.     

Properties of some defective strains of tobacco mosaic virus and their behaviour as affected by inhibitors during storage in sap

From the type strain of tobacco mosaic virus, defective strains were isolated that produced chlorotic or ringspot type symptoms in tobacco and were difficult to transmit without carborundum in the inoculum. Their concentration was less than 0–1 μg/ml of sap instead of the usual 2 mg/ml with the type strain. Phenol extracts of infected leaves were a little more infective than extracts in buffer, whereas phenol extracts of leaves infected with type strain were very much less infective than extracts in buffer. Electron microscopy of infective sap rarely showed any virus particles, but preparations concentrated by ultracentrifugation contained virus particles, many of which were broken or seemed inadequately assembled. Changing the ambient temperature at which infected plants were kept from 20 to 35°C did not increase the amount or improve the appearance of the virus. Some of the strains were inactivated during heating for 10 min between 70 and 80 °C. Undiluted sap lost its infectivity in 3 days at 20 °C, as did the type strain when diluted to 0–1 μg/ml in sap from healthy leaves. This is because substances that inhibit infection were produced by microbes in the sap. The ability of sap from healthy leaves to inhibit infection increased by more than twenty-five times when left 3 days at 20 °C. Infectivity of appropriate mixtures of type strain and aged sap was restored by diluting them in buffer. Sodium azide at 0·02% in sap prevented formation of the inhibitor. The infectivity of the defective strains increased when inoculated together with the type strain.     

Peroxidase Activity and Isoperoxidase Pattern in Tobacco Leaves Infected with Tobacco Necrosis Virus and Other Viruses Inducing Necrotic and Non-Necrotic Alterations

The level of peroxidase activity was greatly enhanced in tobacco leaves infected by tobacco necrosis virus (TNV) and other viruses which induce necrotic symptoms (TMV, ToMV and PVY^N). The intensity was related to the age of the leaves infected: absent or neglible in mature leaves and very pronounced in young growing infected leaves. On the contrary, changes in peroxidase activity were negligible when the infection was provoked by viruses which do not produce necrotic reactions (TMV and PVY^O). Analysis of the peroxidase isoenzymes, pattern in tobacco leaves infected by TNV and other necrosis-inducing viruses revealed in all cases, a slight increase in anionic (pl 3.5–3.7) and a considerable increase in moderately anionic isoenzymes particularly the pl 4.6 isoenzyme which in TNV and PVY^N-infected leaves reached levels up to 21 and 72 times the healthy control values. A considerable increase in the cationic (pl9.3–8.8) isoenzymes and the appearance of one moderately cationic isoenzyme (pl 8.2) was also detected. In leaf extracts from-virus-infected tobacco leaves with nonnecrotic response, no, or negligible alterations on the isoenzyme pattern were detected. However, infection by a fungal parasite (Erisyphe cichoracearum), which established a fully compatible, non-necrotic, interaction with tobacco leaves, like the necrosis-inducing viruses, changed the isoperoxidase pattern. The data suggest the necrotic alterations and associated changes in the peroxidase activity and isoperoxidase pattern in virus-infected leaves are not clearly related.