Tissue‐print and squash real‐time PCR for direct detection of ‘Candidatus Liberibacter’ species in citrus plants and psyllid vectors

Huanglongbing (HLB) disease is seriously threatening and/or damaging the citrus industry worldwide. Accurate detection of the three species associated with HLB disease, ‘Candidatus Liberibacter asiaticus’, ‘Candidatus Liberibacter africanus’ and ‘Candidatus Liberibacter americanus’, is essential for the preventive control of the disease. Real‐time PCR is a useful tool for bacterial detection. However, nucleic acid purification steps limit the number of samples that can be processed by PCR. Universal detection of ‘Ca. Liberibacter’ species was achieved by direct tissue‐printing and spotting of plant leaf petiole extracts or squashing of individual psyllids onto paper or nylon membranes. Primers were designed and used with TaqMan chemistry for accurate detection of the bacterium in immobilized targets (prints of 10 overlapping leaf pedicels per tree, or squashed single vectors), by extraction with water and direct use for real‐time PCR. This simplified method was validated and could detect HLB‐liberibacters in 100% of leaves with symptoms and 59% of symptomless leaves collected from HLB‐infected trees. The use of direct assays as template showed good agreement with use of purified DNA (κ = 0·76 ± 0·052). The squash assay allowed detection of the bacterium in 40% of mature Diaphorina citri that fed on leaves of HLB‐infected trees with or without symptoms. A commercial ready‐made kit based on this technology showed 96% accuracy in intra‐laboratory performance studies. The simplified direct methods of sample preparation presented herein can be effectively adopted for use in rapid screening of HLB agents in extensive surveys, certification schemes or for epidemiological and research studies.     

Alteration of photosynthetic performance and source–sink relationships in wheat plants infected by Pyricularia oryzae

Two experiments were carried out to assess the changes associated with photoassimilate production and partitioning in the source–sink relationship of flag leaves and spikes of wheat plants infected with Pyricularia oryzae, the causal agent of blast. Flag leaves and spikes were inoculated at 10 and 20 days after anthesis (daa) with a conidial suspension of P. oryzae. Analysis of chlorophyll a fluorescence using maximal photosystem II quantum efficiency (F_v?F_m), fraction of energy absorbed that is used in photochemistry (YII), quantum yield of non‐regulated energy dissipation (Y(NO)) and quantum yield of regulated energy dissipation (Y(NPQ)), showed an impairment of the photosynthetic performance in both infected flag leaves and spikes, coupled with reduced concentrations of chlorophyll a  + b and carotenoids. Compared to non‐inoculated controls, there was lower capacity for CO₂ fixation by RuBisCO in the infected flag leaves. Similarly, in the infected flag leaves and grains (obtained from infected spikes), there were lower concentrations of soluble sugars, while the hexoses‐to‐sucrose ratio increased in infected flag leaves. Compared to non‐inoculated controls, infected flag leaves showed lower sucrose phosphate synthase (SPS) activity and lower expression of the sucrose synthesis (SuSy) gene, while higher expression and activity of acid invertases also occurred. At the advanced stages of fungal infection, the concentration of starch in grains decreased but remained high for the infected flag leaves. There were reductions in ADP‐glucose pyrophosphorylase activity and the expression of ADP‐glucose pyrophosphorylase genes and a down‐regulation of β‐ and α‐amylase expression at the advanced stages of fungal infection on flag leaves and spikes. In conclusion, the effect of blast on both grain quality and yield can be associated with alterations in both production and partitioning of carbohydrates during the grain filling process.     

Coincidence of maximum severity of powdery mildew on grape leaves and the carbohydrate sink‐to‐source transition

Grapevine leaves infected with powdery mildew are a source of inoculum for fruit infection. Leaves emerging on a single primary shoot of Vitis vinifera cv. Cabernet Sauvignon were exposed to average glasshouse temperatures of 18°C (0·23 leaves emerging/day) or 25°C (0·54 leaves emerging/day). All leaves on 8–10 shoots with approximately 20 leaves each were inoculated with Erysiphe necator conidia to assess disease severity after 14 days in the 25°C glasshouse. Two photosynthetic ‘source’ leaves per shoot on the remaining 8–10 shoots were treated with ¹⁴CO₂ to identify, by autoradiography, the leaf position completing the carbohydrate sink‐to‐source transition. There was a clear association between the mean modal leaf position for maximum severity of powdery mildew (position 3·7 for 18°C; position 4·4 for 25°C) and the mean position of the leaf completing the sink‐to‐source transition (position 3·8 for 18°C; position 4·7 for 25°C). The mean modal leaf position for the maximum percentage of conidia germinating to form secondary hyphae was 4·2 for additional plants grown in the 25°C glasshouse. A higher rate of leaf emergence resulted in a greater proportion of diseased leaves per shoot. A Bayesian model, consisting of component models for disease severity and leaf ontogenic resistance, had parameters representing the rate and magnitude of pathogen colonization that differed for shoots developing in different preinoculation environments. The results support the hypothesis that the population of leaves in a vineyard capable of supporting substantial pathogen colonization will vary according to conditions for shoot development.     

Potential for eradication of the exotic plant pathogens Phytophthora kernoviae and Phytophthora ramorum during composting

Temperature and exposure time effects on Phytophthora kernoviae and Phytophthora ramorum viability were examined in flasks of compost and in a large‐scale composting system containing plant waste. Cellophane, rhododendron leaf and peat‐based inoculum of P. kernoviae and P. ramorum isolates were used in flasks; naturally infected leaves were inserted into a large‐scale system. Exposures of 5 and 10 days respectively at a mean temperature of 35°C in flask and large‐scale composts reduced P. kernoviae and P. ramorum inocula to below detection limits using semi‐selective culturing. Although P. ramorum was undetectable after a 1‐day exposure of inoculum to compost at 40°C in flasks, it survived on leaves exposed to a mean temperature of 40·9°C for 5 days in a large‐scale composting system. No survival of P. ramorum was detected after exposure of infected leaves for 5 days to a mean temperature of ≥41·9°C (32·8°C for P. kernoviae) or for 10 days at ≥31·8°C (25·9°C for Phytophthora pseudosyringae on infected bilberry stems) in large‐scale systems. Fitted survival probabilities of P. ramorum on infected leaves exposed in a large‐scale system for 5 days at 45°C or for 10 days at 35°C were <3%, for an average initial infection level of leaves of 59·2%. RNA quantification to measure viability was shown to be unreliable in environments that favour RNA preservation: high levels of ITS1 RNA were recovered from P. kernoviae‐ and P. ramorum‐infected leaves exposed to composting plant wastes at >53°C, when all culture results were negative.     

Impact of sugarcane yellow leaf virus (ScYLV) on the carbohydrate status of sugarcane: Comparison of virus-free plants with symptomatic and asymptomatic virus-infected plants

The carbohydrate concentration of old and young sugarcane plants, either virus-free or infected by sugarcane yellow leaf virus, was compared. Internode lengths and fresh weights of symptomatic plants were reduced, but not the sucrose concentrations. Starch and sucrose concentration in green leaf tops of mature (18 months old) was the highest in asymptomatic plants followed by symptomatic, and then by virus-free plants. Carbohydrates increased during daytime by 5–20%. The carbohydrate concentration in leaves of young, 6 months old plants was much lower than in mature plants and it increased to 500% during daytime. Asymptomatic leaves had a higher level of carbohydrates, especially starch, from late afternoon until the end of the night, suggesting a reduction of assimilate export.     

Partial Purification of Thai Isolate of Citrus Huanglongbing (Greening) Bacterium and Antiserum Production for Serological Diagnosis

We aimed to improve the purification of citrus Huanglongbing (greening) bacterium (HB), Candidatus Liberobacter asiaticum and to produce an antiserum against HB. Periwinkle plants Catharantus roseum L. graft-inoculated with HB were used to produce an antiserum. All young leaves of new shoots incubated at 20–25°C and 25–30°C, a few mature leaves incubated at 20–25°C, and all mature leaves incubated first at 25–30°C and later transferred to 20–25°C developed yellowing symptoms and were then used to prepare immunogen. The HB was partially purified from these leaves by an improved method that included a macerating enzyme treatment of the midribs of infected leaves and homogenization of infected phloem sieve tissues. An antiserum raised against partially purified HB reacted clearly at a dilution of 1/16 with HB-infected citrus extract prepared at a concentration of 40 times, but did not react with healthy or tristeza virus-infected citrus extract in microprecipitin tests. Received 23 August 2002/ Accepted in revised form 4 December 2002     

Anatomical distribution of abnormally high levels of starch in HLB-affected Valencia orange trees

The citrus disease Huanglongbing (HLB or citrus greening) is characterized, among other symptoms, by extraordinary levels of starch accumulation in leaves. This condition denotes imbalances in carbohydrate source sink relationship which in turn may have direct implications in the overall health of HLB-trees and in future strategies to manage the disease. Using light, scanning, and transmission electron microscopy we investigated the extent of carbohydrate partitioning imbalances throughout the tree. In all aerial tissues, starch accumulation in HLB-affected trees far exceeded that of HLB-negative control trees. Starch accumulated extensively in photosynthetic cells as well as phloem elements and vascular parenchyma in leaves and petioles. In stems, starch was commonly observed in xylem parenchyma and in the phelloderm of HLB-affected trees but absent from control samples. In contrast, roots from HLB-affected trees were depleted of starch whereas roots from control trees contain substantial starch deposits. The data supports the notion that the substantial changes in carbohydrate partitioning observed throughout the citrus tree may not only be a result of HLB infection, but in itself, a cause for the rapid decline and death of infected trees.     

Association of ‘Candidatus Liberibacter asiaticus’ root infection,but not phloem plugging with root loss on huanglongbing‐affected trees prior to appearance of foliar symptoms

Huanglongbing (HLB) is a systemic disease of citrus caused by phloem‐limited bacteria ‘Candidatus Liberibacter’ spp. with ‘Ca. Liberibacter asiaticus’ (Las) the most widespread. Phloem‐limited bacteria such as liberibacters and phytoplasmas are emerging as major pathogens of woody and herbaceous plants. Little is known about their systemic movement within a plant and the disease process in these tissues. Las movement after initial infection was monitored in leaves and roots of greenhouse trees. Root density, storage starch content, and vascular system anatomy in relation to Las presence in field and greenhouse trees, both with and without symptoms, showed the importance of root infection in disease development. Las preferentially colonized roots before leaves, where it multiplied and quickly invaded leaves when new foliar flush became a sink tissue for phloem flow. This led to the discovery that roots were damaged by root infection prior to development of visible foliar symptoms and was not associated with carbohydrate starvation caused by phloem‐plugging as previously hypothesized. The role of root infection in systemic insect‐vectored bacterial pathogens has been underestimated. These findings demonstrate the significance of early root infection to tree health and suggest a model for phloem‐limited bacterial movement from the initial insect feeding site to the roots where it replicates, damages the host root system, and then spreads to the rest of the canopy during subsequent leaf flushes. This model provides a framework for testing movement of phloem‐limited bacteria to gain greater understanding of how these pathogens cause disease and spread.     

Contrasting canopy and fibrous root damage on Swingle citrumelo caused by ‘Candidatus Liberibacter asiaticus’ and Phytophthora nicotianae

Huanglongbing (HLB), associated with the phloem‐limited bacterium ‘Candidatus Liberibacter asiaticus’ (Las), is devastating trees in citrus orchards of Florida. Additionally, Phytophthora nicotianae, omnipresent in citrus soils, causes root rot that reduces water and nutrient uptake by fibrous roots. To investigate fibrous root damage and replacement and canopy size in relation to infection of fibrous roots by Las and P. nicotianae, rootstock seedlings of Swingle citrumelo (Citrus paradisi × Poncirus trifoliata) were inoculated with Las or P. nicotianae in two greenhouse pot trials. Phytophthora nicotianae caused root damage within 5 weeks post‐inoculation, which led to greater reduction of canopy size than for Las‐infected seedlings by the end of the experiment. Las increased accumulation of fibrous root biomass at 5 weeks post‐root trimming (wpt) in the 2014 trial and at 11 wpt in the 2015 trial. New root length was not consistently increased by Las. Reduced total leaf area of symptomless Las‐infected seedlings compared to noninoculated controls might be due to the combined effect of altered carbohydrate allocation between shoots and roots and altered leaf morphology.     

稻瘟病菌和胡麻斑病菌碳氮营养的比较研究

 稻瘟病菌和胡麻斑病菌的碳氮营养有明显的差别。
稻瘟病菌对碳源利用的顺序是:麦芽糖、葡萄糖 > 果糖、蔗糖 > 乳糖、淀粉 > 木糖 > 门冬酰胺、半乳糖、阿拉伯糖,不能利用甘油。胡麻斑病菌利用碳源的顺序是:麦芽糖、乳糖 > 淀粉 > 蔗糖 > 木糖 > 葡萄糖、果糖、半乳糖 > 阿拉伯糖,不能利用门冬酰胺和甘油。     

Carbohydrate composition of sugarcane cultivars that are resistant or susceptible to <Emphasis Type="Italic">Sugarcane yellow leaf virus</Emphasis>

Sugarcane cultivars with a high (susceptible cultivars) and low (resistant cultivars) virus titer of Sugarcane yellow leaf virus were grown in the field. The carbohydrate composition in green leaf tops and in stems was determined. In RT-PCR of leaf extracts, susceptible cultivars had a high SCYLV-titer, whereas resistant cultivars had a very low titer. The cultivars differed in biomass yield, but these differences were not correlated with susceptibility. However, carbohydrate composition did have susceptibility-specific differences. Hexose levels were lower in green leaf tops and stalks of susceptible (strongly infected) cultivars than in those of resistant (weakly infected) cultivars. The stalks of susceptible cultivars also had less starch than those of resistant cultivars. Thus, the viral susceptibility (and infection) affected sugar metabolism. In addition, a positive correlation between hexose and starch in stems and between hexose and sucrose in green leaf tops was observed. The results from susceptible versus resistant cultivars were the opposite of those in the comparison between infected versus virus-free lines of the same cultivar. The breeding process apparently had unintentionally selected clones with modulated carbohydrate metabolism to avoid or compensate for the adverse effects of SCYLV infection.     

Liberibacters associated with orange jasmine in Brazil: incidence in urban areas and relatedness to citrus liberibacters

Two surveys (2005/2006 and 2009) were conducted in the state of São Paulo, Brazil, to investigate the incidence of ‘Candidatus Liberibacter asiaticus’ and ‘Ca. L. americanus’, two liberibacters associated with citrus huanglongbing (HLB) disease and both transmitted by Diaphorina citri, in orange jasmine (Murraya exotica), a widespread ornamental tree in cities and villages. The graft‐transmissibility of the two species, and their DNA relatedness to citrus‐associated liberibacters, were also investigated. Quantitative PCR was applied to PCR‐positive orange jasmine and HLB‐positive citrus growing in backyards and orchards to assess their inoculum source potentials. Liberibacters were detected in 91 of 786 sampled orange jasmine plants in 10 of 76 sampled locations. PCR‐positive trees exhibited yellow shoots and/or dieback symptoms indistinguishable from those on PCR‐negative trees. ‘Candidatus Liberibacter americanus’ was more common in 2005/2006 (96·6%) and ‘Ca. L. asiaticus’ in 2009 (84·8%). rplJ nucleotide sequences were identical within all populations of either species. Graft transmission succeeded only in homologous host combinations, including ‘Ca. L. americanus’ (2/10) from/to orange jasmine and ‘Ca. L. americanus’ (5/18) and ‘Ca. L. asiaticus’ (5/9) from/to citrus. Symptoms were mild and developed less rapidly in orange jasmine than in citrus, probably as a result of lower liberibacter multiplication rates. Respective titres of ‘Ca. L. americanus’ and ‘Ca. L. asiaticus’ in orange jasmine averaged 4·3 and 3·0 log cells g^?1 tissue, compared with 5·5 and 7·3 in citrus. The results indicate that orange jasmine does not favour liberibacter multiplication as much as citrus. However, its importance in HLB epidemics should not be underestimated as it is a preferred host of D. citri and is not under any strict tree‐eradication programme or measures for insect control.     

Influence of temperature on infection and establishment of ‘Candidatus Liberibacter americanus’ and ‘Candidatus Liberibacter asiaticus’ in citrus plants

The objectives of this work were (i) to determine the influence of temperature on infection of citrus by ‘Candidatus Liberibacter asiaticus’ and ‘Candidatus Liberibacter americanus’, the two bacterial species associated with citrus huanglongbing (HLB) in Brazil, and (ii) to determine the influence of temperature on citrus colonization by ‘Ca. L. asiaticus’, which has taken over from ‘Ca. L. americanus’ as the predominant species in Brazil since 2008. Two experiments were carried out with graft‐inoculated Valencia oranges on Rangpur lime rootstocks. Immediately after inoculation the plants were maintained for 423 days in growth chambers under the following night/day temperature conditions: 17/22, 22/27 or 27/32°C, with a dark/light photoperiod of 8/16 h. Infection and colonization of plants were determined using quantitative PCR (qPCR). ‘Candidatus Liberibacter americanus’ did not infect the plants maintained at 27/32°C; however, infection by ‘Ca. L. asiaticus’ occurred at all studied temperatures. Two months after inoculation, ‘Ca. L. asiaticus’ was distributed throughout the inoculated plants, with mean C_t values in the range of 30–31 for leaves and 25–28 for roots. Over time, ‘Ca. L. asiaticus’ reached the highest titres in mature leaves (mean C_t value = 26·7) of citrus plants maintained at 22/27°C. ‘Candidatus Liberibacter asiaticus’ colonization of citrus plants was negatively affected by the daily temperature regime of 27/32°C (mean C_t value in mature leaves = 33·6).     

Correlation of in planta endo‐beta‐1,4‐xylanase activity with the necrotrophic phase of the hemibiotrophic fungus Mycosphaerella graminicola

The association of the cell wall degrading enzyme endo‐beta‐1,4‐xylanase (EC 3.2.1.8) with pathogenicity of Mycosphaerella graminicola was examined in planta. The enzyme production of two M. graminicola isolates (T0372 and T0491), as well as their ability to infect seedlings of susceptible wheat cv. Scorpion, was first compared. No significant difference was found between the two isolates regarding spore germination rates, mycelial growth on the leaf surface or direct and stomatal penetrations. However, restricted hyphal growth was observed inside leaves inoculated with T0372, whereas successful mesophyll colonization with a strong intercellular fungal growth was found in leaves infected with T0491. Likewise, T0372 was unable to induce lesions bearing pycnidia and to produce endo‐beta‐1,4‐xylanase activity until 22 days post‐inoculation (d.p.i.). On the other hand, significant high increases of both diseased leaf area bearing pycnidia and endo‐beta‐1,4‐xylanase activity were observed between 16 and 22 d.p.i. for T0491 (r = 0·98). The investigation of 24 additional isolates, including the IPO323 and IPO94269 reference isolates, highlighted a strong correlation between endo‐beta‐1,4‐xylanase activity and disease development levels (r = 0·94). This study demonstrates that differences in pathogenicity in M. graminicola are not linked to events on the leaf surface or to frequency of leaf penetration, but to the ability of the fungus to colonize the mesophyll and to produce the cell wall degrading enzyme endo‐1,4‐beta‐xylanase during the necrotrophic phase.     

Biological characterization of French Potato virus Y (PVY) isolates collected from PVY‐susceptible or ‐resistant tobacco plants possessing the recessive resistance gene va

Improved tobacco cultivars introgressed with alleles of the recessive resistance va gene have been widely deployed in France to limit agronomical consequences associated with Potato virus Y (PVY) infections. Unfortunately, necrotic symptoms associated with PVY have been reported on these cultivars suggesting that PVY is able to overcome the resistance. A field survey was performed in France in 2007 to (i) estimate the prevalence of PVY in tobacco plants showing symptoms and (ii) characterize PVY isolates present in susceptible and va‐derived tobacco cultivars. A serological typing procedure, applied to 556 leaves collected from different French tobacco growing areas, was performed using polyclonal antisera raised against different viral species including PVY. Viral species were detected in 80·8% of leaves and PVY was present in 83·5% of infected samples. However, statistical analysis confirmed that the probability of a tobacco plant being infected with PVY is reduced in va hosts. Eighty‐six PVY isolates were mechanically inoculated on one susceptible and three va‐derived tobacco cultivars used as indicator hosts to define virulence of these isolates against alleles 0, 1 and 2 of the va gene. Both qualitative and quantitative analyses showed that 55 PVY isolates were able to overcome the three va alleles. Moreover, the monitored biological diversity of PVY isolates was higher in the susceptible tobacco hosts than in the va‐derived ones. This study helps to understand consequences of the deployment of the va gene in tobacco on diversity and virulence of PVY isolates.     

Quantification and ecological study of ‘Candidatus Liberibacter asiaticus’ in citrus hosts,rootstocks and the Asian citrus psyllid

The use of proper management strategies for citrus huanglongbing (HLB), caused by ‘Candidatus Liberibacter asiaticus’ (Las) and transmitted by Asian citrus psyllid (ACP) (Diaphorina citri), is a priority issue. HLB control is based on healthy seedlings, tolerant rootstock cultivars and reduction of ACP populations. Here, dynamic populations of Las in different citrus hosts and each instar of ACP were studied, together with the seasonal growth and distribution of Las in different tissues, using conventional and TaqMan real‐time PCR. Different levels of susceptibility/tolerance to HLB were seen, resulting in different degrees of symptom severity and growth effects on hosts or rootstocks. Troyer citrange, Swingle citrumelo and wood apple were highly tolerant among 11 rootstock cultivars. Regarding distribution and seasonal analysis of Las, mature and old leaves contained high concentrations in cool temperatures in autumn and spring. Las was detected earlier through psyllid transmission than through graft inoculation, and the amounts of Las (AOL) varied in different hosts. Thus, different AOL (10⁴–10⁷ copy numbers μL^?1) and Las‐carrying percentages (LCP; 40–53.3%) were observed in each citrus cultivar and on psyllids, respectively. Furthermore, both AOL and LCP were lower in nymphs than in adult psyllids, whereas the LCP of psyllids were not affected by increasing the acquisition‐access time. The present study has significant implications for disease ecology. The combination of early detection, use of suitable rootstocks and constraint of psyllid populations could achieve better management of HLB disease.     

Changes in carbohydrate composition of cucumber leaves during the development of powdery mildew infection

Within 3 days after inoculation, greater rates of decrease of hexoses, and of increases in inositol and α-glucans, were detected in leaves of cucumber leaves infected with Sphaerotheca fuliginea (now reclassified as Podosphaera xanthii ) than in healthy leaves, together with synthesis of fungal mannitol, arabitol and trehalose. With the onset of sporulation (5 days after inoculation, dai), glucose, polyols, trehalose and glycogen content, as well as fresh and dry weight of leaf tissue, increased more rapidly than in healthy plants, while sucrose, raffinose and starch declined. Starch, glycogen and other α-glucans, extracted from ethanol-insoluble tissue fractions of different water-solubility from healthy and infected leaves, were quantified after amyloglucosidase treatment. Infection-induced alterations in partitioning of photosynthate during a 14 h photoperiod at 7 dai, including a close correspondence in patterns of synthesis of glucose, polyols and glycogen, further clarified the process of diversion of host assimilate to the pathogen. The presence of glucose, but only traces of sucrose in spores, supported evidence from other powdery mildew infections for photosynthate being transferred to S. fuliginea as glucose.     

Temporal variation in contamination of pine engraver beetles with Fusarium circinatum in native Monterey pine forests in California

The relative importance of beetle species associated with Fusarium circinatum‐infected Monterey pines was investigated in three Monterey pine forests along the coast of central California, USA from April to November in 2004 and 2005. Fusarium circinatum was frequently isolated from Ips mexicanus and I. plastographus. The mean percentage isolation based upon numbers of I. mexicanus and I. plastographus carrying propagules of F. circinatum was 17·7 and 10·9% in 2004 and 16·7 and 17·3% in 2005, respectively. The mean percentage isolation was high in the spring and early summer and low in late summer and autumn in all three locations for both species. Isolation was higher from beetles emerging from harvested F. circinatum‐infected pine‐stems than for trapped beetles, 42·4% for I. mexicanus and 45·9% for I. plastographus. The mean (± SE) propagule load of trapped I. mexicanus was 269·5 (± 14·1) in 2004 and 281·7 (± 35·7) in 2005 and was 216·1 (± 28·9) in 2004 and 251·9 (± 28·4) in 2005 for I. plastographus. Mean propagule loads decreased from May to November in all locations for both species. Propagule loads of beetles emerged from infected stems were lower than that of trapped beetles, with means of 89·4 (± 23·2) and 93·0 (± 23·2) for I. mexicanus and I. plastographus, respectively. Thus beetles must acquire fungal propagules from more than one infected host. These results also suggest that higher contamination rates and propagule loads in spring and early summer may indicate a higher risk of pitch canker transmission, relative to late summer or autumn.     

Impact of <Emphasis Type="Italic">Erysiphe alphitoides</Emphasis> on transpiration and photosynthesis in <Emphasis Type="Italic">Quercus robur</Emphasis> leaves

Oak powdery mildew, (Erysiphe alphitoides) causes one of the most common diseases of oaks. We assessed the impact of this pathogen on photosynthesis and water relations of infected leaves using greenhouse-grown oak seedlings. Transpiration of seedlings infected by oak powdery mildew was also investigated. Altogether, E. alphitoides had a low impact on host gas exchange whether at the leaf or whole plant scale. Maximal stomatal conductance of infected leaves was reduced by 20–30% compared to healthy controls. Severely infected seedlings did not experience any detectable change of whole plant transpiration. The reduction in net CO₂ assimilation, A_n, was less than proportional to the fraction of leaf area infected. Powdery mildew reduced both the maximal light-driven electron flux (J_max) and the apparent maximal carboxylation velocity (Vc_max) although Vc_max was slightly more impacted than J_max. No compensation for the infection occurred in healthy leaves of partly infected seedlings as the reduced photosynthesis in the infected leaves was not paralleled by increased A_n levels in the healthy leaves of the seedlings. However, E. alphitoides had a strong impact on the leaf life-span of infected leaves. It is concluded that the moderate effect of E. alphitoides on oak might be related to the small impact on net CO₂ assimilation rates and on tree transpiration; nevertheless, the severe reduction in leaf life-span of heavily infected leaves may lead to decreased carbon uptake over the growth season.