A nonlinear model for temperature‐dependent development of Erysiphe necator chasmothecia on grapevine leaves

Production and development of the chasmothecia of Erysiphe necator on Vitis vinifera leaves were studied using potted plants in controlled and outdoor environments and grapevines in a vineyard. The optimum temperature for ascocarp production was 20°C; fewer chasmothecia were produced at 15°C and even fewer at 25°C; at 10 and 30°C, no or very few chasmothecia were observed, and none reached maturity. Nonlinear equations describing ascocarp development as a function of time and temperature were developed, parameterized with data from experiments at constant temperatures, and evaluated under fluctuating temperatures. Goodness‐of‐fit showed high agreement between observed and predicted data: the model efficacy ranged from 0·74 to 0·97 (1·0 indicates a perfect fit), and the root mean square error ranged from 0·001 to 0·01 (zero indicates a perfect fit). The high proportion of the observed variability accounted for by these equations (R² = 0·83–0·98) supported the hypothesis that temperature has a predominant role in ascocarp development under natural conditions, when all environmental factors interact. The equations tended to overestimate the production of mature chasmothecia (the coefficient of residual mass was ?0·23), but this inconsistency mainly occurred when rainfall apparently washed the mature chasmothecia from leaves during the logarithmic phase of the ascocarp developmental curve. Results from this work will be useful for predicting the development of chasmothecia in a vineyard and for timing the use of natural products, fungicides or biocontrol agents for reducing the population of chasmothecia, which are all more effective when they are applied to immature chasmothecia.     

Efficiency of inorganic fungicides against the formation of Erysiphe necator chasmothecia in vineyards

BACKGROUND

A reduction in chasmothecia, an important inoculum of grape powdery mildew (Erysiphe necator Schwein.), is essential for disease control in vineyards; the use of fungicides during the formation of chasmothecia on vine leaves, late in the growing season, may accomplish this. Inorganic fungicides, such as sulphur, copper, and potassium bicarbonate, are very useful for this purpose because of their multisite mode of action. The aim of this study was to evaluate chasmothecia reduction using different fungicide applications late in the growing season in commercially managed vineyards and in an exact application trial.

RESULTS

Chasmothecia on vine leaves were reduced in commercial vineyards by four copper (P = 0.01) and five potassium bicarbonate (P = 0.026) applications. The positive effect of potassium bicarbonate was also confirmed in the application trial, where two applications showed lower chasmothecia numbers than the control (P = 0.002).

CONCLUSION

The application of inorganic fungicides reduced the amount of chasmothecia as the primary inoculum source. Potassium bicarbonate and copper are of further interest for disease control as these fungicides can be used by organic and conventional wine growers. The application of these fungicides should be carried out as late as possible before harvest to reduce chasmothecia formation and, consequently, the potential for powdery mildew infestation in the subsequent season. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Dynamics of ascospore maturation and discharge in Erysiphe necator, the causal agent of grape powdery mildew

Dynamics of ascocarp development, ascospore maturation, and dispersal in Erysiphe necator were studied over a 4-year period, from the time of ascocarp formation to the end of the ascosporic season at the end of June in the following spring. Naturally dispersed chasmothecia were collected from mid-August to late November (when leaf fall was complete); the different collections were used to form three to five cohorts of chasmothecia per year, with each cohort containing ascocarps formed in different periods. Chasmothecia were exposed to natural conditions in a vineyard and periodically sampled. Ascocarps were categorized as containing mature or immature ascospores, or as empty; mature ascospores inside chasmothecia were enumerated starting from late February. Ascospore discharge was determined using silicone-coated slides that were placed 3 to 4 cm from sections of the vine trunk holding the chasmothecia. Before complete leaf fall, 34% of the chasmothecia had mature ascospores, 48% had immature ascospores, and 18% were empty; in the same period, the trapped ascospores represented 56% of the total ascospores trapped in an ascosporic season (i.e., from late summer until the next spring or early summer). The number of viable chasmothecia diminished over time; 11 and 5% of chasmothecia had mature ascospores between complete leaf fall and bud break and after bud break, respectively. These ascocarps discharged ≈2 and 42% of the total ascospores, respectively. All the ascocarp cohorts released ascospores in autumn, survived the winter, and discharged viable ascospores in spring; neither ascospore numbers nor their pattern of temporal release was influenced by the time when chasmothecia were collected and exposed in the vineyard. Abundance of mature ascospores in chasmothecia was expressed as a function of degree-days (DD) (base 10°C) accumulated before and after bud break through a Gompertz equation (R2 = 0.92). Based on this equation, 90% of the ascospores were mature when 153 DD (confidence interval, 100 to 210 DD) had accumulated after bud break. Most ascospores were trapped in periods with >2 mm of rain; however, a few ascospores were airborne with <2 mm of rain and, occasionally, in wet periods of ≥3.5 h not initiated by rain.     

Identification and distribution of mating‐type idiomorphs in populations of Podosphaera macularis and development of chasmothecia of the fungus

Podosphaera macularis, the causal agent of hop powdery mildew, is known to produce chasmothecia (formerly cleistothecia) in eastern North America and Europe. Ascocarps have not yet been reported from the Pacific Northwestern region of North America. Reasons for the apparent absence of chasmothecia in the Pacific Northwest were unknown. This study established that P. macularis is heterothallic and ascocarp ontogeny, maturation, dehiscence and ascospore infection proceed similarly to other powdery mildew fungi. Genome sequencing of a MAT1‐1 isolate revealed the structure of the MAT1 locus and presence of MAT1‐1‐3, demonstrating further similarities to other powdery mildew fungi. PCR assays with primers designed from conserved domains of the MAT1 idiomorphs were developed to characterize the frequency of idiomorphs in populations of P. macularis. Amongst 317 samples of P. macularis collected during 2012 and 2013 from the Pacific Northwest only the MAT1‐1 idiomorph was found. In contrast, among 56 samples from the eastern United States and Europe, MAT1‐1 and MAT1‐2 idiomorphs were detected at equivalent frequencies. At temperatures representative of late season conditions in the Pacific Northwest, chasmothecia formed readily when a Pacific Northwest MAT1‐1 isolate was paired with a MAT1‐2 isolate collected from outside the region. Although these findings do not encompass all climatic, geographic or temporal barriers that could inhibit the formation of chasmothecia, the current absence of the ascigerious stage of P. macularis in the Pacific Northwest could be explained by the absence of the MAT1‐2 mating type idiomorph.     

加那利海枣叶斑病病原鉴定*

对加那利海枣叶斑病的症状进行了观察。结果表明,加那利海枣叶斑病主要发生在叶片和叶柄。初期症状是在叶片和叶柄出现水渍状褪绿色的褐色小点,病斑扩大成浅褐色椭圆形或不规则病斑,病斑周围有黄色的晕圈,后期变褐色至灰褐色,中央产生小黑点。按照柯赫氏法则对该病害的病原菌进行分离和致病性测定。通过形态学观察和rDNA ITS序列分析鉴定,将病原菌鉴定为异色拟盘多毛孢［Pestalotiopsis versicolor (Speg.) Steyaert］。     

Ochratoxin A-producing fungi in Spanish wine grapes and their relationship with meteorological conditions

Forty vineyards from four wine making regions of Spain were sampled at three different growth stages in 2002 and 2003. The aim was to study the fungi associated with grapes and their ability to produce ochratoxin A (OTA) on synthetic media. Among the total mycoflora, 464 (7.7%) and 648 (10.8%) Aspergillus section Nigri (black aspergilli) strains were isolated in 2002 and 2003, respectively, and were classified into three groups: isolates with uniseriate heads, A. niger aggregate and A. carbonarius. The latter presented the highest percentage of OTA-positive strains (82% in 2002 and 76% in 2003) and produced the highest levels of toxin (2.5–25 μg g⁻¹). The sampling year, sampling date, the region and their interactions presented significant differences in the number of black aspergilli isolated. Most black aspergilli were found in 2003 and at harvest. A positive correlation between the number of black aspergilli found in grapes and the temperature in the field was found. Grapes from 2003, the warmest year, and from Costers del Segre, the warmest region, were significantly the most contaminated. No significant correlation between black aspergilli presence and other meteorological factors such as relative humidity or rainfall could be established. Musts from all the vineyards were also analysed in both years, although no OTA was found in either year.     

Combining sanitation and disease modelling for control of grapevine powdery mildew

Chasmothecia of Erysiphe necator form in one season, survive winter and discharge ascospores that cause primary infections and trigger powdery mildew epidemics in the next season. A strategy for powdery mildew control was developed based on (i) the reduction in overwintering chasmothecia and on (ii) spring fungicide applications to control ascosporic infections timed based on estimate risk (two to five sprays per season). Several fungicides, the hyperparasite Ampelomyces quisqualis, and a mineral oil product were first tested as separate applications in a greenhouse and in vineyards. In the greenhouse, A. quisqualis suppressed chasmothecia formation by 41 %; fungicides and mineral oil suppressed chasmothecia formation by 63 % and ascospore viability by 71 %. In vineyards, application of boscalid + kresoxim-methyl or meptyldinocap once after harvest, as well as application of A. quisqualis pre- and post-harvest, delayed disease onset and epidemic development in the following season by 1 to 3 weeks and lowered disease severity (up to the pea-sized berry stage) by 56 to 63 %. Risk-based applications of sulphur and of synthetic fungicides provided the same control as the grower spray program but required fewer applications (average reduction of 47 %). Sanitation strategies were then tested by combining products and application times (late-season, and/or pre-bud break, and/or spring). Adequate disease control with a reduced number of sprays was achieved with the following combination: two applications of A. quisqualis (pre- and post-harvest), one application of mineral oil before bud break, and model-based applications of sulphur fungicides between bud break and fruit set.     

吐鲁番地区葡萄生长期果实腐烂病研究初报

通过葡萄果实腐烂病田间发病情况调查与症状观察,并对发病果穗进行病原分离、培养和回接试验,结果表明,葡萄果实腐烂病的病原是 Aspergillus niger v. Tiegh,其菌落呈黑褐色,分生孢子头球形,直径200～500 μm,分生孢子梗壁光滑,带黄色。顶囊球形或近球形,直径35～70 μm,褐黄色,全部表面可育;产孢结构双层,瓶梗8～10（~12）μm×2～3 μm。分生孢子球形或近球形,直径3～5 μm,孢子壁表面粗糙,有的生有小疣。     

海南省温郁金炭疽病的病原鉴定

温郁金炭疽病是一种重要的真菌性病害,该病主要危害叶片,初期产生水渍状的淡黄色病斑,后逐渐扩大成圆形、椭圆形或不规则形的灰褐色病斑,上面有黑色小点,病斑外面环绕一层黄色晕圈,严重时多个病斑联合造成叶片卷缩和枯死,2016年我们从海南省的温郁金种植基地采集炭疽病样本,经病原菌的分离纯化、柯赫氏法则验证、病原菌的形态特征观察及多基因谱系分析方法,确定其病原菌为郁金炭疽菌Colletotrichum curcumae(Syd.)E.J.ButlerBisby。     

烟草不同发育阶段对赤星病的抗性表现与综合防治技术

人工接种结果表明，随着烟叶自下而上逐渐成熟、衰老，抗病性下降。不同抗性类型的品种，其绿色叶片对病害均表现免疫或近免疫，近成熟叶片高抗，成熟衰老叶片表现为感病，与品种田间抗病性结果一致。田间结果表明，采用增施钾肥、适时打顶和下部叶片近成熟期摘除1～3片底脚叶等措施，提高烟草品种抗病性、配合有效的农药品种综合防治烟草赤星病，在重病区布局抗病品种能有效地推迟病害的发生时期，减慢病情的发展速度，且能减少农药施用量和施用次数，减少环境污染。     

Hyperspectral signal decomposition and symptom detection of wheat rust disease at the leaf scale using pure fungal spore spectra as reference

This study establishes a method to detect and distinguish between brown rust and yellow rust on wheat leaves based on hyperspectral imaging at the leaf scale under controlled laboratory conditions. A major problem at this scale is the generation of representative and correctly labelled training data, as only mixed spectra comprising plant and fungal material are observed. For this purpose, the pure spectra of rust spores of Puccinia triticina and P. striiformis, causal agents of brown and yellow rust, respectively, were used to serve as a spectral fingerprint for the detection of a specific leaf rust disease. A least-squares factorization was used on hyperspectral images to unveil the presence of the spectral signal of rust spores in mixed spectra on wheat leaves. A quantification of yellow and brown rust, chlorosis and healthy tissue was verified in time series experiments on inoculated plants. The detection of fungal crop diseases by hyperspectral imaging was enabled without pixel-wise labelling at the leaf scale by using reference spectra from spore-scale observations. For the first time, this study shows an interpretable decomposition of the spectral reflectance mixture during pathogenesis. This novel approach will support a more sophisticated and precise detection of foliar diseases of wheat by hyperspectral imaging.     

我国甘薯新病害-茎腐病的研究初报

 2006年以来广东省主要甘薯产区发现1种甘薯新病害,主要症状表现为叶片变黄,茎基部呈黑色水浸状腐烂,并逐渐沿茎枝向顶端腐烂,后整株倒伏、死亡。从甘薯病茎部分离得到了病菌,经柯赫法则验证为致病病原菌。根据病原菌的形态特征、培养性状、生理生化及16S rDNA序列分析,鉴定其为菊欧文氏菌(Erwinia chrysanthemi)。采用茎枝、菌液共培养法接种8个甘薯品种,结果表明8个品种均无抗病性,病原菌致病性较强。这是我国首次发现该病害。      

Phylogenetic and recombination analysis of sorghum isolates of Sugarcane yellow leaf virus

Recombination has played an important role in evolution and genetic diversity of Sugarcane yellow leaf virus (SCYLV) isolates sequenced to date. This study found that three newly sequenced SCYLV sorghum isolates from the USA underwent intraspecies recombination. No statistical significance on probable progeny–parent relationships involving SCYLV sorghum isolates were found in possible interspecies recombination with 18 members of the Luteoviridae family. Sorghum isolates deposited in the GenBank database under accession numbers KT960995, KT960996 and KT960997 were phylogenetically closely related to SCYLV genotypes IND, CUB and CHN1, all members of phylogroup II. Networked relationships among the sorghum isolates showed that numerous incompatibilities occurred in the sequences. These conflicting signals were probably due to recombination, especially in KT960997, which was heavily impacted by recombination. The KT960997 accession was positioned on a distinct branch compared to other members of phylogroup II, suggesting that it has probably emerged as a new genotype. Future studies on molecular evolution may reveal further insights into the adaptation capacity of these SCYLV lineages to new environments.     

Dry-dispersal and rain-splash of brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rust spores from infected wheat leaves exposed to simulated raindrops

The dispersal of spores from lesions of brown ( Puccinia recondita f.sp. tritici ) or yellow ( P. striiformis ) rusts of wheat by impacting drops was studied. Using a generator of uniform-size drops, drops of 2.5, 3.4, 4.2 and 4.9 mm in diameter were released from rest at heights of 5, 50 and 100 cm above horizontal and primary leaves uniformly covered with sporulating lesions. Dry-dispersal and rain-splash occurred simultaneously in response to drop impaction. A coloration technique allowed separate counting of dry-dispersed and rain-splashed spores caught on slides. More spores were rain-splashed than dry-dispersed. Neither removal mechanism affected in-vitro germination of spores, which was higher in brown than in yellow rust. For both rusts, the number of both dry-dispersed and rain-splashed spores, as well as their travel distance, increased with drop diameter and fall height. The fall speed of incident drops in relation to diameter and fall height was obtained by solving numerically the equation of vertical drop motion. The number of spores removed by a given impacting drop was found to be a power function of the calculated kinetic energy of the impacting drop. Based on this experimental relationship, a simulation study showed the relevance of rain type in the removal of spores.     

Co‐infection by Botryosphaeriaceae and Ilyonectria spp. fungi during propagation causes decline of young grafted grapevines

Decline of newly planted, grafted grapevines is a serious viticultural problem worldwide. In the Riverina (New South Wales, Australia), characteristic symptoms include low fruit yields, very short shoots and severely stunted roots with black, sunken, necrotic lesions. To determine the cause, roots and wood tissue from affected plants in 20 vineyards (Vitis vinifera cv. Chardonnay grafted to V. champini cv. Ramsey rootstock) were assayed for microbial pathogens. Ilyonectria spp. (I. macrodidyma or I. liriodendra, producers of phytotoxin brefeldin A, BFA, and cause of black foot disease of grapevines) and Botryosphaeriaceae spp. (predominantly Diplodia seriata) were isolated from rootstocks of 100 and 95% of the plants, respectively. Togninia minima and Phaeomoniella chlamydospora (cause of grapevine Petri disease) were isolated from 13 and 7% of affected plants, respectively. All Ramsey rootstock stems of grafted plants sampled from a supplier nursery were infected with Ilyonectria spp. and D. seriata. Diplodia seriata, but not Ilyonectria spp., was also isolated from 25% of canes sampled from the rootstock source block. Root inoculation of potted, disease‐free Chardonnay plants with Ilyonectria isolates from diseased vineyards caused typical disease symptoms, while co‐inoculation with Botryosphaeriaceae spp. increased disease severity. This is the first study to show that a major cause of young grapevine decline can be sequential infection by Botryosphaeriaceae from rootstock cuttings and Ilyonectria spp. from nursery soil. Although the Petri disease fungi were less common in young declining grafted grapevines in the Riverina, they are likely to contribute to the decline of surviving plants as they mature.     

Alternaria Brown Spot of Minneola in Greece; Evaluation of Citrus Species Susceptibility

During the last three years, a new disease was observed in northwestern Greece on Minneola trees, hybrid of mandarin and grapefruit. On May small brown necrotic leaf spots surrounded by yellow halo areas of various sizes appeared and covered a major portion of the leaves with extension of necrosis into the veins. On young fruits small, slightly depressed black spots were the first symptoms, which later became 2–7 mm in diameter. Brown spots were observed on the leaves and fruits in several orchards in the same area, causing leaves and fruits to drop. In some orchards over 50% of the fruits were affected. From the fruit and leaf spots the typical small-spore species Alternaria alternata was isolated. Pathogenicity tests were performed by artificially inoculating fruits of Minneola, common mandarin and Clementine. The symptoms of the disease were reproduced only on fruits of Minneola hybrids by the specific strain of the fungus Alternaria alternata pv. citri. Different citrus susceptibility tests indicated that mandarins Minneola, Nova and Page were very susceptible to tested isolates while Clementine SRA and Poros Clementine were not. All lemons and lime Seedless were not susceptible. Grapefruit New Hall was not susceptible, while the Star Ruby was. Orange Lane Late, Navel Late, Oval Poros, Olinda, Navel Athos were not susceptible and only Moro showed reaction being slightly susceptible only to one isolate.     

Species prevalence and disease progression studies demonstrate a seasonal shift in the Alternaria population composition on potato

To assess the incidence of early blight/brown spot (EB/BS) in Flanders (Belgium), potato fields in 22 locations were monitored and scored over the duration of the growing seasons 2014 and 2015. The average disease incidence was shown to be higher in 2014 than in 2015. Soil type, rainfall and temperature were also analysed in relation to disease incidence. In 2014, potato plants grown in sandy soils had more EB/BS disease than those grown in clay or loamy soils. However, the low disease incidence in 2015 meant that differences in disease levels between soil types could not be discerned for that growing season. A windowpane analysis demonstrated that rainfall and humidity accounted for the differences in disease incidence between the two growing seasons. During the course of the survey, the species composition in leaves with symptoms was assessed using real‐time PCR. Remarkably, small‐spored Alternaria species, such as A. alternata and A. arborescens, rather than the more virulent A. solani were the predominant species on potato leaves throughout the growing season. As the disease progressed, the proportion of A. solani increased. In view of these results, the virulence of a collected set of Alternaria isolates was assessed by an in vitro assay. Despite A. solani being more virulent than A. alternata or A. arborescens, the most abundant species isolated from potato leaves with symptoms was A. arborescens.     

High throughput real‐time RT‐PCR assays for specific detection of cassava brown streak disease causal viruses,and their application to testing of planting material

Cassava brown streak disease (CBSD) caused by Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) is causing severe losses in cassava production in Kenya, Tanzania and Uganda. Two real‐time RT‐PCR assays based on TaqMan chemistry capable of detecting and distinguishing these two viruses are described. These assays were used to screen 493 cassava samples collected from western and coastal Kenya, the main cassava regions of Uganda and inland Tanzania. Both viruses were found in all three countries and across regions therein. Association of CBSD leaf symptom status with CBSV and UCBSV assay results was weak, confirming the need for a diagnostic assay. For leaf samples that were observed with CBSD‐like leaf symptoms but shown as CBSV and UCBSV negative by the RT‐PCR assay, deep sequencing using a Roche 454 GS‐FLX was used to provide additional evidence for the absence of the viruses. The probability of the CBSD associated diagnostics detecting a single CBSV or UCBSV positive sample amongst other non‐CBSD samples was modelled. The results of this study are discussed in the context of the application of diagnostics of CBSD‐associated viruses under the Great Lakes Cassava Initiative and the need to minimize the risk of further spread of the viruses with cassava multiplication material. It is shown that high throughput testing undertaken at Fera of 300 cassava leaves taken from fields for seed multiplication, when analysed in pools of 10, has given a 95% probability of detecting 1% infected plants in the field.     

New Leaf Spot Disease of Cymbidium Species Caused by Fusarium subglutinans and Fusarium proliferatum

Fusarium species were consistently isolated from yellow, swollen spots with reddishbrown centers and small black spots on leaves of Cymbidium plants in the greenhouse. Fusarium subglutinans caused the yellow spots and Fusarium proliferatum caused either the yellow or the black spots. We propose the name “yellow spot” for the new disease. To denote differences in their pathogenicity to orchid plants, we designate the population causing yellow spot as race Y and that causing black spot as race B of F. proliferatum. Received 29 October 1999/ Accepted in revised form 10 March 2000