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Cadophora luteo‐olivacea is the causal agent of the skin‐pitting disease of kiwifruit, a syndrome that appears after 4–5 months of cold storage. However, it is assumed that the infection takes place in the field during fruit development. The present work takes into consideration the production of volatile organic compounds (VOCs) of Actinidia deliciosa 'Hayward' at different phenological phases as potential C. luteo‐olivacea infection biomarkers. In vitro assays were conducted to gain knowledge on the effect of kiwifruit VOCs on pathogen conidial germination and mycelial growth. VOCs produced by kiwifruit either inoculated or not with C. luteo‐olivacea were analysed at different phenological phases by SPME/GC–MS analysis. In particular, ethanol, o‐xylene, d‐limonene and acetic acid showed a significant increase in the presence of fungal inoculation. Ethanol and d‐limonene were also detected as volatile metabolites of the pathogen. The effect of each compound (ethanol, o‐xylene, d‐limonene and acetic acid) was tested on the fungal conidial germination at different concentrations, showing a growth stimulation at lower amounts. These results show how the production of some VOCs can contribute to the knowledge of fruit–pathogen interaction in the field with the aim of developing future tools for early disease detection and consequent effective control. [ABSTRACT FROM AUTHOR]

Brown rot in apples is caused by fungi belonging to the genus Monilinia and occurs annually in orchards with varying severity. The symptoms of the disease on apple fruits may occur during harvest and can also develop much later in storage. Therefore, from a practical point of view, it is very important to be able to test a sample of asymptomatic apples before putting them in storage or sending them to distant markets. Thus, there is a need to develop a quick and sensitive method of detecting disease causal agents in asymptomatically infected fruit. For this purpose, a procedure for DNA isolation from the skin of apples was developed, and a set of primers for the loop‐mediated isothermal amplification (LAMP) reaction was designed based on a fragment of the hsp60 gene encoding heat shock protein 60 of Monilinia fungi that are pathogenic to apple: M. fructigena, M. fructicola and M. polystroma. The method was validated by reactions with DNA from infected apple fruit exhibiting brown rot symptoms and DNA from infected apple fruit without visible brown rot symptoms (latent infection). To increase the sensitivity of detection before the LAMP reaction, preamplification of the target DNA was carried out. The sensitivity of detection in the LAMP assay for the DNA of all three tested species of fungi was 2 pg. The DNA preamplification significantly increased the sensitivity of Monilinia DNA detection to 0.1 pg. This study shows that the developed LAMP method can be used to detect Monilinia fungi in asymptomatically infected apples. [ABSTRACT FROM AUTHOR]

It is critical to quantify latent Plasmopara viticola infections in commercial vineyards rapidly and accurately to estimate the potential for grapevine downy mildew (GDM) epidemics. In this study, a well‐established quantitative PCR (qPCR) assay was employed to quantify the amount of P. viticola in asymptomatic grapevine leaves from various commercial vineyards in the Ningxia Hui Autonomous Region (Ningxia), China. The qPCR revealed efficient detection of P. viticola, with 56.6% and 95.8% of the asymptomatic leaves testing positive to the pathogen in 2018 and 2019, respectively. The molecular disease index (MDI), which offers insights into the progression of latent infection preceding the appearance of visible GDM symptoms, showed relatively modest values in the initial phases. However, it increased variably as grapevines grew. The disease index progress curve and the relative area under the disease progress curve revealed that all assessed vineyards experienced different degrees of epidemics after the initial GDM observation, with most vineyards experiencing more severe GDM epidemics in 2019 than in 2018. Bayesian and non‐linear regression analyses demonstrated that MDI data matched well with occurrence of GDM at early, mid and late stages of disease, and the overall accuracy of predictions exceeded 50%, 64% and 70%, respectively. Furthermore, latent infection levels were positively correlated with subsequent assessments of disease progression (p < 0.001). This study provides evidence for the interrelation between latent P. viticola infections and development of GDM epidemics under field conditions. This insight has substantive implications for the formulation of effective disease management strategies. [ABSTRACT FROM AUTHOR]

The grey mould pathogen Botrytis cinerea forms systemic associations in some hosts, spreading into plant organs produced a considerable time after initial infection. These infections may have no macroscopic symptoms during much of the hosts' lifetime and are at least partially within the host tissue. The aim of the studies reported here was to locate and visualize these infections at a cellular level in Lactuca sativa (lettuce) and Arabidopsis thaliana. Symptomless but infected plants were produced by dry spore inoculation of plants growing in conditions previously shown to result in fungal spread from the initial inoculation site to newly developing plant organs. Tissue taken from inoculated plants was examined using confocal laser scanning microscopy. Two B. cinerea isolates were used: B05.10 and its GFP‐labelled derivative Bcgfp1‐3. Spore germination on leaf surfaces was followed by development of subcuticular inclusions and plant cell damage in single infected epidermal cells and sometimes a few nearby cells. Sparsely branched long hyphae arose and spread from the inclusions, mostly on the outer surface of the epidermal layer but occasionally below the cuticle or epidermal cells, where further inclusions formed. This was consistent with the pattern in time of recovery of B. cinerea from surface‐sterilized leaf tissue. In the late symptomless phase, mycelium arising from internal fungal inclusions formed mycelial networks on the surface of leaves. Symptomless exterior mycelium grew on the roots in A. thaliana. [ABSTRACT FROM AUTHOR]

Control of grey mould, caused by Botrytis spp., is a major challenge in open field strawberry production. Botrytis was isolated from plant parts collected from 19 perennial strawberry fields with suspected fungicide resistance in the Agder region of Norway in 2016. Resistance to boscalid, pyraclostrobin and fenhexamid was high and found in 89.1%, 86.0% and 65.4% of conidia samples, respectively. Multiple fungicide resistance was common; 69.6% of conidia samples exhibited resistance to three or more fungicides. Botrytis group S and B. cinerea sensu stricto isolates were obtained from 19 and 16 fields, respectively. The sdhB, cytb, erg27 and mrr1 genes of a selection of isolates were examined for the presence of mutations known to confer fungicide resistance to boscalid, pyraclostrobin, fenhexamid and pyrimethanil plus fludioxonil, respectively. Allele‐specific PCR assays were developed for efficient detection of resistance‐conferring mutations in cytb. Among B. cinerea isolates, 84.7%, 86.3% and 61.3% had resistance‐conferring mutations in sdhB, cytb and erg27, respectively. A triplet deletion in mrr1, resulting in ΔL497, commonly associated with the multidrug resistance phenotype MDR1h, was detected in 29.2% of Botrytis group S isolates. High frequencies of resistance to several fungicides were also detected in Botrytis from both imported and domestically produced strawberry transplants. Fungicide resistance frequencies were not different among fields grouped by level of grey mould problem assessed by growers, indicating factors other than fungicide resistance contributed to control failure, a fact that has important implications for future management of grey mould. [ABSTRACT FROM AUTHOR]

Anthracnose is the main disease of persimmon and is caused by Colletotrichum spp. The study of field epidemiology is essential for the development of efficient management of this disease. In this study, we investigated infection by Colletotrichum spp. throughout the persimmon growing season to understand the host–pathogen interactions better. We observed the production of primary inoculum of persimmon anthracnose and described how epidemics progress from secondary infections during the fruit crop season. The field study was carried out in an organic orchard with three susceptible persimmon cultivars, Fuyu, Kakimel and Jiro, for three consecutive seasons. Our results indicate that the pathogen survives in 1‐year‐old shoots, which are the sources of primary inoculum. Later that growing season, the inoculum reaches flowers and new shoots, developing symptoms and producing the secondary inoculum. Fruit drop was also observed, with or without symptoms of anthracnose, throughout the plant cycle. In some of the symptomless fruit, collected from the plant and from the ground where they had fallen, latent infections of Colletotrichum spp. were detected. Shoots, flowers, immature and ripened fruit remained infected throughout the growing season, producing conidia that could lead to new secondary infections within and among plants. The incidence of anthracnose in fruit at harvest and postharvest proved to be less relevant for disease management. Practices for chemical and cultural control of the disease throughout the persimmon growing season are discussed. [ABSTRACT FROM AUTHOR]

Black dot and silver scurf are potato blemish diseases whose economic impact has increased in recent years. Because their symptomatology on tubers is visually similar, disease assessment does not usually differentiate between the two pathogens, which share the same ecological niche. The epidemiology of black dot has been extensively studied, especially in the UK, but the factors that influence silver scurf have been less investigated. In this study, the influence of cultivar, source of inoculum, and environmental conditions on both diseases was studied in field trials over a three‐year period (2016–2018) in Switzerland. Planting minitubers did not prevent either disease in daughter tubers, indicating the contribution of soil as an inoculum reservoir. An arbitrary threshold of Colletotrichum coccodes soil inoculum could be set to discriminate between low and high disease risk. For the first time, Helminthosporium solani DNA was detected in stolons, and infections appeared earlier in stolons than in tubers. H. solani stolon and tuber infections usually appeared later in the season than those of C. coccodes. Black dot severity correlated positively with precipitation, while silver scurf severity correlated positively with temperature. Table potato cultivars commonly grown in Switzerland exhibited significant differences in susceptibility to both diseases, and cultivars with low susceptibility to both silver scurf and black dot were identified. These results gave new insights into understanding the factors driving the epidemiology of potato blemish diseases and may contribute to building a risk assessment scheme to manage both diseases simultaneously. [ABSTRACT FROM AUTHOR]

Potato blackleg is a seedborne disease that can cause significant economic losses for growers. Disease development depends mainly on two drivers, namely seed inoculum and local climatic conditions. To better establish the relationship between these two drivers, blackleg development was monitored in Swiss field trials at multiple locations from 2010 to 2013 involving three sets of naturally infected seed lots planted in each of three locations. The seed lot itself was thereby the most important factor explaining differences in disease development, rather than environmental factors. In a further on‐farm project conducted at various locations in Switzerland and southern Germany from 2013 to 2015, the implementation of a seed‐testing procedure was investigated. A total of 177 seed lots were tested for natural latent infection with soft rot Pectobacteriaceae and the corresponding blackleg incidence was tracked in 242 fields. The reliability of the relationship between latent infection and field incidence was found to be strongly linked to the bacterial species. Dickeya spp. field infection could be predicted with an acceptable reliability, whereas Pectobacterium carotovorum subsp. brasiliense, even when detected as latent tuber infection, was not consistently expressed as visual blackleg. Moreover, commonly found mixed latent infections with several bacterial species made it even harder to predict which bacteria would cause blackleg symptoms. Finally, variability in the reliability of seed testing may also be explained by differences in local farming practices. These trials over several years with naturally infected potato seed highlight the usefulness and limits of seed testing to manage blackleg. [ABSTRACT FROM AUTHOR]

Citrus tatter leaf virus (CTLV) causes chlorotic leaves with bud union incompatibility in citrus scion and its current incidence in Taiwan is 65%. However, the burden of infection has long been underestimated due to its common latency, and the serious threat of tatter leaf disease should be investigated and addressed without delay. Here, the biological characteristics and genome sequences of Taiwanese isolates are evaluated. Full‐length sequencing and phylogenetic analysis of three local isolates revealed similarities with isolates from neighbouring countries, indicating their possible origins. Two mild isolates obtained from Liuchen sweet orange (LCd‐NA‐1) and kumquat (Kq‐6‐2‐46) were inoculated on indicator plants and exhibited differences in foliar symptom expression and temperature tolerance compared with foreign isolates causing severe disease. Improved quantitative PCR‐based detection methods using redesigned primer pairs indicated high sensitivity and specificity for both local and foreign isolates. Similar but delayed systemic symptoms were observed when wildtype virus in vitro transcripts generated from a full‐length infectious clone of the Ponkan mandarin isolate (pCTLV‐Pk‐8) were inoculated on Chenopodium quinoa. These symptoms were unevenly distributed and highly concentrated on the inoculated sides of leaves and top leaves. A single codon mutation (U5642C) in the coat protein transcription start site in the pCTLV‐Pk‐8 sequence resulted in defective systemic infection, indicating a role for the coat protein in viral movement. The combination of biological assays, improved detection and infectious clone construction provides comprehensive information and a foundation for citrus virus management and further research. [ABSTRACT FROM AUTHOR]

Based on partial sequence analysis of the β-tubulin gene, 19 isolates of fungi causing bull's eye rot on apple in Poland were classified into species: Neofabraea alba, N. perennans and N. kienholzii. To the authors' knowledge, the detection of N. kienholzii is the second in Europe and the first in Poland. Species affiliation of these fungi was confirmed by a new species-specific multiplex PCR assay developed on the basis of previously published methods. The new protocol allowed for the specific identification of bull's eye rot-causing species, both from pure cultures and directly from the skin of diseased or apparently healthy apples. In 550 samples of diseased fruits collected from nine cold storage rooms located in three regions of Poland, in 2011 and 2012, N. alba was detected as the predominant species causing bull's eye rot, occurring on average in 94% of the tested samples. Neofabraea perennans was found in a minority of apple samples, N. kienholzii was found only in two apple samples, while N. malicorticis was not detected in any sample tested. In tests on 120 apparently healthy fruits, only N. perennans was detected in a single sample. The results of genetic diversity analyses of bull's eye rot-causing fungi based on the β-tubulin gene sequence and an ISSR (inter-simple sequence repeat) PCR assay with two primers were consistent, showing the expected segregation of tested isolates with respect to their species boundaries. However, the genetic distance between N. perennans and N. malicorticis was very low, as reported previously. [ABSTRACT FROM AUTHOR]

Botrytis cinerea, the main pathogen of strawberry, has the ability to remain quiescent in unripe tissue and develop disease symptoms in ripe fruit. As strawberry ripening is characterized by an increase of aroma compounds, the role of volatile emission in the development of infection was investigated. Thirty-five strawberry volatile organic compounds ( VOCs) were tested on B. cinerea in vitro and volatile emission was analysed in strawberry harvested at four ripening stages by headspace solid-phase microextraction/gas chromatography-mass spectrometry and proton transfer reaction-time of flight-mass spectrometry. The coupling of such data sets made it possible to conclude that key strawberry aroma compounds stimulate B. cinerea conidial germination and some typical wound-volatiles stimulate pathogen conidial germination or mycelial growth. This study is the first report of fungal stimulation by some VOCs naturally occurring in strawberry: the esters ethyl butanoate, cis-3-hexenyl acetate, trans-2-hexenyl acetate, methyl butanoate and hexyl butanoate, the furanones furaneol and mesifurane, and the alcohol trans-2-hexenol. The results of this work provide advances in understanding the functional role of fruit VOCs and suggest, for the first time, that fruit VOCs may influence the development of B. cinerea from the latent phase and that they could favour the invasive growth of B. cinerea after wounding. In particular, ethyl butanoate and furaneol could signal strawberry ripening, and the green leaf volatiles trans-2-hexenol, trans-2-hexenyl acetate and cis-3-hexenyl acetate could signal the presence of damaged tissues that are easier sites for penetration by B. cinerea. [ABSTRACT FROM AUTHOR]

Botrytis bunch rot ( BBR), caused by Botrytis cinerea, degrades wine grapes during ripening, even though infection can occur as early as flowering. Effective BBR management requires knowledge of whether some stages of fruit development are more important than others in relation to infection and BBR severity at harvest. Bunches of Vitis vinifera ' Sauvignon blanc' and/or ' Pinot noir' were inoculated in two vineyard trials and one glasshouse trial with nitrate non-utilizing ( nit) mutant strains at three phenological stages: early flowering, pre-bunch closure ( PBC) and veraison. Isolates recovered from symptomless berries at veraison and from bunches with symptoms at harvest were screened to measure the incidence of the nit strains used in the inoculations. It was found that latent infections, which resulted in BBR at harvest, could become established at all three phenological stages and no single stage was associated with greater latent incidence or harvest severity than any other stage. It was concluded that a proportion of BBR at harvest resulted from the expression of latent infections that had accumulated throughout the season. However, the time between infection and BBR symptom expression in near-ripe grape berries was sufficiently short for polycyclic secondary infection to also contribute to epidemic development. [ABSTRACT FROM AUTHOR]

Dickeya species (formerly Erwinia chrysanthemi) cause diseases on numerous crop and ornamental plants world-wide. Dickeya spp. (probably D. dianthicola) were first reported on potato in the Netherlands in the 1970s and have since been detected in many other European countries. However, since 2004-5 a new pathogen, with the proposed name ' D. solani', has been spreading across Europe via trade in seed tubers and is causing increasing economic losses. Although disease symptoms are often indistinguishable from those of the more established blackleg pathogen Pectobacterium spp., Dickeya spp. can initiate disease from lower inoculum levels, have a greater ability to spread through the plant's vascular tissue, are considerably more aggressive, and have higher optimal temperatures for disease development (the latter potentially leading to increased disease problems as Europe's climate warms). However, they also appear to be less hardy than Pectobacterium spp. in soil and other environments outside the plant. Scotland is currently the only country in Europe to enforce zero tolerance for Dickeya spp. in its potato crop in an attempt to keep its seed tuber industry free from disease. However, there are a number of other ways to control the disease, including seed tuber certification, on-farm methods and the use of diagnostics. For diagnostics, new genomics-based approaches are now being employed to develop D. dianthicola- and ' D. solani'-specific PCR-based tests for rapid detection and identification. It is hoped that these diagnostics, together with other aspects of ongoing research, will provide invaluable tools and information for controlling this serious threat to potato production. [ABSTRACT FROM AUTHOR]

Real-time PCR assays for Colletotrichum acutatum, one of the most important pathogens of strawberry worldwide, were developed using primers designed to the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) and the β-tubulin 2 gene. Using TaqMan technology, the ITS-based assay could reliably detect as little as 50 fg genomic DNA, 100 copies of target DNA, or 25 conidia. The β-tubulin-based assay was c. 66 times less sensitive, and therefore less suitable for detection purposes. The TaqMan-ITS assay recognized all C. acutatum isolates tested from various intraspecific molecular groups, while no amplification was observed with several other Colletotrichum species or other strawberry pathogens, indicating the specificity of this assay. Detection and quantification of C. acutatum was demonstrated in artificially and naturally infected strawberry leaves. First, C. acutatum was detected in plant mixes of which only 0·001% of the tissue was infected by C. acutatum. Secondly, real-time PCR analysis of leaf samples taken at various times after inoculation indicated that the assay allowed monitoring of growth progression of C. acutatum. This real-time PCR-mediated monitoring of the pathogen was well-correlated with microscopic data, and confirmed that leaf age may play a role in the extent of C. acutatum infection. Finally, the assay allowed detection of C. acutatum in naturally infected and symptomless strawberry leaves collected from production fields and planting material. [ABSTRACT FROM AUTHOR]

Xanthomonas fragariae, the causal agent of angular leaf spot on strawberry, is a quarantine organism in strawberry propagation material in the European Union. For the reliable screening of planting material for latent infections, a real-time PCR assay based on Taqman® chemistry for the detection of X. fragariae was developed. Primers and probe sequences were based on a DNA fragment amplified by a previously reported X. fragariae-specific technique. The sequence of this genomic fragment had no significant similarity with any published GenBank sequence. Specificity of the designed assay was tested with an extended range of X. fragariae collection strains and isolates, with other Xanthomonas spp. and with unidentified bacterial isolates from strawberry plants. A nested PCR, which until now was the reference method for sensitive detection in planta, cross-reacted with the reference strain of Xanthomonas campestris pv. campestris. In combination with an elaborated DNA extraction procedure, the Taqman® PCR enabled reliable detection down to 300 colony forming units in a 100 mg strawberry leaf sample. The assay offers a new tool for epidemiological research and for sanitary control of plant material with low level or latent infections of X. fragariae. [ABSTRACT FROM AUTHOR]

Resistance of cultivars of Anthurium andraeanum to systemic infection by Xanthomonas axonopodis pv. dieffenbachiae, the causal organism of bacterial blight disease of anthurium, was investigated using a bioengineered bacterial strain containing p519ngfp plasmid. Successful infection establishment in anthurium was found to be cultivar and inoculum density dependent, but independent of plant age. Injection of cut petioles (stage-2 leaf) with 100  µL inoculum (109 CFU mL−1) resulted in 100% infection establishment in susceptible cultivars on a repeatable basis, and differentiated between various levels of observed field resistance. Time to death (weeks) and proportion of dead plants best differentiated between levels of resistance and cultivars were placed in four groups based on these criteria. The susceptible group (32 cultivars) rapidly declined within 6–12 weeks of inoculation (WAI) and resulted in 100% plant death; the moderately resistant group (10 cultivars) declined within 12 WAI, but resulted in less than 100% plant death; the resistant category had less than 100% plant death with a slow decline taking over 20 weeks; and the highly resistant category (15 cultivars) showed 0% infection. The correlation coefficient between green fluorescent protein (GFP)-fluorescence and eventual death of plants was 0·90, indicating that the final death of individual plants can be reasonably well predicted based on GFP-fluorescence data at 5 WAI. Hence GFP data at 5 WAI can be used for early detection of latently infected plants and may assist screening for resistance in segregating populations of anthurium. [ABSTRACT FROM AUTHOR]

This research investigated the effects of wounding, fruit age and wetness duration on the development of cherry brown rot. Both Monilinia laxa and M. fructigena infected wounded detached cherry fruits, but M. laxa caused more infections than M. fructigena and only M. laxa efficiently infected intact detached fruits. Results from field monitoring and controlled inoculation in a polyethylene tunnel showed that the susceptibility of fruits to infection by M. laxa increased with fruit maturity. Infection of attached intact fruits by M. laxa was not affected by the length (3–24 h) of the wet period tested. [ABSTRACT FROM AUTHOR]

Three soft rot erwinias, Erwinia carotovora ssp. carotovora , E. carotovora ssp. atroseptica and E. chrysanthemi are associated with potatoes causing tuber soft rot and blackleg (stem rot). Latent infection of tubers and stems is widespread. As opportunistic pathogens, the bacteria tend to cause disease when potato resistance is impaired. Pathogenesis or disease development in potato tubers and stems is discussed in terms of the interaction between pathogen, host and environment, microbial competition and recent findings on the molecular basis of pathogenicity. Emphasis is placed on the role of free water and anaerobiosis in weakening tuber resistance and in providing nutrient for erwinias to multiply. Blackleg symptoms are expressed when erwinias predominate in rotting mother tubers, invade the stems and multiply in xylem vessels under favourable weather conditions. Soft rot erwinias tend to out-compete other bacteria in tuber rots because of their ability to produce larger quantities of a wider range of cell wall-degrading enzymes. However, despite extensive studies on their induction, regulation and secretion, little is known about the precise role of the different enzymes in pathogenesis. The putative role of quorum-sensing regulation of these enzymes in disease development is evaluated. The role certain pathogenicity-related characters, including motility, adhesion, siderophores, detoxifying systems and the hrp gene complex, common to most bacteria including symbionts and saprophytes, could play in latent and active infections is also discussed. [ABSTRACT FROM AUTHOR]

Transmission of pectinolytic Erwinia species from infected mother tubers to daughter tubers has been studied mainly through detection tests, carried out at harvest, on limited samples of tubers produced by plants grown from artificially inoculated mother tubers. However, detection has not been performed on samples collected at different stages of crop development, in order to follow the contamination progress in different organs through the plants to the progeny tubers. In this study the bacterial contamination of progeny tubers was investigated by detecting Erwinia carotovora ssp. atroseptica in different symptomless plant organs (stolons, stems, progeny tubers) and in the parts with or without symptoms of diseased stems, collected at various stages of crop development. Infection levels in below- and above-ground organs of plants of two cultivars differing in their resistance to Erwinia, infected by either vacuum infiltration or sand wounding, were monitored throughout the growing season and at harvest using DAS-ELISA and PCR. Detection tests showed that healthy organs from symptomless plants were less frequently contaminated than symptomless organs from diseased plants, and that stolons were precociously and more frequently contaminated than stems and daughter tubers, irrespective of the health of the plant. Stem infections were shown to progress latently in the stem, bacteria usually being recovered 10–15 cm past visible lesions. In many cases, typical aerial stem-rot symptoms could be related to this upward movement of bacteria from the infected mother tuber. Daughter tubers without symptoms were shown to be frequently contaminated, usually at heel ends, suggesting internal contamination from mother tuber to progeny. [ABSTRACT FROM AUTHOR]

This article examines the use of Chenopodium (C.) quinoa as an indicator plant for carnation latent virus. The reactions of C. quinoa to carnation latent virus are usually sufficiently distinct from those of other carnation viruses to provide a diagnostic indicator plant for routine testing of carnation. These reactions might sometimes be confused with those of vein mottle virus, but the latter is seldom seen in British carnation stocks. In sap-rubbing inoculation, C. quinoa reacted to carnation latent virus with indefinite local yellowish spots, followed by systemic interveinal yellowish spotting-mottle, puckering, and rugosity.

An abstract of the article "First report of Narcissus latent virus in New Zealand," by T. Wei, M. N. Pearson and D. Cohen is presented.