Tropical cover crops for the management of the yam nematode,Scutellonema bradys

Yam tuber dry rot caused by Scutellonema bradys results in the reduction of tuber quality. Studies were conducted on the effect of intercropping cover crops with yams on nematode damage to tubers. Eleven cover crops were intercropped with yams in pot screen house and field experiments. Plants were inoculated with S. bradys in water suspension in pots and by incorporating infected yam peels into soil around plant roots in field experiments with un-inoculated plants serving as control. The experiments were factorial with six replications per treatment in pot and four in field experiments. In pot experiments, populations of S. bradys recovered from tubers and nematode damage were lower in yam tubers intercropped with Tagetes erecta, Aeschynomene histrix, Stylosanthes guianensis, Mucuna pruriens and Pueraria phaseoloides compared to where no cover crops were intercropped with yams. Conversely, S. bradys populations from pots with Crotalaria ochroleuca, Crotalaria juncea, Centrosema pubescens, Cajanus cajan and Lablab purpureus were similar to Vigna unguiculata, a known host of S. bradys. The use of A. histrix, T. erecta and S. guianensis, M. pruriens and P. phaseoloides offers promise for inclusion in yam-based cropping systems as a measure for S. bradys management in yam production.     

Detection of the root lesion nematode Pratylenchus penetrans in potato tubers

The root lesion nematode Pratylenchus penetrans parasitizes a wide range of economically important crops, including potato (Solanum tuberosum). Damage by P. penetrans impacts not only the potato yield but can also reduce the tuber quality. Detailed information on tuber infection by P. penetrans is scarce for most cultivars and molecular detection of nematodes from infected tubers is needed. The objective of this study was to assess tuber symptomatology due to P. penetrans infection in 10 potato cultivars and to provide an accurate molecular methodology for nematode detection using tuber peels. Sprouts of certified potato seed from cultivars Agata, Agria, Camel, Désirée, Dirosso, Kennebec, Laura, Picasso, Royata, and Stemster were planted in 2 L pots, and soil was inoculated with 4 P. penetrans/g of soil. Sixty days after inoculation, tubers were harvested, inspected for lesions, and the number of nematodes/g of potato peel assessed. Observations of tubers with symptoms showed the presence of P. penetrans in superficial layers of peels around the lenticels and injured necrotic tissue. Different nematode stages were detected in tubers of all inoculated cultivars, varying from 4 to 46 nematodes/g of potato peel. Species-specific primers showed suitable sensitivity and reproducibility for the detection of P. penetrans in tuber potato peel samples. The molecular detection of P. penetrans directly from tuber peels can facilitate routine nematode inspections of potato seed tubers or cull potatoes for nematode detection, and prevent further dissemination of this species.     

Biological control of potato soft rot caused by Dickeya solani and the survival of bacterial antagonists under cold storage conditions

Dickeya and Pectobacterium are responsible for causing blackleg of plants and soft rot of tubers in storage and in the field, giving rise to losses in seed potato production. In an attempt to improve potato health, biocontrol activity of known and putative antagonists was screened using in vitro and in planta assays, followed by analysis of their persistence at various storage temperatures. Most antagonists had low survival on potato tuber surfaces at 4 °C. The population dynamics of the best low-temperature tolerant strain and also the most efficient antagonist, Serratia plymuthica A30, along with Dickeya solani as target pathogen, was studied with TaqMan real-time PCR throughout the storage period. Tubers of three potato cultivars were treated in the autumn with the antagonist and then inoculated with D. solani. Although the cell densities of both strains decreased during the storage period in inoculated tubers, the pathogen population was always lower in the presence of the antagonist. The treated tubers were planted in the field the following growing season to evaluate the efficiency of the bacterial antagonist for controlling disease incidence. The potato endophyte S. plymuthica A30 protected potato plants by reducing blackleg development on average by 58.5% and transmission to tuber progeny as latent infection by 47–75%. These results suggest that treatment of potato tubers with biocontrol agents after harvest can reduce the severity of soft rot disease during storage and affect the transmission of soft rot bacteria from mother tubers to progeny tubers during field cultivation.     

腐烂茎线虫在马铃薯不同组织中的数量动态

腐烂茎线虫病是近年来我国马铃薯生产中的一种新病害, 了解腐烂茎线虫的侵染规律对该病害的预防和控制及对马铃薯产量和品质提高具有重要意义。本研究采用盆栽法, 在苗期接种腐烂茎线虫, 研究该线虫在马铃薯不同组织中的数量动态及危害程度。结果显示, 在发芽-幼苗期(播种0 ~ 75 d), 母薯先受到腐烂茎线虫侵染; 在发棵期(播种75 ~ 90 d), 母薯和匍匐茎均检测到腐烂茎线虫; 在结薯期到成熟期(播种90 ~ 105 d), 线虫开始危害新生块茎, 随着块茎膨大危害逐渐加重。在马铃薯成熟期, 症状表现最明显, 新生块茎中线虫数量最多, 可达767.67条/g。不同时期的接种试验表明, 在匍匐茎形成期(播种后70 d)接种, 马铃薯受害最为严重, 发病率达100%, 块茎中线虫密度最高, 达到99.32条/g, 且块茎鲜重最低, 为14.02 g/个。本研究结果可为马铃薯腐烂茎线虫的适时有效防治提供参考。     

Root infection of potato by Spongospora subterranea: knowledge review and evidence for decreased plant productivity

Information is reviewed on root infection of potato by the plasmodiophorid Spongospora subterranea f. sp. subterranea. This pathogen has long been recognized as the cause of root galls (hyperplasia) and the economically important disease powdery scab on tubers (modified stolons). The significance for plant productivity of the zoosporangium stages of the pathogen in potato roots has only recently begun to be documented. Two experiments are described that assessed effects of S. subterranea root infection on potato plant root function and productivity. A greenhouse experiment measured root function and plant parameters for eight potato cultivars with markedly different susceptibilities to tuber powdery scab. Water uptake and plant growth were reduced by S. subterranea inoculation in all eight cultivars. The magnitudes of these negative effects, and intensities of root hyperplasia, differed among the cultivars, but were not related to respective susceptibilities to tuber powdery scab. A field trial assessed root function and plant productivity for a cultivar (Iwa) that is very susceptible to Spongospora tuber and root diseases. Soil water content beneath uninoculated plants was consistently less than for inoculated plants, indicating that inoculation reduced water uptake (root function). Inoculation reduced shoot and root dry weights, and reduced weight of tubers per plant by 42%. Spongospora subterranea causes three diseases of potato: root membrane dysfunction, root hyperplasia and tuber powdery scab. The root diseases caused by the pathogen are likely to be important both for powdery scab management and for deleterious effects on potato crop yields.     

PCR-based determination of colonization patterns during potato tuber infection by single and multiple pathogens

Potato tubers piled in storage are prone to infection by numerous pathogens. Each pathogen can cause damage alone, but severe losses often arise when more than one pathogen is involved. Currently, only a visual diagnosis is practiced on potato tubers before storing them, which does not allow any prediction of further disease spread. The aim of the present study was to determine differences in patterns of tissue colonization by several tuber decay pathogens and how late blight infection affects further tuber colonization by other important tuber pathogens. This study was conducted using artificial inoculation of potato tubers and PCR to provide an early and accurate diagnosis of disease development for major potato tuber rots, and to assess potential synergism/antagonism between Phytophthora infestans and other pathogens in stored tubers. In order to accurately follow the progress of each pathogen in tuber tissues, samples were collected over time from both the surface (peel, 0–2 mm depth) and internal tissues (flesh, depth > 2 mm) of the tubers at various distances from the inoculation site, at 3, 5, 7, 10, 12, 14, 17, and 19 days after inoculation. Successful detection of single or multiple pathogens was achieved using specific PCR-primers for each pathogen. Pathogens were always detected several centimeters ahead of the visible lesions. This tracking enabled us to determine the extent of colonization both on the tuber’s surface and in internal tissues by each tested pathogen, either after single or multiple infections involving P. infestans as the primary pathogen. The presence of P. infestans was shown to enhance the development of Pectobacterium atrosepticum and to slow down that of P. erythrospetica and Pythium ultimum. No noticeable effect on further tuber colonization by F. sambucinum, V. dahliae or V. albo-atrum was observed in the presence of P. infestans. This approach involving more than one pathogen is more realistic than classical studies considering single pathogens, and may be helpful in monitoring the sanitary status of stored tubers. Our results make the outcome of certain combinations of pathogens in potato tubers more predictable and may result in more efficient preventive measures.     

Stem and bulb nematode, Ditylenchus dipsaci, associated with a dry rot of potato tubers

About 10% of the ware-sized tubers from a 1990 potato crop were found to have dry rot. The affected tubers had well defined circular patches of dry rot, either with the tuber surface depressed and cracked, or with deep, open cavities where the tuber surface had disintegrated. The rotted tissues contained large numbers of the stem and bulb nematode, Ditylenchus dipsaci. This is the first recorded case in the UK; dry rot of potato tubers is usually caused by the potato tuber nematode D. destructor.     

Assessment of recent outbreaks of <Emphasis Type="Italic">Dickeya</Emphasis> sp. (syn. <Emphasis Type="Italic">Erwinia chrysanthemi</Emphasis>) slow wilt in potato crops in Israel

Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification. All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation, and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA.     

Evaluation of a diagnostic microarray for the detection of major bacterial pathogens of potato from tuber samples

Potato can be infected with many bacterial pathogens, the detection of which is necessary in seed certification. In this study, a diagnostic microarray previously tested for specificity of probes for detecting the potato bacteria causing blackleg and soft rot (Pectobacterium atrosepticum, Pectobacterium carotovorum, and Dickeya spp.), ring rot (Clavibacter. michiganensis subsp. sepedonicus), scab (Streptomyces scabies and Streptomyces turgidiscabies) and brown rot (Ralstonia solanacearum) from pure culture was evaluated for analytical sensitivity when testing directly from tuber samples. The microarray readily detected all the bacterial species when 100 ng of the target bacterial DNA from pure culture was mixed with DNA from soil microbes and potato. However, detection was inconsistent when total DNA isolated directly from infected tubers or enriched bacterial culture was used. While the high specificity of the probes could be confirmed from the results of the DNA cocktail experiment used as a control, the study demonstrated that the level of analytical sensitivity of the microarray under the tested condition was not sufficient to detect bacteria directly from tubers. Therefore, in addition to the cost and organizational complexities, the low analytical sensitivity and limited reproducibility of the microarray are constraints for establishing the platform for routine detection of potato bacterial pathogens from tuber samples.     

Effect of foliar applications of phosphite on post-harvest potato tubers

The utilization of phosphites (Phi) could be considered as another strategy to be included in integrated disease management programmes to reduce the intensive use of fungicides and production costs. The aim of the present work was to analyze whether the beneficial effects of phosphite treatment previously observed in potato plants grown under greenhouse conditions, were reflected after harvest of field grown potatoes, both in disease protection and in yield. In addition, biochemical compounds possibly involved in induced defence responses by Phi, like phytoalexins, pathogenesis related proteins and oxidative stress enzymes were measured. Foliar applications of KPhi to field grown crops resulted in post-harvest tubers with a reduced susceptibility to Phytophthora infestans, Fusarium solani and Erwinia carotovora infections, suggesting that this compound induced a systemic defence response. An increase in phytoalexin content in P. infestans inoculated tubers obtained from Phi-treated plants suggests their participation in the defence response. Chitinase content increased 72 h after wounding or inoculation with P. infestans in tubers from KPhi-treated plants compared to wounded or infected tubers from non-treated plants. Contrary to this, the isoforms of β-1,3-glucanases analyzed did not increase in the tubers of Phi-treated plants. The increment in peroxidase and polyphenol oxidase activities indicated that these enzymes could be part of the Phi defence mechanism. No negative effects were observed in potato yield at harvest, measured as total tuber weight and dry matter, after foliar KPhi treatment. This suggests that the energetic cost involved in the defence response activation would not be detrimental to plant growth.     

Role of co‐infection by Pectobacterium and Verticillium dahliae in the development of early dying and aerial stem rot of Russet Burbank potato

Potato early dying (PED) is a disease complex primarily caused by the fungus Verticillium dahliae. Pectolytic bacteria in the genus Pectobacterium can also cause PED symptoms as well as aerial stem rot (ASR) of potato. Both pathogens can be present in potato production settings, but it is not entirely clear if additive or synergistic interactions occur during co‐infection of potato. The objective of this study was to determine if co‐infection by V. dahliae and Pectobacterium results in greater PED or ASR severity using a greenhouse assay and quantitative real‐time PCR to quantify pathogen levels in planta. PED symptoms caused by Pectobacterium carotovorum subsp. carotovorum isolate Ec101 or V. dahliae isolate 653 alone included wilt, chlorosis and senescence and were nearly indistinguishable. Pectobacterium wasabiae isolate PwO405 caused ASR symptoms including water‐soaked lesions and necrosis. Greater Pectobacterium levels were detected in plants inoculated with PwO405 compared to Ec101, suggesting that ASR can result in high Pectobacterium populations in potato stems. Significant additive or synergistic effects were not observed following co‐inoculation with these strains of V. dahliae and Pectobacterium. However, infection coefficients of V. dahliae and Ec101 were higher and premature senescence was greater in plants co‐inoculated with both pathogens compared to either pathogen alone in both trials, and V. dahliae levels were greater in basal stems of plants co‐inoculated with either Pectobacterium isolate. Overall, these results indicate that although co‐infection by Pectobacterium and V. dahliae does not always result in significant additive or synergistic interactions in potato, co‐infection can increase PED severity.     

Infection of potato by Rhizoctonia solani: effect of anastomosis group

Glasshouse and field experiments showed that the pathogenicity and disease type on potato varied between different anastomosis groups (AGs) of Rhizoctonia solani. For example, severe stem and stolon disease developed in plants inoculated with a single isolate of AG3PT and AG5. Severe root disease was observed with single isolates of AG8 and to a lesser extent AG3PT, but rarely with single isolates of the other AGs tested. In both field and glasshouse experiments the AG2‐1 isolate (X81) produced only small lesions (<5 mm). However, this was not representative of two other AG2‐1 isolates. When AG2‐1 isolates of the three different rDNA IGS1 types were tested in a glasshouse trial, one caused more severe stem and stolon infection than AG3PT. In the field experiment, the yield of tubers, by weight, was significantly less (P < 0·05) in all inoculated plants than for uninoculated (control) plants. Yield losses were greatest and tuber numbers smallest in plots inoculated with an AG8 isolate, suggesting that root infection is important in determining quantitative yield loss. The incidence of black scurf was greatest in the progeny tubers in plots inoculated with AG3PT (83·9%), whereas only very small amounts of black scurf developed on tubers from plants infected with AG2‐1 (510 bp) or AG5 isolates. This is supported by laboratory tests, where isolates of AG3PT produced significantly more sclerotia on potato dextrose agar than isolates of AGs 2‐1, 4, 5 and 8.     

Evidence that nematodes may vector the soft rot‐causing enterobacterial phytopathogens

Bacterial soft rot is a globally significant plant disease that causes major losses in the production of many popular crops, such as potato. Little is known about the dispersal and ecology of soft‐rot enterobacteria, and few animals have been identified as vectors for these pathogens. This study investigates whether soil‐living and bacterial‐feeding nematodes could act as vectors for the dispersal of soft‐rot enterobacteria to plants. Soft‐rot enterobacteria associated with nematodes were quantified and visualized through bacterial enumeration, GFP‐tagging, and confocal and electron scanning microscopy. Soft‐rot enterobacteria were able to withstand nematode grazing, colonize the gut of Caenorhabditis elegans and subsequently disperse to plant material while remaining virulent. Two nematode species were also isolated from a rotten potato sample obtained from a potato storage facility in Finland. Furthermore, one of these isolates (Pristionchus sp. FIN‐1) was shown to be able to disperse soft‐rot enterobacteria to plant material. The interaction of nematodes and soft‐rot enterobacteria seems to be more mutualistic rather than pathogenic, but more research is needed to explain how soft‐rot enterobacteria remain viable inside nematodes.     

Trichoderma harzianum T39 and T. virens DAR 74290 as Potential Biological Control Agents for Phytophthora erythroseptica

Trichoderma harzianum isolate T39 and T. virens isolate DAR 74290 were evaluated as potential biological agents for control of pink rot of potato and root and stem rot of tomato caused by Phytophthora erythroseptica. Cell-free metabolites of T. virens DAR 74290 completely inhibited growth of P. erythroseptica in vitro and appeared to be fungicidal. T. virens DAR 74290 and Trichodex, a commercial formulation of T. harzianum T39, were tested for their ability to protect potato and tomato plants from disease caused by P. erythroseptica in glasshouse experiments. Trichodex and T. virens DAR 74290, alone and combined, reduced disease severity in shoots and roots of potatoes 10 weeks after inoculation with the pathogen. The yield of potatoes from plants treated with P. erythroseptica and T. virens DAR 74290 (mean of 12.9g fresh weight/pot) was significantly greater than in controls inoculated with the pathogen alone (mean of 2.1g/pot). Treatment with Trichodex alone increased the yield of tubers compared to the uninoculated controls. T. virens DAR 74290 increased the survival of tomato seedlings inoculated with the pathogen, and both this isolate and Trichodex decreased the severity of disease on tomato.     

Effect of temperature on resistance to Potato virus Y in potato cultivars carrying the resistance gene Rychc

The effect of cultivation temperatures on the resistance reaction to three Potato virus Y strains (PVY^O, PVY^N and PVY^NTN) in potato cultivars carrying Ry_chc was examined. When potato plants carrying Ry_chc were cultivated at 22 °C, a few small necrotic spots developed on inoculated leaves by 5 days after mechanical inoculation (dpi), and systemic infection of a few symptomless plants was confirmed at 28 dpi by IC‐RT‐PCR. At 28 °C, distinct necrotic spots developed on inoculated leaves by 5 dpi, and systemic symptoms occasionally appeared at 28 dpi. Thus, high temperature weakens Ry_chc‐conferred resistance. However, the incidence of systemic infection and the titre of virus in resistant cultivars at 28 °C were lower than in a susceptible cultivar. In graft inoculation under high summer temperatures, some plants developed necrosis on the leaves and stem, but PVY was barely detected by RT‐PCR in leaves on potato carrying Ry_chc. When seedlings from progeny tubers of plants that were inoculated with PVY and grown in a greenhouse at >30 °C in the daytime were examined by ELISA and IC‐RT‐PCR, PVY was not detected in cultivars carrying Ry_chc. These results show that Ry_chc confers an extreme resistance to PVY strains occurring in Japan.     

The MeloTuber Test: a real‐time TaqMan® PCR‐based assay to detect the root‐knot nematodes Meloidogyne chitwoodi and M. fallax directly in potato tubers

Potato is one of the many important hosts for the root‐knot nematodes Meloidogyne chitwoodi and M. fallax that can infest roots as well as tubers. In the latter they may cause surface galls and necrotic spots below the skin. In the EU these pathogens are categorized as quarantine organisms and are therefore regulated. Phytosanitary measures (PMs) are implemented and one aspect involves diagnostic procedures to detect these pathogens. To date, visual screening of external and internal symptoms is combined with the specific identification of these pests, either through microscopy, biochemical or molecular tests. A disadvantage of all these tests is the requirement of the prior extraction of nematodes from the tubers, which is not suitable for high‐throughput screening. This paper describes the MeloTuber Test developed to simultaneously detect M. chitwoodi and M. fallax by triplex real‐time TaqMan^® PCR directly in secondary potato tuber peelings. DNA extraction is carried out in a 96‐well plate of pooled secondary peelings from one hundred tubers. The analytical sensitivity is such that a single female can be readily detected in such a sample size. The validation data, described here, prove the suitability of this molecular test for the detection of M. chitwoodi and M. fallax in large scale screening tests.     

Temporal association of potato tuber development with susceptibility to common scab and Streptomyces scabiei‐induced responses in the potato periderm

Using hydroponics and novel non‐destructive pot culture systems which enable inoculation at specific tuber development stages, the dynamics of common scab infection patterns in potato were studied in order to provide more precise identification of tuber physiological factors associated with susceptibility. At the whole‐tuber level, infection percentages were greatest when Streptomyces scabiei inoculation occurred early; at 2 weeks after tuberization (WAT) 68% of tubers became infected, contrasting with late inoculation (8 WAT), when only 4% infection occurred. The first‐formed internodes were most susceptible to infection, whilst later‐forming and slower‐expanding internodes were less susceptible. Detailed tuber physiological examination of internode 2 showed that pathogen‐induced changes, including increased phellem (periderm) thickness, cell layers and phellem suberization (key physiological features believed critical to S. scabiei infection) were promoted through S. scabiei inoculation. Sequential harvesting showed enhanced phellem suberization (28% greater than the control) within 7 days of pathogen exposure, while phellem thickness and layer responses were also initiated early in the infection process (10–14 days after pathogen exposure) and these responses were independent of symptom expression. Differences in cultivar response were observed, with greater phellem suberization observed 10 days after tuberization (DAT) in the common‐scab‐tolerant cv. Russet Burbank than in the susceptible cv. Desiree. Likewise, Russet Burbank had thicker and more numerous cell layers in the phellem (up to eight cell layers) during early tuber growth (20–30 DAT) than Desiree (up to six cell layers).     

Spongospora subterranea root infection assessed in two potato cultivars differing in susceptibility to tuber powdery scab

Infection by Spongospora subterranea of roots of two potato (Solanum tuberosum) cultivars, either very resistant or very susceptible to powdery scab on their tubers, was studied in a glasshouse experiment. Plants grown in sand/nutrient solution culture were inoculated with S. subterranea sporosori 2 weeks after planting. Plant parameters, the intensity of zoosporangium infection in roots, numbers of Spongospora root galls and amounts of Spongospora DNA in roots, measured using quantitative PCR (qPCR), were assessed at sequential harvests. Inoculation with S. subterranea reduced water use (56 days after planting) by 26% in the tuber resistant cultivar compared with uninoculated plants, and by 60% in the susceptible cultivar. Inoculation did not affect growth of the resistant cultivar, nor shoot mass of the susceptible cultivar, but caused a 38% reduction in root mass of the susceptible cultivar. The intensities of zoosporangium development in both cultivars were similar. The susceptible cultivar had approximately four times more Spongospora root galls g^?1 root mass than the resistant cultivar. Quantitative PCR detected S. subterranea DNA in roots 1 week after inoculation, and indicated a twofold greater amount of pathogen DNA in roots of the susceptible than the resistant cultivar. This study suggests that the S. subterranea zoosporangium stage in host roots is affected differently by host resistance factors than the sporosorus (root gall and tuber scab) stages. The study has also demonstrated the usefulness of qPCR for sensitive and consistent detection of S. subterranea across the duration of potato root infection.     

Potato spindle tuber viroid on potato

Specific scope

This standard describes a national regulatory control system for Potato spindle tuber viroid (PSTVd) that provides guidance on preventing its introduction, surveillance for the pathogen and its containment and eradication if found infecting potato plants or tubers.

Specific approval and amendment

First approved in 2011‐09.