The behaviour of potato mop-top virus in soil, and evidence for its transmission by Spongospora subterranea (Wallr.) Lagerh.

Potato mop-top virus (PMTV) was best detected in field soils by air-drying them for more than a week before remoistening and growing seedlings of Nicotiana tabacum or N. debneyi for a 6–10 week period. Infection of N. tabacum was assessed by inoculating sap from roots and shoots to Chenopodium amaranticolor. Similar inoculations from N. debneyi were far less convenient for detecting PMTV than recording leaf symptoms, but slightly more efficient. Air-dry soil retained PMTV infectivity for 9 months, when passed through a 50 μ sieve or when diluted with 10³ but not 10⁴ parts of steamed soil. Tobacco seedlings were not infected when their roots were steeped in PMTV-containing tobacco sap. Infective soils contained Spongospora subterranea, spore balls of which resisted air-drying for more than a year and passed a 50 μ sieve. Roots of susceptible seedlings were infected with PMTV when exposed to spore balls of S. subterranea taken from powdery scabs on PMTV-infected potato tubers, or to suspensions obtained by steeping, in nutrient solution, roots infected with virus-carrying cultures of S. subterranea. Plants in several families were hosts of S. subterranea, but probabilities of infection when exposed to spore balls differed greatly between families and only species of Solanaceae were good hosts. The ten species infected with PMTV when grown in infective soil or when exposed to spore balls of S. subterranea taken from PMTV-infected potato tubers are all members of this family. PMTV seems to be carried internally in S. subterranea spore balls and survived in them for at least a year. PMTV was transmitted by S. subterranea to Arran Pilot potato, causing yellow blotches in some leaves and spraing in many tubers. However, when newly infected with PMTV in the field, not all Arran Pilot tubers developed spraing. Also, although many spraing-affected or symptomless but PMTV-infected tubers carried PMTV-containing spore balls of S. subterranea, powdery scabs were rarely found near the centres of the rings of primary spraing. PMTV became established in virus-free soil when PMTV-infected tubers carrying S. subterranea were planted as ‘seed’ but not when virus-free tubers bearing powdery scabs were used. 5. subterranea seems the main, and possibly the only, vector of PMTV in the soils examined. S. subterranea did not transmit potato aucuba mosaic virus from potato to N. debneyi or Capsicum annuum.     

Field and glasshouse experiments on the control of potato mop-top virus

Field observations during 3 yr on a stock of potato cv. Red Craigs Royal partially infected with potato mop-top virus (PMTV) confirmed that the virus was passed by an infected mother plant to only a proportion of its progeny tubers, and showed that in this cultivar symptomless plants gave rise only to symptomless progeny. The elimination of PMTV from stocks can therefore be greatly accelerated by removing symptom-bearing plants. Infected potato tubers were not freed from PMTV by treating them at 37 °C for up to 8 wk. Treating ‘seed’ tubers bearing powdery scabs that contain PMTV-carrying resting spores of Spongospora subterranea with formaldehyde or organo-mercurial fungicide greatly decreased PMTV establishment when the tubers were planted in previously uninfective soil, but fumigation with 2-aminobutane was ineffective. Decreasing the pH of infective soil to 5-0 by applying sulphur greatly decreased the infection of potato cv. Arran Pilot with PMTV and S. subterranea in field experiments, but this treatment did not eliminate either; when the pH of treated soil was raised the transmission of PMTV resumed. Treating infective soil with a range of fungicides greatly decreased the infection of Nicotiana debneyi bait seedlings in glasshouse experiments but only calomel at 75 kg/ha controlled spread of PMTV and 5. subterranea to potato in field experiments. In other field experiments, applying zinc frit, zinc sulphate or zinc oxide to infective soil greatly decreased the spread of both to potato. The amount of zinc required increased with increase in clay content of the soil. However, treatment with zinc compounds did not eliminate PMTV-carrying vectors from soil, and when treated soil was diluted with autoclaved soil many of the bait seedlings planted in the mixture became infected. The zinc frit was phytotoxic because of its boron content but zinc sulphate and zinc oxide caused little or no decrease in tuber yield. The zinc content of potato tubers was increased but not doubled in zinc-treated plots, and during the first year after treatment the zinc content of topsoil decreased greatly. The zinc content of ryegrass grown after potatoes was greater than of potato tubers but did not reach a level considered dangerous to livestock. Treatment of soil with sulphur, zinc oxide or calomel may be useful for small plots used in the early stages of propagation of virus-tested potato clones where there is risk of infection with PMTV.     

Detection,distribution and control of Potato mop-top virus,a soil-borne virus,in northern Europe

Potato mop-top virus (PMTV; genus Pomovirus; family Virgaviridae) is transmitted by the soil-borne Spongospora subterranea f.sp. subterranea, a protoctist that causes powdery scab on potato. PMTV is distributed widely in the potato growing areas in South and North America, Japan and northwestern Europe. This article reviews the current knowledge on detection, distribution and control of PMTV with focus on the Baltic Sea region. Since the 1980s, PMTV has caused great economic losses to potato production in the Nordic countries (Norway, Sweden, Denmark and Finland), but its occurrence in other countries of the Baltic Sea region remained unknown. To fill this knowledge gap, harmonised sampling and virus detection procedures including bioassays and serological and molecular methods were employed by 21 research institutions to detect PMTV in potato tubers and soil samples in 2005–2008. Potato growing areas were widely contaminated with PMTV in the Nordic countries. Only the main seed potato production area in northern Sweden and the High Grade seed potato production zone in Finland were negative for PMTV. Intensive and systematic surveys in Poland in 2004–2008 found no evidence of PMTV, except a single PMTV-infected tuber detected in 2008. Surveys in the Baltic countries (Lithuania, Latvia and Estonia) and northwestern Russia (Leningrad province) were negative for PMTV, except infection of minitubers in a screenhouse in Latvia in 2005. Varying percentages of tubers expressing spraing symptoms in Sweden, Norway, Denmark and Poland were infected with Tobacco rattle virus, and bioassays indicated similar results for Russia. Incidence of symptomless infections with PMTV was high in tubers of many potato cultivars. Here, we discuss the contrasting patterns of distribution of PMTV in the Baltic Sea region, factors playing a role in dispersal and establishment of PMTV in new fields and means for controlling PMTV and its spread to new areas. We emphasise the use of the current virus-specific methods for the detection of PMTV in symptomless potato tubers and the high risks of disseminating PMTV to new fields and areas in viruliferous resting spores of S. subterranea in the soil adhering to seed tubers. PMTV-resistant potato cultivars will provide the only sustainable means for preventing yield losses in the infested fields and the prospects of resistance breeding are summarised.     

Factors affecting the field infection of potato tubers of different cultivars by blight (Phytophthora infestans)

The relationships between rain and blight (Phytophthora infestans) were studied in unsprayed crops of cultivars differing widely in foliage and tuber susceptibility. The occasions when tubers were infected depended on rain and not cultivar, but numbers of tubers infected after rain was affected by the blight susceptibility of the cultivar. Infected tubers were first found when less than 5 % (BMS key) of the potato foliage was infected but few fresh infections occurred when 50–75% of the foliage had been destroyed. Some tubers were infected after 8 mm rain (tubers near the surface with even less) but large increases in numbers of tubers infected usually occurred only after 25 mm or more had increased soil moisture to above ‘field capacity’ around the tuber for at least 24 h. The most susceptible cultivars Ulster Ensign and Arran Banner had all plants with some tuber blight, and some plants with all tubers affected and often many lesions per tuber. Cultivars of intermediate susceptibility, King Edward and Up-to-Date, had some plants without blighted tubers, many with a few and very few with all. The more resistant cultivars Majestic and Arran Viking had many plants without infected tubers and many lesions that aborted while still necrotic threads, so that the fungus did not spread. Most infections occurred through tuber eyes, lenticels or sometimes growth cracks. The distribution of blight lesions on tubers differed in the different seasons, for example, lenticels were most commonly infected on Arran Banner and Ulster Ensign and eyes on King Edward, Majestic and Arran Viking. In late or slowly developing attacks, lesions on stems became more numerous and larger than in fast, early attacks and were prolific sources of spores on King Edward and Up-to-Date but not on Majestic and Arran Viking. Because much rain water runs down the stems of Up-to-Date and King Edward, stem lesions can provide an important source of inoculum for tubers.     

Methods of measuring the prevalence of Phoma exigua on potatoes and in soil

Methods are described for assessing inoculum of Phoma exigua by directly wounding potato test tubers and by inoculating soil into Arran Banner test tubers or slices. Large samples of tubers were wounded by dropping onto perforated metal sheet or grading on farm riddles, but standardization was achieved by inflicting four uniform wounds with shaped brass teeth. Tubers were then incubated for 12 wk at 5 ^oC. Soil samples to be tested were inoculated into crush wounds on surface-sterilized test tubers or tuber slices which were incubated for 8 wk at 5 ^oC. Assessments were speeded by immersing wounded slices in a mixture of Agrimycin and isopropylphenyl carbamate (IPC) solutions before inoculating, and then incubating for 2–3 wk at 10 ^oC. Although the sensitivity of test tubers is limited by their latent infection, comparisons of the tests in 3–5 years showed that the test tuber methods were frequently more sensitive than direct wounding, especially when amounts of inoculum were small.     

Selection of individual tubers in potato breeding

Summary Out of 720 field-grown potato first year seedlings plants with best tuber apearance and large and medium size tubers with best shape were respectively selected. The tuber progeny of each group was evaluated in field experiments. It was demonstrated that the selection of individual tubers was very effective in the elimination of clones with irregular tuber shape and deep eyes. The tuber progeny of selected tubers was not inferior to the tuber progeny of selected plants. Possible applications of the results to practical potato breeding are discussed.     

Effects of common scab (Streptomyces scabies) on potatoes

Seed tubers with severe (71% cover) or slight (12% cover) common scab selected from a badly infected crop of cv. Maris Piper were boxed for sprouting in November and February and planted in field experiments at Woburn and Rothamsted in 1980. The severely scabbed seed lost more moisture than the slightly scabbed seed during sprouting and sprouted from a larger number of eyes but total stem numbers were greater on plants from the slightly scabbed seed. Plant growth during the first 6–9 wk after planting was less from the severely scabbed than from the slightly scabbed seed. This was shown by records of ground cover, leaf area index, fresh weights of stems and leaves, total tuber yield (18% lower at 9 wk) and tuber number. Subsequently these differences became non-significant. Seed with slight scab boxed in November produced smaller numbers of progeny tubers than that boxed in February but there were no other consistent effects of boxing date on growth and yield. Scab incidence on progeny tubers was very slight and unrelated to the severity of disease on the seed tubers. It seems unlikely that severe scab infection of seed tubers would significantly decrease tuber yields except perhaps in early potato production.     

The effect of soil- and tuber-borne inoculum on the incidence of potato gangrene

Under optimum growing conditions neither tuber- nor soil-borne Phoma exigua var. foveata inoculum appreciably affected stand or yield of the subsequent potato crop. Seed tubers with gangrene rots caused high levels of stem and tuber symptoms when planted in var. foveata contaminated or uncontaminated land; contaminated seed tubers with no rots also produced progeny with a high gangrene potential. Sufficient soil-borne inoculum was carried over in land that produced a gangrene affected crop in the previous year to override the effect of tuber disinfection. Effective gangrene control was achieved by a combination of tuber disinfection shortly after harvest over successive years with a 1 in 5 yr potato crop rotation. Gangrene rots usually developed through injuries to the tuber periderm, rots in other tubers being associated with pustules of powdery scab (Spon-gospora subterranea).     

Host range and some properties of potato mop-top virus

Potato mop-top virus (PMTV) was transmitted by inoculation of sap to twenty-six species in the Solanaceae or Chenopodiaceae and to Tetragonia expansa; species in eleven other plant families were not infected. The virus was cultured in inoculated leaves of Nicotiana tabacum cv. Xanthi-nc or in N. debneyi. Diagnostic local lesions were produced in Chenopodium amaranticolor. In winter, ten solanaceous species were slowly invaded systemically but the first leaves infected were those immediately above inoculated leaves. When transmitted to Arran Pilot potato by the vector Spongospora subterranea, PMTV induced all the main types of shoot and tuber symptoms found in naturally infected plants. Isolates of PMTV from different sources differed considerably in virulence. PMTV-containing tobacco sap lost infectivity when heated for 10 min at 80 °C, diluted to 10^-4, or stored at 20 °C for 14 weeks. Infectivity was partially stabilized by 0·02% sodium azide. When sap was centrifuged for 10 min at 8000 g, infectivity was mainly in the sediment. Infective sap contained straight rod-shaped particles about 20 nm wide, with lengths up to 900 nm and crossbands at intervals of 2·5 nm. Many of the particles were aggregated side-to-side, and the ends of most seemed damaged. The slight infectivity of phenol-treated leaf extracts was abolished by pancreatic ribonuclease. The present cryptogram of PMTV is R/*:*/*:E/E:S/Fu.     

Evaluation of potato tuber moth (Lepidoptera: Gelechiidae) resistance in tubers of Bt-cry5 transgenic potato lines

The potato tuber moth, Phthorimaea operculella (Zeller), in tropical and subtropical countries, is the most destructive pest of potato, Solanum tuberosum L. The larvae attack foliage and tubers in the field and in storage. The purpose of this study was to evaluate the efficacy of a Bt-cry5 transgene to control the potato tuber moth in tuber tissues. Tuber bioassays using stored (11-12 mo old) and newly harvested tubers of Bt-cry5-Lemhi Russet and Bt-cry5-Atlantic potato lines showed up to 100% mortality of 1st instars. Mortality was lowest in the newly harvested tubers of Bt-cry5-Atlantic lines (47.1-67.6%). Potato tuber moth mortality was 100% in the Bt-cry5-Spunta lines that were transformed with Bt-cry5 gene controlled by the CaMV 35S promoter (pBIML5 vector) and in 2 of 3 lines transformed with Bt-cry5 gene controlled by the Gelvin super promoter (pBIML1 vector). The transgenic Spunta lines expressing Bt-cry5 controlled by the patatin promoter (pBMIL2 vector) showed the lowest tuber moth mortality (25.6 and 31.1%). The Bt-cry5 transgenic lines with high tuber expression of B. thuringiensis have value in an integrated pest management system to control potato tuber moth.     

Comparison of nucleic acid hybridisation and other tests for detecting tobacco rattle virus in narcissus plants and potato tubers

All isolates of tobacco rattle virus (TRV) found in naturally infected narcissus leaves produced nucleoprotein particles, mostly in large concentrations but, because of antigenic diversity, less than half of the isolates were identified by immunosorbent electron microscopy (ISEM) and still fewer by enzyme-linked immunosorbent assay. All were identified by a nucleic acid hybridisation test in which DNA complementary to RNA-1 of strain PRN of TRV was allowed to react with nucleic acid extracted from leaf tissue. Spraing-affected tubers in some potato stocks yielded only NM isolates of TRV. These isolates do not produce virus particles and they were therefore not detected by ISEM. The infectivity of nucleic acid extracts from recently harvested tubers with spraing symptoms was much greater than that of extracts prepared from tubers after 8 months' storage. In other potato stocks, some spraing-affected tubers contained NM isolates and the rest contained particle-producing isolates (M isolates) of TRV. The infectivity of sap and of nucleic acid, extracted 7 months after harvest from tubers infected with M isolates, was much greater than that of nucleic acid extracted from comparable tubers infected with NM isolates. TRV was detected by nucleic acid hybridisation in extracts of almost all tubers containing either M or NM isolates, even when the tubers were not tested until 7–8 months after harvest. The probable sequence of events occurring after tubers are infected with TRV is outlined, and it is suggested that the virus will rarely become established in fields as a result of planting infected tubers.     

Stem canker (Rhizoctoniu solani) on three early and three maincrop potato cultivars: effects of seed tuber size on growth and yield

Large seed tubers (mean 134 g) and small seed tubers (50 g) of three early and three maincrop potato cultivars, spaced respectively 48 and 30 cm apart within rows, were inoculated with Rhizoctonia solani at planting in 1985 and 1986. All seed of early cultivars was sprouted and maincrop seed was either sprouted or not sprouted. In all cultivars, plant emergence was slower from small than large seed and with both was delayed by inoculation. From 11 wk after planting numbers of stems and tubers and weights of foliage and tubers/m² were usually similar from small and large seed when not inoculated, but inoculating delayed plant growth and decreased tuber numbers and yield more from small than large seed. Inoculating decreased mean yields from large and small seed of early cultivars at 11 wk by respectively 24% and 31% (Arran Comet), 12% and 18% (Estima) and by 10% and 28% (Wilja) and losses were greatest with saleable sized tubers (3–4 cm). When grown to maturity inoculating decreased yields by 7% (Arran Comet), 5% (Estima) and 14% (Wilja). With maincrop cultivars, yields in October from large and small seed were decreased by respectively 4% and 10% (DCsirke), 9% and 12% (Maris Piper) and by 14% and 22% (Pentland Squire). In all cultivars yields of tubers < 44 mm and 44–70 mm were decreased and, with Pentland Squire, tubers > 82 mm were increased. The incidence of stem canker and of black scurf on progeny tubers was not affected by seed size but in all cultivars the percentage of greened tubers was slightly increased by inoculation.     

Application of enzyme-linked immunosorbent assay to the detection of potato leafroll virus in potato tubers

Factors affecting the detection of potato leafroll virus (PLRV) by enzyme-linked immunosorbent assay (ELISA) in tubers of field-grown potato plants with primary or secondary infection were studied. The reactions of extracts of virus-free potato tubers were minimised by pre-incubating the extracts at room temperature and by careful choice of the dilution of enzyme-conjugated globulin. PLRV was reliably detected in tubers produced by secondarily infected plants of all six cultivars tested. PLRV concentration was greater in heel-end than in rose-end vascular tissue of recently harvested tubers but increased in rose-end tissue when tubers stored at 4°C for at least 5 months were placed at 15–24°C for 2 wk. PLRV occurred at greater concentration in tubers from plants of cv. Maris Piper with natural or experimentally induced primary infection than in tubers from secondarily infected plants; again PLRV concentration was greater in heel-end than in rose-end vascular tissue. Plants whose shoots were infected earliest in the growing season were invaded systemically and produced the greatest proportion of infected tubers; plants infected late in the season also produced infected tubers but PLRV was not detected in their shoot tops. PLRV concentration in tubers from the earliest-infected plants was less than in tubers from later-infected plants. PLRV was detected reliably by ELISA in tubers from progenies that were totally infected but was not detected in all infected tubers from partially infected progenies. ELISA is suitable as a routine method of indexing tubers for PLRV, although the virus will not be detected in all infected tubers produced by plants to which it is transmitted late in the growing season.     

The effects of soil moisture at the time potato tubers are forming on the incidence of common scab (Streptomyces scabies)

Two complementary field experiments at Rothamsted in 1965, with the potato variety Majestic, related the incidence of common scab (Streptomyces scabies) to rainfall, soil moisture and time of tuber formation. In plots where the soil was maintained at field capacity (less than 10 cm. Hg moisture tension) by watering, tubers had little or no scab; infection increased in amount on plots allowed to dry to 30 cm., 50 cm. or more Hg moisture tension during late June. The time tubers formed was varied by planting sprouted (chitted) and non-sprouted seed tubers, and tuber development and scab incidence were observed on sample plants lifted at frequent intervals from unwatered plots. Scab lesions were first seen on 12 July when the distribution of lesions on the surface of tubers was affected by size of tuber both within and between the different seed-tuber treatments. These differences were correlated with estimates of tuber size on 28 June, the beginning of the first dry period. Tubers 1.0 cm. or more in diameter on 28 June had few or no lesions at the stolon attachment (heel) end of tubers on 12 July, whereas tubers smaller than this on 28 June had many lesions in this region on 12 July. The larger the tuber was on 28 June, the greater was the area free from scab lesions on 12 July.     

Laboratory evaluation of lufenuron on immature stages of potato tuber moth (Lepidoptera: Gelechiidae)

The effects of the chitin synthesis inhibitor lufenuron against potato tuber moth, Phthorimaea operculella (Zeller), eggs were determined by topically exposing different age groups of eggs (1-4 d old) to treated potato tubers (Solanum tuberosum L.) under laboratory conditions. Larval hatch from both treated (4 and 12 g [AI]/100 liter) and untreated tubers was >95%, but mortality of first instars was high in treated tubers (>90%) compared with untreated tubers. Examination of the treated tubers showed that most of the larvae were unable to penetrate or cause any noticeable damage to the potato tubers. However, the few first instars that survived were able to penetrate the tubers and continue their development to the pupal or adult stages. At 12 g (AI)/100 liter, adult emergence was <2% and most of the emerged adults had morphological deformities such as reduced wing size and they were unable to free themselves from the pupal sacs. These data suggest that topical application of lufenuron to eggs before larval hatch would reduce the amount of damage caused by potato tuber moth as part of integrated pest management program.     

Detection of potato leafroll and potato mop-top viruses by immunosorbent electron microscopy

Attachment of virus particles to antiserum-coated electron microscope grids (immunosorbent electron microscopy) provided a test that was at least a thousand times more sensitive than conventional electron microscopy for detecting potato leafroll (PLRV) and potato mop-top (PMTV) viruses. The identity of the attached virus particles was confirmed by exposing them to additional virus antibody, which coated the particles.
PLRV particles (up to 50/μm² of grid area) were detected in extracts of infected potato leaves and tubers, infected Physalis floridana leaves, and single virus-carrying aphids. On average, Myzus persicae yielded 10–30 times more PLRV particles than did Macrosiphum euphorbiae .
PMTV particles (up to 10/μm² of grid area) were detected in extracts of inoculated tobacco leaves, and of infected Arran Pilot potato tubers with symptoms of primary infection. Particles from tobacco leaves were of two predominant lengths, about 125 nm or about 290 nm, and fewer particles of other lengths were found than in previous work, in which partially purified or purified preparations of virus particles were examined, using grids not coated with antiserum.     

Methods of measuring the prevalence of pathogenic fungi on potato tubers

Methods are described whereby the prevalence of pathogenic fungi on seed potato tubers can be measured quickly and quantitatively. Eye-bearing plugs of tuber tissue, 8 mm diameter by 10 mm long, are excised, incubated over water at 15 °C for 5 days and then examined microscopically. The occurrence of fungi and macroscopic tuber symptoms are recorded directly on punched tape and tabulated by computer.     

HRE-type genes are regulated by growth-related changes in internal oxygen concentrations during the normal development of potato (Solanum tuberosum) tubers

The occurrence of hypoxic conditions in plants not only represents a stress condition but is also associated with the normal development and growth of many organs, leading to adaptive changes in metabolism and growth to prevent internal anoxia. Internal oxygen concentrations decrease inside growing potato tubers, due to their active metabolism and increased resistance to gas diffusion as tubers grow. In the present work, we identified three hypoxia-responsive ERF (StHRE) genes whose expression is regulated by the gradual decrease in oxygen tensions that occur when potato tubers grow larger. Increasing the external oxygen concentration counteracted the modification of StHRE expression during tuber growth, supporting the idea that the actual oxygen levels inside the organs, rather than development itself, are responsible for the regulation of StHRE genes. We identified several sugar metabolism-related genes co-regulated with StHRE genes during tuber development and possibly involved in starch accumulation. All together, our data suggest a possible role for low oxygen in the regulation of sugar metabolism in the potato tuber, similar to what happens in storage tissues during seed development.     

Ecological studies on potato mop-top virus in Scotland

Plants with symptoms of potato mop-top virus (PMTV) occurred in many commercial seed stocks of Arran Pilot and Red Craig's Royal potato in Scotland, but their incidence rarely exceeded 5%. In nuclear stocks of seed potatoes, most varieties examined in 1967 and 1968 were infected at one or more locality, but infected plants did not occur in all clones or at all stages of propagation of any one variety. infection of nuclear stocks resulted both from propagation on virus-infested land and from unwitting selection of infected plants to start new clones. PMTV was detected in farm soils ranging from light sands to heavy loams, in five Scottish counties. Soil was infested throughout the ploughed layer but the severity of infestation varied greatly within any one field; some sites of former potato clamps were heavily infested. PMTV was detected in field soil 12 years after potatoes were grown. In glasshouse tests many British crop and wild plants were colonized by Spongospora subterranea. Within some families all species tested were moderate to good hosts. (Solanaceae, Chenopodiaceae and Cruciferae), in others, species differed greatly in susceptibility (Compositae and Umbelliferae), and in a few, species were poor hosts or were not infected (Caryophyllaceae and Gramineae). Of the British crop and weed species that were moderate to good zoosporangial hosts of S. subterranea, only Solanum nigrum, potato, spinach and sugar beet were hosts of vector-borne PMTV. Potato probably survives between potato crops mainly in the resting spores of S. subterranea. PMTV was probably first brought to Europe with potatoes from South or Central America.