Acquisition of potato leafroll virus byMyzus persicae from secondarily-infected potato plants of different genotypes

Summary The acquisition of potato leafroll virus (PLRV) byMyzus persicae nymphs from the top leaves of potato plants was studied throughout a growing season in relation to the antigen titre in those leaves and the feeding behaviour of the aphid. Secondarily-infected plants of eight potato genotypes with different levels of field resistance served as virus sources. Early in the growing season, plants were efficient sources for virus acquisition. The amount of viral antigen detected inM. persicae nymphs fed on the top leaves was strongly correlated with the titres of viral antigen in these leaves. Virus acquisition from the top leaves of older potato plants was markedly impaired and could not be correlated with their virus titre. With increasing age of the potato plants and the development of virus symptoms, the virus titre in the leaves declined and the initial weak correlation between the virus titre and field resistance ratings disappeared. Thus, screening secondarily-infected potato plants for field resistance to PLRV based on the concentration of viral antigen in leaves or in aphids fed on them should be avoided later in the growing season. The feeding rate ofM. persicae, measured by the number of honeydew droplets excreted, did not account for the reduced uptake of virus from older plants since it was not influenced by the age of the plant. Throughout the growing season, the feeding rate ofM. persicae nymphs on PLRV-infected plants was higher on genotypes with low levels of field resistance to PLRV than on genotypes with high ones.     

Evidence that resistance to potato leafroll virus accumulation in tetraploidsolarium tuberosum L. is controlled by one or few major genes that are not complementary

Summary Previous investigations into the inheritance of resistance to accumulation of potato leafroll virus indicated a dominant major-gene effect, but the segregation ratios in progenies of crosses were a closer fit to a model involving two complementary genes (both required for resistance and one present in the susceptible parent cultivar, Maris Piper) than to a single gene model. In this study, we tested the complementary gene hypothesis by backcrossing susceptible segregants from one of these progenies to their susceptible parent, Maris Piper. No resistant segregants were found in the five backcross progenies examined, so the complementary gene hypothesis was not supported. There was significant variation between susceptible progeny-members in these backcrosses. The progeny of another, highly resistant parent clone, G. 8107(1), selfed, was also examined: all members were resistant. Whilst there is evidently a dominant major-gene effect involved, this is not the whole picture and there are other unidentified genetic effects.     

Inheritance of extreme resistance to potato virus Y in potato

Summary Segregation for extreme resistance to PVY was evaluated in progenies derived from crossing two extremely resistant potato clones with parents differing in resistance. Resistance was evaluated after mechanical inoculation with PVY^O and PVY^N, and after graft inoculation with PVY^O. Biological and serological tests (ELISA) were used for virus detection. The extreme resistance is governed by a single dominant gene, but observed segregations deviated from the expected ratios. Considerable modifying effects were detectable, depending on the potato genotype and virus isolate, for a significant excess of susceptible genotypes was observed in some progenies. Moreover, genotypes with non-parental types of resistance to PVY were observed.     

General resistance against potato virus Y introduced into a commercial potato cultivar by genetic transformation with PVYN coat protein gene

Summary Transgenic cv. Folva potato plants expressing the coat protein gene of potato virus Y strain N (PVY^N) were produced usingAgrobacterium tumefaciens mediated transformation. Forty independent transformants were selected for resistance screening. Four clones showed complete resistance to mechanical inoculation with all the five PVY isolates tested: the PVY^N isolate from which the coat protein gene was derived, two PVY^O isolates, and two PVY^NTN isolates. Two of the fully resistant clones contained only one copy of the transgene, demonstrating that it is possible by genetic engineering to obtain highly virus resistant potato clones that can also be useful in future breeding programmes.     

The infection pressure of potato leafroll virus and potato virus Y in relation to aphid populations in Cyprus

Summary The infection pressure of two viruses, potato leafroll (PLRV) and potato virus Y (PVY), both common in seed potatoes grown in Cyprus, was determined in three experiments in 1982–83. Virus-free bait plants, of potato and four other species, were exposed weekly to field infection during the growing season (March–June), and then returned to an aphid-free glasshouse for symptom expression. Only tobacco plants produced clear symptoms enabling reliable assessment of PVY infection pressure. When assessed with ELISA or by tuber indexing, the potato plants were efficient baits for both viruses whose infection period commenced at emergence (mid March to early April) and ended within 6–7 weeks. The seasonal trend of aphid populations, determined with Moericke traps or 100-leaf counts, correspond to that of virus spread. Correlation and regression analysis of aphid and virus data implicated the alate form ofMyzus persicae as the principal vector of both viruses.     

The development of mature plant resistance in four potato cultivars against aphid-inoculated potato virus YO and YN in four potato cultivars

Summary Field-grown potato plants of cvs King Edward, Record, Maris Piper and Désirée were inoculated on seven different dates during the growing season of 1987 and 1988 with either potato virus Y^O (PVY^O) or PVY^N, using three viruliferous peach-potato aphids (Myzus persicae) per plant. In each cultivar, the proportion of progeny tubers infected with PVY^O or PVY^N was high in plants inoculated during the four weeks following emergence, the proportion declining to zero or close to zero in the subsequent 4–6 wks.     

Diploid genotype DW.84-1457, highly resistant to potato leafroll virus (PLRV)

Summary The diploid clone DW.84-1457 which has outstanding resistance to potato leafroll virus (PLRV), has been selected at the Mlochów Centre of the Institute for Potato Research. It has in its pedigree PLRV-resistant clones from the Max Planck Institute nos. MPI 44.1016/10, MPI 44.335/130 and MPI 49.540/2. Its behaviour in the field and response to aphid inoculation indicate high resistance to infection, and the low concentration of the virus in graft-inoculated plants indicates high resistance to multiplication. This combination within one genotype of two aspects of resistance is not connected with hypersensitivity, and is heritable. Clone DW.84-1457 has other desirable characters such as extreme resistance to potato virus X (PVX), high resistance to potato virus M (PVM) and good table and processing quality. It is being utilized in the development of parental lines, both at the diploid and tetraploid level.     

Extreme resistance to infection by potato virus X in genotypes of wild tuber-bearingSolanum species

Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X¹ (also called RX_act).     

Potato leafroll virus distribution in potato meristem tips and production of virus-free plants

Summary Uridine-H³ was incorporated into meristem tips of both healthy and PLRV-infected potato plants, of the cultivars, Majestic and Primura. Autoradiograms of tips pretreated with actinomycin D showed that 13 of 14 and 11 of 14 respectively, were virus free in the dome and first 4 leaf primordia, 1 and 3 were infected in the 4th leaf primordium, and 1 of Primura also in its 3rd primordium. The highest plantlet yield from cultures in vitro of meristem tips that included 4 leaf primordia (7 to 9 plantlets per meristem), was obtained by growing them in sequence on 3 sequential media, each based on Murashige and Skoog basic medium complemented with various hormones. The 2nd medium, containing benzylaminopurine (0.5 mg/l) and gibberellic acid (0.5 mg/l), elicited callus and the subsequent formation of adventitious buds. Of the plantlets of the cvs Vivaks, Primura and Majestic, 88, 91 and 100 %, respectively, were PLRV-free. Propagation in vitro, by the single-node cutting technique and tuberlet production, were successful.     

Progeny testing for resistance to potato virus Y: a comparison of susceptible potato cultivars for use in testcrosses with resistant parents

Summary For potato breeding or genetical research purposes, the number of copies of a dominant major gene for resistance to potato virus Y in a parent clone can be determined by test-crossing with a susceptible parent and observing the segregation ratio of resistant to susceptible seedlings in the progeny. In a comparison of different susceptible cultivars for use in test-crosses for this purpose, their progenies differed in the clarity of symptoms and parental phenotype proved an unreliable guide. Cvs Maris Piper, Arran Peak and Dr MacIntosh were found to be suitable tester parents, Pentland Squire less so and cv. Désirée was confirmed as having a major gene conferring incomplete resistance. It was found advisable to test-cross with more than one susceptible parent.     

Bacillus thuringiensis cry1Ab gene confers resistance to potato againstHelicoverpa armigera (Hubner)

Summary Helicoverpa armigera is one of the important insect pests adversely affecting the yield of potatoes in India. A synthetic gene encoding the insecticidal crystal protein (Cry1Ab) ofBacillus thuringiensis (Bt) has been introduced into five genotypes of potato usingAgrobacterium tumefaciens. Southern analysis of DNA from transgenic plants confirmed the integration and copy number of the transgene. Double-antibody quantitative sandwich ELISA analysis demonstrated high levels of Cry1Ab protein expression in transgenic plants. Insect bioassays on the leaves of transgenic plants showed considerable protection against the larvae ofH. armigera in terms of leaf area consumed and larval weight reduction.     

Field resistance to late blight of four indian potato cultivars

Summary The nature and extent of field resistance to late blight, and the effect of plant age and inoculum level on resistance components and host attributes, were studied on the Indian potato cultivars Kufri Chandramukhi, Kufri Jyoti, Kufri Badshah and Kufri Sherpa. Latent period, infection efficiency, colonization rate and sporulation were found to operate in cvs Kufri Jyoti, and Kufri Badshah. Based on component analysis cv. Kufri Jyoti was similar to or sometimes more resistant than Kufri Badshah. No correlation was observed between any of the host attributes and resistance to late blight, except that the erect canopy of Kufri Badshah was associated with its slow blighting rate. At high inoculum, colonization rate and total sporulation were increased but infection efficiency was decreased.     

Transgenic potato plants expressing oxalate oxidase have increased resistance to oomycete and bacterial pathogens

Summary Potato (cv. Bintje) was transformed with a gene encoding an oxalate oxidase from wheat under the control of the CaMV35S promoter. Transgenic potato plants produced high constitutive levels of H₂O₂ as visualized by 4-chloro-l-naphtol staining. The resistance of these plants was tested againstPhytophthora infestans. An increased level of resistance to the disease was marked by a reduced number of lesions as well as by a decreased number of sporangia formed per lesion. In addition, oxalate oxidase overexpressing plants also exhibited improved resistance toStreptomyces reticuliscabiei, the causal agent of netted scab. Increased expression of oxalate oxidase had no effect on the interaction withErwinia carotovora. These experiments show that overexpression of oxalate oxidase represents a potentially interesting approach for protection of potato to pathogens.     

Influence of the duration of acquisition and inoculation feeding on the effectiveness of potato leafroll virus transmission byMyzus persicae Sulz

Summary In laboratory experiments PLRV transmission byMyzus persicae Sulz was shown to be possible even during a very brief feeding period onPhysalis floridana Rydb, in both acquisition and inoculation feeding. When the acquisition feeding time (AFT) was varied between 30 sec and 48 h the inoculation feeding time (IFT) was constant (48h) and vice versa: when the AFT was constant (48 h), the IFT was variable. The rate of infection was 2.2% following 30 sec AFT and 1.1% after 30 sec IFT. After 1 min feeding these infection rates increased to 7.9% and 1.8% respectively. The capacity for virus transmission was closely correlated with the increase in feeding time for both acquisition and inoculation feeding.     

Assessing progenies of potato for resistance to early blight

Summary Progenies from crosses between cultivars varying widely in resistance to early blight (Alternaria solani Sor.), were assessed for resistance as true seedlings in a glasshouse in Scotland. The resistance of a representative sample of surviving genotypes from each progeny was compared with samples of the same progenies not previously exposed to the fungus, both in the glasshouse in Scotland and in the field in Israel. The exposed population was more resistant. Resistance was identified more effectively in adult plants from tubers in the glasshouse than in true seedlings and agreement between glasshouse and field assessment was better when progenies were compared rather than individual genotypes. The mid parent and progeny mean scores of the unexposed population were correlated at both sites, thus confirming that the resistance is heritable. Selecting resistant individuals at the seedling stage is suggested as a useful tool for resistance breeding, having first chosen the best parents for crossing.     

Laboratory screening for resistance to gangrene and toFusarium dry rot in potato

Summary For laboratory screening for resistance to gangrene andFusarium dry rot, the ‘scrape and paint’ method was compared to a ‘prick and paint’ method. In the latter method the potatoes were wounded before inoculation by rolling them over a group of nail points. The disease scores increased when the length of the nail points was increased from 2 to 6 mm, but the scores were lower than for scraped potatoes. For the 10 cultivars used, the results for the pricked and the scraped potatoes correlated equally well with the results from a ‘field inoculation’ method. The two laboratory methods discriminated equally well between resistant and susceptible cultivars. However, pricking is much less laborious than scraping. In routine tests with the ‘prick and paint’ method, differences between cultivars were highly significant over three years for resistance to gangrene but less significant forFusarium dry rot.

---

Zusammenfassung Für die Laborprüfung auf Resistenz gegenPhoma-Faule (Phoma exigua var.foveata) undFusarium-Trockenf?ule (Fusarium solani var.coeruleum) wurde die ‘scrape and paint’-Methode von Wiersema (1977a) mit einer ‘prick and paint’-Methode verglichen, wobei Kartoffeln vor Inokulation durch Rollen über eine Gruppe von Nagelspitzen, 1 cm auseinander und von einer h?lzernen Basis aufw?rtsweisend, besch?digt wurden. Für beide Krankheiten stieg der Krankheitsindex an, wenn die L?nge der Nagelspitzen von 2 auf 4 und 6 mm anstieg, war im Ganzen jedoch niedriger als bei geschürften Kartoffeln (Tabellen 2 und 4). Für die zehn für jeden Erreger in den Versuchen verwendeten Sorten korrelierten die Ergebnisse bei ‘prick’ (Rollen über Nagelspitzen) und ‘scrape’ (Schürfen) gleichermassen mit einem vermutlich besser zu verwirklichendem Feld-Inokulationstest (Bjor, 1980), dessen Ergebnisse in Tabelle 1 dargestellt sind. Die Unterscheidung von Sorten unterschiedlicher Resistenz war mindestens so deutlich beim ‘prick’ — wie beim ‘scrape’-Test, wie die F-Werte für Sorten in der Varianzanalyse zeigen (Tabellen 2 und 4). Darüberhinaus ist die ‘prick’-Methode betr?chtlich weniger arbeitsaufwendig als die ‘scrape’-Methode. Die ‘prick and paint’-Methode mit 4 mm-Nagelspitzen ergab Unterschiede zwischen 14 Standard-Sorten in Routinetests aus drei verschiedenen Jahren beiPhoma-F?ule, aber nicht so gut beiFusarium-Trockenf?ule (Tabelle 6). Die Korrelation zwischen den Ergebnissen nach der ‘prick and paint’-Methode und dem Feld-Inokulationstest bei den 14 Sorten warr=0,86 undr=0,71 beiPhoma-bzw.Fusarium-F?ule. Allerdings war die Variation innerhalb der Sorten bei Resistenz gegenPhoma-F?ule betr?chtlich gr?sser als die bei Resistenz gegenFusarium-Trockenf?ule.

---

Résumé Le criblage au laboratoire pour la résistance à la gangrène (Phoma exigua var.foveata) et à la pourriture sèche fusarienne (Fusarium solani var.coeruleum) est réalisé selon deux méthodes. La méthode du ‘grattage et du badigeonnage’ est comparée à celle de la ‘piq?re et du badigeonnage’ où les pommes de terre sont blessées avant l'inoculation en les faisant passer sur un groupe de 10 clous distants de 1 cm, orientés vers le haut et fixés sur un support de bois. Pour les deux maladies, la note de maladie augmente quand la longueur de la pointe des clous passe de 2 à 4 et 6 mm, mais les tubercules blessés par piq?res sont moins malades que ceux ayant subi des égratignures (tableaux 2 et 4). Pour les 10 cultivars inclus dans les expériences pour chacune des maladies, les résultats des tubercules blessés par chacune des 2 méthodes sont également corrélés avec ceux d'un test ‘inoculation au champ’, plus s?r (Bjor, 1980) figurant au tableau 1. Le classement des cultivars de différents niveau de résistance est aussi clair à partir des tubercules blessés par piq?re que ceux blessés par grattage comme le montre la valeur du F pour les cultivars, des analyses de variance (tableaux 2 et 4). De plus, la méthode des piq?res est beaucoup moins fastidieuse que la méthode par égratignures. La méthode des piq?res avec des pointes de clous de 4 mm reproduit les différences entre 14 cultivars de référence inclus dans des tests de routine depuis 3 ans pour la gangrène, mais de fa?on moins nette pour la fusariose (tableau 6). La corrélation entre les résultats obtenus avec la méthode des piq?res et le test ‘d'inoculation au champ’ pour les 14 cultivars estr=0,86 etr=0,71, respectivement pour la gangrène et la fusariose. En outre, la variabilité des cultivars pour la résistance à la gangrène est considérablement plus grande que celle vis-à-vis de la fusariose (Figure 1).

     

Testing the resistance to silver scurf in commercial potato cultivars under controlled environmental conditions

Summary Optimum storage conditions to identify resistance to silver scurf among commercially grown potato cultivars were determined in a series of experiments. Inoculation of field-grown tubers with a conidial suspension ofH. solani (concentration 10⁴ conidia per ml of water) and incubation at 15°C with 95% RH for 1 month followed by 2 months at 85% RH produced the greatest differences in disease severity between potato cultivars. Results with glasshouse-grown tubers were comparable with those from field-grown tubers, indicating that the test could be used to identify resistance in wild species of potato and to understand the inheritance of this resistance.     

The involvement of phenolics and phytoalexins in resistance of potato to soft rot

Summary Phenolics, for example chlorogenic, caffeic and ferulic acid, and phytoalexins, such as rishitin and phytuberin, were identified in potato tubers cv. Kufri Chandramukhi. The tissue of healthy tubers contained no detectable phytoalexins but did contain phenolics. The levels of these compounds were correlated with soft rot development. The rotting tissue either was free of these groups of compounds or had low concentrations. The wound periderm formed as a result of recovery from injury and infection contained high levels of the compounds. Much higher concentrations were detected at lower storage temperatures when oxygen supply was adequate. Antibacterial properties of the phenolics identified were tested againstErwinia carotovora which was inhibited by chlorogenic, caffeic and ferulic acids. The three phenolics were more effective together, in proportions in which they occurred in wound periderm, than individually. It was observed that none of these phenolics could inhibit pectolytic enzymes ofE. carotovora.     

Correlation between resistance to late blight in foliage and tubers in potato clones from parents of contrasting resistance

Summary The relationship between resistance to late blight in foliage and tuber was examined on 50 clones of each of five progenies from crosses where one parent was resistant in both foliage and tuber and the other susceptible. Foliage resistance was assessed in a field trial and tuber resistance in a laboratory test on glasshouse-grown tubers. The genetical and environmental components of variance and the phenotypic and genetical correlations between foliage and tuber blight scores were determined for each progeny. Foliage and tuber resistance were correlated, indicating either that both are determined by the same genes or by different linked genes. The environmental component of variation was greater for tuber blight than foliage blight. We suggest that the most effective way of selecting for resistance to both aspects of the disease in a breeding programme is to select those resistant in the foliage and then screen them for tuber resistance.