Early selection for extreme resistance to potato virus Y and tobacco etch virus in potato using a β-glucuronidase-tagged virus

The Ry_sto gene from Solanum stoloniferum introduced into potato cultivars (Solanum tuberosum L. ssp. tuberosum) confers resistance to potato virus A, potato virus V and potato virus Y (PVY). In addition to PVY, tobacco etch virus (TEV) and a TEV construct that encodes β‐glucuronidase (TEV‐GUS) were inoculated to determine the inheritance of resistance to these viruses in progenies obtained from potato cultivars containing the Ry_sto gene. While cultivars ‘Karlena’ and ‘Delikat’ were susceptible, ‘Bettina’ and clone 927eY were resistant to PVY, TEV and TEV‐GUS, as determined by enzyme linked immunosorbent assay, biotest and GUS assay, respectively. The segregation ratios obtained from the progenies of ‘Bettina’בDelikat’ and 816eY בKarlena’ indicate that resistances to PVY and TEV are governed by one dominant gene or two genes tightly linked in coupling phase. Evidently, Ry_sto confers broad spectrum resistance to potyviruses. TEV resistance could be reliably detected 4 days after inoculation with the TEV‐GUS construct by GUS assay. Therefore, the GUS‐tagged TEV construct can be used for early selection for resistances based on the gene Ry_sto or closely linked genes.     

Resistance to potato leafroll virus derived from Solanum chacoense: characterization and inheritance

Summary Resistance to potato leafroll virus (PLRV) was detected in an accession of Solanum chacoense. Inoculations with viruliferous aphids and subsequent graft challenges using Datura tatula and potato as PLRV sources determined that resistance appears to be of an extreme type. Virus was not detectable using enzyme-linked immunosorbent assay (ELISA) in S. chacoense, and in resistant F₁ and BC₁ progenies after attempts to transmit the virus through grafting. The segregation ratios of BC₁ progenies for positive and negative ELISA tests are consistent with simple dominant inheritance.     

Transfer of resistance to potato cyst nematode (globodera pallida) into cultivated potato Solanum tuberosum through first division restitution 2n pollen

Genetic resistance to potato cyst nematode is considered as one of the most effective means of increasing yield and reducing nematode infestation levels in potato fields. In this study, resistance to this nematode was successfully transferred from diploid tuber-bearing Solanums to the tetraploid gene pool using a 4x-2x breeding approach. More specifically, resistance from Solanum vernei, S. sparsipilum and haploids of S. tuberosum group Andigena was introgressed into conventional tetraploid clones, using first division restitution (FDR) 2n gametes. Furthermore, some of the FDR diploid parents had similar breeding values as advanced resistant tetraploid clones which were developed only after several cycles of selection against the potato cyst nematode. This revised version was published online in July 2006 with corrections to the Cover Date.     

The relations of broad nematode resistance to quality characteristics as a consequence of marker-assisted selection in potato breeding programs

Cultivating resistant varieties of potato is the most effective and environmentally sound method of protecting potato crops against pests and diseases. Potato cyst nematodes (PCN) are major nematode pests causing severe constraints in potato production worldwide. There are five pathotypes of Globodrea rostochiensis (Ro1–Ro5) and three of G. pallida Pa1–Pa3. Cultivation of potato varieties with broad nematode resistance may influence the growth of the wide spectrum of PCN pathotypes, but there is limited availability of such varieties on the market. The use of molecular markers allows for the effective selection of resistant genotypes at early stages of breeding. However, the impact of early selection for nematode resistance on the agronomic value of the final selected clones is a cause of concern for potato breeders. This study investigates the relationships between the presence of the combined resistance genes H1, Gro1-4 and GpaV_vrn, which confer resistance to the nematodes, and certain agricultural traits. Clones with broad nematode resistance conferred by the genes H1, Gro1-4 and GpaV_vrn presented yields and tuber morphology traits similar to those of the clones without identified resistance genes.     

Breeding for impact resistance in potato: use of a pendulum in progeny testing

Breeding for impact resistance in potato is common at the end of a breeding scheme, where there is no more variability for it. The purpose of this study was to propose an objective progeny test using a pendulum and single-year data. Data were recorded over a 5-year period using 275 progenies, organized in small factorial and hierarchical designs. The variability of response of the additive variance made it impossible to generalize parental choice. However, once the crosses were made, the high degree of variability among the progenies and the high level of broad-sense heritability in every case suggest that selection can be carried out efficiently using single-year data.     

Development of SCAR markers linked to the Ns locus in potato

A random amplified polymorphic DNA marker OPG17₄₅₀ linked to the Ns gene that confers resistance of potato to potato virus S (PVS), was used to develop sequence‐characterized amplified region (SCAR) markers. After cloning and sequencing of OPG17₄₅₀ new polymerase chain reaction (PCR) primers were designed to generate dominant (SCG17₃₂₁) and codominant (SCG17₄₄₈) markers. For SCG17₄₄₈, polymorphism between susceptible and resistant genotypes was recovered after digestion of the marker with the restriction enzyme Muni. In addition to the band corresponding to ‘susceptible’ allele that does not contain the Muni cleavage site, two bands of approximately 251 bp and 197 bp were observed in the resistant genotypes. The usefulness of these SCAR markers was verified in diploid potatoes possessing the Ns locus from clone G‐LKS 678147/60, and in tetraploid potatoes derived from G‐LKS 678147/60 and from clone MPI 65118/3.     

AFLP analysis of Solanum phureja DNA introgressed into potato dihaploids

Introgression of Solanum phureja DNA into S. tuberosum dihaploids was analysed by the use of amplified fragment length polymorphism (AFLP) markers. Five dihaploids, derived from crosses between S. tuberosum cv. Pentland Crown and two different S. phureja pollinators (IVP48 and EC90) were investigated by use of 17 AFLP primer pairs. Also 30 dihaploids, derived from pollination of five different S. tuberosum seed parents with S. phureja IVP101, were investigated for the presence of S. phureja‐specific markers. In total approximately 680 and 850 AFLP products were detected in the diploid S. phureja clones and in the tetraploid S. tuberosum genotypes, respectively. A total of 68 S. phureja IVP48‐specific markers were detected, while the total number of S. phureja IVP101‐specific markers was in the range of 72‐96, depending on the S. tuberosum seed parent. Introgression of DNA in the S. tuberosum cv. Pentland Crown dihaploids, after pollination with S. phureja IVP48 and S. phureja EC90, was demonstrated by the detection of 14 of 68 IPV48‐specific markers in the dihaploids. However, no DNA introgression was found in any of the 30 S. tuberosum dihaploids derived from S. phureja IVP101. Hence, S. phureja IVP101 is regarded as an excellent pollinator in the production of S. tuberosum dihaploids in potato breeding programmes because of the high yield of dihaploids per 100 berries, and because no introgression of DNA into the S. tuberosum dihaploids was evidenced.     

Introgression of resistance to Columbia and Northern root-knot nematodes fromSolanum bulbocastanum into cultivated potato

Summary Resistance toMeliodogyne chitwoodi races 1 (MC1) and 2 (MC2) andM. hapla (MH) derived fromSolanum bulbocastanum was introduced into the cultivated potato gene pool through somatic fusion. The initial F₁ hybrids showed resistance to the three nematodes. Resistance to reproduction on roots by MC1 was accompanied by resistance to tuber damage in F₁ clones. Tuber damage sometimes occurred, however, in hybrids of BC₁ progeny resistant to reproduction on roots when MC2 and MH were the challenging nematodes. Resistance to reproduction was transferred into BC₁ individuals, but a greater proportion of BC₁ progeny was resistant to MC1 than to MC2 or MH. Resistance to MC1 appears to be dominant and discretely inherited. F₁ and BC₁ progeny were pollen sterile, but seed were produced from crosses using cultivated tetraploid pollen sources. Approximately 11 and 33 per cent of pollinations produced berries on F₁ and BC₁ pistillate parents, respectively. Seed yield increased fourfold overall in crosses with F₁ compared to BC₁ individuals.Abbreviations MC1 Meloidogyne chitwoodi race 1 - MC2 Meloidogyne chitwoodi race 2 - MH Meloidogyne hapla - Rf Reproductive factor     

Combination of resistance to potato late blight in foliage and tubers by intraspecific dihaploid protoplast fusion

Five dihaploid Solanum tuberosum genotypes, encompassing different levels of polygenically inherited resistance to potato late blight disease (Phytophthora infestans (Mont.) de Bary) in foliage and tubers, were used in four intraspecific protoplast fusion combinations. Vigorous growing putative hybrid calli were selected four weeks after electrofusion. Intact plants were regenerated from 6% of the selected calli. Verification of hybridity was accomplished by use of RAPD analysis which revealed that 53% of the regenerated plants were true somatic hybrids. The score of true somatic hybrids in the different fusion combinations ranged within 21% – 100%. The hybrid plants were analysed for resistance to foliage blight in a field trial and assessed for tuber blight resistance by use of a laboratory test. Resistance to late blight in foliage and tubers varied between the hybrids. Very high levels of resistance to both foliage and tuber blight were obtained in some hybrids. However, loss of resistance in some hybrids as compared to the parental plants were also observed. Possible reasons for the phenotypic disappearance of resistance to either foliage or tuber blight are suggested. This revised version was published online in July 2006 with corrections to the Cover Date.     

The reaction of potato differentials to Phytophthora infestans isolates collected in nature

Available data have been evaluated on the reaction of potato differentials to over 5000 Phytophthora infestans isolates, collected in various parts of the world. The differentials were able to identify up to 11 virulence factors in P. infestans. The isolates differed in virulence expression, depending on the isolate and testing conditions. All 11 virulence factors were found in both ‘old’ and ‘new’P. infestans populations. The resistance of individual differentials was not overcome with equal frequency. The resistance of differentials R5, R8 and R9 was overcome least frequently. This may be due to instability of expression of the respective virulence factors in P. infestans and/or the kind of resistance present in the differentials. Whatever the reason, such resistance may possibly be used in breeding potato cultivars with durable resistance to P. infestans.     

Effectiveness of in vitro selection for agronomic characters in potato

Effectiveness of in vitro selection for agronomic characters was studied by finding correlation coefficients between in vitro and in vivo performance of 22 potato genotypes. Evaluation was performed under eight in vitro and two in vivo conditions. Genotypic differences were highly significant for various characters under all in vitro and in vivo conditions. Error mean squares were much lower in in vitro experiments than in in vivo experiments. In vitro selection was found to be highly effective for tuber colour, stem pigment and number of eyes, and moderately effective for average tuber weight, plant vigour and foliage senescence under specific conditions. The results also indicated the possibility of in vitro selection for heat tolerance. For tuber yield and number of tubers, effectiveness of in vitro selection was low to very low, because differences in phenotypic expressions of genotypes were much enlarged under in vitro conditions compared to under in vivo conditions. Correlation coefficients for certain characters were better when in vitro conditions were closer to in vivo conditions. To improve the efficiency of in vitro selection, there is perhaps a need to simulate and identify the in vitro conditions under which genotypic differences are reflected in phenotypes more realistically as they are expressed under in vivo conditions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance and mapping of a light green mutant in cultivated diploid potatoes

A light green mutant was found in a population of adapted cultivated diploid potatoes. Genetic analysis indicates that this trait is controlled by a single nuclear gene. The gene symbol lg is proposed. The segregation ratios fit a pattern which strongly suggest that there is a close linkage between the Lg allele and a locus which confers lethality in its homozygous recessive state. Some crossing over between the lg locus and the lethal was found to occur but LgLg genotypes were not observed in progenies from sib-matings. The lg locus mapped to the potato linkage group VI between the restriction fragment length polymorphism (RFLP) loci CP18 and GP24. This revised version was published online in August 2006 with corrections to the Cover Date.     

Expression of nonacclimated freezing tolerance and cold acclimation capacity in somatic hybrids between hardy wild Solanum species and cultivated potatoes

The expression of freezing tolerance was characterized in interspecific somatic hybrids between S. tuberosum (tbr) and two cold-hardy wild species, S. brevidens (brd) and S. commersonii (cmm). The nonacclimated freezing tolerance (NA) and acclimation capacity (ACC, increase in freezing tolerance in response to low nonfreezing temperature), two main genetic components of freezing tolerance, were evaluated separately. In contrast to cmm, which exhibited excellent NA and ACC, the freezing tolerance of brd was mainly due to ACC. However, the ACC of brd was only moderately expressed in the somatic hybrids. The NA of cmm was also suppressed in combination with tbr genomes. However, with acclimation, some of the tbr (+) cmm somatic hybrids achieved freezing tolerance comparable to pure hardy species such as brd used in this study. Analysis of chloroplast DNA type by RFLP markers revealed no significant difference in ACC between somatic hybrids carrying chloroplasts from either tbr or cmm. The reasons for the reduced expression of freezing tolerance from either the brd or cmm parent and the utilization of these somatic hybrids in breeding programs are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of RAPD markers linked to the Ns locus in potato

Using the RAPD method and bulked segregant analysis we identified four RAPD markers linked to a dominant gene Ns, responsible for a hypersensitive reaction of potato (Solanum tuberosum L.) to potato virus S (PVS) infection. The markers OPE15⁵⁵⁰, OPJ13⁵⁰⁰, OPG17⁴⁵⁰ and OPH19⁹⁰⁰ were found to be closely linked to the Ns gene in diploid potato clones. They are situated at 2.6, 3.3, 4.6 and 6.6 cM from Ns, respectively. As a source of the gene, clone G-LKS 678147/60, which is known to carry Ns transferred from S. tuberosum ssp. andigena was used. These RAPD markers were not amplified in resistant tetraploid clones containing Ns derived from the clone MPl65 118/3, also having an andigenum origin. This suggests that there may be two separate loci of Ns in the sources identified, or different alleles with the same specificity at a single locus, or that the genetic background of tetraploids tested results in different RAPD amphlification patterns.     

Widespread introgression of Solatium phureja DNA in potato (S. tuberosum) dihaploids

Dihaploids were assumed to be of parthenogenic origin but cv. ‘Pentland Crown’ dihaploids produced at the Scottish Crop Research Institute (SCRI), Dundee, Scotland, are known to contain DNA from the S. phureja dihaploid inducer. This study investigates whether inducer DNA occurs in dihaploids of cultivars ‘Aminca’, ‘Brio’, ‘Lizen’ and ‘Sirtema’, produced at INRA, Ploudaniel, France. Two microsatellite markers and five simple sequence repeat 5′-anchored polymerase chain reaction (PCR) primers were used to generate markers. Markers originating from the dihaploid inducer were detected in 13 of the 19 INRA dihaploids and in some dihaploids from each cultivar. Greater introgression occurred in ‘Lizen’ dihaploids than in ‘Brio’ dihaploids, suggesting that the female (S. tuberosum) parent influences introgression. The percentage of INRA dihaploids containing inducer DNA was similar to that for ‘Pentland Crown’ dihaploids (c. 65%). The micro-satellite markers provided the first evidence of the transfer of specific, potentially useful, genes from the inducer to the dihaploid offspring. Interspecific introgression during dihaploid induction is more widespread than previously thought and researchers should be aware that it may influence the results of research using dihaploids.     

DNA analysis of potato + Solanum brevidens somatic hybrid lines

Three somatic hybrid lines between potato (cv. While Lady line no. Ke 79, 2n = 2x = 48) + Solanum brevidens (PI 218228, 2n = 2x = 24) were evaluated using randomly amplified polymorphic DNA (RAPD) markers. The lines originated from the same callus but showed different reactions to Erwinia carotovora ssp. carotovora, the cause of potato soft rot. By the use of 48 oligomer primers producing 99 scorable bands, DNA polymorphism were detected on 7 of 12 S. brevidens chromosomes. Loss of certain DNA segments on chromosome 5, 6, 9 and 11 were observed. Some of the variations could have taken place in early callus stage of development; others may have occurred after initiation of individual shoot regeneration. The possible involvement of missing RAPD products specific to one somatic hybrid that shows decreased resistance to bacterial soft rot is discussed.     

Production of somatic hybrids between S. tuberosum L. and late blight resistant Mexican wild potato species

Interspecific somatic hybrids between dihaploid breeding clones of potato, S. tuberosum and two accessions of wild Mexican species S. pinnatisectum and the hybrid line S. pinnatisectum × S. bulbocastanum were regenerated following electrofusion of mesophyll protoplasts to combine important agricultural traits of S. tuberosum and a high level of late blight resistance from selected wild accessions. In two fusion combinations 239 calli were regenerated; 162 from 195 calli analysed were identified as hybrids by means of isozyme analysis of peroxidases and, for some hybrid clones, by RAPD analysis. Depending on the fusion combination, 47–89 percent of the somatic hybrids had the expected ploidy level and 7–16 percent were mixoploids. Somatic hybrids were phenotypically intermediate as compared to their parents and some of them were able to be backcrossed sexually with potato. Fertility and crossability depended on combination and ploidy level of the somatic hybrids. In tests with detached leaves the wild partner clones had a high late blight resistance score of 8,6 and 8,9; the susceptible tuberosum-partners of 2,8 and 3,5, respectively. Nearly 25 percent of somatic hybrids had a resistance level of 6 or higher in the first year of assessment. The average resistance value of most somatic hybrids was lower than the average parental level. The reasons for variation in resistance values are discussed in connection with the practical application of fusion hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Combining ability and correlations for yield components in early generations of potato breeding

A partial diallel set of crosses was made between 14 potato cultivars chosen for their fertility, from those included in a potato breeding programme at the NEIKER – Basque Institute for Agricultural Research. The progeny were grown in completely randomized trials from 1997 to 1999. Performance for yield, tuber number and average tuber weight was analysed in seedling and two clonal generations. Variance estimates due to both general combining ability (GCA) and specific combining ability (SCA) were significant in all generations for all traits under study. However, SCA was more important than GCA in almost all cases. Correlation coefficients among characters, generations, GCA and SCA effects were examined. For tuber yield no relation was obtained between generations; however, average tuber weight and yield were positively associated in all generations. The results indicate that appropriate selection criteria depend strongly on the particular cross. The implication for a breeding strategy are discussed.