DNA marker-assisted evaluation of potato genotypes for potential resistance to potato cyst nematode pathotypes not yet invading into Japan

One of major objectives of crop breeding is conferring resistance to diseases and pests. However, large-scale phenotypic evaluation for many diseases and pests is difficult because strict controls are required to prevent their spread. Detection of disease resistance genes by using DNA markers may be an alternative approach to select potentially resistant accessions. Potato (Solanum tuberosum L.) breeders in Japan extensively use resistance gene H1, which confers nearly absolute resistance to potato cyst nematode (Globodera rostochiensis) pathotype Ro1, the only pathotype found in Japan. However, considering the possibility of accidental introduction of the other pathotypes, breeding of resistant varieties is an important strategy to prevent infestation by non-invading pathotypes in Japan. In this study, to evaluate the prevalence of resistance genes in Japanese genetic resources, we developed a multiplex PCR method that simultaneously detects 3 resistance genes, H1, Gpa2 and Gro1-4. We revealed that many Japanese varieties possess not only H1 but Gpa2, which are potentially resistant to other pathotypes of potato cyst nematode. On the other hand, no genotype was found to have the Gro1-4, indicating importance of introduction of varieties having Gro1-4. Our results demonstrate the applicability of DNA-marker assisted evaluation of resistant potato genotypes without phenotypic evaluation.     

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.     

Inheritance of resistance to potato cyst-eelworm (Heterodera rostochiensis Woll.) in the genus Lycopersicon

Summary The presence of resistance to potato cyst-eelworm in wild Lycopersicon species was confirmed.The resistance of two plant accessions, L. pimpinellifolium, B6173, and L. peruvianum, B6001, was compared in screening tests against three eelworm isolates, using the susceptible L. esculentum Ailsa Craig as a control. Both accessions were shown to possess high levels of resistance to the Wren isolate (pathotype A) of H. rostochiensis, and resistance to the Castle Donington (pathotypes A, B, and E) and Hare Lane (pathotype B) isolates. The level of resistance in L. pimpinellifolium was greater than that in L. peruvianum. Because of its greater resistance, and the ease with which it can be hybridised with tomato cultivars, L. pimpinellifolium was chosen as a donor parent for breeding programmes designed to incorporate potato cyst-eelworm resistance into a commercial tomato variety.The resistance of L. pimpinellifolium, B6173, to the Wren isolate of H. rostochiensis was shown to be controlled by a single dominant gene for which the symbol Hero is proposed.     

Molecular markers for late blight resistance breeding of potato: an update

Late blight is the most devastating disease of the potato crop that can be effectively managed by growing resistant cultivars. Introgression of resistance (R) genes/quantitative trait loci (QTLs) from the Solanum germplasm into common potato is one of the plausible approaches to breed resistant cultivars. Although the conventional method of breeding will continue to play a primary role in potato improvement, molecular marker technology is becoming one of its integral components. To achieve rapid success, from the past to recent years, several R genes/QTLs that originated from wild/cultivated Solanum species were mapped on the potato genome and a few genes were cloned using molecular approaches. As a result, molecular markers closely linked to resistance genes or QTLs offer a quicker potato breeding option through marker‐assisted selection (MAS). However, limited progress has been achieved so far through MAS in potato breeding. In near future, new resistance genes/QTLs are expected to be discovered from wild Solanum gene pools and linked molecular markers would be available for MAS. This article presents an update on the development of molecular markers linked to late blight resistance genes or QTLs by utilization of Solanum species for MAS in potato.     

The importance of wild potato species resistant to the potato cyst nematode,Globodera pallida,pathotypes P4A and P5A,in potato breeding. I. Resistance studies

Summary Seven wild diploid potato species, Series Tuberosa, representing 1023 clones were screened for resistance to the potato cyst nematode, Globodera pallida. Over 25% of the clones were resistant to pathotype P₄A and almost 30% were resistant to pathotype P₅A. The resistance in hybrid progenies of these and other resistant species with cultivated potatoes was evaluated, and over 2200 seedlings were screened. High frequencies of resistance (>50%) to P₄A were found in progenies with Solanum leptophyes, S. vernei, S. gourlayi and S. capsicibaccatum, whereas resistance to P₅A was found in these species as well as S. sparsipilum. The importance of nematode resistant wild species for potato breeding is discussed.     

Pathotype-specificity in potato cyst nematodes,a reconsideration

Summary Within the two potato cyst nematode (PCN) species, Globodera rostochiensis and G. pallida, eight pathotypes are presently discerned by seven differential Solanum clones, whereby resistance is defined as a Pf/Pi ratio <1.0, and susceptibility as a ratio above 1.0. This pathotyping is felt to be unsatisfactory. Reconsidering the original data and data published by others resulted in the conclusion that there are only three reliably recognizable Ro-pathotypes Ro1/Ro4, Ro2/Ro3 and Ro5. The differential clones 60.21.19 and 65.346.19 differentiate only between the two species. Within G. pallida it does not seem possible to identify pathotypes reliably. The resistance-virulence pattern is of a quantitative nature fitting a polygenic system. There appear to be pathotype-specific effects, which are not large enough to identify and classify pathotypes reliably. It is therefore suggested to rename the Ro-pathotypes into Ro1 (old Ro1 and Ro4), Ro3 (old Ro2 and Ro3) and Ro5 (old Ro5) and to speak of more and less virulent Pa-populations.The rigorous requirements for resistance in various countries present great problems to the breeders and result in the removal of much valuable partial resistance. Especially with G. pallida these rigorous requirements are probably an obstacle rather than an encouragement to solve the pallida problem.     

Breeding for resistance to potato root eelworm

Summary A study was made on the inheritance of resistance to Heterodera rostochiensis, the potato root eelworm. Crosses between resistant andigenum seedlings and between these seedlings and tuberosum varieties revealed the existence of a dominant gene H governing the inheritance of resistance.The segregation ratios in the andigenum crosses (Table 2) are in close agreement with the assumption of a tetraploid scheme without double reduction. However, in the andigenum-tuberosum crosses the susceptible seedlings were in excess.Segregation ratios in some progenies indicate the existence of two more genes K and L causing resistance only when both are present.The resistant plants possess the hatching factor and the larvae accordingly penetrate the roots in large numbers, but they do not reach sexual maturity.The introduction of the resistance genes seems to be quite easy, but combining resistance with all other desirable features will be a very arduous task.     

Genomic variation and genetic relationships in Ipomoea spp.

Random amplified polymorphic DNA (RAPD) markers were used to develop genetic fingerprints and analyse genetic relationships among 29 Ipomoea accessions from different geographical locations around the world, including unique wild species, and reproducible profiles were obtained for all accessions using random decamer primers. The primers generated 46 polymorphic markers, one primer alone having 10 products, enabling the discrimination of all 29 accessions. A high level of genetic variability in sweet potato collections was suggested by the degree of polymorphism. Half of the Japanese land races were closely related while accessions from Papua New Guinea and The Philippines were distinct and exhibited the greatest genetic diversity. The wild species Ipomoea gracilis and Ipomoea tiliacea formed a group distinct from the cultivated sweet potato. The wild tetraploid accession K233 and the species Ipomoea trifida were progressively more related genetically to the cultivated sweet potato and are the probable progenitors of Ipomoea batatas, and may be suitable as germplasm for genetic enhancement. RAPDs proved to be useful for sweet potato systematics and should be valuable for germplasm management, gene tagging and efficient choice of parents in breeding programmes.     

Further sources of resistance toHeterodera rostochiensis woll. in the andigena potatoes

Summary The Andigena potatoes (Solanum tuberosum ssp.andigena) of the Commonwealth Potato Collection (C.P.C.) were screened for resistance to non-pathotype A populations ofHeterodera rostochiensis. Three populations were used: Duddingston (pathotype B), Nocton (pathotype C) and Changed Little Ouse (pathotype C). Resistance was found in C.P.C. 2775, 2802 and 2805. C.P.C. 2775 also had resistance to pathotype A (Feltwell population) and appeared to have two dominant genes,H ₁ giving resistance to pathotype A andH ₃ giving resistance to pathotypes B and C. C.P.C. 2802 was susceptible to pathotype A and appeared to have only anH ₃-type gene. C.P.C. 2805, which has not been investigated thoroughly because the plants lacked vigour, may also have anH ₁ and anH ₃ gene. The value of C.P.C. 2775, 2802 and 2805 for breeding nematode-resistant potatoes will only be known when they have been tested with further non-pathotype A populations and when the genetics of resistance have been investigated thoroughly.     

Resistance to late blight in potato. A putative gene that suppresses R genes and is elicited by specific isolates

Eleven progenies, ten bi-parental and one selfed, of tetraploid potato were assessed for phenotypic segregation ratios of resistance to susceptible individuals against two isolates of P. infestans, one avirulent to all 11 known potato R genes (avirulent pathotype) and one virulent to eight R genes (virulent pathotype). Assuming Mendelian inheritance of independent R genes, the numbers of individuals segregating for resistance to both races could not be explained sufficiently for all progenies. However, the assumed presence of dominant, specific suppressors of the R gene function that were elicited by specific isolates of the fungus and that segregated in the host independently from R genes explained all frequencies of resistant and susceptible individuals in each of the progenies studied. This revised version was published online in July 2006 with corrections to the Cover Date.     

The development of females of the potato root eelworm, (Heterodera rostochiensis Woll.) on ‘tuber-pieces’ of susceptible and resistant potatoes

A method is described for determining the degree of resistance shown by potatoes to a non-aggressive biotype of the potato root eelworm, Heterodera rostochiensis Woll., using tuber slices inoculated with H. rostochiensis larvae. The degree of resistance assessed by this method was found to agree well with the results obtained by root screening. The tuber piece method of testing for resistance could be used when root screening is impracticable or in conjunction with it.     

New sources of resistance to potato cyst-nematodes in the Commonwealth Potato Collection

Summary Accessions obtained from the Commenwealth Potato Collection were evaluated for resistance to European populations of potato cyst-nematodes (PCN). With over 36% of the current collection assessed, resistance to pathotypes of both Globodera rostochiensis and G. pallida were identified in 16 species from Argentina, Bolivia or Peru. Resistance to all PCN pathotypes was particularly prevalent in species from the Andean regions of Bolivia and North Argentina, supporting the hypothesis of co-evolution of the potato and PCN in these regions.     

Breaking the crossability barriers between disomic tetraploid Solanum acaule and tetrasomic tetraploid S. tuberosum

Summary A combination of compatible second pollinations and embryo rescue was applied for systematic production of true tetraploid hybrids from crosses between disomic tetraploid Solanum acaule and tetrasomic tetraploid potato, S. tuberosum. Several genotypes of tetraploid potatoes were pollinated with S. acaule, and the compatible second pollinations were made on the following day, with a genotype of S. phureja, IvP 35 to promote fruit development. Embryo rescue was carried out in 21 families, 14 to 27 days after the first pollination. A total of eight plants were obtained from the embryo rescue and their chromosome numbers were counted in the root tips. Three of the eight plants were identified as tetraploid, and five others as diploid. Morphology, isozyme banding patterns, and pollen stainability, as well as potato spindle tuber viroid (PSTVd) resistance, indicated the hybrid nature of the three plants. This is the first report of successful tetraploid hybrid production between disomic tetraploid S. acaule (4x) and tetrasomic tetraploid potatoes. Seed set from the crosses between one of hybrids and diploid potatoes indicated workable levels of both male and female fertility for introgression of valuable genes from S. acaule into the cultivated potato gene pool. The methodology used may be applied to other disomic tetraploid tuber-bearing Solanum species and with some modifications also to distantly related solanaceous species and genera.     

A new resistance gene to powdery mildew identified in <Emphasis Type="Italic">Solanum neorossii</Emphasis> has been localized on the short arm of potato chromosome 6

A new resistance (R) gene to powdery mildew has been identified and characterized in a population derived from the wild potato species, Solanum neorossii under natural infection in the greenhouse. The segregation of resistance has revealed that this R gene is controlled by a single monogenic and dominant gene designated Rpm-nrs1. Analysis of the DNA sequence on an internal transcribed spacer (ITS) region of the pathogen genome suggests that the pathogen causing the powdery mildew disease is either Golovinomyces orontii or G. cichoracearum. The resistance locus was localized to the short arm of chromosome 6 where several disease R genes already identified in potato and tomato are known to reside. The resistance locus cosegregated in 96 progeny with three AFLP markers and one PCR marker. The sequences of the two cosegregating AFLP markers are highly homologous to Mi-1 conferring resistance to nematode, potato aphid and whitefly and Rpi-blb2 conferring resistance to late blight. The results in this study will facilitate the cloning of this gene conferring resistance to powdery mildew.     

Validation of molecular markers for pathogen resistance in potato

DNA markers have a large potential to improve efficiency and precision of conventional plant breeding programmes based on marker‐assisted selection (MAS). In our study, we have evaluated the predictive abilities of the SCAR marker RYSC3 and the CAPS marker GP122₅₆₄ with regard to the PVY resistance genes Ry_adg and Ry_sto, respectively, and of marker TG689 linked to H1 conferring resistance to Globodera rostochiensis and marker HC associated with high levels of G. pallida resistance. The evaluations were made in 28 cultivars and accessions and in 219 progeny genotypes descending from ten different crosses. We observed in all evaluated cultivars and accessions the expected marker patterns according to their phenotypic classification into resistant and susceptible genotypes. However, in part considerable discrepancies were observed when analysing progeny of controlled crosses involving these resistance sources, particularly with respect to H1. Based on these results, practical aspects for the efficient implementation of marker‐assisted selection are discussed, which consider the genetic origin of the material, costs aspects and methodology applied.     

New sources of resistance to the potato cyst nematodes Globodera pallida and G. rostochiensis in the Commonwealth Potato Collection

The Commonwealth Potato Collection contains 77 species following the incorporation of the personal collection of Professor JG Hawkes. The material originates from several expeditions to the Americas and represents a potentially valuable source of resistance to pests and pathogens, including potato cyst nematodes (PCN). The germplasm from the Hawkes collection screened in this study included taxa mainly from Mexico, Bolivia, Argentina and Peru. Seedlings of 198 accessions, from 63 different species were screened for reaction to the PCN species G. pallida and G. rostochiensis. Of the 198 accessions tested, 56% contained G. pallida-resistant clones and 53% contained clones resistant to G. rostochiensis. Twelve species in this survey are believed to contain novel, previously unreported sources of resistance. The geographical distribution of resistance indicated a contrast between the two PCN species. Although, as in previous studies, resistance to G. rostochiensis was centred on Bolivia and Argentina, G. pallida resistance was found across the geographical range of tuber-bearing Solanum. The possibility of cross-resistance arising from selective pressure from the more northern species G. mexicana is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Aegilops ventricosa segment on chromosome 2AS of the wheat cultivar 'VPM1' carries the cereal cyst nematode resistance gene Cre5

Previous studies showed that the intermediate level of resistance in bread wheat line ‘VPM1’ to pathotype Ha12 of the cereal cyst nematode could be conferred by an Aegilops ventricosa‐derived gene, CreX, in chromosome arm 2AS, which also carries the rust resistance genes Yrl7, Lr37 and Sr38. Near isogenic lines (NILs) differing for the presence and absence of the Ae. ventricosa‐derived linked genes Yrl7/Lr37/Sr38 were tested with cereal cyst nematode. Lines carrying Yr17 produced significantly fewer nematode cysts than the controls. An infested soil experiment produced better differentiation among resistant and susceptible genotypes. Susceptibility of ‘Trident’ indicated that linkage between CreX and Yr17 is incomplete. Microsatellite markers did not differentiate between ‘Trident’ and CreX‐carrying genotypes. However, Xgwm636 (104) was associated with the presence of Yr17 in all six genetic backgrounds. Since none of the reported cereal cyst nematode resistance genes is located in chromosome 2AS, CreX was designated as Cre5.     

Two distinct potato late blight resistance genes from <Emphasis Type="Italic">Solanum berthaultii</Emphasis> are located on chromosome 10

For breeding potato varieties resistant to late blight, identification of resistance genes to Phytophthora infestans (Rpi genes) is essential. Introduction of Rpi genes from wild Solanum species into cultivated potato is likely to be a good method to achieve durable resistance to P. infestans. In this study, we identified two Rpi genes (Rpi-ber1 and Rpi-ber2) derived from two different accessions of Solanum berthaultii. These two genes are closely linked on the long arm of chromosome 10. There are similarities between the predicted genetic locations of the previously identified Rpi-ber and Rpi-ber1, which given the common origin of these genes, may indicate that they are the same. However, the genetic positions of Rpi-ber1 and Rpi-ber2 are different. Rpi-ber1 is positioned between CT214 and TG63, whereas Rpi-ber2 is located below both of these two markers. In addition, the sequences of four linked markers to both R genes showed different polymorphisms indicating the two Rpi genes could be transmitted from different haplotypes (chromosomes).     

Resistance to Phytophthora infestans in Potato Genotypes Originating from Wild Species

The reaction to P. infestans in the foliage and tubers of potato was evaluated in resistant genotypes and in their progeny. The genotypes originated from wild species known to carry genes for specific resistance. Evidence was obtained that three types of resistance to P. infestans were presented in the genotypes: (1) Resistance due to major, dominant genes was predominant. The presence of these genes was expressed in both foliage and tubers and did not influence maturity. Some of the genes were R genes, providing specific resistance, but for others this could not be determined as they were effective against the fungus inoculum with the highest virulence available: 1.2.3.4.5.7.10.11. (2) Resistance was present, associated with late maturity. (3) Tuber resistance was present, expressed after inoculation of whole tubers but not after inoculation of tuber slices. Testing of the progeny was essential for the determination of the type of resistance present in the genotypes evaluated.