The use of hand-held terminals and micro-computers for the recording and analysis of potato cyst nematode counts

Summary An automated system of data capture and summarisation is described. Although it is of widespread applicability, it is illustrated by a specific example involving the screening of potato tubers for resistance to potato cyst nematode.     

Breeding for quantitative resistance to potato cyst nematode (Globodera pallida) in tetraploid potatoes using dihaploids and unreduced gametes

Summary Dihaploids were produced from tetraploids resistant to potato cyst nematode (Globodera pallida (Stone)). High levels of resistance were found in the dihaploids and three were used to produce tetraploid progenies by crossing them with susceptible tetraploid cultivars. One dihaploid, PDH505, produced more highly resistant offspring than the other two, PDHs 417 and 418. The latter gave progenies whose levels of resistance were similar to those obtained from susceptible dihaploids crossed with resistant tetraploids.The differences between the progenies of the resistant dihaploids were probably due to different modes of unreduced gamete formation (PDH505 producing gametes by first division restitution (FDR) and PDHs 417 and 418 by second division restitution (SDR)) although cytological studies would be necessary to confirm this. The methods by which dihaploids could be utilised in a tetraploid potato breeding programme are discussed in relation to the mode of unreduced gamete formation.     

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.     

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.     

Durability of resistance against potato cyst nematodes

A requirement for evaluating the effectiveness of major resistance genes (R-genes) is detailed knowledge of the genetic variability of the nematode populations in an area. Obtaining insight in the genetic variation in an area is a tour de force, because qualitative data are not sufficient allele frequency data at well studied loci are required. Another important step in predicting the durability of major R-genes is to study the parasite at the molecular level. Also for nematodes it seems that R-genes are part of a molecular surveillance system recognising foreign molecules. For potato cyst nematodes it is becoming clear that numerous proteins are secreted to manipulate the host plant and it may be assumed that some of these secretory proteins are also recognised by the host. Knowledge of the molecular nature and function of these avirulence gene products may reveal clues to predict the durability of an R-gene. R-genes that recognise avirulence gene products that have a crucial function in the nematode maybe very durable. When the current molecular models for gene-for-gene interactions are correct, the durability of an R-gene may be predicted by studying the dispensability factor and functional constraints of the avirulence gene products. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

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

Summary Over 2400 pollinations were made to investigate the crossability relationships between cultivated potatoes, and wild diploid species from Series Tuberosa and Circaeifolia, as well as wild polyploid species in Series Tuberosa and Longipedicellata resistant to potato cyst nematode, Globodera pallida pathotypes P₄A and P₅A. Wild diploids in Series Tuberosa crossed easily with cultivated diploid species, except with Solanum lignicaule where most pollinations failed, and seed set was extremely low (0.2 seeds per pollination or less). It is suggested that this species is 1EBN. S. capsicibaccatum is clearly isolated from Series Tuberosa, but can form hybrids with S. lignicaule, which can act as a bridging species to S. tuberosum haploids. S. gourlayi and S. oplocense can be crossed with both subspecies of S. tuberosum, but S. papita, Series Longipedicellata is reproductively isolated from the tetraploid cultigens. The crossability data are discussed in the light of germplasm utilisation for breeding potato varieties resistant to potato cyst nematode.     

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.     

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.     

Chilean potato germplasm with resistance to potato viruses Y and S

Summary Four clones of the Chilota Potato Collection from the germplasm bank of the Universidad Austral de Chile, with potential combined resistance to PVX, PVY and PVS, were mechanically inoculated with these viruses in order to demonstrate wether they were truly resistant or not. Virus coat protein in the inoculated clones was determined by means of NCM-ELISA, DAS-ELISA, Western Blot and NASH.Two of the four inoculated clones were resistant to both PVY and PVS. This information will allow the utilization of these two clones in plant breeding programs.Abbreviations NCM-ELISA Nitrocellulose membrane enzyme-linked immunosorbent assay - DAS-ELISA double antibody sandwich enzyme-linked immunosorbent assay - NASH nucleic acid spot hybridization     

Breeding for resistance to cereal cyst nematode in wheat

Summary The progress of a backcross breeding programme to introduce resistance against the cereal cyst nematode into wheat is described. Methods of resistance screening and criteria for selection are detailed and the results discussed with reference to alternative procedures for the introduction of new resistance genes into major breeding programmes.     

Performance and prediction of potato genotypes tested in international trials

Summary A series of international trials with the potato crop was conducted in eight countries representing widely differing environments. The purpose of the experiments was: (1) to develop a method for obtaining environmental indexes which measure productivity of diverse potato-growing areas, and (2) to construct performance models for individual genotypes based on the environmental indexes. Yield data of 32 genotypes obtained from a series of 13 experiments conducted in the eight countries in 1983 and 1984 were used in the study. The response pattern of the genotypes was studied by a multivariate analysis. Five genotypes were chosen as representatives for different response patterns to the environments and their data were used for calculating a series of independent indexes for each of the experimental sites based on principal component analysis. Performance models for each of the genotypes were constructed by stepwise regression analysis of yields of a genotype on the environmental indexes based on 1983 data. Yield data of 1984 experiments were used to verify the models. Satisfactory correspondence between observed and predicted yields was obtained for most of the genotypes.     

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     

A re-examination of apparent selection in Globodera pallida on Solanum vernei hybrids

Summary This paper re-examines the data in support of recent findings by Turner et al. (1984), that selection for virulence in Globodera pallida takes place on Solanum vernei hybrids. It is concluded that there are a number of factors that must be taken into account and that these data must be interpreted with caution.     

Evaluation of resistance to Pseudomonas solanacearum in potato under controlled conditions

Summary Potato plantlets derived from in vitro propagation of three cultivars known for their field resistance (Cruza 148 and BR-63.65) or susceptibility (Désirée) to Pseudomonas solanacearum E.F. Smith were artificially inoculated under controlled conditions. The aim of this work was to determine the optimal inoculum concentration and the best observation period in which the cultivars would show different reactions to bacterial infection as expected on the basis of their field performance.A suitable statistical analysis of disease indices is proposed to distinguish between resistant and susceptible responses, with a particular care for the applicative needs and a univocal interpretation of the results. In order to evaluate the significance of sources of variation related to the observed mean differences, the analysis of variance and a convenient clustering procedure of disease index means were applied.The statistical analysis revealed that, under our conditions, an inoculum concentration of 5×10⁶ cfu/plant was suitable for separating resistant from susceptible responses, in accordance with the reactions already observed in field experiments by other authors. Also, differences among the three cultivars were best observed nine to twelve days after inoculation with the pathogen.     

Breeding for resistance to cereal cyst nematode in wheat II. Use of test tube cultures in selection

Summary The use of soil. naturally infested with Heterodera avenae, to select resistant heterozygotes in backcross progenies of wheat, was tested for reliability. Selfed progenies from plants selected as resistant were cultured monoxenically in test tubes with nematodes hatched from single cysts, while backcross progenies from the same parent plants were grown in pots of naturally infested soil. Cyst counts were made after two months' growth. The results showed that over 50% of the backcross lines, screened in previous generations with naturally infested soil, had been erroneously selected as resistant. The test tube cultures clearly differentiated lines carrying resistance from those which were susceptible and corroborated results from pot tests.     

The effect of seed tuber weight on characters in the first and the second clonal generation of potato populations

Summary Fifty-five populations from crosses of three Solanum tuberosum ssp. tuberosum cultivars and seven selected S. tuberosum ssp. andigena clones adapted to the northern temperate zone have been studied in the seedling and the first and second clonal generation. This paper only deals with the effect of seed tuber weight on various important agronomic characters which are selected for in early clonal generations, independently of the type of crosses.The results showed that a distinction has to be made between the effects of differences in seed tuber weight within and between seedlings, i.e. genotypes. The effects on the first clonal generation of differences in weight of the tubers within seedlings and those of the first heaviest tubers between seedlings were similar and comparable to seed tuber weight effects within cultivars. In contrast to the within seedlings effects the between seedlings effects proved to be heritable. This is concluded from the fact that, on average, independent of seed tuber weight seedlings with larger first heaviest tubers performed better in the second clonal generation than those with smaller ones. Notably plant height, general impression, date of maturity and particularly tuber yield and mean tuber weight in the second clonal generation were related to the weight of the first heaviest tubers of the seedlings. The potentialities of these findings for an early generation selection are discussed.     

Genetic resistance to cereal cyst nematode Heterodera avenae Woll. in wild oat Avena sterilis I. 376

Summary Three dominant genes A, B and C are involved in the resistance of Avena sterilis I. 376 to Heterodera avenae. When associated, genes A and B are responsible for the high level of resistance in I. 376. In the absence of the first two genes, gene C confers intermediate resistance characterized by the presence of a limited number of cysts on the roots. Only the completely recessive genotype allows the nematode to develop normally.     

Multiple resistance to diseases and pests in potatoes

Summary The potato has more characters of economic importance that need to be considered by the breeder than any other temperate crop. In Europe these include resistance to at least twelve major diseases and pests. Highest priority has been given to resistance to late blight (Phytophthora infestans), virus diseases (particularly those caused by potato leafroll virus and potato virus Y) and potato cyst nematode (Globodera rostochiensis andG. pallida). Useful sources of resistance are available and early generation screening techniques have been developed to allow positive selection for multiple resistance and the breeding value of clones used as parents to be determined. Progress in restriction fragment length polymorphism technology should result in more efficient selection in the future.     

Search for sources of early blight (Alternaria solani) field resistance not associated with vegetative late maturity in tetraploid potato germplasm

Summary A total of 934 potato cultivars and clones from different breeding programmes across the world were tested for their reaction to early blight (Alternaria solani) in Brasilia-DF, Brazil. This field trial was set up in order to identify tetraploid potato genotypes with useful levels of resistance to early blight (EB) that were not correlated with late maturity or lateness. Artificial inoculation (spread ofA. solani infected leaves over plant canopy) was employed even though EB occurs naturally at epidemic levels in the test area. Four disease assessments were made every 10 days, beginning 45 days after planting. Based on the evaluation made 65 days after planting, only 27 clones were identified that had less lesioned leaf area than the resistant standard (cv. Aracy). Area under disease progress curve (AUDPC) was also used as a criterion of resistance. Using AUDPC as a parameter, 22 genotypes were identified presenting values lower than or identical to cv. Aracy. Differences in the commercial yield were recorded in genotypes with identical response to EB, suggesting that distinct levels of tolerance to EB also exist. In spite of the strong correlation between EB resistance and late maturity, some genotypes derived from the parental materials NDD 277-2, Kufri Jyoti, CIP 377888-7, Maine-28, and the clones CFS 69.1 and I-853 were identified as having good levels of EB field resistance not linked with the negative phenotypic trait of lateness. These materials presented medium to medium-early maturity with a vegetative cycle of around 90–95 days. Some selected genotypes presented more acceptable agronomic characteristics including resistance toPhytophthora infestans, potato leaf roll luteovirus (PLRV) and potato virus Y (PVY). The EB resistance sources identified in this work can be recommended as parental materials for widening the genetic base for EB disease resistance in breeding programmes for highland tropical and subtropical areas.