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     

Introgression of the low browning trait from the wild Mexican species Solanum hjertingii into cultivated potato (S. tuberosum L.)

Internal discoloration of tubers resulting from impact damage (blackspot bruise) is a serious quality problem in potato production and utilization, reducing profits to growers and increasing costs for processors. Resistance to blackspot bruise has been identified in the wild species Solanum hjertingii and is therefore a potential germplasm resource for genetic resistance to this problem. A bridging cross between S. hjertingii and a cultivated diploid clone was used to produce a triploid hybrid population that exhibited very low tuber browning potential, indicating a dominant pattern of inheritance for this trait. The triploid progeny were subjected to in vitrochromosome doubling and the resulting hexaploid clones were screened for browning potential. A hexaploid clone selected for low browning was reciprocally crossed with cultivated S. tuberosum cultivars exhibiting high susceptibility to blackspot bruise. Tubers obtained from the seed progeny of these 4x-6x crosses (hereafter referred to as the BC₁ populations) were evaluated for browning potential and polyphenol oxidase (PPO) activity. Tubers from the BC₁ populations displayed a very low potential for melanin production, while PPO activity was quite variable. The low Pearson correlation coefficient (r² = 0.45), between browning potential and PPO activity suggests that the mechanism of blackspot bruise resistance derived from S. hjertingii cannot be explained simply as a reduction in the initial PPO activity. The expression of substantial resistance to browning and dominant expression pattern in these BC₁ progeny indicate that utilizing genetic elements derived from S. hjertingii provides a robust approach for developing blackspot bruise resistant potato varieties. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

Introduction of an amylose-free (amf) mutant into breeding of cultivated potato,Solanum tuberosum L.

Summary Using an amylose-free (amf)mutant of diploid potato (Solanum tuberosum), diploid and tetraploid clones with different genotypes at the amf-locus were produced. In order to make use of the diploid material in analytic breeding of amf-potatoes, clones were selected that produced a considerable frequencies of 2n-pollen and 2n-eggs. Successful attempts were made to select normal synaptic as well as desynaptic clones producing 2n-gametes. Based on the phenotype of starch in the microspores, tetraploid clones with nulliplex, simplex and duplex genotypes at the amf-locus were selected. Prospects of using this material in conventional as well as in analytic breeding of potato are discussed.     

Variation in resistance level of potato genotypes and virulence level of potato cyst nematode populations

Summary In two experiments, using different testing methods, the number of newly formed cysts was determined on nine potato genotypes with resistance from various sources. Ten potato cyst nematode (PCN) populations were used in these experiments. Rank correlation between numbers of cysts over potato genotype-PCN population combinations for both experiments was high (r_s = 0.90). Dendrograms for PCN populations and potato genotypes were constructed, based on a simultaneous hierarchical clustering procedure for potato genotype-PCN population interaction terms. Several virulence groups could be identified within Globodera rostochiensis as well as within G. pallida. Host genotypes, derived from the same sources of resistance, were clustered in different resistance groups.     

Transgenic potato plants resistant to viruses

Summary Traditional potato breeding is a laborious process in which, by intercrossing, valuable traits from different parental clones are combined in a progeny genotype. Depending on the availability of genes, molecular techniques can be used to add specific genes to existing cultivars that, although otherwise satisfactory, lack a lew commercially important traits. For virus resistance the gene for the coat protein of a given virus transplanted into the genome of the plant renders the plant resistant to that virus. In conferring such resistance to potato varieties it proved to be possible to preserve their intrinsic properties.     

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.     

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.     

Characterization of morphological variation and cold resistance in interspecific somatic hybrids between potato (Solanum tuberosum L.) and S. brevidens Phil.

Summary Somatic hybrids between Solanum tuberosum L. cv. Gracia (2n=4x=48) and Solanum brevidens Phil. (2n=2x=24) were produced via fusion of mesophyll protoplasts. Selection of the protoplast derived putative hybrid calli was based on their vigorous growth. Additive isozyme patterns and chromosome numbers as well as the expression of parental morphological characters have proved the hybrid origin of the selected regenerants. Extensive chromosome loss during the regeneration process resulted in aneuploid hybrids with high frequency. Genomic instability could not be detected in these plants during the period of vegetative propagation. Regenerants from hybrid tissues exhibited wide morphological variation especially in tuber formation. The detailed morphological analysis based on the use of multivariate method (principal component analysis, PCA) enabled to identify morphological groups among the hybrid clones. The positioning of hybrid clones in the PCA space could not be correlated with chromosome numbers. The genomic ratio represented by the tetraploid and diploid parents influenced the morphology of somatic hybrid population according to the applied analytical system. Two selected hybrid clones have exhibited an intermediate degree of frost tolerance compared to the parents, based on the recovery of plants from lower buds after freezing of potted plants.     

Enhancing the resistance of the potato to Southern Root-knot Nematodes by using Solanum sparsipilum germplasm

Hypersensitive resistance to RKN was observed in infested roots from ex- S. sparsipilum material hybridised with S. tuberosum in F1 and in their BC F2 progeny. This resistance acts against M. incognita, M. javanica and M. arenaria and their isolates virulent against the Mi gene. It does not act against M. mayaguensis. This resistance protects the potatoes from galling on the roots and on the tubers. Like in the case of the Mi gene, this resistance decreases with high temperatures. Results from the F1 segregation confirm the hypothesis of a single dominant gene, here after called Mh. A pre-breeding program was carried out in order to cumulate both heat stable resistance and commercial traits. This program produced 48 genotypes selected for resistance and tolerance to high temperatures which were evaluated in the Souss Sahara in naturally infested conditions. Resistance was confirmed in year 2000 and 15 genotypes selected for agronomic traits in year 2001. This revised version was published online in August 2006 with corrections to the Cover Date.     

Introgression of Cajanus platycarpus genome into cultivated pigeonpea, C. cajan

Summary Cajanus platycarpus, an incompatible wild species from the tertiary gene pool of pigeonpea (C. cajan (L.) Millspaugh), has many desirable characteristics for the improvement of cultivated varieties. To necessitate such transfers, embryo rescue techniques were used to obtain F₁ hybrids. The F₁ hybrids were treated with colchicine to obtain tetraploid hybrids, that were selfed to obtain F₂, F₃ and F₄ progenies. All of the hybrids and subsequent progenies had an intermediate morphology between the two parents. Backcrossing of the tetraploid hybrids with cultivated pigeonpea was not possible given embryo abortion, with smaller aborted embryos than those obtained in the F₀ parental cross.As a route of introgression, diploid F₁ hybrids were backcrossed with cultivated pigeonpea and BC₁ progeny obtained by in vitro culture of aborting embryos. BC₂ plants were obtained by normal, mature seed germination. Although embryo rescue techniques had to be used to obtain F₁ and BC₁ plants, it was possible to produce BC₂ and subsequent generations through direct mature seed. Every backcross to cultivated pigeonpea increased pollen fertility and the formation of mature seeds.Special project assistant till December, 2003.     

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.     

Introgression from wild Cicer reticulatum to cultivated chickpea for productivity and disease resistance

Interspecific hybridization is known to improve productivity and resistance to diseases in many crops. Therefore, an attempt was made to introgress productivity and disease resistance into chickpea from wild Cicer species. The true F₁ hybrids of cultivated chickpea genotypes ‘L550’ and ‘FGK45’ with C. reticulatum were backcrossed twice to their cultivated female parents to minimize the linkage drag of undesirable wild traits. The pedigree method was followed to advance the segregating populations from straight crosses (without backcross) and BC₁/BC₂ generations to F₅–F₇. The interspecific derivatives recorded up to a 16.9% increase over the check cultivars and a 25.2% increase over the female parent in a preliminary yield evaluation trial. Of the 22 interspecific derivatives thus derived, four desi and two kabuli lines were further evaluated for seed yield in replicated trials at three diverse locations. These lines possess a high degree of resistance to wilt, foot rot and root rot diseases, and recorded a 6.1–17.0% seed yield increase over the best check cultivars.     

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.     

Coagulable protein in potato: Screening method and prospects for breeding

Summary From tubers of 34 varieties of Solanum tuberosum, extracts were prepared and analysed for crude protein and coagulable protein by the Kjeldahl method. Content of coagulable protein was 0.37–1.24%. From the same material, juice was prepared with a juice centrifuge and analysed for coagulable protein by the Kjeldahl and microbiuret method. The average amount of coagulable protein in the juice was 81.8% of the total coagulable protein. The correlation coefficient between coagulable protein in the juice and total coagulable protein was 0.956. The correlation coefficient between Kjeldahl and microbiuret data for coagulable protein in the juice was 0.956. Analysis of tuber juice by the microbiuret method is recommended as a rapid screening technique for coagulable protein.Relationships between protein data of the 34 varieties and earliness, yield and content of dry matter were analysed statistically. Content of coagulable protein in fresh material correlated with content of dry matter (rs=–0.756), yield of fresh potatoes (r=–0.615) and earliness (r=–0.361) but not significantly with yield of dry matter (rs=–0.309). Coagulable protein in dry matter correlated with fresh yield (r=–0.525), but not significantly with content of dry matter (r=–0.260), yield of dry matter (r=–0.131) and earliness (r=–0.054). Path coefficient analysis showed that 67% of the variation in coagulable protein in fresh material was statistically determined by earliness, content of dry matter, fresh yield and content of uncoagulable protein in fresh material, whereas only 34% of the variation in content of coagulable protein in dry matter was determined by these components. A high content of coagulable protein in fresh or dry matter can be combined with early maturity and with high yield of dry matter. Coagulable protein in dry matter seems to be a more suitable criterion of selection than coagulable protein in fresh material.     

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.     

A comparison between single plant plots and five plant plots for the initial selection stage of a potato breeding programme

Summary The efficiency of single plant selection in the initial selection stage of a potato breeding programme was examined. A random sample of potato clones was grown in single plant plots and also in tworeplicates of five plant plots. After harvest, each plot was visually assessed by three potato breeders; total tuber weight and number of tubers per plant were also recorded, from which mean tuber weight was calculated. The error variances of the single plant plots were found to be significantly greater than from the five plant plots for total tuber weight, mean tuber weight and number of tubers per plant, but not significantly different for breeders' preference. Coefficients of correlation between single and five plants plots were significantly greater than zero for all traits examined, but they were lower than the corresponding correlations between the two replicates of five plant plots. Although a random sample of clones was examined, each clone had been assessed for breeders' preference the previous year. When the clones were grouped according to the previous years preference scores, it was found that the correlation coefficients between single and five plant plots for breeders' preference were inversely related, in magnitude, to the mean preference score of each group. It was concluded that single plant selection was generally ineffective, particularly when only the better clones, from the previous years assessment, are examined.     

The breeding of the cultivated potato species solanum x juzepczukii and S. x curtilobum II. The resynthesis of S. x juzepczukii and S. x curtilobum

Summary The possibility of combining anew the genomes of wild and cultivated progenitors of triploid S. x juzepczukii and pentaploid S. x curtilobum by following the known evolutionary pathway of these species was investigated.The resynthesis of S. x juzepczukii was easy, and a wide range of synthetic forms was bred. Among these were forms with higher frost resistance (-5°C) than has been found in natural S. x juzepczukii. The total tuber glycoalkaloid content of several synthetic hybrids was lower than or as low as that of natural clones. Most synthetic hybrids were more vigorous than natural S. x juzepczukii and produced about the same types of tubers as are found in the natural range of variation. The best diploid parents were found in the species S. goniocalyx.The attempt to resynthesize pentaploid S. x curtilobum has not been successful but tetraploid plants were obtained in the process. An explanation for the occurrence of tetraploids resulting from triploid x tetraploid and/or diploid crosses is offered.The newly bred tetraploids contain at least one genome from S. acaule (possibly two) and hybridize easily with ssp. andigena. They thus provide a means for the transfer of S. acaule germ plasm into the tetraploid cultivated gene pool which would profit from the frost resistance of S. acaule.     

Female fertility of potato (Solanum tuberosum ssp. tuberosum) dihaploids

Summary Results of testcrosses and seed set data of the dihaploid breeding program at the Potato Research Institute in Gross Lüsewitz, Germany, indicate that the low level of female fertility could hinder success in crossing work. With the objective to reveal major components of female fertility, the behavior of 47 outstanding potato dihaploids (S. tuberosum ssp. tuberosum, 2n=2x=24) was assessed in testcrosses over a period of 3 years. Analysis of variance was carried out for the traits: number of seed per berry (S/B), number of intact seed per berry (IS/B), and number of non-germinable seed per berry (NGS/B). Significant sources of variation included cross combinations, years, and cross combination-year interactions. More than 20 S/B were obtained in 25.4% of all cross combinations performed over 3 years using up to three pollinators. Berry set was not correlated to seed set, but was in general determined by the pollinator used. The number of seedless berries (SLB/F) was related to neither total berry set nor seed production. The trait S/B remained stable over different years, and thus it deemed to be most useful as a measure of the level of female fertility in practical breeding work. Selection for female fertility in early generations of a breeding program is recommended to avoid limitations in the production of variability via sexual recombination.     

Resistance to Pseudomonas solanacearum in the potato: I. Effects of sources of resistance and adaptation

Summary F1 hybrid true potato seed progenies with multiple sources of specific resistance to Pseudomonas solanacearum and adaptation were evaluated under tropical greenhouse conditions for resistance to a race 1 isolate of P. solanacearum. Results indicated that genes for adaptation are involved in conferring resistance to bacterial wilt. The effect of a particular resistant parent clone on the resistance level of its progeny depended on the resistance, adaptation or the combination of both characteristics of its mate. A heat tolerant parent gave a higher frequency of resistant offspring in combination with an ascertained source of resistance. Combining ability was an apparent feature of resistance to bacterial wilt and resistance was highest in the majority of the crosses that had a wide genetic background for both resistance and adaptation. The possible genetic nature of the resistance to bacterial wilt is discussed.