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.     

A comparison of progeny from diallel crosses of diploid potato with regard to the frequencies of 2n-pollen grains

Summary Selection of diploid potato genotypes that produce high frequencies of 2n-pollen grains in different environments is an important requirement for analytic breeding as well as for true potato seed production. In this context, quantitative estimates of 2n-pollen formation were made in two type of diploid potato populations. One population set consisted of the progeny from diallel crosses of five well-known diploid potato clones (A=US-W 5293.3, B=US-W 5295.7, C=US-W 5337.3, D=US-W 7589.2 and E=772102-37). Another population set was obtained by pollinating three genotypes of Solanum phureja, IvP35, IvP48 and IvP101 with clones A, C and E. It was observed that the behaviour of the parental clones with respect to 2n-pollen grains formation in different environments, such as in Europe and China, was stable. The correlation coefficient between means of 2n-pollen grains frequency of combinations and percentage of genotypes with more than 5% 2n-pollen grains in a given combination were estimated and that between mean and maximum frequency of 2n-pollen grains formation in each combination. The results showed that the former coefficients were significant wheareas the latter were not. The combining ability analysis revealed variation in 2n-pollen grains formation between families. Specific combining ability (SCA) effects contributed much more than general combining ability (GCA) to the variation in 2n-pollen grains formation. The broad-sense heritability of 2n-pollen grains formation was estimated to vary between 0.45 and 0.53 and the narrow-sense heritability was calculated to be 0.15 and 0.20 in the two types of populations. This means that additive variance was relatively low. Two to four genes were estimated to control the inheritance of 2n-pollen grains formation.     

Evaluation of seedling selection for salinity tolerance in potato (Solanum tuberosum L.)

Summary Salt-tolerance in potato (Solanum tuberosum L.) was selected on the basis of germination and survival of seed sown in trays of perlite suspended in either 75 or 150 mM NaCl. Salinity reduced the germination of seed. Genetic differences in salt-tolerance were apparent with salt reducing germination more in seed collected from cv. Cara than in that collected from cv. Maris Piper. Progeny from the seedling selection were then grown to maturity to produce tubers.The relative tolerance of the parental cultivars and of unselected and selected progeny to long-term exposure to salinity was examined in a pot experiment in which plants were irrigated with either fresh water or 50 mM NaCl solution from one week after plant emergence. In this experiment, salinity significantly reduced leaf conductance, total dry matter production and partitioning of assimilate to tubers. Salinity reduced dry matter production and assimilate partitioning to tubers to a greater extent in Cara than in M. Piper. Progeny selected for short-term salt-tolerance did not exhibit greater long-term salt-tolerance than unselected progeny, and both were more sensitive than M. Piper.These results demonstrate genetic variation in salt-tolerance in potato. However, although there was a correlation between the performance of the parent to long-term salinity and survival of progeny in the seedling selection, there was no correlation between short- and long-term salt-tolerance. This suggests that characters underlying short-term tolerance may contribute to long-term tolerance but do not of themselves confer long-term tolerance. Future progress in selecting for improved salt-tolerance depends on understanding the effects of salinity on the physiological processes underlying growth and carbon partitioning.     

An assessment of morphogenic and transformation efficiency in a range of varieties of potato (Solanum tuberosum L.)

Summary To provide a truly genotype-independent transformation system, it is necessary to be able to transform a wide range of potato genotypes. The ability to regenerate shoots in vitro was determined for 34 potato varieties using tuber disc explants. Following a culture regime used extensively in previous studies with the variety Desiree, half of the varieties could be regenerated from tuber discs and half could not. From a sample of varieties that could be regenerated from tuber discs, all but one variety gave transgenic plants. Twelve varieties were evaluated for the capacity to regenerate shoots from leaf and internode explants excised from in vitro grown plants. All of the varieties tested regenerated adventitious shoots. Leaf and internode explants from 5 varieties were subsequently used for transformation, and transgenic plants were produced from two potato varieties that did not give transgenic plants from tuber disc explants. Some varieties could not be transformed by either method, and will require modification of the in vitro regeneration and transformation system to be successful.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid     

Genetics of self-compatibility in dihaploids of Solanum tuberosum L. 5. The S-genotype of tetraploid potato cultivar gineke

Summary Investigations of the genetics of self-compatibility and self-incompatibility in dihaploids and diploid derivatives from cv. Gineke revealed the presence of S ₁, S₂ and S ₃ at the S-locus of Gineke and in addition an S ₁-allele on a translocation. By means of a complete tester set involving the S-alleles S ₁, S₂ and S ₃ (all from Gineke) and S ₄ (from Black 4495) it was demonstrated that some Gineke dihaploids were compatible with all six testers. This indicated a fourth S-allele in Gineke, which differs from those in the tester series and was therefore assigned S ₅. Additional evidence was obtained from an analysis of F₁'s from crosses of two S ₅-bearing dihaploids and one of the testers. So the S-genotype of cv. Gineke was identified as S ₁S₂S₃S₅/S₁, the second S ₁ being the S-allele on a translocated fragment.     

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.     

Regarding the novelty and breeding value of protoplast-derived variants of Russet Burbank (Solanum tuberosum L.)

Summary Striking similarities appear to exist between selected protoplast-derived variants (PD-clones) of the potato cultivar Russet Burbank, and commonly occurring somatic potato sports, called bolters. A limited survey of individuals involved in the certification of Russet Burbank, revealed that bolters do commonly occur in this variety, are highly variable, and sometimes occur at high frequency. It is therefore suggested that bolters and the more promising selected PD-clones of Russest Burbank may be one and the same, arising from the same type of macromutation.One selected PD-clone, three different Russet Burbank bolters, and two controls were analyzed for 15 isozymes in the hope of determining if bolters and promising PD-clones belong to the same class of variants. No isozyme variation between Russet Burbank and its subclones was observed. The breeding value of protoplast-derived variation is discussed in this light.Approved by the Director of the New York State Agricultural Experiment Station for publication as Journal paper No. 3554, Oct. 19, 1983.     

Transfer of resistance to Verticillium dahliae Kleb. from Solanum torvum S.W. into potato (Solanum tuberosum L.) by protoplast electrofusion

Summary Interspecific somatic hybrid plants were regenerated after electrofusion of mesophyll protoplasts with the objective of transferring resistance to Verticillium dahliae from Solanum torvum into potato. Early selection of the putative hybrids was based on differences in cultural behaviour of the parental and hybrid calli (particularly the ability of the latter to regenerate early) in combination with morphological markers. Four putative hybrids were recovered from hundreds of calli, probably resulting from complementation of the two parental genomes. The regenerates were tetraploids (2n=4×=48 chromosomes) and exhibited intermediate traits including leaf form, plant morphology and the presence of anthocyanin. The hybrid nature of the four selected plants was confirmed by examining isoenzyme patterns for isocitrate dehydrogenase (Idh), malate dehydrogenase (Mdh), phosphoglucoisomerase (Pgi) and 6-phosphogluconate dehydrogenase (6-Pgd). While the hybrid plants rooted readily and grew vigorously under in vitro conditions, in the greenhouse their development and growth were retarded by difficulties in rooting. When grafted on potato or S. torvum rootstocks, the hybrid plants recovered normal development and growth. Again, they exhibited intermediate morphological traits. Tests for resistance realized in vitro with medium containing 50% Verticillium wilt filtrate showed that all the somatic hybrids were resistant to the fungus filtrate.     

Parthenogenetic monohaploids (2n=x=12) from Solanum tuberosum L. and S. verrucosum Schlechtd. and the production of homozygous potato diploids

Summary Dihaploid and dihaploid derived clones of Solanum tuberosum and diploid genotypes of S. verrucosum produced 85 viable monohaploids by female parthenogenesis. All were induced using diploid S. phureja clones, homozygous for embryo spot, as pollinator. Frequency of S. tuberosum monohaploids per 100 berries was rather constant in three successive years (14, 17 and 17 respectively). No male and female fertility was found in flowering monohaploids.Colchicine-induced chromosome doubling yielded homozygous s. tuberosum diploids with low pollen quality but good seed fertility.Two diploid self-incompatible species (S. multidissectum and S. berthaultii) produced no monohaploids. The presence of genes for female parthenogenesis in some dihaploids is discussed.     

Heterosis and combining ability in hybrids among 12 commercial varieties of sesame (Sesamum indicum L.)

Heterosis and combining-ability effects were studied in the parents and 66 F₁ hybrids from a diallel cross of 12 elite sesame varieties in five different environments in Venezuela. The mean yield of the hybrids was clearly superior to the parental mean in all five locations. There were three heterotic hybrids that showed a higher yield than controls in all environments. The best hybrid in each place yielded significantly better than the best cultivar, the differences ranging from 28 to 109%. Specific combining-ability (SCA) effects were more important than general combining-ability (GCA) in four of the environments. A large genotypexlocation interaction was found for all traits, suggesting that hybrids should be produced for specific locations. ‘Arawaca’ and ‘Piritu’ should be chosen as parents for any sesame-breeding programme in view of their large GCA effects.     

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.     

Optimising potato breeding. I. The genotypic,environmental and genotype-environment coefficients of variation for tuber yield and other traits in potato (Solanum tuberosum L.) under different experimental conditions

Summary Coefficients of variation for genotypic differences, genotype-location, genotype-year and genotype-location-year interaction were computed from reported data of potato trials in order to use them in a decision support system. The obtained coefficients of variation not only varied considerably between traits, but also between trials within traits. It is argued that only parameter values representative for the pertaining breeding situation should be used for optimising a selection programme.     

A comparative assessment of DNA fingerprinting techniques (RAPD, ISSR, AFLP and SSR) in tetraploid potato (Solanum tuberosum L.) germplasm

Several DNA marker systems and associated techniques are available today for fingerprinting plant germplasm but information on their relative usefulness in particular crops is limited. The study investigated PCR based DNA fingerprinting in a set of 39 potato cultivars using RAPDs (20 primers), ISSRs (6 primers), AFLPs (2 primers) and SSRs (5 primer pairs). Results show that each of the four techniques can on their own, individually identify each cultivar, but that techniques differ in the mean number of profiles generated per primer (or primer pair) per cultivar, referred to as Genotype Index (GI). The order of merit based on this criterium and in this material was AFLPs (GI = 1.0), a multi-locus SSR (GI = 0.77),RAPDs (GI = 0.53), ISSRs (GI = 0.47) and single locus SSRs (GI = 0.36). Problems in relating banding patterns to individual loci and alleles for polyploid genomes, using these techniques as they are currently employed, are also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Variation and heritability of economically important traits in black currant (Ribes nigrum L.) evaluated in a diallel cross design

Summary The inheritance of commercially important characters in black currant (Ribes nigrum L.) was investigated using a complete 5×5 varietal diallel cross. Estimates of the genetic components of variance and broad and narrow-sense heritabilities revealed that additive genetic effects were greater than nonadditive genetic effects for resistance to powdery mildew (Sphaerotheca mors-uvae Berk.), white pine blister rust (Cronartium ribicola Fisher) and leaf spot (Drepanopeziza ribis Kleb.) as well as for bush defoliation and length of strigs. Apart from resistance to leaf spot, narrow-sense heritabilities of these characters exceeded 0.5.Nonadditive genetic effects were greater than additive effects for plant habit, plant vigor, bush size, length of one-year-old shoots, uniformity of berry ripening and fruit yield per bush. Narrow-sense heritabilities of these characters were rather low (0.05–0.42).     

Field resistance to late blight (Phytophthora infestans (Mont.) De Bary) in potato (Solanum tuberosum L.) somaclones associated with instability and pleiotropic effects

Summary Adventitious regenerants (somaclones) of Bintje and their vegetative progeny were screened for field resistance to Phytophthora infestans as follows: the area under the disease progress curve was computed and correlated with resistance rating in Bintje and reference varieties. The resistance rating of the somaclones was determined from this relationship.Clones with stable improved field resistance in successive years' trials were detected, however, most of such clones were also maturation mutants. Variation in resistance rating in clone replicates and between years was detected in most clones.The possible basis of the field resistance and reasons for its instability are discussed.Abbreviations AUDPC area under the disease progress curve     

In vitro adventitious bud techniques for vegetative propagation and mutation breeding of potato (Solanum tuberosum L.). II. Significance for mutation breeding

Summary It was investigated what potentialities for mutation breeding of potato are offered by using adventitious sprouts that arise in vitro from leaf explants (rachis, petiole, leaflet-disc) after X-irradiation. Mutation frequency and chimerism were studied in subterranean and aerial parts in three vegetative generations (vM₁, vM₂, vM₃). Plants obtained from irradiated series produced a very high mutation frequency, a wide mutation spectrum and a very low rate of chimerism. Mutations were observed also in control series, especially in plants derived from the rachis and petiole explant group.     

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.     

Integration of transgenes into sexual polyploidization schemes for potato (Solanum tuberosum L.)

Most potato transgenic research has focused on development of resistance to pathogens and modification of potato physiology. Many transgenes, particularly those conferring pathogen resistance, could substantially lower potato production costs in developing countries. However, transgenes have not been reported in sexually propagated 4x-2x potato hybrids commonly grown in developing countries. Two transgenes,the Bacillus thuringiensis cry3Aa endotoxin protein gene and the PVY°coat protein gene, were engineered intodiploid and tetraploid potato using Agrobacterium tumefaciens-mediated transformation. Cry3Aa was produced at high levels in several lines while the PVY° coat protein was not expressed. Diplandroid and tetraploid genotypes were crossed to produce transgenic 4x-2x hybrids. Genetic transformation had no discernable effect on fertility ofthe primary transformants, germination of4x-2x seed derived from the transformants and agronomic performance(tuber set, average tuber weight and total tuber yield) of the 4x-2xhybrids. The transgenic 4x-2xhybrids produced non-viable pollen and could only be crossed as female parents. Results suggest that transgenes, such ascry3Aa, could be expressed in 4x-2x hybrids to lower costs of production with no significant effect on plant phenotype. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance to potato late blight (Phytophthora infestans (Mont.) de Bary) in Solanum nigrum,S. villosum and their sexual hybrids with S. tuberosum and S. demissum

Summary Solanum nigrum and S. villosum, and their sexual hybrids with S. tuberosum and S. demissum respectively, were inoculated with a complex race of Phytophthora infestans. No visible reaction was seen on S. villosum and one genotype of S. nigrum. Another genotype of S. nigrum and the hybrids showed a hypersensitive response on most inoculated leaves. In one experiment, some sporulation was observed on detached leaves of a hybrid derived from S. nigrum. Microscopical examination of infections in S. nigrum and in a hybrid from S. nigrum, showed that penetration of epidermal cells and subsequent intercellular growth of the pathogen into the spongy mesophyll occurred, but without the formation of haustoria, and that invaded and neighbouring cells became necrotic. Callose appositions were found in epidermis and mesophyll cells of all inoculated genotypes, and also in epidermal cells of the unrelated nonhost species Brassica campestris.     

Genetics of self-compatibility in dihaploids of Solanum tuberosum L. 4. Linkage between an S-bearing translocation and a locus for virescens

Summary Three dihaploids of Solanum tuberosum (two self-compatible, one self-incompatible) were found to be heterozygous for a monogenic recessive virescent mutant. Intercrossing resulted in the expected 3 : 1 ratio only in crosses involving one self-compatible and one self-incompatible parent. Self-compatible x self-compatible matings produced F₁'s in which 6:1 was found. The same ratio was observed in the self progeny of the two self-compatible dihaploids. This significant deviation could be explained by assuming linkage (25% crossing-over) between v and an S-bearing translocation. This translocation causes self-compatibility in the dihaploids used and early lethality when homozygous.