Genetic heterogeneity estimated by RAPD polymorphism of four tuber-bearing potato species differing by breeding system

Most wild potato germplasm in genebanks is collected, preserved, and evaluated as botanical seed populations that may be highly heterozygous and heterogeneous. However, some species are selfers so potentially very homozygous, perhaps also homogeneous. Intrapopulation heterogeneity increases sampling error that can undermine consistency in seed regeneration in the genebank, screening results, germplasm collecting, and estimates of taxonomic relationships. Thus, knowledge of genetic heterogeneity (GH) can predict the need to commit more resources for larger sample sizes or replication when populations of a given species are being regenerated, evaluated, collected, and classified. This study investigated withinpopulation GH in 32 potato populations comprising four different breeding systems observed inSolatium species:S. fendleri (2n=4x=48, disomic selfer),S. jamesii (2n=2x=24, outcrosser),S. sucrense (2n=4x=48, tetrasomic outcrosser), andS. verrucosum (2n=2x=24, selfer). RAPD markers were used to estimate heterogeneity among 24 individuals per population. Populations ofS. verrucosum were quite homogeneous with an average GH of 6.0%. Similarly low heterogeneity was detected among the eight populations ofS. fendleri (average GH=7.1%). In contrast,S. jamesii andS. sucrense had a much higher GH of 29.4% and 44.1%, respectively. These results demonstrate and quantify the great difference in intrapopulation heterogeneity among wild potato species. Calculations based on intrapopulation heterogeneity indicate that samples should be composed of 25 to 30 random plants for low sample variation that is uniform for all species.     

Foliar resistance toPhytophthora infestans (Mont.) de Bary (US-8) in 13 Mexican and South AmericanSolarium species having EBNs of 1, 2, and 4 and implications for breeding

The recent US-8 clonal lineage ofPhytophthora infestans, the pathogen causing late blight (LB) disease, is one of the most widespread and difficult to control. Sources of foliar resistance to US-8 were evaluated in 1927 seedling genotypes by sampling 49 plant introductions (PIs) representing 13 Mexican and South AmericanSolanum species having an Endosperm Balance Number (EBN) of 1, 2, or 4, and one species with an unknown EBN. Species evaluated were 2x(1EBN)S. bulbocastanum, S. cardiophyllum, S. commersonii, S. pinnatisectum, andS. trifidum; 2x(2EBN)S. berthaultii, S. megistacrolobum, S. microdontum, andS. verrucosum; 4x(2EBN)S. fendleri andS. stoloniferum; 6x(4EBN)S. guerreroense;2x unknown EBNS. polyadenium; as well as4x S. tuberosum families as controls. Foliar resistance in the field was monitored in 1998 and 1999. Resistance differed relative to species geographic origin, EBN, species themselves, PIs within a species, and genotypes within a PI. In 1998 Mexican germplasm had a lower area under the disease progress curve (AUDPC, 1140) and greater resistance than South American germplasm (1601), while the 1EBN (1119) and unknown EBN (1075) species had greater resistance than 2EBN (1470) and 4EBN (1888) species. AUDPC ranged among species from 555 (S. cardiophyllum) to 1932 (S. megistacrolobum) and among PIs within a species most widely forS. bulbocastanum (267–1007) and least widely forS.polyadenium (1019–1179). In 1999 LB symptoms were not observed on the species before an early freeze; however, all 1999S. tuberosum families showed LB symptoms and segregated for resistance, pointing to greater resistance in the species than in cultivated germplasm. Resistance found in diploid 2EBN germplasm is more accessible due to greater crossability with cultivated germplasm; however, though more difficult to introgress, resistance found in some of the more reproductively isolated 1EBN germplasm is worth pursuing.     

Lack of association between genetic and geographic origin characteristics for the wild potatoSolarium sucrense Hawkes

Collecting germplasm to broaden breeding resources is an essential activity of genebanks. Research to understand how genetic diversity is partitioned in nature might help to identify collections rich in diversity. Previous studies among wild populations ofSolanum fendleri (a disomic polyploid selfer) andS. jamesii (a diploid outcrosser) revealed no significant associations between genetic and ecogeographic variation. Even physical separation did not predict genetic differences. In this study, 28 populations ofS. sucrense Hawkes (2n=4x=48), a Bolivian species with another breeding system (polysomic polyploid oucrosser), were evaluated. The objective was to assess whether genetic differences between populations are predicted by differences in geographic parameters at the natural site of origin. Genetic differentiation was estimated by using 216 RAPD markers. The average genetic distance (GD) found between pairs of populations was 31% (ranging from 8% to 44%). Correlations of GD with latitude, longitude, altitude and distance were not significant. Multiple regression analysis also confirmed that GD was not explained by the geographic parameters used. We conclude that geographic origin data is not very useful in gauging interpopulation genetic diversity in the genebank.     

Interspecific crossability and cytogenetic analysis of sexual progenies of Mexican wild diploid 1EBN speciesSolanum pinnatisectum andS. cardiophyllum

Mexican wild diploid species,Solanum pinnatisectum (S. pnt) (2n=2x=24, 1EBN: endosperm balance number) is a useful germplasm source of late blight and Colorado potato beetle (CPB) resistance in potato improvement. However, it is very difficult to cross this species with other 1EBNSolanum species. Sexual hybrids among three accessions ofS. pnt and two accessions ofS. cardiophyllum (S. cph) (2n=2x=24, 1EBN) were studied. There were large differences in the cross-compatibility among the genotypes and accessions from these two 2x-1EBN species. Interspecific incompatibility existed in the crosses betweenS. pnt andS. cph, in whichS. cph functioned only as the male parent. The crosses with accessions PI 275236 ofS. pnt produced 35 hybrids following the conventional propagation procedures. Seven hybrids were obtained with theS. pnt accession PI 275233 with the aid of embryo rescue, while hybridization involving accession PI 253214 resulted in no fruit. The cytogenetic analysis indicated that all of the hybrids derived from crosses of accession PI 275233 ofS. pnt withS. cph had 2n = 24 chromosomes and were pollen fertile. Crosses involving accession PI 275236 ofS. pnt produced both 2x (2n=24) and 3x (2n=36) hybrids. Meiotic analysis at metaphase I of pollen mother cells (PMCs) showed a fairly high rate of chromosome pairing that averaged between 10.10 and 11.10 bivalents per cell in most of the 2x hybrids, indicatingS. pnt andS. cph have the similar genome. A high frequency of trivalents and quadrivalents were observed in 3x hybrids derived fromS. pnt 2 ×S. cph 1, indicating homology exists among these chromosomes. These results suggest that specific gene(s) and not either differences between genomes or EBN controlled interspecific crossability and embryo development. The disease and insect tests on the 2x and 3x hybrids revealed that all have high levels of resistance to both late blight and CPB. Thus it could appear that selection of genotypes is a key for successful interspecific hybridization when using Mexican wild diploid species as a source of economic important traits.     

Selfing rates in two wild polyploid solanums

The rate of selfing in the wild, tuber-bearing speciesS. gourlayi Hawkes (2n = 4x = 48) andS. oplocense Hawkes (2n = 6x = 72) was estimated from progeny of plants nulliplex for the “parallel spindles (ps) allele. The estimated values for plants grown in experimental plots were 22% forS. gourlayi and 1% forS. oplocense. The quantitative method used appears to be a simple and efficient way to calculate selfing rates in polyploids. Knowledge of selfing rates is useful for basic population genetic studies and for plant breeding and seed production of economic crops. There is a need for genetic markers inSolanum species if selfing rates are to be taken into consideration for the selection of an appropriate breeding method for the production of true potato seed.     

Variation for 2n pollen production and male fertility in wildSolanum germplasm resistant toPhytophthora infestans (Mont.) de Bary (US-8)

Wild potato species possess genetic variability for valuable traits including resistance toPhytophthora infestans, the causal agent for potato late blight disease. Breeding schemes using 2n gametes are a powerful tool for introgressing these traits. Forty-five plant introductions (PIs) across 12Solanum species representing various Endosperm Balance Numbers (EBN) and having resistance toP. infestans (US-8, isolate ND 95-2) were screened for 2n pollen production and male fertility. Species evaluated were 2x(lEBN)S. bulbocastanum, S. cardiophyllum, S. pinnatisectum, andS. trifidum; 2x(2EBN)S. berthaultii, S. megistacrolobum, S. microdontum, andS. verrucosum; 4x(2EBN)S. fendleri andS. stoloniferum; 6x(4EBN)S. guerreroense; and 2x(unknown EBN)S. polyadenium. Acetocarmine staining and cytological analyses were used to determine 2n pollen production and pollen stainability from genotypes grown across three locations. Based on frequency, genotypes were assigned to 2n pollen production categories (0≤1%, Low=1%–10%, Medium=11%–60%, and High≥60%) and pollen stainability categories (0≤1%, 1=196–5%, 2=6%–10%, 3=11%–20%, 4=21%–50%, and 5≥50%). Eleven of the 12 species and 37 of the 45 PIs examined contained genotypes producing ≥1% 2n pollen with ranges of 0%–63.6% and 0%–81.8%, respectively. Dependence was found between location and pollen stainability, EBN and pollen stainability, but not for location and frequency of genotypes with 2n pollen. Stability for 2n pollen production was observed across environments. Among the 1EBN germplasm, dependence was found between 2n pollen and pollen stainability. Resistance toP. infestans was found to be independent of 2n pollen production; therefore, identifying genotypes combining 2n pollen production, fertility, and resistance toP. infestans was possible.     

Resistance to <Emphasis Type="Italic">Meloidogyne chitwoodi</Emphasis> Identified in Wild Potato Species

Meloidogyne chitwoodi (Columbia root-knot nematode, CRKN) can cause serious damage in potato production systems, decreasing tuber value in the fresh market and processing industries. Genetic resistance to CRKN was first identified from the wild diploid potato species Solanum bulbocastanum accession SB22 and was successfully introgressed into tetraploid potato breeding material. To expand the base of genetic resistance, 40 plant accessions representing nine wild potato species were screened for their resistance to M. chitwoodi. Greenhouse screening identified fifteen clones from S. hougasii, one clone from S. bulbocastanum, and one clone from S. stenophyllidium with moderate to high levels of resistance against three isolates of M. chitwoodi. Geographical mapping showed that the resistance sources identified in this and previous studies primarily originated in the states of Jalisco and Michoacán in west-central Mexico. These new sources of resistance will be introgressed into elite potato populations to facilitate the development of potato cultivars with durable resistance to M. chitwoodi.     

Evaluation of wildSolanum species for resistance to the US-8 genotype ofPhytophthora infestans utilizing a fine-screening technique

Greenhouse assays were carried out to characterizeSolanum accessions previously reported to be late blight resistant and to identify individuals within the accessions with high levels of resistance to late blight. Foliage of wildSolanum species accessions or hybrid cultivated potato x wild species (diploid or tetraploid) from Mexico, Russia, or South America was inoculated withPhytophthora infestans (US-8) and was scored for severity of foliar symptoms at 7 and 14 days after inoculation (DAI). Mexican accessions (S. brachycarpum,S. pinnatisectum,S. guerreroense, andS.fendleri) were significantly more resistant (1.1% infection at 14 DAI) than either Russian (59.1% infection at 14 DAI) or South American (53.4% infection at 14 DAI) accessions. Moreover, the genotypes within the Mexican accessions tended to be more uniformly resistant whereas genotypes within the Russian and South American accessions tended to segregate for resistance. The more resistant genotypes of Russian and South American accessions (176/546) were retested, and 56 genotypes were identified as having potential for use in potato breeding programs for resistance to late blight. Fifty percent of the selected genotypes were fromS. microdontum accessions PI498124, PI595510, and PI595511. The potential of these species for incorporation into breeding programs for late blight resistance is discussed.     

Mexico, 1988 potato germplasm collecting expedition and utility of the Mexican potato species

A joint Danish, Mexican, United States wild potato (Solanum sect.Petota) germplasm collecting expedition was conducted in Mexico between Aug. 21-Oct. 20, 1988. The trip resulted in 93 seed and 25 tuber collections of 18 species and one putative natural hybrid,S. xmichoacanum. Rare species collected include:Solanum darum,S. hintonii,S. lesteri, andS. xmichoacanum. First germplasm collections were made of the disjunct populations ofS. fendleri from Baja California. The potential and realized breeding value of these species is discussed.     

Male fertility and 2n pollen production in haploid-wild species hybrids

Hybrids (2n = 2x = 24) from crosses between Group Tuberosum haploids (2n = 2x = 24) and 24-chromosome, wild species must be fertile and produce 2n gametes if they are to be used in breeding schemes involving sexual polyploidization. Male fertility of the 2x hybrids depends primarily on the species parent. Hybrids involvingS. berthaultii, S. chacoense, S. kurtzianum, S. spegazzinii, andS. tarijense as parents are male fertile. The haploid parents are male sterile, so the species not only contribute genetic diversity and desirable traits, but also genes necessary for male fertility.S. infundibuliforme, S. raphanifolium, andS. sanctae-rosae parents produced male sterile hybrid progeny, while use ofS. boliviense, S. bukasovii, S. canasense, S. microdontum, andS. sparsipilum results in families containing both fertile and sterile plants. Sterility may be due to interactions between Tuberosum cytoplasm and dominant nuclear genes from the species. Genetic male sterility and environmental conditions may also be responsible for low pollen stainability in some hybrids. Many fertile, 2x hybrids produce 2n pollen. Selection for 2n pollen in the species is fast and efficient, but meiotic analysis is required in male sterile haploids. Fortunately, selection in the species parent alone results in a large number of 2n pollen-producing hybrids. These 2x hybrids have been used to generate tetraploids via 4x × 2x crosses.     

ND2858-1: A useful source of resistance to the Colorado potato beetle

This paper describes a new source of host-plant resistance to the Colorado Potato Beetle (CPB;Leptinotarsa decemlineata (Say)) in a tetraploid potato (Solanum tuberosum L.) selection, ND2858-1. This hybrid was recorded as a cross betweenS. fendleri ssp.fendleri (A. Gray) and NDSU selection ND1215-1. Field defoliation of this and progeny clones from a subsequent cross to an cultivar was reduced by 50% to 90% in open-choice field screening plots. Progeny of ND2858-1 are vigorous, with high yield potential, and some clones have tuber glycoalkaloid content equivalent to commercial cultivars. These clones are male and female fertile and may be useful to other potato breeding programs. Further characterization of this resistance source is in progress.     

Response of six wild potato species to chloride and sulfate salinity

Eleven accessions belonging to six species ofSolanum (S. bulbocastanum,S. chacoense, S. gourlayi, S. microdontum, S. papita, and S.sparsipilum) were tested for resistance to salt. Resistance was measured by survival and growth (dry weight of tops) of plants treated with solutions of NaCl and Na₂SO₄. The six species could be grouped into four classes based on good to poor resistance to salt solutions (1.S. chacoense, 2.S. gourlayi, 3.S. microdontum, S. sparsipilum, S. bulbocastanum and 4.S. papita). There was a close relationship (r=0.94**) between the survival of plants and growth of surviving plants treated with salt solutions. There was no relationship between known heat or drought resistance of various accessions and salt resistance. Results indicate there is a large genetic base available to improve the salt resistance of the cultivated potato.     

Distribution and significance of diplandroids among the diploid solanums

Clones of several diploid species capable of generating diplandroids (2n pollen) were identified by screening for the occurrence of 2n pollen. Diplandroid production was detected inSolatium bukasovi, S. canasense,S. cardiophyllum, S. chacoense, S. leptostigma, S. megistacrolobum, S. tarijense, S. tuberosum Group Phureja and Group Stenotomum. Parallel spindles at Ana II was the mechanism producing 2n pollen inS. chacoense, Group Phureja, and Group Stenotomum. The widespread distribution both taxonomically and geographically suggests a possible role for diplandroids in evolution. It also encourages exploration of a proposed breeding method in which tetraploids are generated from 2x × 2x matings.     

Resistance toFusarium sambucinum in wild and cultivatedSolanum species

Several species ofFusarium cause both dry rot in storage and seedpiece decay in the field, which results in severe losses in all major potato-growing regions. In temperate climates twoFusarium species are the principle causes of tuber dry rot-F.sambucinum andF. coeruleum. Resistance toF. sambucinum was identified in cultivated and wildSolanum species. Relatively high levels of resistance exist in the following 2EBN species:S. boliviense (PI#’s 265860, 310975, 310974, 545853, 545889, 545964),S. gourlayi ssp.gourlayi (PI#’s 473344, 545865),S. microdontum (PI#’s 310979, 473171),S. sancta-rosae (PI# 498392),S. kurtzianum (PI# 472930),S. fendleri ssp.fendleri (PI# 262895),S. gandarillasii (PI#’s 265866, 283076),S. oplocense (PI# 498281) andS. vidaurrei (PI# 498332). The resistance is highly heritable but appears, at least in some of the species, to be associated with recessive alleles.     

Co-current introgression of economically important traits in a potato-breeding program

Wild potato species contain many traits of economic importance. Late blight (LB) resistance and cold chipping are traits desired in potato cultivars. These traits could be co-currently introgressed if they occurred together in wild potato species. Our research objectives were (1) to determine if variation for cold chipping exists between potato species, accessions within species, and plants within accessions all having foliar LB resistance, and (2) to identify wild potato genotypes combining LB resistance and cold chipping. Materials include 665 genotypes from 43 LB-resistant accessions of 12 potato species having Endosperm Balance Numbers (EBN) of 1, 2, and 4, and 59 LB-resistant genotypes retained from these accessions for breeding. Potato chips were made from greenhouse-grown tubers following storage at 4 C for 6 months. Chip color was scored 1–10, ≤ 4 is acceptable by industry standards. Most of the variation for chip color was due to differences between species. Species ranged in the percentage of acceptably chipping genotypes (0% – 67%) with nine of 12 species having cold-chipping genotypes. Appreciable variation was present within accessions as well. The best chipping accessions wereS. verrucosum plant introduction (PI) 161173 – 4.33 / 0.67 (mean / proportion acceptable genotypes),S. stoloniferum PI 250510 -4.36 / 0.64,S. pinnatisectum PI 347766 -4.65 / 0.35 and 275233 -4.73 / 0.44, andS. megistacrolobum PI 195210 -5.14 / 0.29. Eleven 1EBN genotypes fromS. pinnatisectum andS. trifidum and five 2EBN genotypes fromS. verrucosum, S. fendleri,S. stoloniferum, andS. microdontum were identified that combined LB resistance and cold chipping. Co-current introgression would require fewer breeding cycles than other breeding methods to identify hybrid genotypes possessing both traits.     

A new wild potato mutant inSolanum stoloniferum Schltdl. Lacking purple pigment

A sample ofSolanum stoloniferum Schltdl. (reclassified fromSolanum fendleri A. Gray), PI 620874, was collected by the authors from a previously undocumented site in the Patagonia Mountains of southern Arizona in September 2001. When original seed was grown at the US Potato Genebank, two of 25 seedlings produced white flowers. Upon closer inspection they were found to be completely without purple pigmentation on the tubers, stems, and underside of leaves—a characteristic previously unknown in this species and rare in wild potato overall.Solanum stoloniferum, like other species of series Longipedicellata (LON), is a disomic tetraploid. Results of crossing trials were consistent with the most simple genetic explanation for this phenotype: a recessivep allele at the P locus, making the mutant (pp pp), or “Purple-less” abbreviated “P-less.” P-less plants were testcrossed with nearly 200 other LON populations originating across the U.S.A. and Mexico to make BC1 generations that had either one-half or one-quarter P-less seedlings. Surprisingly, about one-third of the populations, and from widely distributed origins, produced one-half P-less progeny, indicating that they also possess the recessive allele on one genome. Since this mutant has not been confirmed as an allele at the original P locus, its designation with the same allele symbols in this report (P andp) is provisional. This discovery of the widespreadp allele in LON and P-less mutants could provide a simple, unambiguous seedling marker for study of gene flow and dispersion, pollinator behavior in the wild, gene expression interactions between homeologous genomes, and chromosome pairing control.     

Interspecific somatic hybridization betweenSolanum tuberosum L. andS. bulbocastanum dun. as a means of transferring nematode resistance

Interspecific somatic hybrids were produced between tetraploidSolanum tuberosum and a nematode-resistant accession of the diploid speciesSolanum bulbocastanum by protoplast fusion. Hybrid cells were selected using dual fluorescent labeling of protoplast preparations prior to fusion. Hybridity of regenerated plants was confirmed with a combination of morphological assessment, chromosome counting and isozyme analysis. Somatic hybrids had the same level of resistance to infection by race 1 of the nematodeMeloidogyne chitwoodi as theS. bulbocastanum parent used in the fusion. Some of the somatic hybrids were fertile as females when crossed with tetraploidS. tuberosum breeding lines. Thus, these hybrids can be used in a potato improvement program to incorporate a valuable pest resistance     

Variation in Bactericera cockerelli (Hemiptera: Triozidae) Oviposition,Survival, and Development on Solanum bulbocastanum Germplasm

The potato psyllid, Bactericera cockerelli, is a key pest of potato and important vector of the pathogen that causes zebra chip disease. Control of zebra chip relies entirely on the use of insecticides to reduce populations of this vector. The development of potato varieties resistant to B. cockerelli would contribute to cost-effective control of this insect. Wild potato germplasm are key sources for desirable traits including pest resistance to develop new potato cultivars. Our objective was to screen Solanum bulbocastanum germplasm for resistance to B. cockerelli. The combined use of choice and no-choice assays demonstrated considerable variability among S. bulbocastanum populations in their susceptibility to psyllids. At least six S. bulbocastanum populations exhibited resistance to B. cockerelli: PI 243512, PI 243513, PI 255518, PI 275194, PI 275197, and PI 283096. The documentation of the variability among S. bulbocastanum germplasm populations in their susceptibility to B. cockerelli can aid the development of potato cultivars that are naturally resistant to the potato psyllid.     

Development and evaluation of potato breeding lines with introgressed resistance to Columbia root-knot nematode (Meloidogyne chitwoodi)

Columbia root-knot nematode (Meloidogyne chitwoodi) (CRN) is a serious pest of potato in the Pacific Northwest of the USA. Because this nematode can reproduce rapidly within a single growing season, small initial populations are capable of causing crop loss in the Columbia Basin of Washington or Oregon. Presently, soil fumigation is the main treatment for controlling CRN on potato. Developing potato varieties with resistance to CRN is highly desirable to reduce the cost of control and to alleviate concerns about the effects of fumigants on the environment. Resistance to CRN race 1 was found in two wildSolanum species. Resistance fromS. bulbocastanum was introduced via protoplast fusion and fromS. hougasii via sexual hybridization. Subsequent breeding consisted of repeated backcrossing and selection. The dominant monogenic inheritance was expressed in undiminished fashion across several backcross generations. When tested in replicated trials in three locations, selected resistant clones from the BC₄ and BC₅ of theS. bulbocastanum introgression populations had total marketable yields and yields of >113-g (4 oz) tubers as good or better than standard potato varieties tested in replicated yield trials in three locations. Percentage of tubers weighing more than 113 g in the highest yielding clones was not significantly different from commercial standards. The resistance phenotype, typified by failure of the nematode to reproduce on the root systems, was sufficiently effective to prevent economic damage in a field exposure. All CRN-resistant clones are pollen sterile. Germplasm listed is available upon request.