Segregation of leptines and other glycoalkaloids inSolanum tuberosum (4x) ×S. chacoense (4x) crosses

A clone, 8380-1, selected fromSolanum chacoense (PI 458310) for its high foliage content of the leptine glycoalkaloids, a factor in resistance to Colorado potato beetle, was doubled in chromosome number from 2n=2x=24 to 2n=4x=48. Three 4x clones were crossed with sixS. tuberosum (4x) clones. Foliage glycoalkaloid contents were measured for 452 F1 hybrids from 15 crosses. The 4xchacoense parental clones were not different in respect to glycoalkaloid contents and were similar to the original 2x clone. All F1 hybrids synthesized foliage leptines ranging from 9 to 369 mg/100 g fresh weight (fw) with a mean content of 113 mg/100 g fw. The proportion of leptines in the total glycoalkaloid content ranged from 1% to 62% with a mean of 25%. The 4xchacoense parent mean leptine content was 1482 mg/ 100 g fw which was 90% of the total glycoalkaloid content. Tubers from 136 hybrids and the threechacoense parental clones were tested for glycoalkaloid contents. The tuber solanine + chaconine contents of the 136 hybrids ranged from 30 to 180 mg/100 g fw with a mean of 79. The mean tuber content of the threechacoense parental clones was 157 mg/100 g fw. Leptines were not found in any of the tubers.     

Glycoalkaloid content in foliage of hybrid and backcross populations from aSolanum tuberosum X S. chacoense cross

The glycoalkaloid contents of foliage were measured in populations derived from a cross ofSolanum chacoense Bitter (PI 472810) with a composite sample of genotypes from a randomly matedSolanum tuberosum L. population. The mean total glycoalkaloid contents of the parent PI 472810 andtuberosum populations, and the F₂ and F₄ populations were 856, 121, 286, and 279 mg/100g fresh weight, respectively. The total glycoalkaloid content of the first backcross population, F₂ xtuberosum, was 290 mg/100 g, close to the content of the F₂. In a second backcross totuberosum, the mean total content was 148 mg/100 g, close to the content of the parenttuberosum population. Only the glycoalkaloids α-solanine and α-chaconine were found in PI 472810, whereas in the parenttuberosum, hybrid, and backcross populations, the glycoalkaloids α & β-solamarine also were found in a small number of genotypes. The ratios of chaconine to solanine contents were significantly (P≤.05) different in the parent PI 472810 andtuberosum populations, 2.55 and 2.12, respectively. The ratios in the F₂ and F₄ populations were similar to that of PI 472810, whereas the ratios in the backcross populations were closer to that of thetuberosum parent. The levels of foliage glycoalkaloids in the hybrid and backcross populations paralleled the levels of potato leafhopper,Empoasca fabae Harris, resistance measured in a previous study.     

Glycoalkaloid content in tubers of hybrid and backcross populations from aSolanum tuberosumx s. chacoense cross

The glycoalkaloid contents of tubers were measured in hybrid and back-cross populations derived from a cross ofSolarium chacoense Bitter (PI 472810, 2n=2x=24) withSolanum tuberosum L. (2n=4x=48). In tubers sampled from near the median size in each population and parent, the mean total glycoalkaloid (TGA) contents of theS. chacoense andS. tuberosum parent genotypes, and the F₂ and F₄ hybrid genotypes were 245, 4, 39, and 35 mg/ 100 g fresh weight (fw), respectively. Backcrossing the F₂ genotypes toS. tuberosum reduced the mean TGA content to 15 mg/100 g fw. A second backcross further reduced the content to 9 mg/100 g fw. However, some genotypes in both backcross populations (25% and 10%) had TGA contents above the generally accepted upper limit of 20 mg/100 g fw. Tuber size was significantly smaller in theS. chacoense parent and the hybrid populations compared with the size in theS. tuberosum parent and backcross populations. In a second sampling of tubers from each population, intentionally selected to equalize tuber size among populations, the mean tuber size for populations was reduced by 79%. The mean population TGA content increased by 44%. However, the differences and rankings among parent, hybrid, and backcross populations were the same for both tuber samplings.     

Segregation of leptine glycoalkaloids and resistance to Colorado potato beetle (Leptinotarsa decemlineata (Say)) in F2Solanum tuberosum (4x) ×S. chacoense (4x) potato progenies

Solanum chacoense Bitter is resistant to the Colorado potato beetle (CPB),Leptinotarsa decemlineata (Say). Resistance has been associated with the presence of a rare class of glycoalkaloids, the leptines. In this study, seven tetraploid, F2S. tuberosum xS. chacoense families were evaluated for foliar production of leptines I and II, leptinines I and II, and α-solanine and α-chaconine; and screened for resistance to CPB in the laboratory and field. Resistance was correlated with the concentrations of glycoalkaloids on a family and an individual basis. Leptine concentrations ranged from undetectable to a high of 18.0 mg/g dry weight. All of the progeny produced solanine and chaconine. Family 9623 had the highest mean leptine concentration and the lowest mean leaf disk feeding and CPB defoliation levels. Family 9616 had the lowest mean glycoalkaloid concentration and ranked as one of the most susceptible families. Regression analyses of solanine + chaconine, leptine I and II, and leptinine I and II foliar concentrations versus leaf disk consumption and field defoliation revealed that only increased foliar levels of leptines resulted in decreased CPB feeding. The regression models for leptines versus leaf disk consumption and field defoliation were highly significant, accounting for 17% and 26% of the variation in consumption and defoliation, respectively. To the best of our knowledge, this is the first work reporting the impact of leptine and leptinine concentrations on CPB feeding in tetraploid,S. tuberosum xS. chacoense potato hybrids. Results are discussed within the context of breeding for resistance to CPB.     

Glycoalkaloid content inSolanum species and hybrids from a breeding program for resistance to late blight (Phytophthora infestans)

As part of an effort to study the relationship between the “glycoalkaloid trait” and genetic resistance to late blight (Phytophthora infestans), glycoalkaloid content in tuber and foliar tissues from a wide genetic background withinSolanum, includingS. demissum, S. iopetalum and 15 hybrids selected from a backcrossing breeding scheme was determined. Analysis of variance indicated significant genotypic effect on total glycoalkaloid, solanidine, α-solanine, and α-chaconine content in both tubers and leaves. Tubers from wild potato species commonly contain glycoalkaloids in concentrations that exceed international health regulations for human consumption (20 mg/100 g fresh weight). In this study,S. demissum andS. iopetalum were highest in total tuber glycoalkaloids among all materials tested, with 70.4 and 76.2 mg/100 g fresh weight, respectively. In contrast, both commercial cultivars had the lowest concentration, both below the safety limit. Solanine was more abundant than chaconine in all but one genotype. All hybrids were intermediate to low in total glycoalkaloids. Except for the two wild species, glycoalkaloid concentration in leaves of all genotypes studied was at least twice that in tubers, with glycosylated forms accounting for more than 80% total glycoalkaloid content. Correlation between tuber and foliage alkaloids was poor. In view of the observed field resistance to late blight, it was concluded that tuber glycoalkaloid content may not be responsible for such resistance.     

Foliar leptine production in segregating F1, Inter-F1, and backcross families ofSolanum chacoense bitter

Levels of foliar leptine glycoalkaloids were measured in segregating F₁, inter-F₁, and backcross families derived fromS. chacoense (PI 320287) sibs 55-1 and 55-3, which axe (respectively) high and nil leptine producers, and from crosses of these two genotypes with 8380-1, which produces high levels of leptine, to study the inheritance of leptine biosynthesis inS. chacoense. A total of five F₁ families was produced from reciprocal crosses between 55-3 and 55-1 and between 8380-1 and 55-3, and from the cross 8380-1 x 55-1. The F₁ crosses produced two leptine classes, high (≥62% of total glycoalkaloids; TGA) and low (≤17% of TGA). Twenty-three families were derived from F₁ intercrosses and backcrosses of high x high, low x low, high x low, and low x high F₁ individnals. These families produced all high, all low, or both high and low leptine classes, depending upon type of cross. Three of the reciprocal (high x low) F₁ families segregated 1:1 for high:low leptine, and progeny from the (high x high) 8380-1 x 55-1 cross were all high, consistent with control by a single recessive gene in this population. One of the reciprocal families (9501) differed slightly but significantly (p=0.05) from 1:1. All of the low x high and high x high inter-F₁ and backcross families segregated as expected based on a single recessive gene model; however, five of eight high x low and four of six low x low families differed significantly from expectation, suggesting the presence of additional factors affecting the inheritance and expression of leptine glycoalkaloids in this population orS. chacoense.     

Glycoalkaloids and resistance to the Colorado potato beetle inSolanum chacoense Bitter

Resistance of 20 F₂ Solarium chacoense Bitter clones to the Colorado potato beetle,Leptinotarsa decemlineata (Say), was measured in a field test. Levels of total glycoalkaloids (TGA) and composición of the glycoalkaloid mixtures in foliage of the clones were also determined. Clones with either commersonine or dehydrocommersonine as the major foliar glycoalkaloid were significantly more resistant (lower damage ratings, fewer larvae and adult insects) than clones with solanine and chaconine. Damage ratings were negatively correlated (r = -0.67, p = 0.01) with foliar TGA levels. The results indicate that the types of glycoalkaloids present in the foliage ofS. chacoense may be as important as the level of TGA in limiting damage and numbers of insects.     

The effect of light intensity on Colorado potato beetle resistance and foliar glycoalkaloid concentration of fourSolanum chacoense clones

Controlled environment experiments were used to determine the effect of light intensity on levels of glycoalkaloids and resistance to the Colorado potato beetle (CPB),Leptinotarsa decemlineata Say, inSolanum chacoense Bitter clones that synthesized acetylated glycoalkaloids, the leptines. Plants were grown under two light intensities, 50 or 250 ?mol m^-2S^-1, and bioassayed for CPB resistance by placing neonate larvae on excised foliage. Larval development stage and mortality were recorded after four days. The leptine-synthesizing clones responded to the increase in light intensity with a 2–4 fold increase in level of foliar leptines. Larval development rate on foliage from plants grown under high light intensity was reduced and larval mortality increased. Results showed a significant (p>0.05) effect of light on glycoalkaloid synthesis and CPB resistance.     

Evaluation of wild tuber-bearingSolanum accessions for foliar glycoalkaloid level and composition

Six hundred forty-five accessions of 70Solanum species and six natural hybrids were screened for foliar glycoalkaloids. Total foliar glycoalkaloid (TFGA) level and glycoalkaloid composition of one or more accessions of each species were determined by TLC and GLC analyses. Of the more than 12 different glycoalkaloids found in the wild species in this survey, solanine and chaconine account for more than 60% of the TFGA found in the foliage of these plants. There was wide variation in the total glycoalkaloid levels among the species.S. neocardenasii had the highest average TFGA value of the 70 species, 222 mg/100 g fresh wt (222 mg %) and an accession ofS. chacoense had the highest TFGA value recorded in the survey, 486 mg %. Of the 70 species, only 11 had average TFGA levels of more than 100 mg %. However, one or more accessions in 27 of the species synthesized more than 100 mg %. Forty-five species synthesized an average level of less than 50 mg %. The level usually found in the foliage of commercial potato cultivars is about 50 mg %. Therefore, most of theSolanum species in the potato germplasm collection can be considered low glycoalkaloid species.     

Glandular pubescence,glycoalkaloid composition,and resistance to the green peach aphid,potato leafhopper,and potato fleabeetle inSolanum berthaultii

Fifteen accessions ofSolarium berthaultii Hawkes andS. berthaultii xS. tarijense Hawkes were assessed for resistance to field infestations of the green peach aphid,Myzus persicae (Sulzer), potato leafhopper,Empoasca fabae (Harris), and potato fleabeetle,Epitrix cucumeris (Harris). Accessions bearing both Type A and Type B glandular trichomes were much more resistant to the green peach aphid and potato leafhopper than accessions bearing Type A hairs alone. All accessions had significantly smaller populations of these 3 pests than S.tuberosum cultivars. Total glycoalkaloid (TGA) content of foliage and tubers was not correlated with insect populations. Foliar TGA levels of field-grown plants varied among accessions, ranging from < 2–240 mg/100 g fresh wt. Solasonine and solamargine were the major foliar glycoalkaloids while solamarines predominated in tubers.     

Glycoalkaloid contents in tubers fromSolanum tuberosum populations selected for potato leafhopper resistance

Solanum tuberosum L. Gp. Tuberosum populations that had been improved for resistance to potato leafhopper,Empoasca fabae (Harris), by recurrent selection, were analyzed for tuber contents of the glycoalkaloids α-solanine and α-chaconine. The mean content of both glycoalkaloids was significantly higher in the more resistant populations compared with the unselected population. In a population generated from seven cycles of selection for resistance, the mean content of solanine + chaconine was 9.5 mg/100 g fresh weight compared with 4.0 mg/100 g in the original, unselected population; a 137% increase. Kennebec, in the same experiment, averaged 10.9 mg/100 g; Katahdin averaged 4.6 mg/100 g.     

Horizontal resistance insolanum tuberosum to colorado potato beetle (leptinotarsa decemlineata say)

Horizontal resistance (HR) was assessed as a strategy to reduce potato defoliation by the Colorado potato beetle (CPB). Hybrids amongSolanum tuberosum varieties grown in Iowa were first evaluated with a detached leaflet feeding bioassay. Beginning in 1998, resistance was increased annually over three years by intermating and selecting the most resistant l%–2% of each generation of 300–400 plants using the bioassay. When C (= mg leaf tissue consumed_* neonate larva^-1 over 48 h) was measured for the 1% most-resistant genotypes of all generations grown side by side in 2000, it ranged from 6.1 mg_* larva^-1 in the parents to 2.3 for progeny of the F₃ generation compared to 2.7 in ND2858-1, a resistant check. Field trials in Wisconsin showed less defoliation in the F₁ and F₂ selected genotypes than in the parents. A second breeding program using moreS. tuberosum parents with greater genetic diversity, and much larger breeding populations, resulted in significantly faster accumulation of resistance, further suggesting that recurrent mass selection is effective. Foliar glycoalkaloid analyses revealed low levels of solanine and chaconine in the F₃ selections; no leptines or leptinines were detected. Because the parents wereS. tuberosum varieties in which vertical resistance to CPB is unknown, and because the resistance to feeding from parents to the F₃ resistant plants was linear and gradual while ranging from initially very low to considerably higher in the third year, horizontal resistance is strongly implicated. The development of a significant degree of resistance after three rounds of breeding using relatively small breeding populations suggests that HR can be developed from varieties ofSolanum tuberosum lacking initial high levels of resistance to CPB.     

Diploid and tetraploidSolanum chacoense genotypes that synthesize leptine glycoalkaloids and deter feeding by Colorado potato beetle

A selection (8380-1) fromSolatium chacoense Bitter (2n=2x=24) accession PI 458310 that synthesizes the leptine glycoalkaloids was compared in growth chambers with tetraploid (2n=4x=48) genotypes derived from tissue culture of 8380-1 leaf expiants for plant growth habit, leaf glycoalkaloid content, and effect on the development of Colorado potato beetle [Leptinotarsa decemlineata (Say)] larvae. The plants of the 4x regenerant genotypes were more vigorous with larger, more oval shaped leaflets than 8380-1 plants. The leaf concentrations of leptines and total glycoalkaloids were significantly lower (about 34%) in the 4x genotypes than in 8380-1. The proportion of leptines in the total glycoalkaloid content was nearly the same (about 80%) in both ploidy groups. In leaf-disk feeding tests, the development of Colorado potato beetle neonate larvae was not significantly different for the 2x and 4x genotypes. Both groups significantly slowed development compared with development on cv. Kennebec leaf disks. The 8380-1 selection and a group of 4x 8380-1 regenerant genotypes are maintained in the Vegetable Laboratory, Beltsville Agricultural Research Center, Beltsville, MD 20705 and are available for distribution.     

Selection of monoploids for protoplast fusion and generation of intermonoploid somatic hybrids of potato

The union of potato monoploid genotypes (2n=1x =12) through protoplast fusion may result in vigorous somatic hybrids due to a reduction of the “genetic load” normally present in this highly heterozygous tetraploid (2n=4x=48) crop. More than 100 androgenic monoploids derived from diploid (2n=2x=24)Solanum phureja Juz. & Buk. andS. chacoense Bitt. xS. phureja clones were evaluated in field trials during 1996 and 1997 to identify the most promising genotypes for protoplast fusion experiments. Compared to the total population, the 1996 selected genotypes had higher means for tuber number (30.1 vs 11.2 tubers/plant), average tuber weight (3.0 vs 1.8 g/tuber) and total yield (66.1 vs 20.4 g/plant). Similarly, the 1997 selected genotypes had higher means for tuber number (42.8 vs 25.4 tubers/plant), average tuber weight (3.6 vs 2.5 g/tuber) and total yield (114.0 vs 63.4 g/plant) compared to the total population. The 31 selected monoploid genotypes from 1996-97 varied in their response to protoplast isolation and culture from no growth (9), cell enlargement (5), limited cellular divisions (8), callus formation (5) to plant regeneration from callus (4). Chemical fusion and electrofusion produced three groups of intermonoploid somatic hybrids. Polymorphic simple sequence repeat (SSR) loci enabled distinction of somatic hybrids from parental somaclones. Rapid DNA extraction with SSR analysis enabled screening of calluses to identify somatic hybrid tissue prior to plant regeneration. The somatic hybrids were highly polyploid, mostly hexaploid (2n=6x=72), possibly due to fusion of endopolyploid protoplasts and/or chromosomal doubling during plant regeneration.     

Tuber quality and soft rot resistance of hybrids betweenSolarium tuberosum and the incongruent wild relativeS. commersonii

Tetraploid BC₃ hybrids between sexually incompatible speciesSolanum commersonii andS. tuberosum were characterized for tuber quality (stolon length, tuber eye depth, chipping color from 3-month storage at 5 C, and specific gravity) and soft rot resistance traits. For all analysed morphological traits, except stolon length, genotypes resembled theS. tuberosum phenotype. On average, the specific gravity of selected genotypes was quite high (1.087), and 12 of them had a value >1.090. As for chipprocessing ability, 4.8% of genotypes chipped well directly from cold storage, and 14.3% after reconditioning. All but one of these genotypes also possessed a high specific gravity of tubers. Variability was found for resistance toErwinia carotovora subsp.carotovora. In particular, 24% of hybrids had a rotted area diameter ≤ 4 mm and were classified as resistant. The average percentage ofcommersonii-specific markers still present in the BC₃ hybrids was 22%, ranging from 9% to 37%. Genotypes combining good qualitative tuber traits with a wild genome content lower than the average value of the population were selected for further breeding efforts. The importance of molecular-assisted selection is discussed.     

Reaction of some wild and cultivated potato varieties to charcoal rot

With a view to select potato varieties possessing resistance to charcoal rot, laboratory tests were carried out at the Central Potato Research Institute, Simla (India). Five hundred and twenty six clones comprising tuber-bearing species ofSolanum and varieties and hybrids of potato were screened for their reaction toMacrophomina phaseoli (Maubl.) Ashby. A high degree of resistance was observed in six clones ofSolanum chacoense and four hybrids withS. chacoense germ plasm, while moderate susceptibility was observed in another six clones ofS. chacoense, three clones ofSolanum tuberosum ssp.andigena and nineteen hybrids produced at the Institute.     

Inheritance study of tuber glycoalkaloids in a wild potato,Solanum chacoense bitter

Studies were made of segregating families from crosses between accessions of Solanumchacoense Bitter differing in steroid glycoalkaloids. The major glycoalkaloids were isolated from tubers and identified by thin-layer chromatography. Simple segregation ratios of 3:1 in an F₂ and 1:1 in backcrosses were obtained for the presence:absence of solanine, chaconine, and commersonine. To explain the simultaneous segregation of the three glycoalkaloids, we tentatively hypothesize that: (1) solanine and commersonine are determined by alternative codominant alleles at a single gene locus; (2) a major gene for chaconine segregates independently of the solanine/commersonine locus; and (3) a recessive gene linked with the commersonine allele is epistatic to the chaconine gene, causing the production of β-chaconine instead of the usual α-chaconine. The F₂ distribution could be fit to the ratio 3:6:3:1:2:1, seen as combining the 1:2:1 segregation of solanine: solanine + commersonine: commersonine and the 3:1 segregation of presence: absence of chaconine.     

Pollen-pistil incompatibility in a diploid hybrid potato population with cultivated and wild germplasm

The basis of low seed set was investigated in a diploid hybrid population with germplasm from the cultivated speciesSolanum tuberosum spp.tuberosum andS. tuberosum ssp.phureja and the wild speciesS. chacoense. Controlled crosses were performed following an incomplete diallel mating design which included the hybrid population and the parental species. Pollen grain germination and pollen tube growth were observed with fluorescence microscopy in 174 intrapopulation and interspecific combinations of genotypes. Fifty percent of the combinations of genotypes within the hybrid population and 46% of those between this population and the parental species were incompatible; overall, 71% of the interspecific combinations of genotypes were compatible when plants of the hybrid population were used as females, vs. 5% when used as males. Although the site of reaction could vary in a given intrapopulation or interspecific cross with the combinations of genotypes, pollen tube inhibition occurred in the first third of the style in 42% of them. Since the gene pool of the hybrid population had been widened with germplasm from other geographic areas and the genotypes ofS. tuberosum ssp.tuberosum, S. chacoense andS. tuberosum ssp.phureja used in this study were not the ones involved in the original crosses, it is concluded that the S-locus is not controlling the incompatibility reaction but rather that a cross-incompatibility system, possibly governed by more than one locus, is acting.     

Identification of stem soft rot (Erwinia carotovora subspeciesatroseptica) resistance in potato

Development of potato cultivars resistant to Erwinia species (Erwinia carotovora subspeciesatroseptica, E. carotovora subspeciescarotovora, andErwinia chrysanthemi) causing soft rot of stems and tubers involves identification and transfer of the resistance to breeding lines possessing desirable agronomic traits. In total, 363 accessions of six wildSolanum species, three somatic hybrids and nine sexual progeny of somatic hybrids were screened for stem soft rot resistance. Thirty-one, 50, and 67 percent of accessions ofSolanum boliviense, Solanum chacoense, and Solanumsanctarosae, respectively, were resistant or highly resistant, whereas, 100, 97, and 95 percent of accessions ofSolanum canasense, Solanum tarijense, andSolanum spegazzinii were susceptible or highly susceptible, respectively. Two tuber soft-rot resistant somatic hybrids, A937 and T355-11 produced earlier by the fusion ofSolanum brevidens (PI 218228) andSolanum tuberosum (PI203900) orS. tuberosum cv. Russet Burbank, respectively, and the sexual progeny of A937 andS. tuberosum cv. Katahdin were also highly resistant. Transfer of resistance fromS. brevidens to somatic hybrid (A937) and subsequently to the progeny suggests that the resistance is simply inherited. In addition to the identification of 65 resistant or highly resistant clones of wildSolanum species, this study has identified three tuber soft rot-resistant clones that are also resistant to stem soft rot. Potato cultivars showed a range of responses to the pathogen.     

Performance of populations derived by selecting for resistance to potato leafhopper in A 4XSolanum Tuberosum X 2XSolanum Chacoense cross

Intercross and backcross populations derived from aSolanum tuberosum L. (2n = 2x = 48) ×Solanum chacoense Bitt. (2n = 2x = 48) cross by selecting for resistance to potato leafhopper (PLH) (Empoasca fabae Harris), were tested in the field. In F2 and F4 populations, the levels of nymphal infestation and leaf necrosis from PLH feeding were considerably lower and tuber yields were much lower compared with those measured on the susceptible cultivar Kennebec. No differences were found between the two populations. Backcrosses of the F2 and F4 populations toS. tuberosum decreased resistance to PLH. Resistance to infestation was still greater than that of Kennebec; severity of leaf necrosis equaled that of Kennebec. Tuber yields increased, but were less than Kennebec’s yield. Foliage types shifted closer to those ofS. tuberosum. Three generations of selection and intercrossing within the backcross populations did not change the level of resistance to PLH but did tend to lower tuber yields. Foliage types remained primarilyS. tuberosum in appearance although distinctive types segregated with increased frequency. A second backcross toS. tuberosum did not change the level of resistance to PLH or the level of tuber yield relative to those of the first backcross or Kennebec.