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.     

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.     

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.     

Total foliar glycoalkaloids and resistance of wild potato species toEmpoasca fabae (Harris)

Levels of total glycoalkaloids (TGA) in foliage of 10 wild, tuberbearingSolanum (Tourn.) L. species differentially resistant to infestation by the potato leafhopper,Empoasca fabae (Harris), were determined. Levels of TGA ranged from a high of 688 mg/100 g fresh wt. in a resistant species,S. polyadenium Greenm. to a low of 13 mg/100 g fresh wt. in a susceptible species,S. bulbocastanum Dun. Foliar concentration of TGA and nymphal infestation by the potato leafhopper were highly correlated (r = ?0.75, p = 0.01). The significant correlation of TGA levels and potato leafhopper resistance suggests that foliar TGA may be a significant factor in the defense of wild potato species against this pest.     

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.     

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.     

Glycoalkaloids as pest resistance factors

Potato glycoalkaloids, a class of steroid glycosides, possess antimicrobial and pesticidal properties. These phytochemicals have been implicated as natural resistance factors inSolanum species against the Colorado potato beetle,Leptinotarsa decemlineata Say, and the potato leafhopper,Empoasca fabae (Harris). Adverse effects of these secondary metabolites on insect behavioral and developmental biology are described, as well as considerations in manipulation of foliar glycoalkaloids in breeding for varietal resistance to insects.     

Glycoalkaloid composition of potatoes: Diversity and biological implications

Glycoalkaloids are important potato quality factors. Tuber-bearingSolanum species exhibit wide quantitative and qualitative diversity in glycoalkaloid contents. Some species, of interest to breeders, contain extremely high total glycoalkaloid (TGA) levels and/or unusual types of glycoalkaloids of ill-defined or unknown toxic and teratogenic properties. Characterisation of glycoalkaloid content is a dynamic and, in some cases, controversial subject. Problems associated with analytical methodology and, to a lesser extent, with environmental effects and intraspecific heterogeneity have probably been the major causes of disagreements in the literature. Recent methodological developments have facilitated more accurate and sensitive measurements of TGA levels and of individual glycoalkaloid compositions. Such measurements should be used to analyze parental material and selected hybrid progeny in breeding programs. This is particularly important when certain wildSolanum species are being exploited as undesirable levels and types of glycoalkaloid can be genetically transmitted from the wild species to hybrid progeny. Absence of glycoalkaloid assessment from a breeding program can result in wasted effort, time and money.     

Mortality of potato leafhopper adults on synthetic diets containing seven glycoalkaloids synthesized in the foliage of variousSolanum species

The glycoalkaloids solanine, chaconine, leptine I, leptine II, solasonine, solamargine, and tomatine, synthesized in the foliage of variousSolanum species, were tested in synthetic diets for their effects on potato leafhopper adults,Empoasca fabae (Harris), at concentrations of 0.03%, 0.09%, and 0.27%. Percent mortality was recorded after a 72 hr feeding period. At the lowest concentration (0.03%), tomatine in the diet resulted in the highest adult mortality (95%) with mortalities ranging from 10% to 41% in the other glycoalkaloid diets. At the highest concentration (0.27%) mortalities ranged from 69% to 100% with no significant (P>.05) differences among the diets. Mortality varied the most among the glycoalkaloid diets at the 0.09% concentration. Chaconine, ubiquitously found with solanine inSolanum tuberosum L., caused significantly (P≤.05) higher mortality (59%) than did solanine (8%). Mortalities with solasonine and solamargine in the diets (55% and 54%, respectively) were not significantly (P>.05) different from the mortality with chaconine in the diet. Including leptines I and II in diets resulted in lower mortalities (28% and 30%, respectively). Mortality was significantly (P≤.05) the highest (97%) with tomatine in the diet.     

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.     

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.     

Use of reciprocal grafts to elucidate mode of resistance to Colorado potato beetle (Leptinotarsa decemlineata (Say)) and potato aphid (Macrosiphum euphorbiae (Thomas)) in six wildSolanum species

Reciprocal grafts were done between potato (Solanum tuberosum) and six wildSolanum species known to be resistant to the Colorado potato beetle (Leptinotarsa decemlineata (Say)). Potato or wildSolanum foliage from each reciprocal graft was fed to first instar beetles or adult potato aphids (Macrosiphum euphorbiae (Thomas)). The proportion of second instar larvae after 48 h and survival and fecundity (number of nymphs per female) of the aphids were used to i estimate the resistance level of the foliage. Survival and fecundity of potato aphid were reduced on wildSolanum foliage (except withS. circaeifolium ssp.capsicibaccatum andS. okadae) used as scion or rootstock, but were not different from the control (potato-potato graft) on potato foliage. Development of Colorado potato beetle onSolanum foliage was reduced. In some cases, potato foliage grafted to wildSolanum became resistant to the beetle. Results suggest that a chemical factor or factors providing resistance to Colorado potato beetle are translocated from foliage of some wildSolanum species to potato foliage. They also indicate that mode of resistance to the two insects differs quantitatively or qualitatively.     

Steroid glycoalkaloids and related compounds as potato quality factors

The steroid glycoalkaloids are triterpenoid derivatives which are found in all tissues of the potato plant including the tubers. The compounds are largely localized in the peel of tubers, but tissue beneath the peel rapidly accumulates the steroid glycoalkaloids to levels equal to or greater than those in the peel as a result of injury or environmental stress. The accumulation is restricted to the outer 1–2 mm of injured or stressed tuber. Potatoes containing over 0.02% steroid glycoalkaloids are considered toxic to man, and at this concentration they would impart a distinctly bitter flavor. The accumulation of steroid glycoalkaloids is suppressed and the accumulation of sesquiterpenoids is elicited in tubers infected by various pathogens and nonpathogens including the late blight pathogen,Phytophthora infestans. Arachidonic acid and eicosapentaenoic acids, two polyunsaturated fatty acids isolated fromP. infestans, are potent inhibitors of steroid glycoalkaloid accumulation. Both acids elicit the localized accumulation of sesquiterpenoids including rishitin, lubimin, phytuberin, phytuberol and solavetivone. Rishitin and lubimin generally comprise 85–90% of the total sesquiterpenoids which accumulate. The steroid glycoalkaloids and sesquiterpenoids appear to have a role in disease resistance to some fungal pathogens. Both groups of compounds are synthesized via the acetate-mevalonate pathway. Arachidonic and eicosapentaenoic acids appear to inhibit steroid glycoalkaloid accumulation at the level of the conversion of farnesyl pyrophosphate to squalene and they activate the biosynthesis of sesquiterpenoids. The reduction of steroid glycoalkaloids in potato foliage and tubers for health and flavor considerations should be considered relative to the ability of tubers and foliage to accumulate sesquiterpenoids in response to infection and its influence on disease and insect resistance.     

Determination of the glycoalkaloids α-solanine and α-chaconine levels in 18 varieties of potato (Solarium tuberosum L.) grown in Oman

Summary The glycoalkaloid content in 18 potato varieties grown in Oman was measured by HPLC using peel and flesh tissues. The total glycoalkaloid content of the tubers was found to be variety dependent and varied greatly between the 18 varieties. The total glycoalkaloid in the tubers of seven varieties was within the safe limit of ≤200 mg kg^{− 1} FW recommended for food safety, whereas the remaining 11 varieties exceeded this upper limit. The principal glycoalkaloids α-chaconine and σ-solanine were present in higher concentration in the peel than in the flesh of all varieties with the former being the principal alkaloid in the peel. The flesh of all of the 18 potato varieties contained total glycoalkaloid levels lower than the upper safety limit. The leaves of the 18 potato varieties were also analyzed for glycoalkaloids. The levels of glycoalkaloids in expanding leaves were higher than that of expanded leaves with the exception of six varieties. The principal glycoalkaloid in expanding and expanded leaves was α-chaconine. No correlation was observed between the glycoalkaloids levels of in the leaves and tubers.     

Resistance to potato viruses M,S, X and the spindle tuber virus in tuber-bearingSolanum species

On two separate occasions, some available accessions of tuber-bearingSolanum species were tested for resistance to potato viruses. Challenge inoculations were made mechanically with infective sap; and assessment was by sub-inoculation to differential host plants, supplemented by serological tests. In 1956–57, four accessions remained free from potato virus S (PVS), and nine remained free from PVM. In 1968–70, one or more clones of 11 accessions showed resistance to PVS—one of them,S. megistacrolobum, a diploid, apparently being hypersensitive. Resistance to PVX was found in seven accessions, and to PVM in two. There appeared to be resistance to the potato spindley tuber virus in five accessions.     

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.     

The Inter-genebank potato database and the dimensions of available wild potato germplasm

The Association of Potato Inter-genebank Collaborators (APIC) constructed a database of all wild potato holdings of the most important potato genebanks in Europe, the United States, Peru, and Argentina. The Inter-genebank Potato Database (IPD) now contains data of 11,819 wild potato accessions conserved in seven potato genebanks. The collector’s number is the key identifier used to merge all databases into the IPD. A total of 7,112 different wild potato accessions were identified, which comprise 5,306 accessions with known collector’s numbers. The IPD passport database showed that almost 30% of accessions held in APIC genebanks are from Argentina, a country that comprises less species diversity than Peru and Bolivia. These latter countries are represented by 24% and 20% of accessions, respectively. APIC genebanks maintain 188Solarium taxa out of more than 230 recognized by the latest comprehensive treatment of potatoes(Solarium sect.Petota) by Hawkes (1990). About 60% of the accessions comprise only 20 taxa represented by 785 to 92 accessions each. Conversely, 2% of the accessions comprise 72 taxa and are represented by five or fewer accessions each. About 70 taxa are not available in any genebank. The IPD evaluation database comprises 5,603 records with data from more than 33,000 evaluations of wild potato accessions. Fifty-five traits are summarized, including the reactions of the accessions to 12 pathotypes or races or strains of 12 fungi, four bacteria, 12 viruses, one viroid, 13 nematodes, and seven insects; response to heat and cold stress; and content of dry matter, starch, vitamin C, amylase, reducing sugars, and glycoalkaloids. About 30% of the wild potato populations screened showed various levels of resistance to most of the diseases and pests evaluated. The IPD database is available on the Internet at www.potgenebank.org.     

The dimensions of existingSolanum germ plasma collections

Analysis of the stocks available from threeSolanum germ plasm collections (Commonwealth Potato Collection, D. D. R. Sortiment and U. S. Potato Collection) shows that an adequate number of accessions are available for only a few species. Seventy-five of the 92 wild species being maintained are representd by fewer than 20 accessions. Development of activeSolanum germ plasm collections in the centers of diversity for these species eventually may provide the broader base of germ plasm needed for potato research.     

Resistance to late blight and soft rot in six potato progenies and glycoalkaloid contents in the tubers

Glycoalkaloids are anti-nutritional compounds commonly found in wildSolanum species used as resistance sources to major potato pathogens. It is therefore important for breeding purposes to know whether selecting for resistance using such species necessarily selects also for high glycoalkaloid contents in the tubers. To test this hypothesis, we used six partial progenies from crosses betweenSolanum tuberosum and accessions ofS. andigena, S. berthaultii, S. phureja, andS. vernei to investigate the possible correlation between resistance toPhytophthora infestans and/or toErwinia carotovora subsp.atroseptica and the concentration of glycoalkaloids in tubers. Concentrations of α-solanine and α-chaconine in the tubers segregated in each progeny, as did resistance to each pathogen. Some, but not all, clones from each progeny showed hypersensitive reactions to the isolate ofP. infestans used. Furthermore, clones within each progeny also differed for components of partial resistance toP. infestans, suggesting that all four wild species could be used as sources of both race-specific and partial resistance to late blight. With the exception of low, but statistically significant, correlations between concentration of α-solanine and two late blight resistance components (incubation period and spore production per unit lesion area) in progenies derived fromS. vernei, and despite a trend towards higher glycoalkaloid concentrations in the tubers of the clones most resistant to soft rot within progenies derived fromS. berthaultii andS. vernei, no consistent relationship between resistance to either disease and concentrations of α-solanine and/or α-chaconine was observed. These results indicate that neither race-specific nor partial resistance to late blight and soft rot in the accessions used as progenitors of resistance depend on high solanine or chaconine concentrations. These resistance sources could thus prove useful in breeding programs for improved behaviour againstP. infestans and/orE. carotovora.     

Determination of Glycoalkaloids in Potatoes and Potato Products by Microwave Assisted Extraction

Potato glycoalkaloids can reach levels that are harmful to human health. A rapid and reliable microwave assisted extraction method for quantitative analysis of α-solanine and α-chaconine content in raw potato and potato based products is presented. A chemical microwave was used to determine optimal temperature and pressure conditions for the extraction of α-solanine and α-chaconine from Idaho grown tubers and six commercially available mashed potato products. Recovery efficiency of glycoalkaloids was 37% greater by microwave assisted extraction (19.92 mg/kg glycoalkaloid) as compared to conventional solid/liquid methods (12.51 mg/kg glycoalkaloid). Optimal extraction of glycoalkaloids from potato samples dissolved in methanol was achieved using a microwave reactor set to 90 °C for ten minutes. The interior of Idaho grown tubers was determined to contain lower levels of glycoalkaloids (19.92 mg/kg dry weight; 6.5 ± 1.78 mg α-solanine and 13.40 ± 1.65 mg α-chaconine), as compared to commercial potato products (33.86–81.59 mg/kg).