Taste evaluation of potato glycoalkaloids by the Aymara: A case study in human chemical ecology

A chemical ecological model can be the basis for defining testable hypotheses concerning human interactions with plants. Selection by Aymara subsistence cultivators against toxic glycoalkaloids in the ongoing domestication of the Bolivian potato cultigen Solanum X ajanhuiriwas used as a specific case study of human interactions with phytochemicals. In taste perception tests, Aymara subjects were able to discriminate between concentrations of pure glycoalkaloids in solution only above 20 mg/100 ml. Taste panel tests of potato clones indicated that glycoalkaloid levels are important to the Aymara in determining quality only as part of a decision-making process involving two character states: too high or acceptable. Glycoalkaloids in potatoes are regarded as toxic to humans above 20 mg/100 g fresh weight. Among the Aymara, a breakpoint in the curve for glycoalkaloid preference appears to occur between 20–38 mg/100 g. This distinction is evident in the Aymara potato taxonomy which distinguishes bitter (luq'i ch'oke) from nonbitter (ch'oke) potatoes.     

Optimization of Glycoalkaloid Analysis for Use in Industrial Potato Fruit Juice Downstreaming

Annually, within the European Union about 1.7 million tons of starch is produced by processing over 8 million tons of potato tubers, Solanum tuberosum. In recent years, the potato protein content has gained tremendous industrial interest, since these proteins have excellent nutritional value. As naturally occurring, secondary plant metabolites steroidal potato glycoalkaloids are formed in potatoes. The two major glycoalkaloids in potatoes are α‐solanine and α‐chaconine. Because of the significant toxicity of the glycoalkaloids for human and for animal nutrition it was essential to develop efficient extraction processes. The need for an easy, fast, sensitive and reliable glycoalkaloid assay at the very beginning of the production chain is obvious. In this study an efficient analytical assay for potato glycoalkaloids from powdery protein samples under industrially relevant conditions is described: sample extraction, analyte pre‐purification, and final HPLC analysis. An acetic acid extraction/homogenization process was used for glycoalkaloid extraction from potato protein samples. The extracts were purified by means of solid phase extraction cartridges using the different washing steps developed in this study. The final determination was performed through an HPLC method using a Reprosil‐Pur NH₂ column. The limit of detection was 5 μg/mL for α‐solanine and α‐chaconine, respectively, corresponding to concentrations of 20 ppm in potato protein powder.     

Comparison of rapid liquid chromatography-electrospray ionization-tandem mass spectrometry methods for determination of glycoalkaloids in transgenic field-grown potatoes

Two rapid methods for highly selective detection and quantification of the two major glycoalkaloids in potatoes, alpha-chaconine and alpha-solanine, were compared for robustness in high-throughput operations for over 1000 analytical runs using potato tuber samples from field trials. Glycoalkaloids were analyzed using liquid chromatography coupled to tandem mass spectrometry in multiple reaction monitoring mode. An electrospray interface was used in the detection of glycoalkaloids in positive ion mode. Classical reversed phase (RP) and hydrophilic interaction (HILIC) columns were investigated for chromatographic separation, ruggedness, recovery, precision, and accuracy. During the validation procedure both methods proved to be precise and accurate enough in relation to the high degree of endogenous biological variability found for field-grown potato tubers. However, the RP method was found to be more precise, more accurate, and, more importantly, more rugged than the HILIC method for maintaining the analytes' peak shape symmetry in high-throughput operation. When applied to the comparison of six classically bred potato cultivars to six genetically modified (GM) lines engineered to synthesize health beneficial inulins, the glycoalkaloid content in potato peels of all GM lines was found within the range of the six cultivars. We suggest complementing current unbiased metabolomic strategies by validating quantitative analytical methods for important target analytes such as the toxic glycoalkaloids in potato plants.     

Sterol Side Chain Reductase 2 Is a Key Enzyme in the Biosynthesis of Cholesterol,the Common Precursor of Toxic Steroidal Glycoalkaloids in Potato

Potatoes (Solanum tuberosum) contain α-solanine and α-chaconine, two well-known toxic steroidal glycoalkaloids (SGAs). Sprouts and green tubers accumulate especially high levels of SGAs. Although SGAs were proposed to be biosynthesized from cholesterol, the biosynthetic pathway for plant cholesterol is poorly understood. Here, we identify sterol side chain reductase 2 (SSR2) from potato as a key enzyme in the biosynthesis of cholesterol and related SGAs. Using in vitro enzyme activity assays, we determined that potato SSR2 (St SSR2) reduces desmosterol and cycloartenol to cholesterol and cycloartanol, respectively. These reduction steps are branch points in the biosynthetic pathways between C-24 alkylsterols and cholesterol in potato. Similar enzymatic results were also obtained from tomato SSR2. St SSR2-silenced potatoes or St SSR2-disrupted potato generated by targeted genome editing had significantly lower levels of cholesterol and SGAs without affecting plant growth. Our results suggest that St SSR2 is a promising target gene for breeding potatoes with low SGA levels.     

Growth,photosynthetic activity and tuber quality of two potato cultivars in controlled environment as affected by light source

In order to elucidate the effect of genetic material and light source and their interaction on plant performance of potato (Solanum tuberosum L.), we studied the influence of two light sources, white fluorescent tubes (WF) and red blue LEDs with ratio 8:1 (RB) and two cultivars, ‘Avanti’ and ‘Colomba’ grown in phytotron, on growth, leaf photosynthesis and photochemistry and tuber quality. Under WF, net photosynthesis (NP) increased from the vegetative phase until flowering, then decreased during tuber bulking, with no differences between the cultivars. Lighting with RB increased the NP and the PSII maximum quantum use efficiency compared to WF. RB reduced stem elongation in both cultivars, as well as the number and area of leaves, and the aerial biomass per plant in ‘Colomba’, compared to WF. Conversely, tuber yield was higher in plants under RB light in both ‘Avanti’ and ‘Colomba’. Light source did not influence the tuber content of starch and total glycoalkaloids, while it affected differently in the cultivars the protein content and the glycoalkaloid profile. Our results highlight how interactions between light source and genotype need to be considered for potato cultivation in controlled environment under artificial lighting.     

Potato glycoalkaloids: true safety or false sense of security?

As one of the major agricultural crops, the cultivated potato is consumed each day by millions of people from diverse cultural backgrounds. A product of global importance, the potato tuber contains toxic glycoalkaloids (GAs) that cause sporadic outbreaks of poisoning in humans, as well as many livestock deaths. This article will discuss some aspects of the potato GAs, including their toxic effects and risk factors, methods of detection of GAs and biotechnological aspects of potato breeding. An attempt has been made to answer a question of vital importance - are potato GAs dangerous to humans and animals and, if so, to what extent?     

Reduction of cholesterol and glycoalkaloid levels in transgenic potato plants by overexpression of a type 1 sterol methyltransferase cDNA

Transgenic potato (Solanum tuberosum cv Désirée) plants overexpressing a soybean (Glycine max) type 1 sterol methyltransferase (GmSMT1) cDNA were generated and used to study sterol biosynthesis in relation to the production of toxic glycoalkaloids. Transgenic plants displayed an increased total sterol level in both leaves and tubers, mainly due to increased levels of the 24-ethyl sterols isofucosterol and sitosterol. The higher total sterol level was due to increases in both free and esterified sterols. However, the level of free cholesterol, a nonalkylated sterol, was decreased. Associated with this was a decreased glycoalkaloid level in leaves and tubers, down to 41% and 63% of wild-type levels, respectively. The results show that glycoalkaloid biosynthesis can be down-regulated in transgenic potato plants by reducing the content of free nonalkylated sterols, and they support the view of cholesterol as a precursor in glycoalkaloid biosynthesis.     

Commersonine,a new glycoalkaloid from two Solanum species

Selected plant introduction lines of S. chacoense and S. commersonii contain two major glycoalkaloids, demissine and a new compound called commersonine. In contrast, other plant introduction lines of S. chacoense contain only solanine and chaconine as the major glycoalkaloids. The isolation and characterization of the new glycoalkaloid is described.     

Dual specificity of sterol-mediated glycoalkaloid induced membrane disruption.

In this study the effects of the glycoalkaloids alpha-solanine, alpha-chaconine, alpha-tomatine and the aglycone solanidine on model membranes composed of PC in the absence and presence of sterols have been analysed via permeability measurements and different biophysical methods. The main result is that glycoalkaloids are able to interact strongly with sterol containing membranes thereby causing membrane disruption in a way which is specific for the type of glycoalkaloid and sterol. For this dual specificity both the sugar moiety of the glycoalkaloid and the side-chain of the sterol on position 24 turned out to be of major importance for the membrane disrupting activity. The order of potency of the glycoalkaloids was alpha-tomatine > alpha-chaconine > alpha-solanine. The plant sterols beta-sitosterol and fucosterol showed higher affinity for glycoalkaloids as compared to cholesterol and ergosterol. The mode of action of the glycoalkaloids is proposed to consist of three main steps: (1) Insertion of the aglycone part in the bilayer. (2) Complex formation of the glycoalkaloid with the sterols present. (3) Rearrangement of the membrane caused by the formation of a network of sterol-glycoalkaloid complexes resulting in a transient disruption of the bilayer during which leakage occurs.     

The effect of genetic transformations for pest resistance on foliar solanidine-based glycoalkaloids of potato (Solatium tuberosuni)

Foliage of potato cv. Desiree was harvested from glasshouse‐cultivated plants of five experimental transgenic lines expressing three different insecticidal proteins (snowdrop lectin, Galanthus nivalis agglutinin (GNA); jackbean lectin, Concanavalin A (Con A), cowpea trypsin inhibitor; (CpTi)), tissue‐cultured control plants and standard control (non‐tissue cultured) plants. The foliage was subdivided into stems, upper, middle and lower leaves and analysed separately by HPLC for the solanidine‐based glycoalkaloids a‐solanine and a‐chaconine. The results demonstrate that one or more stages in the plant transformation process (i.e. insecticidal‐ and marker‐gene insertions, gene expression and tissue culture) resulted in a lower level of leaf glycoalkaloids than that found in either the tissue‐cultured controls or standard controls, based on the selected potato lines transformed for insecticidal protein expression. However, the distribution of glycoalkaloids throughout the plant foliage was unaffected by genetic transformation and tissue culture, with the highest glycoalkaloid levels being observed in the top third of the plant. The importance of investigating unexpected effects of genetic engineering on plant secondary metabolism is discussed from an ecological viewpoint.     

Steroidal glycoalkaloids in cell and shoot teratoma cultures ofSolanum dulcamara

Bittersweet (Solanum dulcamara L., Solanaceae) is of interest as a source of steroidal alkaloids for the commercial production of hormones. Since glycoalkaloid production is positively correlated to differentiation, tumor and teratoma cultures of the soladulcidine chemotype were established by transformation withAgrobacterium tumefaciens. A newly developed HPLC-system, which allowed separation and sensitive quantitation of the glycoalkaloids soladulcidine-tetraoside, solamargine and solasonine, was used to analyse glycoalkaloid profiles in plants and cultures. Tumors and teratoma were charcterized by a shift in their alkaloid pattern from soladulcidine tetraoside to the solasodine glycosides solamargine and solasonine. Shoot teratoma showed a total glycoalkaloid content of 1% dw, which is about fivefold higher than in the source plant. A regenerated plant retained the altered alkaloid spectrum; the levels, however, equalled those of the source plant. From the alteration of alkaloid pattern in the transformed cultures suggestions can be made concerning the biosynthetic pathway. Completion of the biosynthesis of the aglycone is likely to be complete before glycosylation occurs.     

A major QTL and an SSR marker associated with glycoalkaloid content in potato tubers from <Emphasis Type="Italic">Solanum tuberosum </Emphasis>× <Emphasis Type="Italic">S. sparsipilum</Emphasis> located on chromosome I

New potato (Solanum tuberosum) varieties are required to contain low levels of the toxic glycoalkaloids and a potential approach to obtain this is through marker-assisted selection (MAS). Before applying MAS it is necessary to map quantitative trait loci (QTLs) for glycoalkaloid content in potato tubers and identify markers that link tightly to this trait. In this study, tubers of a dihaploid BC(1) population, originating from a cross between 90-HAF-01 (S. tuberosum(1)) and 90-HAG-15 (S. tuberosum(2) x S. sparsipilum), were evaluated for content of alpha-solanine and alpha-chaconine (total glycoalkaloid, TGA) after field trials. In addition, tubers were assayed for TGA content after exposure to light. A detailed analysis of segregation patterns indicated that a major QTL is responsible for the TGA content in tubers of this potato population. One highly significant QTL was mapped to chromosome I of the HAG and the HAF parent. Quantitative trait loci for glycoalkaloid production in foliage of different Solanum species have previously been mapped to this chromosome. In the present research, QTLs for alpha-solanine and alpha-chaconine content were mapped to the same location as for TGA content. Similar results were observed for tubers exposed to light. The simple sequence repeat marker STM5136 was closely linked to the identified QTL.     

Synergism between the potato glycoalkaloids alpha-chaconine and alpha-solanine in inhibition of snail feeding

Snails (Helix aspersa L.) were fed filter paper treated with the potato glycoalkaloids, alpha-solanine and alpha-chaconine, singly or together. In pure form, both glycoalkaloids deterred feeding, with chaconine being the more active compound. In combination, authentic solanine and chaconine interacted synergistically in their inhibition of feeding. The antifeedant activities of methanolic extracts of tuber peel of the potato varieties Majestic and Sharpe's Express presented via filter paper discs did not differ significantly from those of authentic glycoalkaloid solutions of comparable concentration and ratio. In contrast, feeding inhibition by diluted tuber peel extracts of the variety Homeguard was greater than that elicited by comparable authentic glycoalkaloid solutions suggesting additional inhibitory compound(s) in the peel of this variety. Comparison of data from peel extracts of all three potato varieties and authentic glycoalkaloids indicated that the level of feeding inhibition by the extracts was, at least in part, a consequence of a synergism between solanine and chaconine.     

Biosensors for assay of glycoalkaloids in potato tubers

The possibility of practical application of biosensors based on pH-sensitive field-effect transistors and butyrylcholinesterase to glycoalkaloid analysis in potato tubers was studied. The main analytical features of the designed biosensors and measurement conditions were optimized. The biosensor was applied to quantitative analysis of glycoalkaloids in tubers of different potato varieties. The results proved to be in good agreement with those obtained by conventional protocols. Experiments on glucose assay were performed. An inverse correlation between the contents of glucose and glycoalkaloids in potato tubers was demonstrated.     

Biosensors for assay of glycoalkaloids in potato tubers

The possibility of commercial application of biosensors based on pH-sensitive field-effect transistors and butyrylcholinesterase to glycoalkaloid analysis in potato tubers was studied. The main analytical features of the designed biosensors and measurement conditions were optimized. The biosensor was applied to quantitative analysis of glycoalkaloids in tubers of different potato varieties. The results proved to be in good agreement with those obtained by conventional protocols. Experiments on glucose assay were performed. An inverse correlation between the contents of glucose and glycoalkaloids in potato tubers was demonstrated.     

Glycoalkaloid increase in Solarium tuberosum on exposure to light

Tubers of six commercially available potato cultivars were placed in bright light (140 jUmol“¹ m”²), approximately equivalent to dull daylight, for a continuous period of up to seven days. The tubers were sampled at intervals, scored for degree of greening, freeze-dried and subsequently assessed for glycoalkaloid and chlorophyll content. There were significant differences between the cultivars in their rates of greening and increase in glycoalkaloid content. There was an apparent relationship between the two characters. Increases in the individual glycoalkaloids a–chaconine and a-solanine were also assessed. The results are discussed in terms of the implications for the potato industry.     

Photo-induced synthesis of tomatidenol-based glycoalkaloids in Solanum phureja tubers

The effect of light exposure on the steroidal glycoalkaloid content of Solanum phureja tubers has been investigated and compared with that in domesticated potato (Solanum tuberosum) tubers. The results indicated that the increase in the concentration of solanidine-based glycoalkaloids, alpha-solanine and alpha-chaconine was broadly similar in both species. However, in the S. phureja tubers, light exposure also induced the synthesis of tomatidenol-based glycoalkaloids. These have been identified as alpha- and beta-solamarine. These glycoalkaloids were not detected in tubers continually stored in darkness.     

Quantitative trait loci analysis of seed‐specialized metabolites reveals seed‐specific flavonols and differential regulation of glycoalkaloid content in tomato

Given the potential health benefits (and adverse effects), of polyphenolic and steroidal glycoalkaloids in the diet there is a growing interest in fully elucidating the genetic control of their levels in foodstuffs. Here we carried out profiling of the specialized metabolites in the seeds of the Solanum pennellii introgression lines identifying 338 putative metabolite quantitative trait loci (mQTL) for flavonoids, steroidal glycoalkaloids and further specialized metabolites. Two putative mQTL for flavonols and one for steroidal glycoalkaloids were cross‐validated by evaluation of the metabolite content of recombinants harboring smaller introgression in the corresponding QTL interval or by analysis of lines from an independently derived backcross inbred line population. The steroidal glycoalkaloid mQTL was localized to a chromosomal region spanning 14 genes, including a previously defined steroidal glycoalkaloid gene cluster. The flavonoid mQTL was further validated via the use of transient and stable overexpression of the Solyc12g098600 and Solyc12g096870 genes, which encode seed‐specific uridine 5′‐diphosphate‐glycosyltransferases. The results are discussed in the context of our understanding of the accumulation of polyphenols and steroidal glycoalkaloids, and how this knowledge may be incorporated into breeding strategies aimed at improving nutritional aspects of plants as well as in fortifying them against abiotic stress.     

Varietal susceptibility of potatoes to wireworm herbivory

1 Wireworms, the soil dwelling larvae of click beetles, Agriotes spp., have recently become a more prevalent pest of potatoes. The present study investigated whether potato varieties showed variable susceptibility to wireworm herbivory, and also tested whether increased susceptibility was associated with lower concentrations of glycoalkaloids. Twelve varieties were originally screened across a range of experimental scales, including laboratory and tunnel experiments and a large‐scale field trial involving over 2000 tubers. 2 In laboratory no‐choice tests, Maris Peer, Marfona and Rooster varieties were significantly more susceptible to wireworm attack, with 63% of tubers showing damage, compared with just 15% of the less susceptible varieties of King Edward, Nadine and Maris Piper. There was also greater tissue consumption and weight gain when wireworms were reared on the most susceptible varieties. 3 In choice tests, wireworms showed a significant preference for those varieties previously identified as being the most susceptible to wireworm herbivory (4.2 holes per tuber) compared with the least susceptible (1.2 holes per tuber). Similar patterns of susceptibility were seen in the field trial, although there was generally more variation in susceptibility. 4 In a tunnel experiment, Marfona and Maris Peer were significantly more susceptibile to wireworm attack (47% of tubers showing damage) compared with Nadine, King Edward and Maris Piper (27% of tubers showing damage). Although Nadine, in particular, had the highest glycoalkaloid concentrations (309.33 mg/kg) and lowest amounts of wireworm herbivory, the relationship between susceptibility and glycoalkaloid concentrations was weak, suggesting that this is unlikely to be the sole mechanism underpinning varietal susceptibility.     

The relationship between glycoalkaloids and resistance to the white potato cyst nematode, Globodera pallida in potato clones derived from Solanum vernei

No relationship was found between the degree of resistance to Globodera pallida and total glycoalkaloid content of the roots or tubers of a number of potato clones derived from Solanum vernei × S. tuberosum. Nematode infestation of the roots did not lead to increases in the glycoalkaloid content of susceptible or resistant potatoes.