Effect of light exposure on the glycoalkaloid content of Solanum phureja tubers.

The individual glycoalkaloid contents of tubers from eleven Solanum phureja genotypes have been determined prior to and following exposure to light. In all genotypes, light exposure resulted in a statistically significant increase in total glycoalkaloid content. In nine of the genotypes studied, this was not only due to an increase in the levels of the solanidine-based glycoalkaloids, alpha-solanine and alpha-chaconine, but also due to the light-induced synthesis of a tomatidenol-based glycoalkaloid, alpha-solamarine. Those genotypes that accumulated alpha-solamarine in their tubers also contained tomatidenol-based glycoalkaloids in their leaves, but only solanidine-based glycoalkaloids were detected in the sprouts.     

Inheritance of morphological characters and glycoalkaloids in potatoes of somatic hybrids between dihaploid Solanum acauleand tetraploid Solanum tuberosum.

Steroidal glycoalkaloids occur in potatoes and are reported to impart resistance to phytopathogens including bacteria, fungi, and insects. Because glycoalkaloids can be passed to progenies during breeding programs designed to develop improved potatoes, it is of importance to determine the quality of desired characteristics and the composition of glycoalkaloids of new somatic hybrids. The objective of this study was to determine the appearance, size, and shape (morphological characters) as well as the glycoalkaloid content of potato tubers of somatic hybrids between tetraploid Solanum tuberosum cv. Dejima (2n = 4x = 48 chromosomes) and the dihaploid clone ATDH-1 (2n = 2x = 24 chromosomes) induced by anther culture from Solanum acuale-T (acl-T, 2n = 4x = 48 chromosomes). Tuber size and shape in somatic hybrids were in accord with those of cv. Dejima, whereas the tuber skin color resembled that of ATDH-1. Thin-layer chromatography, high-performance liquid chromatography, and gas-liquid chromatography/mass spectrometry studies showed that the two steroidal glycoalkaloids (alpha-chaconine and alpha-solanine) were present in the tubers of S. tuberosum, whereas acl-T and ATDH-1 tubers were found to contain alpha-tomatine and demissine. The concentrations of total glycoalkaloids in both acl-T and ATDH-1 was >100 mg/100 g of fresh weight tuber cortex, much higher than in S. tuberosum. All somatic hybrids, except one clone, contained four glycoalkaloids (alpha-chaconine, alpha-solanine, alpha-tomatine, and demissine) derived from the fusion parents. The lack of alpha-tomatine in the remaining clone may be due to somaclonal variation. The results show that character expression is influenced by ploidy level and that total glycoalkaloid levels in most somatic hybrids were intermediate between those of the fusion parents. The possible significance of these findings for plant breeding and food safety is discussed.     

Geographical variation of solanidane aglycone glycoalkaloids in the wild potato species Solanum chacoense Bitter

The Colorado Potato Beetle is a serious pest of the cultivated potato. Natural resistance has been found in a few wild species, including Solanum chacoense Bitter, in which the resistance is attributed to the presence of leptine glycoalkaloids. Production and accumulation of these compounds within S. chacoense varies widely and appears to be inherited in a quantitative fashion, but high leptine-producing clones occur rarely. In the present study, 15 different accessions from various locations and altitudes of origin within central to northern Argentina and Paraguay were analyzed for foliar glycoalkaloid (leptine, leptinine, solanine, chaconine) content. The objective was to infer the frequency of leptine production in ecogeographically distinct S. chacoense accessions, and to ascertain any possible association between leptine levels/concentrations and ecogeographical location. Leptines were detected in 8 of the 15 accessions, and the amounts within each accession varied widely. Most of the leptine-containing accessions originated from western Argentina except two in province Córdoba in central Argentina. There was no relationship between elevational level and leptine, but there was a negative trend with total glycoalkaloids (TGA) and elevation, due to solanine and chaconine levels which decreased with increasing elevation. In addition, nine unidentified, putative glycoalkaloids were detected, in very high proportions in some individuals and accessions. This study raises interesting questions about glycoalkaloid distribution, helps provide direction for new avenues of leptine and glycoalkaloid research, and proposes a systematic, ecogeographically based method for bioprospecting genes controlling rare plant secondary compounds.     

Estimation of glycoalkaloids as solasodine in Solanum laciniatum.

Solasodine is isolated from Solanum laciniatum leaves by a rapid, one-step procedure which involves extraction and hydrolysis of the glycoalkaloids. The resulting steroidal aglycone is directly estimated by the formation of a colored complex with bromocresol green. The method is applicable to concentrations of 25-125 mug solasodine/7ml.     

Solanidine hydrolytic extraction and separation from the potato (Solanum tuberosum L.) vines by using solid-liquid-liquid systems

Solanidine is a steroidal aglycon of potato (Solanum tuberosum L.) glycoalkaloids and a very important precursor for the synthesis of hormones and some pharmacologically active compounds. Glycoalkaloids are hydrolyzed by mineral acid, yielding solanidine. This paper deals with the kinetics of solanidine hydrolytic extraction in different solid-liquid-liquid systems. The dried and milled potato (S. tuberosum L.) vines were used as a source of glycoalkaloids and as the solid phase. The solutions of hydrochloric acid in 2 and 10% (w/v) aqueous acetic acid, in 50% (volume) aqueous methanol, and in 50% (volume) aqueous ethanol were first liquid phase, and the medium for glycoalkaloid extraction from potato vines and their hydrolysis to solanidine. The chloroform, trichloroethylene, or carbon tetrachloride were the second, organic, liquid phase and the medium for solanidine extraction. This procedure combines three different processes: extraction of glycoalkaloids from potato vines, their hydrolysis to solanidine, and the extraction of solanidine, in a single step. The term hydrolytic extraction of solanidine was used for these processes. The purpose of the paper was to choose an optimal solid-liquid-liquid system for solanidine extraction and to define the procedure for its isolation from the organic liquid phase. The best degree of solanidine hydrolytic extraction (DHE) of more than 98% was achieved when 10% (w/v) hydrochloric acid in 50% (volume) methanol were the first liquid phase and chloroform was the second liquid phase, after 90 min. The yield of solanidine (q(S)) under these conditions is calculated to be 0.24 g/100 g of potato vines. Approximately 78% of the maximal possible yield of solanidine was isolated from chlorofom liquid phase. The IR and MS spectra of isolated solanidine were recorded.     

Glycoalkaloid content and chemical composition of potatoes improved with nonconventional breeding approaches

This paper reports the results of chemical analyses performed on two distinct groups of new potato genotypes. The first group contained five clones transformed with the gene ech42 encoding for an endochitinase. The second included 21 interspecific hybrids between the cultivated potato Solanum tuberosum and the wild species S. commersonii, obtained either by somatic fusion or by sexual hybridization. Tubers from transgenic plants were analyzed for several morphological and biochemical parameters to ascertain the substantial equivalence between the transgenic genotypes and the original cultivar Désirée. The interspecific hybrids were analyzed for the same parameters in order to identify genotypes with novel improved chemical characteristics and with low levels of glycoalkaloids deriving from the wild species and potentially hazardous to human health. For transgenic tubers, the results provided evidence that indicates the substantial equivalence between the transgenic genotypes and the cultivated control for the considered traits. The results suggest that chitinase gene insertion did not alter other metabolic pathways of potato tubers and did not cause unintentional pleiotropic effects. As far as interspecific hybrids are concerned, wide variability for all of the parameters analyzed was found. For some useful traits (e.g., soluble solids and proteins, dry matter content) the interspecific hybrids performed better than both the cultivated control and the wild species. In a number of genotypes, glycoalkaloid levels were close to or lower than those of the control varieties, suggesting that selection for low glycoalkaloid content is possible. The results also indicated that glycoalkaloids from S. commersonii may be lost rapidly. Indeed, some hybrids were found to have the same glycoalkaloid profile as S. tuberosum. Finally, the results showed that among the parameters considered, glycoalkaloid content is the most sensitive to variation. Therefore, glycoalkaloid determination should be used for routine control of genotypes produced by interspecific hybridization.     

Glycoalkaloids and metabolites inhibit the growth of human colon (HT29) and liver (HepG2) cancer cells

As part of an effort to improve plant-derived foods such as potatoes, eggplants, and tomatoes, the antiproliferative activities against human colon (HT29) and liver (HepG2) cancer cells of a series of structurally related individual compounds were examined using a microculture tetrazolium (MTT) assay. The objective was to assess the roles of the carbohydrate side chain and aglycon part of Solanum glycosides in influencing inhibitory activities of these compounds. Evaluations were carried out with four concentrations each (0.1, 1, 10, and 100 microg/mL) of the the potato trisaccharide glycoalkaloids alpha-chaconine and alpha-solanine; the disaccharides beta(1)-chaconine, beta(2)-chaconine, and beta(2)-solanine; the monosaccharide gamma-chaconine and their common aglycon solanidine; the tetrasaccharide potato glycoalkaloid dehydrocommersonine; the potato aglycon demissidine; the tetrasaccharide tomato glycoalkaloid alpha-tomatine, the trisaccharide beta(1)-tomatine, the disaccharide gamma-tomatine, the monosaccharide delta-tomatine, and their common aglycon tomatidine; the eggplant glycoalkaloids solamargine and solasonine and their common aglycon solasodine; and the nonsteroidal alkaloid jervine. All compounds were active in the assay, with the glycoalkaloids being the most active and the hydrolysis products less so. The effectiveness against the liver cells was greater than against the colon cells. Potencies of alpha-tomatine and alpha-chaconine at a concentration of 1 microg/mL against the liver carcinoma cells were higher than those observed with the anticancer drugs doxorubicin and camptothecin. Because alpha-chaconine, alpha-solanine, and alpha-tomatine also inhibited normal human liver HeLa (Chang) cells, safety considerations should guide the use of these compounds as preventative or therapeutic treatments against carcinomas.     

Rapid screening of ascorbic acid, glycoalkaloids, and phenolics in potato using high-performance liquid chromatography

Evaluation of phenolic metabolism in potato tubers (Solanum tuberosum) would be facilitated by faster analytical methods. A high-throughput HPLC method was developed for the qualitative and quantitative determination in potato of numerous phenolic compounds, the sum of the glycoalkaloids chaconine and solanine, plus ascorbic acid. Following a fast extraction, HPLC run times of 12 min were achieved with the use of a monolithic RP C18 column. UV and MS analyses were used to characterize the phenolic complement in extracts from two white-fleshed varieties. Over 30 compounds were identified, some of which are thought to possess either nutritional value or are involved in plant disease resistance. This method is expected to be useful for germplasm mining and for varietal development programs in which large numbers of lines are generated.     

Tomato glycoalkaloids: role in the plant and in the diet

Tomatoes, a major food source for humans, accumulate a variety of secondary metabolites including phenolic compounds, phytoalexins, protease inhibitors, and glycoalkaloids. These metabolites protect against adverse effects of hosts of predators including fungi, bacteria, viruses, and insects. Because glycoalkaloids are reported to be involved in host-plant resistance, on the one hand, and to have a variety of pharmacological and nutritional properties in animals and humans, on the other, a need exists to develop a better understanding of the role of these compounds both in the plant and in the diet. To contribute to this effort, this integrated review presents data on the history, composition, and nutrition of tomatoes, with special focus on the assessment of the chemistry, analysis, composition, nutrition, microbiology, and pharmacology of the tomato glycoalkaloids comprising alpha-tomatine and dehydrotomatine; their content in different parts of the tomato plant, in processed tomato products, and in wild and transgenic tomatoes; their biosynthesis, inheritance, metabolism, and catabolism; plant-microbe relationships with fungi, bacteria, viruses, insects, and worms; interactions with ergosterol and cholesterol; disruption of cell membranes; tomatine-induced tomatinases, pantothenate synthetase, steroid hydroxylases, and cytokines; and inhibition of acetylcholinesterase. Also covered are tomato-human pathogen relationships and tomatine-induced lowering of plasma cholesterol and triglycerides and enhancement of the immune system. Further research needs in each of these areas are suggested. The overlapping aspects are discussed in terms of general concepts for a better understanding of the impact of tomato glycoalkaloids in the plant in general and in food in particular. Such an understanding can lead to the creation of improved tomatoes and to improved practices on the farm and in the consumption of tomatoes.     

Tracer studies on the incorporation of [2-14C]-DL-mevalonate into chlorophylls a and b, alpha-chaconine, and alpha-solanine of potato sprouts

Chlorophyll and glycoalkaloids are synthesized in different parts of the potato plant including leaves, tubers, and sprouts. Although light stimulates the biosynthesis of both constituents, the question of whether the two biosynthetic pathways are under the same genetic control has not been resolved. This study investigated the dynamics of incorporation of labeled [2-(14)C]-DL-mavalonate into chlorophyll a, chlorophyll b, and the glycoalkaloids alpha-chaconine and alpha-solanine in potato sprouts after 7 and 14 days of storage in the light and in the dark. No chlorophyll synthesis occurred in the dark. Fractionation of the "glycoalkaloid" extract followed by high-performance liquid chromatography produced four peaks. The fractions were collected and analyzed for radioactivity. About 80% of the radioactivity resided in fraction 1, the composition of which is unknown. Two of the fractions, with 1-14% of the original label, were alpha-chaconine and alpha-solanine. The radioactivity derived from mevalonate largely resides in unidentified compound(s) eluting as a single peak on the HPLC column before the peaks associated with the glycoalkaloids. The specific radioactivity of alpha-chaconine and alpha-solanine increased approximately 2-fold in going from 7 to 14 days of exposure in the light and in the dark. These and additional observations point to the near identity of the dynamics of biosynthesis of the two glycoalkaloids. These data also implicate a non-mevalonate pathway for the synthesis of both chlorophylls and the glycoalkaloids and are consistent with independent genetic control of the concurrent formation of the two classes of compounds during greening of potatoes.     

Sulfur compounds reduce potato toxins during extrusion cooking.

Free sulfhydryl groups in sulfur compounds have been reported to act directly on natural toxins to reduce toxicity. The objective of this study was to reduce protease inhibitors and glycoalkaloids in simulated snack foods by the addition of sulfur-containing compounds prior to extrusion. Thiamine, methionine, and benzyl disulfide were added to potato flakes at levels of 0.5% or 1.0% prior to twin-screw extrusion. Total and free thiols and protease inhibitors were monitored before and after extrusion by colorimetric assays. Potato glycoalkaloids were analyzed by HPLC and by immunoassay. Extrusion reduced potato flake disulfide bonds; disulfide bonds were higher in samples containing added sulfur compounds. Trypsin inhibitor activity was reduced by as much as 79% by extrusion plus methionine. Extrusion significantly reduced carboxypeptidase inhibitor, but only when benzyl disulfide and 0.5% methionine were not added. One percent methionine and thiamine resulted in 60% reductions in glycoalkaloids.     

LC-MS analysis of solanidane glycoalkaloid diversity among tubers of four wild potato species and three cultivars (Solanum tuberosum)

Secondary metabolites in potato tubers include both phytonutrients and plant defense compounds. The extent these small molecules vary among different potato genotypes is not well characterized. LC-MS analysis of tuber extracts from seven potato genotypes showed that one large source of small molecule variation is the glycoalkaloids. Glycoalkaloids are involved in the resistance of potatoes to pathogens and pests, but they also have implications for human health and nutrition. This study focused on glycoalkaloids with solanidane or solanidane-like aglycones, of which over 50 were tentatively identified, many of which appeared to be novel glycoalkaloids. Results suggested the variety of glycoalkaloids in potatoes is considerably greater than previously thought. Dissecting the role of these many glycoalkaloids in human health or pest and pathogen resistance will be a formidable undertaking.     

Potato glycoalkaloids and metabolites: roles in the plant and in the diet

Potatoes, members of the Solanaceae plant family, serve as major, inexpensive low-fat food sources providing energy (starch), high-quality protein, fiber, and vitamins. Potatoes also produce biologically active secondary metabolites, which may have both adverse and beneficial effects in the diet. These include glycoalkaloids, calystegine alkaloids, protease inhibitors, lectins, phenolic compounds, and chlorophyll. Because glycoalkaloids are reported to be involved in host-plant resistance and to have a variety of adverse as well as beneficial effects in cells, animals, and humans, a need exists to develop a clearer understanding of their roles both in the plant and in the diet. To contribute to this effort, this integrated review presents data on the (a) history of glycoalkaloids; (b) glycoalkaloid content in different parts of the potato plant, in processed potato products, and in wild, transgenic, and organic potatoes; (c) biosynthesis, inheritance, plant molecular biology, and glycoalkaloid-plant phytopathogen relationships; (d) dietary significance with special focus on the chemistry, analysis, and nutritional quality of low-glycoalkaloid potato protein; (e) pharmacology and toxicology of the potato glycoalkaloids comprising alpha-chaconine and alpha-solanine and their hydrolysis products (metabolites); (f) anticarcinogenic and other beneficial effects; and (g) possible dietary consequences of concurrent consumption of glycoalkaloids and other biologically active compounds present in fresh and processed potatoes. An enhanced understanding of the multiple and overlapping aspects of glycoalkaloids in the plant and in the diet will benefit producers and consumers of potatoes.     

Anticarcinogenic effects of glycoalkaloids from potatoes against human cervical, liver, lymphoma, and stomach cancer cells

Methods were devised for the isolation of large amounts of pure alpha-chaconine and alpha-solanine from Dejima potatoes and for the extraction and analysis of total glycoalkaloids from five fresh potato varieties (Dejima, Jowon, Sumi, Toya, and Vora Valley). These compounds were then evaluated in experiments using a tetrazolium microculture (MTT) assay to assess the anticarcinogenic effects of (a) the isolated pure glycoalkaloids separately, (b) artificial mixtures of the two glycoalkaloids, and (c) the total glycoalkaloids isolated from each of the five potato varieties. All samples tested reduced the numbers of the following human cell lines: cervical (HeLa), liver (HepG2), lymphoma (U937), stomach (AGS and KATO III) cancer cells and normal liver (Chang) cells. The results show that (a) the effects of the glycoalkaloids were concentration dependent in the range of 0.1-10 mug/mL (0.117-11.7 nmol/mL); (b) alpha-chaconine was more active than was alpha-solanine; (c) some mixtures exhibited synergistic effects, whereas other produced additive ones; (d) the different cancer cells varied in their susceptibilities to destruction; and (e) the destruction of normal liver cells was generally lower than that of cancer liver cells. The decreases in cell populations were also observed visually by reversed-phase microscopy. The results complement related observations on the anticarcinogenic potential of food ingredients.     

Sampling leaves of young potato (Solanum tuberosum) plants for glycoalkaloid analysis

Earlier attempts to measure potato (Solanum tuberosum) leaf glycoalkaloids indicated variability among similar plants, suggesting that a single small sampling of a young plant might not be a reliable measure of composition. It was also suggested that freeze-dried leaf samples might be less variable than fresh ones. In the present work, variability was minimized by comparing single leaves from the same stem position of each plant. Comparisons involving other leaves indicated that the glycoalkaloid content was not constant with respect to either time or position on the stem. In addition, the possible influence of differences in growing conditions at different times suggests that repeated plantings should include a known variety as a control to which other plants are compared. Variability was reduced by calculating glycoalkaloid concentrations on a dry weight rather than fresh weight basis. The method of drying the samples, however, had no influence on the variability of data. These considerations should be generally applicable to the sampling of replicate leaves of any plant for analysis of any components.     

Microsatellite-based assessment of five Solanum nigrum complex species and their progeny

Abstract

The aim of this study was to investigate the potential of simple sequence repeats as genetic markers in five species of the Solanum nigrum complex found in South Africa as well as their progeny. Some of the primers developed for sorghum, tomato, pepper and potato were used. Twenty-nine random sets of primers were selected for screening and seven were used to amplify each of the parents and their progeny. The optimum conditions were not the same for the various primer pairs with annealing temperatures of 45, 50 and 55 °C for different primers. The genetic-distance value amongst the species and their progeny varied from 0.33 to 0.55, showing the close relationship between entries and confirming the fact that the Solanum nigrum complex is a group of plants very closely related, yet the parents and their progeny could be clearly distinguished. This marker system therefore was very effective in distinguishing closely related accessions arising from a similar pedigree and could be used for molecular breeding in this species.     

Dehydrotomatine and alpha-tomatine content in tomato fruits and vegetative plant tissues

Tomato plants (Lycopersicon esculentum) synthesize the glycoalkaloids dehydrotomatine and alpha-tomatine, possibly as a defense against bacteria, fungi, viruses, and insects. We used a high-performance liquid chromatography method with UV detection at 208 nm for the analysis of these compounds in various tissues. An Inertsil ODS-2 column with a mobile phase of acetonitrile/20 mM KH2PO4 (24/76, v/v) afforded good separation of the two glycoalkaloids in mini-tomato extracts, fruit harvested at different stages of maturity, and calyxes, flowers, leaves, roots, and stems. The two peaks appeared at approximately 17 and approximately 21 min. Recoveries from tomato fruit extracts spiked with dehydrotomatine and alpha-tomatine were 87.7 +/- 6.8 and 89.8 +/- 3.4% (n = 5), respectively. The detection limit is estimated to be 0.39 microg for dehydrotomatine and 0.94 microg for alpha-tomatine. The dehydrotomatine and alpha-tomatine content of tomatoes varied from 42 to 1498 and 521 to 16 285 microg/g of fresh weight, respectively. The ratio of alpha-tomatine to dehydrotomatine ranged from 10.9 to 12.5 in tomatoes and from 2.3 to 7.8 in the other plant tissues. These results suggest that the biosynthesis of the glycoalkaloids is under separate genetic control in each plant part. Degradation of both glycoalkaloids occurred at approximately the same rate during maturation of the tomatoes on the vine. An Inertsil NH2 column, with acetonitrile/1 mM KH2PO4 (96/4, v/v) as the eluent, enabled the fractionation of commercial tomatidine into tomatidenol and tomatidine, the aglycons of dehydrotomatine and alpha-tomatine, respectively. The information should be useful for evaluating tomatoes and vegetative tissues for dehydrotomatine/alpha-tomatine content during fruit development and their respective roles in host-plant resistance and the diet.     

RAPD markers efficiently distinguish heterogenous populations of wild potato (Solanum)

Genetic characterization of germplasm is important for setting objective guidelines for conservation. One common problem found in genebanks is determining the value of populations with insufficient or unreliable data regarding their geographic origin. In this study, a genetic analysis based on RAPD markers was conducted to characterize a `mystery' population of Solanum sucrense, a polysomic tetraploid potato (2n=4x=48), for which adequate documentation was lacking. The comparative analysis of genetic similarities between this mystery population and each one of 30 other S. sucrense populations in the genebank revealed that all populations within this species, including the mystery population, are significantly different from being duplicates, and are therefore worthy of separate conservation. RAPD markers also distinguished the mystery population from closely related tetraploid species S. oplocense, S. gourlayi and S. tuberosum ssp. andigena, suggesting that it is also not a duplicate of a population of these species. If RAPDs can clearly differentiate populations within highly heterogeneous tetraploids like S. sucrense, they should be generally useful for determining germplasm organization within potato species.     

Sensitivity and specificity improvement of an ion sensitive field effect transistors-based biosensor for potato glycoalkaloids detection

Butyryl cholinesterase of different origin along with variations of the time of enzyme immobilization on the potentiometric transducer surface is offered to control the ion sensitive field effect transistor (ISFET)-based biosensor sensitivity. Because butyryl cholinesterase has been already used to develop the sensors for heavy metals, organophosphorus/carbamate pesticides, and steroidal glycoalkaloids analysis, the present study has been focused on the investigation and adjustment of the ISFET-based biosensor specificity exclusively to the glycoalkaloids. Utilization of ethylendiaminetetracetate (a complexon of heavy metal ions) and phosphotriesterase (a highly efficient catalyst for the hydrolysis of organophosphorus compounds) enabled the highly specific determination of glycoalkaloids at the background of lead and mercury (up to 500 microM of ions concentration) and paraoxon (up to 100 microM of pesticide concentration). The developed biosensor has been validated for glycoalkaloids detection in potato varieties cultivated in Ukraine, and the results obtained are compared to those measured by the methods of HPLC and TLC.