五种茄科糖苷生物碱及其混合物的抗真菌活性研究

本文研究了五种茄科糖苷生物碱(茄碱、查茄碱、边缘茄碱、澳洲茄碱和番茄碱)对两种植物病原真菌白菜白斑病菌和葱紫斑病菌的抑制活性.结果表明番茄碱的抗真菌活性最强,其后依次是查茄碱、边缘茄碱和澳洲茄碱,茄碱的活性最弱;不同浓度茄碱和查茄碱(马铃薯中的两种糖苷生物碱)的混合物均具有协同抗真菌作用,且低浓度的混合物产生的协同作用效果较大;边缘茄碱和澳洲茄碱(龙葵中的两种糖苷生物碱)的混合物基本没有协同抗真菌作用;边缘茄碱和查茄碱的混合物以及澳洲茄碱和茄碱的混合物(均为来自不同植物的糖苷生物碱的混合物)在抗真菌活性上都呈现了相加关系.     

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.     

Synergistic interaction between the potato glycoalkaloids α-solanine and α-chaconine in relation to lysis of phospholipid/sterol liposomes

At pH 7.2, the steroidal glycoalkaloid α-chaconine disrupted phosphatidylcholine/cholesterol liposomes whereas α-solanine was virtually without effect. A glycoalkaloid mixture extracted from potato sprouts and comprising approximately equal amounts of solanine and chaconine had, at 150 μM, a lytic effect the same as a 150 μM solution of chaconine only. The apparent synergistic interaction between the two compounds was confirmed using 1:1 mixtures of authentic solanine and chaconine from different sources and of different batches. Combinations (1:1) of solanine or chaconine and tomatine or digitonin (both of which lysed liposomes) or β₂-chaconine (which is non-lytic) did not produce synergistic effects. The synergism between solanine and chaconine was observed only when the two compounds were present together, although the order of addition into the test system did not appear crucial. Pretreatment of liposomes with one glycoalkaloid and its subsequent removal did not permanently sensitize the membranes to the second glycoalkaloid. The magnitude of the synergism was dependent on the relative amounts of solanine and chaconine with maximal effects where chaconine comprised 40% or more of the mixture.     

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.     

Effects of glycoalkaloids from Solanum plants on cucumber root growth

The phytotoxic effect of four glycoalkaloids and two 6-O-sulfated glycoalkaloid derivatives were evaluated by testing their inhibition of cucumber root growth. The bioassays were performed using both compounds singly and in equimolar mixtures, respectively. Cucumber root growth was reduced by chaconine (C), solanine (S), solamargine (SM) and solasonine (SS) with IC₅₀ values of 260 (C), 380 (S), 530 (SM), and 610 μM (SS). The inhibitory effect was concentration-dependent. 6-O-sulfated chaconine and 6-O-sulfated solamargine had no inhibitory effects, which indicated that the carbohydrate moieties play an important role in inhibiting cucumber root growth. The equimolar mixtures of paired glycoalkaloids, both chaconine/solanine and solamargine/solasonine, produced synergistic effects on inhibition of cucumber root growth. By contrast, mixtures of unpaired glycoalkaloids from different plants had no obviously synergistic effects. The growth inhibited plant roots lacked hairs, which implied that inhibition was perhaps at the level of root hair growth.     

Identification of molecular markers associated with leptine production in a population of Solanum chacoense Bitter

  Solanum chacoense Bitter, a wild relative of the cultivated potato, produces several glycoalkaloids, including solanine, chaconine, and the leptines. The foliar-specific leptine glycoalkaloids are believed to confer resistance to the Colorado Potato Beetle (CPB). Using two bulked DNA samples composed of high- and low-percent leptine individuals from a segregating F₁ population of S. chacoense, we have identified two molecular markers that are closely linked to high percent solanine+chaconine and, conversely, to nil/low percent leptine. One of these, a 1,500-bp RAPD product (UBC370-1500), had a recombination value of 3% in the F₁ progeny, indicating tight linkage. UBC370-1500 mapped to the end of the short arm of potato chromosome 1, in the region of a previously mapped major QTL for solanidine, from a S. tuberosum (solanidine)×S. berthaultii (solasodine) cross. Taken together, these results suggest that either (1) a major locus determining solanidine accumulation in Solanum spp. is on chromosome 1 in the region defined by the RFLP markers TG24, CT197, and CT233, or (2) this region of chromosome 1 may harbor two or more important genes which determine accumulation of steroidal aglycones. These findings are important for the genetics of leptine (as well as other glycoalkaloid) accumulation and for the development of CPB-resistant potato varieties. Received: 5 March 1998 / Accepted: 28 July 1998     

Effect of steroidal glycoalkaloids of the potato on the permeability of liposome membranes

At pH 7.2, the potato glycoalkaloid α-chaconine caused release of entrapped peroxidase from phosphatidylcholine liposomes containing different free sterols but was ineffective against sterol-free liposomes. The alkaloid was able to complex with all the tested sterols in vitro although there was no close correlation between the extent of sterol binding and liposome disruption. α-Solanine also complexed with sterols in vitro but had no effects on sterol-containing liposomes under these conditions. Both sterol concentration and alkaloid concentration were limiting factors in the action of chaconine but did not markedly affect that of solanine. Solanine destabilized liposome membranes only at pH values of 8 and above but was less effective than chaconine. The importance of the carbohydrate moiety of glycoalkaloids was further demonstrated by the inability of β₂-chaconine to complex with sterols or disrupt liposomes.     

马铃薯糖苷生物碱对人血胆碱酯酶的抑制

马铃薯糖苷生物碱是一类有异味有毒性的含氨甾族化合物,其致毒机理与有机磷杀虫剂相似,表现出对人体内胆碱酯酶活性的抑制,使胆碱能神经兴奋增强,引起一系列中毒症状。从块茎萌发芽中提取的总糖苷生物碱的毒性较高,对人血胆碱酯酶的体外抑制率为６３．０８％,单个糖苷生物碱（α茄碱和α卡茄碱）的毒性较低,抑制率分别为５２．０８％和４１．１５％,总糖苷生物碱的水解产物糖苷配基基本无毒性,抑制率仅为１１．３６％。     

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.     

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.     

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.     

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.     

Congenital deformities produced in hamsters by potato sprouts

Dried gound potato sprout preparations from seven varieties produced congenital deformities in one strain of hamsters. Incidence of affected litters varied from 8 to 25%, depending on potato variety. Certain steroidal solanum and veratrum alkaloids produced similar defects. Neither peel nor tuber material was teratogenic from one of the potato varieties with highly teratogenic sprouts.     

Potato Glycoalkaloids: Chemistry,Analysis, Safety,and Plant Physiology

Potatoes, members of the Solanaceae plant family, serve as a major, inexpensive food source for both energy (starch) and good-quality protein, with worldwide production of about 350 million tons per year. U.S. per capita consumption of potatoes is about 61 kg/year. Potatoes also produce potentially toxic glycoalkaloids, both during growth and after harvest. Glycoalkaloids appear to be more toxic to man than to other animals. The toxicity may be due to anticholinesterase activity of the glycoalkaloids on the central nervous system and to disruptions of cell membranes affecting the digestive system and other organs. The possible contribution of glycoalkaloids to the multifactorial aspects of teratogenicity is inconclusive. Possible safe levels are controversial; guidelines limiting glycoalkaloid content of potato cultivars are currently being debated. This review presents an integrated, critical assessment of the multifaceted aspects of the role glycoalkaloids play in nutrition and food safety; chemistry and analysis; plant physiology, including biosynthesis, distribution, inheritance, host-plant resistance, and molecular biology; preharvest conditions such as soil composition and climate; and postharvest events such as effects of light, temperature, storage time, humidity, mechanical injury, sprouting inhibition, and processing. Further research needs are suggested for each of these categories in order to minimize pre- and postharvest glycoalkaloid synthesis. The overlapping aspects are discussed in terms of general concepts for a better understanding of the impact of glycoalkaloids in plants and in the human diet. Such an understanding can lead to the development of potato varieties with a low content of undesirable compounds and will further promote the utilization of potatoes as a premier food source for animals and humans.     

α and β-solamarine in kennebec Solanum tuberosum leaves and aged tuber slices

Two major steroid glycoalkaloids, in addition to α-solanine and α-chaconine, were isolated from leaves and aged tuber slices of potato, Solanum tuberosum L. var Kennebec. They are glycosides of tomatidenol and have been identified as α- and β-solamarine. The compounds were not found in tuber peel or freshly sliced Kennebec tubers or in 20 other cultivars.     

Host plant defenses of black (Solanum nigrum L.) and red nightshade (Solanum villosum Mill.) against specialist Solanaceae herbivore Leptinotarsa decemlineata (Say)

Black nightshade (Solanum nigrum, S. nigrum L.) and red nightshade ( Solanum villosum, S. villosum Mill.) are medicinal plants from the Solanaceae family that synthesize glycoalkaloids and other secondary metabolites. To recognize the potential insecticide activity of these compounds, leaf extracts (containing glycoalkaloid and methanol fractions) were tested for enzyme inhibition, antifeedant activity and toxicity. For in‐vitro glutathione S‐transferase (GST) inhibition activity, we used insecticide‐resistant Colorado potato beetle, Leptinotarsa decemlineata ( L. decemlineata; Say) midgut and fat‐body homogenate. In‐vivo toxicity and the antifeedant activity were performed using larval bioassays. The methanol extracts had greater GST inhibitory activity compared to the glycoalkaloids, as well as greater 2nd instar larvae mortality and antifeedant activity. Furthermore, the green leaf volatile compound, cis‐hex‐3‐enyl acetate, at the concentration of 5 ppm, caused 50% mortality of 2nd instar larvae. Our findings suggest the potential usefulness of S. nigrum and S. villosum extracts to control L. decemlineata.     

Amelioration by glucose-6-phosphate and NADP of potato glycoalkaloid inhibition in cell, enzyme and liposome assays.

Lysis of human erythrocytes by 20 microM chaconine was reduced by 0.5 mM glucose-6-phosphate (G6P) and NADP. Both compounds caused approximately 50% inhibition of haemolysis at 1 mM. Glucose, glucose-1-phosphate, rhamnose, galactose and galactose-6-phosphate were ineffective; NAD was effective, although not to the extent of NADP. Of the tested sugars, only G6P reduced solanine-induced haemolysis. G6P also reduced the synergistic haemolytic action of solanine and chaconine in combination. G6P and NADP at or above 5 mM antagonised chaconine-induced betanin loss from excised red beet root discs; NADP was more effective than G6P. Disruption of PC/cholesterol liposomes by chaconine and inhibition of acetylcholinesterase by chaconine or solanine, were unaffected by up to 10 mM NADP or 50 mM G6P.     

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.     

Construction of a potato fraction library for the investigation of functional secondary metabolites

A potato fraction library was constructed to investigate functional secondary metabolites from 8 cultivars: Kitahime, Pilka, Sakurafubuki, Atlantic, Toyoshiro, Snowden, Kitamurasaki, and Northern Ruby, which were divided into flower, leaf, stem, roots, tuber peel, and tuber. Each fraction was a semi-purified extract and about 800 fractions were prepared for the library. They were analyzed by DAD-LC/MS to obtain structural information and were evaluated for various biological activities. LC/MS data showed that each part had a specific characteristic for their constituents supported by principal component analysis (PCA). Approximately 40% of fractions showed significant biological activities at 30 μg/mL, especially the flower fractions showed strong cytotoxicity. PCAs based on the activity and LC/MS data suggested that the strong cytotoxicity of flowers was derived from a complex mixture of potato glycoalkaloids. In addition, tuber peel fractions showed strong antimalarial activity, which had not been reported before. Also, some fractions showed significant antibacterial activities.