Unsaponifiable lipids from haulm and tuber sprouts of potato (Solanum tuberosum L.)

Summary Fractions containing Δ⁵- and Δ⁷-sterols, 4-methyl-sterols, triterpenic alcohols, tocopherols and hydrocarbons were isolated by TLC from petroleum ether extracts of the haulm and tuber sprouts of cv Désirée. Sterol and triterpenic alcohol fractions of unsaponifiable lipids of the haulm and tuber sprouts were found to contain twelve sterols and four triterpenic alcohols, respectively. 24R-4-stigmasten-3-on, Δ⁷, Δ⁷, lanosterol, cycloeucalenol and obtisufoliol have not been identified previously in unsaponifiable lipids from haulm and sprouts. In the hydrocarbon fractions of the haulm extract, C₂₃-to C₃₃-n-parafins, C₁₉-and C₃₁-cyclohexyl hydrocarbons, C₂₂-to C₃₈-olefins and squalene were identified, and in the hydrocarbon fractions of the tuber sprouts C₁₂-to C₂₆-olefins and squalene were identified.     

A rapid method for the quantification of steroidal glycoalkaloids by reversed phase HPLC

Summary Methods were developed for clean-up of potato tuber extracts on solid phase extraction CN-cartridges and for analysis of steroidal glycoalkaloids by reversed phase HPLC. The alkaloids α-solanine, α-chaconine, α-solasonine and α-solamargine could be separated on C₈ or C₁₈ reversed phase columns using a mobile phase of acetonitrile/Tris-buffer (3∶2, v/v). The analysis time of tuber extracts containing these alkaloids was less then 4 min, if the pH of the mobile phase was adjusted to 7.4 and run at a flow rate of 0.5 ml/min.     

A method to assess cultivar differences in rate of physiological ageing of seed tubers

Inter-cultivar differences in rate of physiological ageing of potato tubers were assessed in a laboratory test and field tests. The physiological status of the seed tubers was varied by storing them at 4°C and a range of higher temperatures for 28 weeks (= excluding curing and pre-sprouting periods), after which tubers were desprouted. In a laboratory test, growth vigor was assessed by determining number and weight of sprouts after 28 days of incubation. In a field test, percentage emergence, total plant height, haulm and tuber weights were determined and a stand score was allocated 4 weeks after emergence of the 4°C storage treatment. Ageing indices were calculated by dividing the values for the warm-stored treatments by values for the tubers stored at 4°C. Using the tests the cultivars were scored from 0–10 and ranked accordingly. The indices were more evenly distributed over the range of 0–10 in the field test than in the laboratory test. Fresh haulm yield proved very suitable for calculating ageing indices but it could be replaced by a visual score of the haulm development without a great loss in accuracy     

Einfluss von Lagerung und Sorte auf die Verteilung der Glykoalkaloide in der Kartoffelknolle

Zusammenfassung Fünf Speisekartoffelsorten wurden w?hrend einer sechsmonatigen Lagerung fünfmal auf α-Solanin und α-Chaconin untersucht. Es wurden vier Gewebebereiche der Knolle getrennt überprüft. Der Gesamtglykoalkaloidgehalt betrug im Augenbereich 164, in der restlichen Schale 101 und in der Gef?ssbündelzone mit ?usserem Speichergewebe 8mg/100g. Trockensubstanz. Im restlichen Innenk?rper waren nur noch Spuren (0.164mg/100g TS) nachzuweisen. Zwischen den Sorten und diesen Gewebepartien traten statistisch gesicherte Wechselwirkungen auf, die im wesentlichen von einer Sorte verursacht wurden. W?hrend der Lagerung sind die Alkaloidgehalte leicht, aber statistisch nicht gesichert gesunken. α-Solanin und α-Chaconin kamen generell im Verh?ltnis von etwa 1∶1.5vor; die Korrelation zwischen beiden Formen ist sehr eng (r=0.865). Bezogen auf die gesamte Knolle lagen die Gesamtglykoalkaloide je nach Sorte zwischen 2.4 und 8mg/100g Frischsubstanz und damit im gesundheitlich unbedenklichen Bereich.

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Summary Five table potato cultivars (Erna, Granola, Gusto, Ulse, Margit) from the same source were examined for their content of α-solanine and α-chaconine during a sixmonths storage period. For the testing and subsequent analysis the tubers were dissected into four kinds of tissues: eye-zone, peel, vascular bundle region, medulla. The total glycoalkaloid content (TGA) in the total tuber mass varied between 2.4 and 8mg per 100g fresh matter depending on cultivar (Fig. 1). Despite significant differences between cultivars (Table 1), all potato cultivars remained within the safe range during the entire storage period. Remarkable differences in TGA per 100g dry matter could be noted in the tuber parts. For the eye-zones a TGA of 164mg was found; the peel region showed 101mg and the vascular bundle zone 8mg. The medulla of the tubers contained only traces of TGA (0.164mg/100g dry matter, Fig. 2). The statistical analysis only revealed significant interactions between cultivars and tuber parts (Table 1), mainly caused by the cultivar Gusto which showed higher contents of α-solanine and α-chaconine in its vascular bundle regions (Figs 3–5). The ratio between amounts of α-solanine and α-chaconine was approximately 1∶1.5, and after eliminating effects of cultivar, storage and tuber parts, the correlation coefficient between amounts was 0.865. This close correlation can be used for breeding. The non-significant, slightly negative effect of the six-months storage period on alkaloid content was noteworthy. It is assumed that only after long storage periods or through inappropriate storage (sprouting of tubers) a significantly higher glycoalkaloid content may be expected.

     

Hydrolysis of glycoalkaloids fromSolanum tuberosum L. haulm by enzymes present in plant material and by enzyme preparation

Summary This paper deals with the kinetics of enzymatic hydrolysis of glycoalkaloids from potato (Solanum tuberosum L.) haulm. The hydrolysis was carried out by the action of the enzymes present in fresh haulm, juice of fresh haulm and in haulm dried at various temperatures. The highest degree of enzymatic hydrolysis of 90% was obtained during fermentation of haulm dried at 40 °C after 30 h incubation time. The enzyme preparation was obtained from the juice of fresh potato haulm by using capillary dialysator HM 16 (AQM 1681, 1.6 m² Hemofan 8 υ). The best degree of enzymatic hydrolysis by enzyme preparation, 68%, was achieved after 20 h time of incubation. The enzyme preparation from juice of fresh haulm was characterized by K_m of 0.70 mM at pH 5.5 and 35 °C.     

Glycoalkaloid accumulation during tuber development of early potato cultivars

Summary Plants of early potato cultivars were sampled at successive harvests during tuber development and individual tubers were analysed for α-solanine and α-chaconine using high-performance liquid chromatography. Mean tuber total glycoalkaloid concentration (α-solanine plus α-chaconine) per plant decreased with time and statistically significant differences between cultivars were observed in the patterns of decline during tuber growth. The mean tuber ratio of α-chaconine to α-solanine increased during tuber development and was also affected by genotype. Total glycoalkaloid concentration for individual tubers of marketable sizes (up to 50 g fresh weight) exceeded the safety limit of 20 mg per 100 g fresh weight for cultivars Home Guard and British Queen but not for Rocket. These differences were attributable to differences in both rates and patterns of glycoalkaloid accumulation during tuber development, although in all cultivars results were consistent with glycoalkaloid accumulation occurring in the expanding peripheral cell layers of tubers for a considerable period after tuber initiation.     

Relation between growth conditions and dormancy of seed potatoes. 2. Effects of temperature

Summary In two indoor experiments under short day conditions, the effect of temperature during tuber bulking on dormancy of tubers was investigated for cvs Diamant and Désirée. Temperature treatments started after tuber initiation and lasted for 4 weeks, after which the haulm was removed. In Experiment 1, the day/night temperature regimes 18/12, 22/22, 26/18 and 32/12 °C (T18/12 etc.) were compared. In Experiment 2, three day temperatures (18, 24 and 30 °C) were combined with three night temperatures (12, 18 and 24 °C), resulting in nine treatments. The dormancy of cv. Diamant was shortest after very high day temperatures (30–32 °C), but intermediate day temperatures (22–26 °C) had no shortening effect compared to T18/12. Dormancy of cv. Désirée was not shortened, but rather tended to be prolonged by high temperatures (22–32 °C) during growth. High temperatures during growth resulted in more sprouts per tuber after dormancy had ended.     

Changes in the content of L-ascorbic acid,glucose, fructose,sucrose and total glycoalkaloids in potatoes (cv. Bintje) stored at 7, 16 and 28°C

Summary We investigated the changes in the content of L-ascorbic acid (L-AA), glucose, fructose, surose and total glycoalkaloids (TGA) during storage of potatoes, cv. Bintje, for 12 weeks at 7,16 and 28°C. The initial amount of L-AA was 8.2 mg/100g fresh weight; after 12 weeks' storage at 7°C it had decreased, while at 16 and 28°C it had increased. The sugar content went up most in potatoes stored at 28°C, but remained below the level at which potatoes taste sweet. The sugar content rose slightly at 16°C, while it decreased in tubers stored at 7°C. The initial TGA content, in equivalent amounts of α-solanine, was 3.6 mg/100 g fresh weight; after 12 weeks' storage it had decreased slightly at all three storage temperatures.     

Morphological changes in tuber buds during dormancy and initial sprout growth of seed potatoes

Summary Potato tuber dormancy is usually defined as lasting from tuber initiation until a sprout of 2 mm long has been formed under storage conditions optimal for sprouting. We tried to find out whether there is a period during which buds of seed tubers do not grow and whether different batches of seed take the same time to grow sprouts 2 mm long. We measured changes in number of leaf primordia and length of tuber buds of cvs Diamant and Désirée over two years. After early haulm pulling, buds did not grow for at least 60 days (‘Diamant’) or 95 days (‘Désirée’). Buds in both cultivars and two tuber weights of ‘Diamant’ took about 20 days from the estimated onset of sprouting to grow 2 mm long. We question whether this period is always similar and thus whether the moment sprouts 2 mm long have formed is a good criterion for the end of dormancy.     

马铃薯氮素的吸收、积累和分配规律

植株体内氮素浓度的高低反映了其生长势的强弱,马铃薯生育期间各器官氮素浓度的变化始终表现为叶片>地上茎>块茎,叶片中的氮素浓度高低反映了叶片光合活性的大小。马铃薯对氮的吸收与营养生长和块茎的增长密切相关,植株对氮的需求量受其生长状况所控制。而且,氮在植物体内很容易流动,块茎形成后,大量的氮素转移到块茎中,用于块茎的建成和营养贮存。马铃薯植株在淀粉积累开始后,各器官中氮素加快了向块茎的转移,使叶片和地上茎的衰老进一步加剧。因此,在马铃薯高产栽培实践中,须注重氮、磷、钾的适量与配合施用,使之既能满足块茎的形成与生长的需要,又可防止植株生长过旺或后期发生早衰。本试验表明,在因素中量(适量)组合下,每生产500kg块茎需要纯N2 65kg。     

Amylolytic activity in stored potato tubers. 1. Estimation usingp-nitrophenyloligosaccharides

Summary Methods for the estimation of amylolytic activity are reviewed. A procedure for the routine extraction of amylolytic activity from freeze-dried powder prepared from potato tubers is described. The extraction medium is buffered at pH 7.0 and contains glycerol, dithiothreitol, calcium chloride and the non-ionic detergent, nonidet P-40. α-Amylase activity and exoamylolytic activity were estimated in crude extracts of potato tubers using the substrates, blockedp-nitrophenyl-maltoheptaoside andp-nitrophenyl-maltopentaoside respectively. These substrates are included in kits supplied by Biocon, (UK) Ltd to measure the α- and ?-amylase activity in cereals. The validity of using these kits for the determination of α- and ?-amylase activity in potato tuber tissue is discussed.     

Amylase activity in potato tubers

Summary Amylase activity in extracts of sprouted tubers was optimised at final concentrations of soluble starch in the incubation medium of 0.6–2.0 mg cm⁻³. Optimum pH was 6. The exclusion of calcium ions from extraction and incubation media did not result in reduced enzyme activity. This, together with a shift in the absorption maximum of the starch-iodine complex almost identical to that observed with pure β-amylase, indicates the predominance of β-amylase in the extracts. Over a 15-min incubation period the linearity of the response was dependent upon the volume of tuber extract included in the assay medium. Gel filtration of extracts did not influence this response.     

Relation between growth conditions and dormancy of seed potatoes. 3. Effects of light

Summary The effects on the dormancy of progeny tubers of the timing and level of shading during plant growth were investigated in three field experiments. The tubers were harvested while immature. Shading (50–75%) the crop shortly before haulm pulling shortened dormancy by 5–7 days in two experiments. However, dormancy was hardly affected by 50% shading in one experiment, carried out in a sunny dry period. The effects of the photoperiod during tuber bulking on dormancy were studied in two indoor experiments. Plants were grown at 18/12°C (day 12h/night) and a 12 h photoperiod. Shortly after tuber initiation, the photoperiod was extended to 18 h for 4–6 weeks by photosynthetically inactive incandescent light, or kept at 12 h. The effects of the photoperiod on dormancy were up to 9 days, but differed between the cultivars and experiments. Generally, the effects on tuber dormancy of differences in growth conditions, as reported in this series of papers, were limited.     

Preparative isolation ofSolanum tuberosum L. glycoalkaloids by MPLC

Summary A new, efficient and economic method employing Medium Pressure Liquid Chromatography (MPLC) for the isolation of the two majorSolanum tuberosum L. glycoalkaloids (α-solanine and α-chaconine) is described. Potato peelings are homogenised with 5% acetic acid, the glycoalkaloids purified by filtration through an XAD-2 column and then by precipitation from the aqueous solution. The resulting glycoalkaloid fraction was purified by MPLC using a Silica Gel column and a CHCl₃:MeOH:2% NH₄OH mixture (70∶30∶5) as mobile phase to yield pure α-chaconine and a-solanine. This methodology can be used to obtain glycoalkaloids for enthomology and toxicological research where large amounts of these compounds are required.     

Effect of potato seed tuber glycoalkaloid content on subsequent infection byRhizoctonia solani

Since glycoalkaloids have been shown to have fungitoxic properties, it is important to know what impact this might have on potato plant breeding programs which select for low levels of total glycoalkaloids (TGA) in the tuber. Potato clones with TGA levels ranging between 1.6 and 32.8 mg/100 g fresh tissue were planted in two trials to evaluate the relationship between seed tuber TGA and incidence ofRhizoctonia solani infection in the developing plant. An inoculation procedure was followed in the greenhouse and field to establish uniform infection of potato seedlings. Plants were examined for lesion development on stems and stolons and assigned disease ratings. In the field, yield of malformed tubers was recorded and included in the overall disease ratings. The results reported herein indicate that TGA levels in the seed tuber are unrelated to the severity ofR. solani infection in the seedling.     

Effect of weight of potato minitubers on sprout growth,emergence and plant characteristics at emergence

Summary The behaviour of minitubers in five weight classes, having mid-point values between 0.19 and 3.00 g, was studied during sprouting and emergence under controlled conditions. Lighter tubers took longer to produce sprouts of 2 mm, and their sprouts grew more slowly between 2 and 4 mm and 4 and 6 mm. As sprouts lengthened their rate of growth increased. The influence of tuber weight was less for heavier tubers and also decreased as the sprouts grew longer. When tubers with sprouts of the same length were planted in pots, sprouts from lighter tubers took longer to emerge. Emergence was later and differences between weight classes were larger when tubers were planted deeper (6 or 9 cm) or when they had shorter sprouts at planting (2 or 4 mm). At emergence, plants from lighter tubers had thinner stems and lower stem and root weights, but higher stem weights proportional to tuber weights and higher shoot:root ratios.     

Enzymatic treatment to enhance carotenoid content in dehydrated marigold flower meal

The effect of enzymatic treatments using a commercial enzyme (Econase-cep at pH 5.0 and 0.1% w/w concentration) at different levels of dehydrated marigold meal (5, 10, 15 and 20% dry weight), to enhance carotenoid extraction, was evaluated. The AOAC method consisting of a simultaneous extraction and saponification by using hexane-ethanol-acetonetoluene (10:6:7:7 v/v) and metanolic KOH (40% w/v) was used to evaluate the carotenoid content. The measurement of carotenoids was also carried out in samples in which the water soluble compounds were previously eliminated (AOAC-H₂O). Total carotenoids ranged from 11.4 to 17. 4 g/kg and 18 to 24.7 g/kg of control and treated marigold meal, respectively. Highest amount of carotenoids were noted when 5% level of treated meal was used.     

辣木多糖酶促提取技术优化及不同部位含量研究

为研究辣木多糖的酶促提取工艺和辣木不同部位的多糖含量,在单因素试验的基础上,以多糖提取率为指标,选择纤维素酶用量、提取温度、提取时间、液料比为试验因素,采用二次回归正交旋转组合设计对纤维素酶辅助提取辣木多糖的工艺参数进行优化,并在此基础上研究辣木不同部位的多糖含量差异。4种因素对辣木多糖提取率的影响顺序依次为:纤维素酶用量>提取温度>液料比>提取时间。建立辣木多糖提取率(Y)与纤维素酶用量(X1)、提取温度(X2)、提取时间(X3)、液料比(X4)的二次正交回归模型:Y=18.6602+0.8134X1+0.7572X2–0.4312X3+0.6909X4–0.5181X12–0.4935X22–0.6277X42,该模型拟合度好,4个因素均对多糖提取率有显著影响(p<0.05)。通过回归模型获得优化的提取工艺为:纤维素酶用量1.60%,提取温度53℃,提取时间68 min,液料比52∶1,在此条件下辣木多糖提取率为19.83%,实际值与预测值一致。辣木不同部位的多糖含量结果表明,辣木根、花、嫩叶、茎中均含有丰富的多糖,其中根中含量最高且极显著高于其他部位(p<0.01),可进一步开发利用。采用二次回归正交旋转组合设计优化得到的辣木多糖酶促提取工艺条件准确可靠,可有效提高辣木多糖提取率并可在生产中推广应用。     

用蛋白酶组合对大豆分离蛋白改性的研究

用蛋白酶组合对大豆分离蛋白进行有限水解,所得水解液不苦,具有良好的风味.改性后的大豆分离蛋白的氮溶指数由未水解的82提高到95,20℃时溶解度在25%以上.经正交试验得到的大豆分离蛋白水解的最佳工艺参数为:加酶量,0.55%(w/w);固液比,1∶8(w/w);酶解时间,60 min.     

Detection of potato spindle tuber viroid by nucleic acid spot hybridization: Evaluation with tuber sprouts and true potato seed

A diagnostic test for the potato spindle tuber viroid (PSTV) that is based on hybridization of highly radioactive, recombinant DNA complementary to PSTV with PSTV bound to a nitrocellulose membrane and autoradiographic detection of the resulting DNA-RNA hybrids has been evaluated with tubers from 20 potato clones maintained at the International Potato Center (CIP) and with true seed obtained from healthy or PSTV-infected potato plants. The nucleic acid spot hybridization test detected PSTV not only in tuber sprouts from 10 clones that had previously tested positive in polyacrylamide gel electrophoretic (PAGE) analysis, but also in tuber sprouts from 9 clones that had previously tested negative in PAGE analysis. Further tests confirmed the presence of PSTV in these clones. The spot hybridization test detected PSTV in mixtures of seed extracts equivalent to as few as one seed from a PSTV-infected plant and 80 seeds from healthy plants. The spot hybridization test was shown to be more sensitive and reliable than PAGE analysis; it is suitable for the testing of large numbers of samples with a minimum expenditure of labor and materials.