Invertase inhibitors from sweet potato (Ipomoea batatas): purification and biochemical characterization

Two proteinaceous invertase inhibitors, designated ITI-L and ITI-R, were purified to electrophoretic homogeneity. ITI-L was purified from acetone powder of sweet potato leaves through sequential steps entailing buffer extraction, acid treatment, DEAE-Sephacel ion-exchange chromatography, and Sephacryl S-100 gel filtration. ITI-R was purified from sweet potato tuberous roots by sequentially applying buffer extraction, Con A-Sepharose affinity chromatography, DEAE-Sephacel ion-exchange chromatography, Sephacryl S-200, and Superose 12 gel filtration. The optimal pHs for interaction between ITI-L and ITI-R and acid invertase from sweet potato leaves were 5.5 and 5.0, respectively. The molecular masses of ITI-L and ITI-R were 10 and 22 kDa, respectively, as estimated by both gel filtration and SDS-PAGE. Both inhibitors were thermostable (90% of the activity remained after incubation at 100 degrees C for 20 min), and Western blotting showed them to be immunologically related.     

Potential chemopreventive properties of extract from baked sweet potato (Ipomoea batatas Lam. Cv. Koganesengan)

The extract from baked sweet potato (Ipomoea batatas Lam. cv. Koganesengan) showed potential cancer-preventing effects. The extract was partially fractionated to four fractions (I, II-a II-b, and III) by Sephadex G-25 gel chromatography. The cytotoxicity against human myelocytic leukemia HL-60 cells, the suppression of TPA-induced transformation in mouse skin JB6 C141 cells, the apoptosis inducing activity in HL-60 cells, and the scavenging capacity against DPPH radical were tested on the four fractions. Fractions II-a and III showed markedly strong radical scavenging effects on the DPPH radical, coinciding with the high content of total phenolic compounds in the fractions. Both of these fractions suppressed strongly the proliferation of HL-60 cells with apoptosis induction in a dose-dependent manner. Moreover, the two fractions markedly blocked TPA-induced cell transformation in the JB6 cell line. Taken together, these data suggest that the water extract from baked sweet potato had potential chemopreventive properties.     

步行式甘薯碎蔓还田机的设计与试验

为缓解中国丘陵坡地小田块甘薯碎蔓机械短缺问题,研究设计了步行式甘薯碎蔓还田机.该文在分析整机结构的基础上具体阐述了甘薯碎蔓还田机工作原理,阐明了碎蔓装置、刀座防磨损设计、导向轮调节机构和传动系统等关键部件的设计.甘薯秧蔓粉碎合格率、留茬高度和伤薯率是评价甘薯碎蔓还田机的主要指标,该文在单因素试验基础上运用Box-Benhnken的中心组合试验方法对甘薯碎蔓还田机的工作参数进行试验研究,以刀辊转速、离地间隙、刀片间距进行三因素三水平二次回归正交试验设计.建立了响应面数学模型,分析了各因素对作业质量的影响,同时,对影响因素进行了综合优化.试验结果表明:粉碎合格率影响显著顺序为刀辊转速＞离地间隙＞刀片间距;留茬高度影响显著顺序为离地间隙＞刀辊转速＞刀片间距;伤薯率影响显著顺序为离地间隙＞刀辊转速＞刀片间距;田间试验结果表明:最优工作参数组合为刀辊转速为1 950 r/min、离地间隙为25 mm、刀片间隙为40 mm,此时秧蔓粉碎合格率为94.88％、留茬高度为47.08 mm、伤薯率为0.23％,与理论优化值对比误差小于5％.研究结果可为步行式甘薯碎蔓还田机的结构完善和作业参数优化提供参考.     

Structural characterization and hypoglycemic effects of arabinogalactan-protein from the tuberous cortex of the white-skinned sweet potato (Ipomoea batatas L.)

An arabinogalactan-protein (WSSP-AGP) was isolated from the tuberous cortex of the white-skinned sweet potato (WSSP; Ipomoea batatas L.). It consists of 95% (w/w) carbohydrate and 5% (w/w) protein with high contents of hydroxyproline, alanine, and serine. Its sugar composition is α-L-Rha:α-L-Ara:β-D-Gal:β-D-GlcA in a molar ratio of 1.0:4.1:7.6:1.3. Its weight-average molecular weight was estimated to be 126,800 g/mol by high-performance size exclusion chromatography coupled with multiangle laser light scattering. Structural analysis indicated that WSSP-AGP is a (1→3)-β-D-galactan highly branched at O-6 with (1→6)-β-D-galactan, in which the branched chains are substituted at the O-3 position with α-L-Araf-(1→ and α-L-Araf-(1→5)-α-L-Araf-(1→ and at the O-6 position typically with α-L-Rhap-(1→4)-β-D-GlcAp-(1→ as terminating groups. Continuous administration of WSSP-AGP to KKAy mice significantly lowered fasting plasma glucose levels. This indicates that WSSP-AGP plays an important role in the hypoglycemic effects of WSSP.     

Assessing genetic diversity of sweet potato (Ipomoea batatas (L.) Lam.) cultivars from tropical America using AFLP

The sweet potato genebank at the International Potato Center (CIP) maintains 5,526 cultivated I. batatas accessions from 57 countries. Knowledge of the genetic structure in this collection is essential for rational germplasm conservation and utilization. Sixty-nine sweet potato cultivars from 4 geographical regions (including 13 countries) of Latin America were randomly sampled and fingerprinted using AFLP markers. A total of 210 polymorphic and clearly scorable fragments were generated. A geographic pattern of diversity distribution was revealed by mean similarity, multidimensional scaling (MDS), and analysis of molecular variance (AMOVA). The highest genetic diversity was found in Central America, whereas the lowest was in Peru-Ecuador. The within-region variation was the major source of molecular variance. The between-regions variation, although it only explains 10.0% of the total diversity, is statistically significant. Cultivars from Peru-Ecuador, with the lowest level of within region diversity, made the most significant contribution to the between region differentiation. These results support the hypothesis that Central America is the primary center of diversity and most likely the center of origin of sweet potato. Peru-Ecuador should be considered as a secondary center of sweet potato diversity.     

Synergistic effects of vesicular-arbuscular mycorrhizal fungi and diazotrophic bacteria on nutrition and growth of sweet potato (Ipomoea batatas)

Summary Sweet potatoes were micropropagated and then transplanted from axnic conditions to fumigated soil in pots in the greenhouse. Spores of Glomus clarum were obtained from Brachiaria decumbens or from sweet potatoes grown in soil infected with this fungus and with an enrichment culture of Acetobacter diazotrophicus. Three experiments were carried out to measure the beneficial effects of vesicular-arbuscular mycorrhizal (VAM) fungi-diazotroph interactions on growth, nutrition, and infection of sweet potato by A. diazotrophicus and other diazotrophs obtained from sweet potato roots. In two of these experiments the soils had been mixed with ¹⁵N-containing organic matter. The greatest effects of mycorrhizal inoculation were observed with co-inoculation of A. diazotrophicus and/or mixed cultures of diazotrophs containing A. diazotrophicus and Klebsiella sp. The tuber production was dependent on mycorrhization, and total N and P accumulation were increased when diazotrophs and G. clarum were applied together with VAM fungal spores. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM fungi or when present within G. clarum spores. More pronounced effects on root colonization and intraradical sporulation of G. clarum were observed when A. diazotrophicus was co-inoculated. In non-fumigated soil, dual inoculation effects, however, were of lower magnitude. ¹⁵N analysis of the aerial parts and roots and tubers at the early growth stage (70 days) showed no statistical differences between treatments except for the VAM+Klebsiella sp. treatment. This indicates that the effects of A. diazotrophicus and other diazotrophs on sweet potato growth were caused by enhanced mycorrhization and, consequently, a more efficient assimilation of nutrients from the soil than by N₂ fixation. The possible interactions between these effects are discussed.     

Dehydroascorbate reductase cDNA from sweet potato (Ipomoea batatas [L.] Lam): expression, enzyme properties, and kinetic studies

A cDNA encoding a putative dehydroascorbate reductase (DHAR) was cloned from sweet potato. The deduced protein showed a high level of sequence homology with DHARs from other plants (67 to approximately 81%). Functional sweet potato DHAR was overexpressed and purified. The purified enzyme showed an active monomeric form on a 12% native PAGE. The protein's half-life of deactivation at 50 degrees C was 10.1 min, and its thermal inactivation rate constant K(d) was 6.4 x 10(-2) min(-1). The enzyme was stable in a broad pH range from 6.0-11.0 and in the presence of 0.8 M imidazole. The K(m) values for DHA and GSH were 0.19 and 2.38 mM, respectively.     

An arsenate reductase homologue possessing phosphatase activity from sweet potato (Ipomoea batatas [L.] Lam): kinetic studies and characterization

A cDNA encoding a putative arsenate reductase homologue (IbArsR) was cloned from sweet potato (Ib). The deduced protein showed a high level of sequence homology (16-66%) with ArsRs from other organisms. A 3-D homology structure was created based on AtArsR (PDB code 1T3K ) from Arabidopsis thaliana. The putative active site of protein tyrosine phosphatase (HC(X)(5)R) is conserved in all reported ArsRs. IbArsR was overexpressed and purified. The monomeric nature of the enzyme was confirmed by 15% SDS-PAGE and molecular mass determination of the native enzyme via ESI Q-TOF. The IbArsR lacks arsenate reductase activity but possesses phosphatase activity. The Michaelis constant (K(M)) value for p-nitrophenyl phosphate (pNPP) was 11.11 mM. The phosphatase activity was inhibited by 0.5 mM sodium arsenate [As(V)]. The protein's half-life of deactivation at 25 °C was 6.1 min, and its inactivation rate constant K(d) was 1.1 × 10(-1) min(-1). The enzyme was active in a broad pH range from 4.0 to 11.0 with optimum activity at pH 10.0. Phosphatase would remove phosphate group from nucleic acid or dephosphorylation of other enzymes as regulation signaling.     

Active recombinant thioredoxin h protein with antioxidant activities from sweet potato (Ipomoea batatas [L.] Lam Tainong 57) storage roots

Recombinant thioredoxin h (Trx2) overproduced in Escherichia coli (M15) was purified by Ni2+-chelated affinity chromatography. The molecular mass of Trx2 is approximately 1.4 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Total antioxidant status, 1,1-diphenyl-2-picrylhydrazyl (DPPH) staining, reducing power method, Fe2+-chelating ability, ferric thiocyanate (FTC) method, and protection of calf thymus DNA against hydroxyl radical-induced damage were studied. The thioredoxin h protein with a concentration of 12.5 mg/mL exhibited the highest activity (expressed as 0.37 +/- 0.012 mM ABTS* radical cation being cleared) in a total antioxidant status test. In the DPPH staining thioredoxin h appeared as white spots when it was diluted to 50 mg/mL (a final amount of 15 microg). Like the total antioxidant status, the reducing power, Fe2+-chelating ability, FTC activity, and protection against hydroxyl radical-induced calf thymus DNA damage were found with the thioredoxin h protein. It was suggested that thioredoxin h might contribute to its antioxidant activities against hydroxyl and peroxyl radicals.     

Analysis of genetic diversity in a sweet potato (Ipomoea batatas L.) germplasm collection from Tanzania as revealed by AFLP

Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.     

Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Saccharum spp.), and sweet sorghum (Sorghum vulgare)

Summary Spores of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus clarum obtained from sweet potatoes grown in soil inoculated with this fungus and with an enrichment culture of Acetobacter diazotrophicus contained A. diazotrophicus and several other bacteria, including a diazotrophic Klebsiella sp. Inoculation of micropropagated sweet potatoes with G. clarum and A. diazotrophicus enhanced spore formation in soil compared to VAM inoculation alone. Plants inoculated with VAM spores containing the bacteria showed additional increases in the number of spores formed within roots. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM or when present within VAM spores. Micropropagated sugarcane seedlings inoculated with the same VAM spores containing the diazotrophs also contained much higher numbers of A. diazotrophicus in aerial parts than seedlings inoculated in vitro with the bacteria alone. When grown in non-sterile soil, the sugarcane seedlings again showed the greatest infection of aerial parts after inoculation with VAM spores containing the diazotrophs. This treatment also increased VAM colonization and the numbers of spores formed within roots. Similar effects were observed in sweet sorghum except that the aerial plant parts were not infected by A. diazotrophicus.     

Determination of pharmacologically active ingredients in sweet potato (Ipomoea batatas L.) by capillary electrophoresis with electrochemical detection

Sweet potato (Ipomoea batatas L.), in which vitamin C, chlorogenic acid, caffeic acid, quercetin, and rutin are abundant, is one of the functional food products aimed at introducing human dietary ingredients that aid specific body functions in addition to being nutritious. A method based on capillary electrophoresis with electrochemical detection (CE-ED) to qualitatively and quantitatively determine the pharmacologically active ingredients in sweet potato has been developed by our group. The effects of working electrode potential, pH and concentration of running buffer, separation voltage, applied potential, and injection time on CE-ED were investigated. Under the optimum conditions, the analytes could be well-separated within 20 min at the separation voltage of 18 kV in a 60 mmol L(-1) Borax running buffer (pH 9.0). A good linear relationship was established between peak current and concentration of analytes over 2 orders of magnitude with detection limits (S/N = 3) ranging from 7.14 x 10(-7) to 2.88 x 10(-7) g mL(-1) for all target ingredients. The satisfactory results show that this method is very successful and effective for the analysis of real samples.     

Distribution of potassium fractions in sweet potato (Ipomoea batatas) garden soils in the Central Highlands of Papua New Guinea and management implications

Previous studies indicated that potassium (K) deficiency is an important soil‐related factor for yield decline of the sweet potato gardens in the Central Highlands of Papua New Guinea, where sweet potato is an important staple food crop. An effort was made to characterize various fractions of K in the diverse soils of this region under sweet potato, to ascertain the probable reasons behind the observed K deficiency and its relationship to decreasing yield trends. Soils from two depths (0–10 cm) and (10–20 cm) in two types of gardens (old and new gardens) were assessed for different fractions of soil potassium in volcanic and non‐volcanic soil groups. Volcanic soils (Hydrandepts and Andaquepts) were significantly lower (P < 0.05) in exchangeable K than the non‐volcanic soils (Dystropepts, Tropoqualfs and Eutropepts). Mean exchangeable K content of the non‐volcanic soils was 95.5 mg/kg, whereas that of volcanic soils was 72.4 mg/kg. Similarly, new gardens had an average exchangeable K content of 94.1 mg/kg, which was significantly greater than 71.6 mg/kg soil of older gardens. Non‐exchangeable K content differed significantly (P < 0.001) between the soil types; mean K content was 85.9 mg/kg for the volcanic soils, whereas in non‐volcanic soils, it was 184.9 mg/kg. Garden types also differed significantly (P < 0.05) with respect to non‐exchangeable K content; new gardens registering higher average values (by almost 20%) than the older gardens. Multiple regression analysis showed that variability in the tuber yield was as a result of variability of water soluble and exchangeable K (up to 22%), non‐exchangeable K (2%), mineral K (4%) and leaf K concentrations (10%). Older gardens, which are in volcanic soil groupings, are more susceptible to the K depletion problem because of continuous sweet potato cultivation, possibly owing to their lower K reserves. Such gardens should be managed either with sufficient fallow periods for regeneration of soil fertility or with suitable application of mineral K fertilizers to enhance productivity.     

Calf thymus DNA-binding ability study of anthocyanins from purple sweet potatoes ( Ipomoea batatas L.)

A total of 10 anthocyanin compounds were identified from five purple sweet potato ( Ipomoea batatas L.) varieties, Qunzi, Zishu038, Ji18, Jingshu6, and Ziluolan, by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to assess their calf thymus DNA-binding ability in vitro. The interaction between anthocyanins and calf thymus DNA in Tris-HCl buffer solution (pH 6.9) was evaluated by fluorescence spectroscopy. Using ethidium bromide (EB) as a fluorescence probe, fluorescence quenching of the emission peak was seen in the DNA-EB system when anthocyanins were added, indicating that the anthocyanins bound with DNA. The acylated groups influenced the ability of the interaction with DNA. Anthocyanins from purple sweet potato with more acylated groups in sorphorose have a stronger binding ability with DNA.     

New polyphenol derivative in Ipomoea batatas tubers and its antioxidant activity

Four different polyphenolic compounds were isolated by chromatographic methods from methanolic and hydromethanolic extracts of Ipomoea batatas tuber flour. On the basis of UV, mass, and NMR analysis procedures, the structure of the isolated compounds were determined as 4,5-di-O-caffeoyldaucic acid (1), 4-O-caffeoylquinic acid (2), 3,5-di-O-caffeoylquinic acid (3), and 1,3-di-O-caffeoylquinic acid (4). To the best of our knowledge, this is the first report of isolation and characterization of compound 1. Then, we evaluated the antioxidant activity of daucic acid derivative by using DPPH and FRAP methods together with authentic antioxidant standards, l-ascorbic acid, tert-butyl-4-hydroxy toluene (BHT), and gallic acid. The activity of compound 1 in both methods was higher than that of all standards used at the same molar concentration.     

Oxidative comparison of emulsion systems from fish oil-based structured lipid versus physically blended lipid with purple-fleshed sweet potato (Ipomoea batatas L.) extracts

The effects of the purple-fleshed sweet potato extract (PFSPE) on oxidation stabilities of a model oil-in-water emulsion prepared with enzymatically synthesized fish oil-soybean oil structured lipid (SL) versus physically blended lipid (PBL) without modification were evaluated. The anthocyanins in PFSPE were analyzed and identified by HPLC-MS. The fatty acid composition of SL was similar to that of PBL, except palmitic acid (1.48 in PBL and 9.61% in SL) and linoleic acid (62.47 in PBL and 49.58% in SL). Peonidin 3-caffeoylsophoroside-5-glucoside, peonidin-3-(6',6'-caffeoylferuloylsophoroside)-5-glucoside, peonidin-dicaffeoylsophoroside-5-glucoside, peonidin 3-(6',6"-caffeoyl-p-hydroxybenzoylsophoroside)-5-glucoside were identified as the major anthocyanin compounds in PFSPE. Different levels (200, 500, 1000 ppm) of PFSPE were added into both SL- and PBL-based emulsions, with 200 ppm catechin as comparison. Oxidation was monitored by measuring the peroxide value and thiobarbituric acid reactive substances. The antioxidant activity of PFSPE increased with an increased concentration, the concentration of 1000 ppm showed high antioxidant ability similar to that of catechin in both PBL- and SL-based oil-in-water emulsions. It is notable that the SL-based emulsion appeared to have better oxidative stability than the PBL-based emulsion.     

Changes in polyphenolic content and radical-scavenging activity of sweet potato (Ipomoea batatas L.) during storage at optimal and low temperatures

Polyphenolic content and radical-scavenging activities (RSA) of four sweet potato (Ipomoea batatas L.) cultivars were characterized after storage at optimal (15 degrees C) or low temperature (5 degrees C) for 0, 13, 26, and 37 days. The polyphenolic content increased during storage in three cultivars but not in 'Murasakimasari'. The change in 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity (DPPH-RSA) correlated very well with polyphenolic content. The increases in polyphenolics and the RSA in 'Benimasari' were significantly greater during storage at 5 degrees C than at 15 degrees C. The main polyphenolic components in all cultivars were chlorogenic acid (ChA) and 3,5-di-O-caffeoylquinic acid (3,5-diCQA). ChA level increased more at 5 degrees C than at 15 degrees C, whereas that of 3,5-diCQA was greater at 15 degrees C. Caffeoylquinic acids and RSA in 'Murasakimasari', which contains a large amount of anthocyanin in flesh tissue, were extremely high at the beginning of storage and remained nearly constant or decreased over time. A non-caffeoylquinic acid component that increased during storage, especially in 'J-Red' at 15 degrees C, was purified by successive chromatographic steps. The isolate was identified as caffeoyl sucrose [CSu, 6-O-caffeoyl-(beta- d-fructofuranosyl-(2-->1))-alpha-D-glucopyranoside] by fast atom bombardment-mass spectroscopy (FAB-MS), infrared spectroscopy (IR), and nuclear magnetic resonance spectroscopy (NMR). These results suggest that storage under cultivar-dependent, controlled temperature is one approach for increasing desirable physiologic function associated with RSA of polyphenolic compounds in sweet potato roots.     

Quantity and potential biological activity of caffeic acid in sweet potato [Ipomoea batatas (L.) Lam.] storage root periderm

The caffeic acid content of storage root periderm and cortex tissues of genetically diverse sweet potato [Ipomoea batatas (L.) Lam.] cultivars and breeding clones was quantified by high-performance liquid chromatography. Periderm caffeic acid content of the clones ranged from 0.008 to 7.97 mg/g dry weight, whereas the highest cortex content was 0.047 mg/g. Clones varied greatly in periderm caffeic acid content in all experiments, but there were also differences between experiments in content averaged for all clones. This indicates that periderm caffeic acid content is subject to genetic and environmental influences. Caffeic acid inhibited the growth of four sweet potato pathogenic fungi and germination of proso millet seeds in bioassays. Inhibitory activity in the bioassays suggests that high periderm caffeic acid levels contribute to the storage root defense chemistry of some sweet potato genotypes.