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Morphohistobiochemical characteristics of embryogenic and nonembryogenic callus cultures of sweet potato (Ipomoea batatas L.).

Ipomoea batatas callus culture raised in a medium supplemented with 2,4-D (2,4-dichlorophenoxy acetic acid) alone or 2,4-D in combination with benzyl adenine, were found to be embryogenic. Supplementation of exogenous chemicals, such as 5 g/l NaCI or 0.7 g/l proline together with a mild dose of 0.2 mg/l 2,4-D, enhanced somatic embryogenesis significantly in all the genotypes tested. Morphological, growth, physiological, histological, and biochemical characteristics of the embryogenic callus were different from the nonembryogenic callus. The former was compact, slow growing, and nodular compared with the fast growing, fragile, nonembryogenic callus. The embryogenic callus tissue had more dry matter, protein and reducing sugar contents compared with the less embryogenic callus. The somatic embryogenic response remained steady in the cultures for up to 96 weeks.

2,4-Dichlorophenoxyacetic Acid↗

Growth suppression of human cancer cells by polyphenolics from sweetpotato (Ipomoea batatas L.) leaves.

Sweetpotato leaves (Ipomoea batatas L.) contain a high content of polyphenolics that consist of caffeic acid, chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, 4,5-di-O-caffeoylquinic acid, and 3,4,5-tri-O-caffeoylquinic acid. We investigated the suppression of the proliferation of selected human cancer cells by phenolic compounds isolated from sweetpotato leaf. The human cancer cells used in this research included a stomach cancer (Kato III), a colon cancer (DLD-1), and a promyelocytic leukemia cell (HL-60). Caffeic acid and di- and tricaffeoylquinic acids dose-dependently depressed cancer cell proliferation, and the difference in sensitivity between caffeoylquinic acid derivatives and each kind of cancer cell was observed. Specifically, 3,4,5-tri-O-caffeoylquinic acid effectively depressed the growth of three kinds of cancer cells, and caffeic acid had an exceptionally higher effect against HL-60 cells than other di- and tricaffeoylquinic acids. In attempting to clarify the mechanism of growth suppression with the addition of the apoptotic inhibitor N-ethylmaleimide, we observed that the nuclear granulation in 3,4,5-tri-O-caffeoylquinic acid-treated HL-60 cells suggested apoptosis induction. This effect was confirmed by DNA fragmentation, an increase of caspase-3 activity, and expression of c-Jun. Growth suppression of HL-60 cells by 3,4,5-tri-O-caffeoylquinic acid was determined to be the result of apoptotic death of the cells. These results indicate that 3,4,5-tri-O-caffeoylquinic acid may have potential for cancer prevention.

Apoptosis↗

[Alkaloids of the hairy roots of Ipomoea batatas Lam].

AIM: To study the alkaloid constituents of the hairy roots of Ipomoea batatas Lam. METHODS: Chromatographic methods were used for isolation and purification. The structure was elucidated on the basis of spectral data. RESULTS: An alkaloid was isolated from the hairy roots of Ipomoea batatas. CONCLUSION: The compound is a novel indole-type alkaloid, which is named as ipomine A.

Indole Alkaloids↗

[Study on flavonoids from leaf of Ipomoea batatas].

OBJECTIVE: To investigate the flavonoid constituents of Ipomoea batatas. METHOD: The leaf was extracted with EtOH and partitioned with CHCl3, EtOAc and n-BuOH respectively. The chemical components of EtOAc fraction were isolated by silica gel and sephadex LH-20 column chromatography. The chemical structures were elucidated on the basis of physic-chemical properties and spectral data. RESULT: Five compounds were isolated and identified as: tiliroside (I), astragalin (II), rhamnocitrin (III), rhamnetin (IV) and kaempferol (V). CONCLUSION: All of these five compounds were obtained from this plant for the first time.

Benzopyrans↗

Seven new aminoacyl sugars in Ipomoea batatas.

An analysis of the polar extracts from sweet potato Ipomoea batatas (Convolvulaceae) led to the isolation of seven unknown aminoacyl sugars. On the basis of 1D, 2D NMR, and mass spectrometry data, the structures of the compounds were elucidated as: beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-valyl]-glucopyranoside (1), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-tyrosyl]-glucopyranoside (2), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-threonyl]-glucopyranoside (3), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-hystidyl]-glucopyranoside (4), 2-beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-alanyl]-glucopyranoside (5), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-tryptophanyl]-glucopyranoside (6), and beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-glycyl]-glucopyranoside (7).

Aminoacylation↗

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.

Animals↗

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.

Buffers↗

Occurrence and cytological mechanism of 2n pollen formation in a tetraploid accession of Ipomoea batatas (sweet potato).

We evaluated a 4x accession of Ipomoea batatas (L.) Lam. and four 2x accessions of Ipomoea triloba for 2n pollen production. Approximately 90% of the genotypes of accession 81.2 (I. batatas, 4x) produced 2n pollen with different frequencies. In contrast, none of the genotypes of I. triloba produced 2n pollen. The diameter of the 2n pollen was approximately 30% ((3) sqrt 2) larger than that of the n pollen, making it easy to identify, measure, and quantify. The correlation (r = 0.93**) between the frequency of giant pollen and the frequency of dyads and triads was highly significant, strongly suggesting that the giant pollen grains were 2n pollen. The 2n pollen producers presented either a parallel or tripolar spindle arrangement (Y shaped) at anaphase II instead of the normal 60 degrees crossed spindle orientation. These two abnormal spindle configurations produced dyads and triads, with different frequencies (13-67%), instead of tetrads. Occasionally a metaphase II spindle variation was found with a single fused spindle, which also forms a dyad. The correlation (r = 0.89**) between the frequency of 2n pollen and the frequency of parallel, fused, and tripolar spindle arrangements was also highly significant, suggesting that these abnormal spindle configurations are involved in the production of 2n pollen in I. batatas. When we evaluated the efficiency of 2n pollen in polyploidization using 4x x 4x (2n) crosses, all progenies were 4x, suggesting the existence of barriers to crossability between 4x genotypes and their 2n pollen-producer counterparts.

Anaphase↗

Uptake and transformation of phenol and chlorophenols by hairy root cultures of Daucus carota, Ipomoea batatas and Solanum aviculare.

Hairy root cultures of Daucus carota L., Ipomoea batatas L. and Solanum aviculare Forst were investigated for their susceptibility to the highly toxic pollutants phenol and chlorophenols and for the involvement of inherent peroxidases in the removal of phenols from liquid media. Roots of D. carota grew normally in medium containing 1000 micromol l(-1) of phenol, whilst normal growth of roots of I. batatas and S. aviculare was only possible at levels up to 500 micromol l(-1). In the presence of chlorophenols, normal root growth was possible only in concentrations not exceeding 50 micromol l(-1), except for I. batatas which was severely affected at all concentrations. Despite the reduction in biomass, the growth of S. aviculare cultures was sustained in medium containing up to 2000 micromol l(-1) of phenol or 2-chlorophenol, and up to 500 micromol l(-1) of 2,6-dichlorophenol. The amounts of phenol removed by the roots within 72 h of treatment were 72.7%, 90.7% and 98.6% of the initial concentration for D. carota, I. batatas and S. aviculare, respectively. For the removal of 2,6-dichlorophenol the values were, respectively, 83.0%, 57.7% and 73.1%. Phenols labelled with 14C were absorbed by the root tissues and condensed with highly polar cellular substances as well as being incorporated into the cell walls or membranes. The results suggest that S. aviculare, an ornamental plant, would be best suited for remediation trials under field conditions.

Chlorophenols↗

Developmental changes induced by gamma irradiation in Ipomoea batatas L. Lam (sweet potato).

Storage roots of Ipomoea batatas L. Lam were exposed to gamma irradiation at 90, 180, and 240 Gy. The two highest doses caused a delay in the initiation of rooting and especially of shooting; they also caused formation of rosette-like, stemless shoots without apical meristems. The storage roots irradiated with 90 Gy did form stems, but 50% of them had no apical meristem. The irradiated storage roots produced absorbent roots which developed shoots after 70 days of growth. Nonirradiated storage roots did not differentiate shoots from the absorbent roots. When grown in vitro, phytomers, stem segments with leaves and an axillary meristem, separated from the shoots of irradiated storage roots and exhibited growth aberrations, very intense rooting, and a delay in shooting. Phytomers from nonirradiated normal plants were irradiated with 10, 20, 30, 40, and 90 Gy and grown in a hormone-free medium. The 40- and 90-Gy doses delayed shooting as well as rooting. Only phytomers exposed to 40 and 90 Gy differentiated shoots from the absorbent roots. A stimulation of growth revealed in the accumulation of dry mass was found in the shoots of phytomers irradiated with 10 to 30 Gy. The long after-effects of irradiation as well as possible causes of growth stimulation are discussed.

Dose-Response Relationship, Radiation↗

Rapid and reliable method of extracting DNA and RNA from sweetpotato, Ipomoea batatas (L). Lam.

A quick, simple and reliable method of extracting DNA from sweetpotato (Ipomoea batatas (L.) Lam.) has been developed. The method was applied successfully for extraction of total DNA from leaves and total RNA from leaves and various tissues. The yield of DNA extracted by this procedure was high (about 1 mg/g leaf tissue). The extracted DNA was completely digested by restriction endonucleases indicating the absence of common contaminating compounds. The absorbancy ratios of A260/A230 and A260/A280 of isolated RNA were approx. 2 and the yield was about 0.2 mg/g fresh wt. CIPK and tublin genes were successfully amplified by RT-PCR, suggesting the integrity of isolated RNA. The total DNA and RNA isolated by this method was of sufficient quality for subsequent molecular analysis.

2-Propanol↗

Characterization and immunostimulatory activity of an (1-->6)-a-D-glucan from the root of Ipomoea batatas.

The polysaccharide PSPP (purified sweet potato polysaccharide), isolated and purified from the roots of Ipomoea batatas, was found to be a glucan with a molecular weight of 53.2 kDa and specific rotation of +115.0 degrees (ca. 0.80, H(2)O). On the basis of methylation analysis, periodate oxidation, Smith degradation, infra-red spectroscopy, and (13)C NMR, the polysaccharide was confirmed as a (1-->6)-alpha-D-glucan. We evaluated the effects of polysaccharide PSPP on the in vivo immune function of mouse. Mice were treated with the polysaccharide PSPP (50, 150, and 250 mg/kg body weight) for 7 days. Phagocytic function, proliferation of lymphocytes, natural killer cell activity, hemolytic activity, and serum IgG concentration of the mice were studied. At the dose of 50 mg/kg, significant increments in proliferation of lymphocytes (P<0.05) and serum IgG concentration (P<0.05) were observed. At the dose of 150 and 250 mg/kg, significant increments (P<0.01 or P<0.05) were observed in all tested immunological indexes. A dose-dependent manner was demonstrated in phagocytic function, hemolytic activity, and serum IgG concentration, but not in proliferation of lymphocytes and natural killer cell activity. This suggests that PSPP improve the immune system and could be regarded as a biological response modifier.

Animals↗

Medium alterations improve regrowth of sweet potato (Ipomoea batatas [L.] lam.) shoot tips cryopreserved by vitrification and encapsulation-dehydration.

In vitro grown sweet potato (Ipomoea batatas [L.] Lam.) shoot tips were successfully cryopreserved by both solution based and encapsulation-dehydration vitrification methods. Improved recovery medium enhanced recovery for both vitrification procedures. The effects of sucrose preculture, cryoprotectant preculture and post-warm recovery media on regrowth following LN exposure were investigated. Sucrose preculture was critical for the survival of sweet potato shoot tips cooled to ca. -200 degrees C. Cryoprotectant preculture with 2 M glycerol + 0.4 M sucrose before dehydration with PVS2 gave the highest recovery following LN exposure. The viability of cooled samples following culture on ammonium-free MS medium for 5 days was increased three-fold over those cultured on MS medium. The improvement in recovery by altering post-warming conditions suggests that cryoinjury is not always lethal and can be ameliorated by suitable culture conditions.

Cryopreservation↗

Purification and characterization of p-coumaroyl-D-glucose hydroxylase of sweet potato (Ipomoea batatas) roots.

p-Coumaroyl-D-glucose hydroxylase in sweet potato (Ipomoea batatas Lam.) has been purified to apparent electrophoretic homogeneity using a combination of anion-and cation-exchange, hydrophobic and gel filtration chromatography. The purified enzyme was a monomer with a molecular weight of 33,000 and pI of 8.3. The purified enzyme showed not only hydroxylase activity but also polyphenol oxidase activity. L-Ascorbic acid was the best electron donor for the hydroxylation reaction, which had an optimum pH of 7.0. The enzyme hydroxylated p-coumaroyl-D-glucose, p-coumaric acid, and p-cresol but did not act on o-coumaric acid, m-coumaric acid, 4-hydroxy-3-methoxycinnamic acid, p-hydroxybenzoic acid or L-tyrosine. While the enzyme utilized p-coumaroyl-D-glucose and p-coumaric acid equally at pH 7.0, it hydroxylated only p-coumaroyl-D-glucose at pH 5.5. The enzyme oxidized diphenols such as D,L-(3,4-dihydroxyphenyl) alanine and caffeic acid, but exhibited no clear pH optimum in this reaction characteristic of polyphenol oxidase. Both the hydroxylase and the polyphenol oxidase activities were strongly inhibited by beta-mercaptoethanol, diethyldithiocarbamate, KCN, and p-coumaric acid (in concentrations higher than 5 mM). Ammonium sulfate and sodium chloride activated the hydroxylase activity but not the polyphenol oxidase activity of the enzyme. The enzyme activity and L-ascorbic acid contents changed in a manner suggesting their involvements in chlorogenic acid biosynthesis during incubation of sliced sweet potato root tissues.

Ammonium Sulfate↗

Studies on aldose reductase inhibitors from natural products. IV. Constituents and aldose reductase inhibitory effect of Chrysanthemum morifolium, Bixa orellana and Ipomoea batatas.

The hot water extracts of Chrysanthemum morifolium, Bixa orellana and Ipomoea batatas, were found to have potent inhibitory activity towards lens aldose reductase (AR). Ellagic acid (4) was isolated from C. morifolium and I. batatas, isoscutellarein (7) from B. orellana and 3,5-dicaffeoylquinic acid (10) from I. batatas, respectively, as potent inhibitors.

Aldehyde Reductase↗

Anthocyanin compositions in sweetpotato (Ipomoea batatas L.) leaves.

The anthocyanin composition of three varieties, Simon No. 1, Kyushu No. 119, and Elegant Summer, in sweetpotato (Ipomoea batatas L.) leaves was examined for promoting new uses. Fifteen anthocyanin compounds were identified and measured. HPLC clearly showed quantitative differences, but not qualitative ones. The anthocyanins were acylated cyanidin and peonidin type. The result suggests that the major anthocyanin composition of sweetpotato leaves is cyanidin type.

Acylation↗

Starch-branching enzyme I gene (IbSBEI) from sweet potato (Ipomoea batatas); molecular cloning and expression analysis.

The cDNA of the starch-branching enzyme I gene (IbSBEI) in the sweet potato (Ipomoea batatas) has been cloned and sequenced. The IbSBEI amino acid sequence was 81% identical to that of potato StSBEI. DNA gel-blot analyses demonstrated that at least two copies of IbSBEI are present in the sweet potato genome. IbSBEI was strongly expressed in tuberous roots. Transcript levels in the roots of single leaf cuttings were extremely low during the first 15-40 d after planting and continuously increased up to 50 d, by which time the tuberous roots had almost completely developed. This indicates that IbSBEI may work in concert with the AGPase large subunit during the primary phase of starch granule formation.

1,4-alpha-Glucan Branching Enzyme↗

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.

Antioxidants↗