Determination of total folate in plant material by chemical conversion into para-aminobenzoic acid followed by high performance liquid chromatography combined with on-line postcolumn derivatization and fluorescence detection

A procedure involving chemical conversion of all forms of folate present in plant material into para-aminobenzoic acid (PABA) and a liquid chromatographic-fluorimetric determination with on-line postcolumn derivatization is reported. All folates are cleaved with liberation of PABA by hydrogen peroxide followed by acid hydrolysis using concentrated hydrochloric acid (37%) at 110 degrees C for 6 h. The reaction yield for individual folates conversion to PABA ranged from 44.4 to 97.3%. PABA could be determined sensitively by on-line postcolumn derivatization with fluorescamine, the detection limit for PABA being 3.02 nM. On the basis of this principle, a method for the determination of total folate in plant material, including a purification step on an affinity column, is presented, which offers a sufficient sensitivity and selectivity for routine analysis of total folate in natural samples. The total folate contents of tomatoes, carrots, white cabbage, and spinach were determined, and the results were quite comparable to the data reported. The recovery of PABA and the comparison of total folate analysis in spinach on different occasions (over 6 months) are also reported. The method is reliable, universal for all folates, including polyglutamate and monoglutamate forms, and eliminates the need for a deconjugation step and multiple conversion reactions.     

Soybean mosaic virus Helper Component-Protease Alters Leaf Morphology and Reduces Seed Production in Transgenic Soybean Plants

ABSTRACT Transgenic soybean (Glycine max) plants expressing Soybean mosaic virus (SMV) helper component-protease (HC-Pro) showed altered vegetative and reproductive phenotypes and responses to SMV infection. When inoculated with SMV, transgenic plants expressing the lowest level of HC-Pro mRNA and those transformed with the vector alone initially showed mild SMV symptoms. Plants that accumulated the highest level of SMV HC-Pro mRNA showed very severe SMV symptoms initially, but after 2 weeks symptoms disappeared, and SMV titers were greatly reduced. Analysis of SMV RNA abundance over time with region-specific probes showed that the HC-Pro region of the SMV genome was degraded before the coat protein region. Transgenic soybean plants that expressed SMV HC-Pro showed dose-dependent alterations in unifoliate leaf morphologies and seed production where plants expressing the highest levels of HC-Pro had the most deformed leaves and the lowest seed production. Accumulation of microRNAs (miRNAs) and mRNAs putatively targeted by miRNAs was analyzed in leaves and flowers of healthy, HC-Pro-transgenic, and SMV-infected plants. Neither expression of SMV HC-Pro nor SMV infection produced greater than twofold changes in accumulation of six miRNAs. In contrast, SMV infection was associated with twofold or greater increases in the accumulation of four of seven miRNA-targeted mRNAs tested.     

Chromosome pairing within genomes in maize-Tripsacum hybrids

When the ten chromosomes of maize were inserted inlto a polyploid (2n = 72) Tripsacum dactyloides background they formed up to five pairs at meiosis. Two plants that each contained 36 Tripsacum and 14 maize chromosomes were deprived from the F(1) of maize x Tripsacum. Chromo. somes of these plants frequently formed 25 bivalents, 18 between Tripsacum chromosomes and seven between maize chromosomes. Maize chromosomes could be distinguished from Tripsacum chromosomes on the basis of size. The withint-genome pairing is probably induced by the genetic background.     

Risk assessment of Aphanomyces euteiches root rot disease: quantification of low inoculum densities in field soils using droplet digital PCR

European Journal of Plant Pathology - Fusarium verticillioides is a soil-borne plant pathogen of maize plants (Zea mays L.) responsible for major yield losses by causing root, stalk, and ear rot...     

Effect of l‐Cysteine on the Rheological Properties of Wheat Flour

Extrudate expansion of cereal‐based products is largely dependent on the molecular interactions and structural transformations that proteins undergo during extrusion processing. Such changes strongly influence the characteristic rheological properties of the melt. It is possible to modify rheological properties of wheat flour during extrusion processing, in particular shear viscosity, with cysteine. The objective of this work was to further develop an understanding of the molecular interactions and structural transformations of wheat flour from dynamic oscillatory rheological measurements. Temperature and frequency sweeps were conducted in the linear viscoelastic range of the material. Changes in the storage modulus (G′), the loss modulus (G″) and the loss tangent (tan δ) of 25% moisture wheat flour disks as a function of cysteine concentration (0–0.75%) were monitored. Molecular weight between cross‐links (M_c) and the number of cross‐links (N_c) per glutenin molecule were determined from frequency sweep data. Increasing cysteine concentration broke cross‐links by decreasing G′ maximum and increasing tan δ values. Molecular weight between cross‐links increased and the number of cross‐links decreased. G′ values from temperature sweeps showed a similar trend. This information leads to a better understanding of the viscoelastic behavior of wheat flour doughs during extrusion cooking and elucidation of protein‐protein reaction mechanisms and other interactions in extruded cereal‐based snack foods.     

Inhibition of tumorigenesis in ApcMin/+ mice by a combination of (-)-epigallocatechin-3-gallate and fish oil

The effect of a combination of (-)-epigallocatechin-3-gallate (EGCG) with fish oil on intestinal tumorigenesis in Apc (Min/+) mice fed a high-fat diet was investigated in the present study. The combined treatment of EGCG and fish oil for 9 weeks reduced the tumor number by 53% as compared to controls while neither agent alone had a significant effect. Apoptosis was significantly increased in all treatment groups. beta-Catenin nuclear positivity in adenomas from the combination group was lower than control mice, implicating the modulation of Wnt signaling by the combination. Fish oil and the combination significantly reduced prostaglandin E 2 (PGE 2) levels in small intestinal tumors as compared to controls, suggesting modulation of aberrant arachidonic acid metabolism by fish oil. Akt phosphorylation in adenomas was significantly reduced in all treatment groups, which may have contributed to the observed increase in apoptosis. The results indicate that a combination of low doses of EGCG and fish oil can inhibit tumor multiplicity in Apc (Min/+) mice.     

Verification of identities in global cacao germplasm collections

The ex situ conservation of cacao genetic resources is in the form of field genebanks. Misidentified trees though, represent a serious problem to (i) curators, who need to capture genetic diversity, and (ii) users of germplasm material, who must be certain of the identities of the accessions for proper evaluation of their results. The presence of mislabelled trees was assessed from data compiled in the International Cocoa Germplasm Database (ICGD) 2000 v4.1CD-ROM, which contains the published records of cacao accessions in global holdings. Circumstantial evidence revealed that many germplasm collections, especially those in Brazil (CEPEC), Colombia (ICA), Costa Rica (CATIE), Côte d'Ivoire, Mexico, Malaysia, Togo, Trinidad (ICG,T) and Venezuela (EEC) probably possessed misidentified material. The severity of the problem ranged from 15–44 % and is suggested as an area for immediate resolution. Based upon the morphological data provided by the ICGD CD-ROM, several accessions were highly suspect, including 'EET 19 [ECU]', 'EET 48 [ECU]', 'EET 162 [ECU]', 'ICS 1', 'ICS 6', 'ICS 43', 'ICS 89', 'ICS 100', 'IMC 67', 'PA 121 [PER]', 'POUND 7 [POU]', 'SCA 6', 'SPA 9 [COL]', 'UF 29', 'UF 613' and 'UF 667'. However, the identification of mislabelled trees should depend ultimately on molecular analysis. Hence, the use of voucher specimens for verification is forwarded and the concept of a public domain database for microsatellite analyses of germplasm holdings is raised.     

Reduced Atmospheric Manganese in Montreal Following Removal of Methylcyclopentadienyl Manganese Tricarbonyl (MMT)

Methylcyclopentadienyl manganese tricarbonyl (MMT) was used between 1990 and 2003 as an antiknock agent and as an octane booster in Canadian unleaded gasoline. Its combustion leads to Mn emissions. The objective of this research was to examine the variations in atmospheric Mn in Montreal (Canada) from 2001 to 2007, covering the period prior (2001?C2003) to and following (2005?C2007) MMT use. Three sampling stations were selected because of their proximity to roads with widely differing and well-known traffic patterns. Filters from 2001 to 2007 were obtained from the Montreal Urban Community. The first sample of each month was selected, and Mn analysis was performed by neutron activation analysis. Total suspended particulates (TSP) was calculated by weighing the filters before and after dust collection. Results show a significant decrease of Mn over time at each station, whereas TSP decreased significantly in two stations. Comparing atmospheric Mn during and after the period of use of MMT 2001?C2003 vs 2005?C2007 showed a significant decrease at all stations. For TSP, only one station showed borderline significant decline between these two periods. Overall, between the two periods, Mn and TSP decreased by 39% and 17%, respectively. These data suggest that the combustion of MMT led to an increase of airborne Mn of approximately 22%. These findings should help in decision-making processes concerning the use of MMT in gasoline in other countries.     

The use of ultrasonography in the reproductive evaluation of boars

The objective was to study the use of ultrasound as a complementary test in the breeding soundness evaluation in male pigs and study the pattern of echogenicity of the testicular parenchyma in boars of different racial groups. Twenty‐six adult boars from four different racial groups were used, 10 from the Piau breed (group 1), four from the commercial and finishing group (group 2), six Pietrain breed (group 3) and six from the Duroc breed (group 4). All animals were evaluated for breeding soundness evaluation and the ultrasound examination of the testicles. The groups of animals that were evaluated showed no difference in the main semen parameters that were evaluated, except for the sperm volume, concentration of the ejaculated sperm and the supravital staining; the lowest figures were for the animals from the Piau breed (group 1). In relation to the testicular biometrics, Duroc animals (group 4) had a greater scrotal width compared to the other groups. But when we assessed the intensity of pixels of the testicles, there was a difference between groups. The groups 2 (finishing animals), 3 (Pietrain) and 4 had no difference between themselves. Group 3 had greater pixel intensity in relation to group 1. Of the 26 animals studied, five showed an abnormality during ultrasound evaluation, like hydrocele, hyperechoic mass in the testicular parenchyma, cyst in the head of the epididymis and the presence of fluid in the head and tail of the epididymis. The various animal groups studied did not differ in the principal reproductive parameters evaluated, showing that despite the great variability of reproductive traits between breeds and within the same breed, the breeding soundness evaluation, the more complete it is, is essential for the selection of breeders and the ultrasonography of the reproductive system becomes an important addition in this examination.