Cytoplasmic substitution inSolanum. I. Seed production,germination, and sterilities of reciprocal backcross generations

Summary Genes ofSolanum tuberosum L. ssp.tuberosum were introduced into cytoplasm ofS. tuberosum ssp.andigena (Juz. & Buk.) Hawkes andS. phureja Juz. & Buk. to reduce cytoplasmic sterilities. Alleles that influenced berry set and seed content produced differences between reciprocal progenies. Their expressions were interpreted in terms of genes of the maternal and/or paternal parents. When the ssp.andigena genotype was introduced from the maternal parent if favoured high berry set and low seed number. Maternal ssp.tuberosum favoured lower berry set and higher seed number. The source ofS. phureja cytoplasm, aS. phureja×S. chacoense F₁, used as maternal parent favoured lower berry and seed set. During successive backcrosses progeny expressions approached those of the recurrent ssp.tuberosum pollen parents. A low level of pollen sterility occurred in BC₂ plants, unrelated to direction of cross or cytoplasmic factors. By BC₄, seed set and fertility were as good as ssp.tuberosum, and recurrent backcrossing could be terminated without loss of seed production or fertility. Pennsylvania Agricultural Experiment Station Journal Series No. 7493.     

Bactericidal kinetics of marine-derived napyradiomycins against contemporary methicillin-resistant Staphylococcus aureus

There is an urgent need for new antibiotics to treat hospital- and community-associated methicillin-resistant Staphylococcus aureus (MRSA) infections. Previous work has indicated that both terrestrial and marine-derived members of the napyradiomycin class possess potential anti-staphylococcal activities. These compounds are unique meroterpenoids with unusual levels of halogenation. In this paper we report the evaluation of two previously described napyradiomycin derivatives, A80915A (1) and A80915B (2) produced by the marine-derived actinomycete, Streptomyces sp. strain CNQ-525, for their specific activities against contemporary and clinically relevant MRSA. Reported are studies of the in vitro kinetics of these chemical scaffolds in time-kill MRSA assays. Both napyradiomycin derivatives demonstrate potent and rapid bactericidal activity against contemporary MRSA strains. These data may help guide future development and design of analogs of the napyradiomycins that could potentially serve as useful anti-MRSA therapeutics.     

New Alkaloids from the Mediterranean Sponge Hamigera hamigera

The Mediterranean sponge Hamigera hamigera (family Anchinoideae) was studied since its total extract showed deterrent activity in a fish feeding assay. Eight compounds were isolated from the biologically active fractions and four of these proved to be new natural products, hamigeroxalamic acid (1), hamigeramine (2), hamigeramide (3) and hamiguanosinol (5). The structures of the new compounds were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry.     

Development and evaluation of an indirect in situ polymerase chain reaction for the detection of porcine circovirus type 2 in formalin-fixed and paraffin-embedded tissue specimens

Taking advantage of the high sensitivity of polymerase chain reaction (PCR) and the cell-localizing ability of in situ hybridization (ISH), an indirect in situ PCR (ISPCR) method was developed for detecting the distribution of porcine circovirus type 2 (PCV2) in formalin-fixed and paraffin-embedded inguinal lymph nodes obtained from clinically healthy PCV2-carrier pigs and postweaning multisystemic wasting syndrome (PMWS)-affected pigs. Comparisons of the relative sensitivity of indirect ISPCR with other routinely used diagnostic methods for PCV2 indicated that nested PCR was the most sensitive method followed by indirect ISPCR, conventional PCR, ISH, and immunohistochemical (IHC) staining. Although indirect ISPCR, ISH, and IHC staining all revealed a similar signal distribution pattern of PCV2, using indirect ISPCR allowed specific amplification and detection of previously uneasily detected PCV2 signal than by routine ISH or IHC staining, particularly in those cells within the germinal center in clinically healthy PCV2-carrier pigs. Furthermore, six different PCV2 signal expression patterns in conjunction with the correlated lymphoid lesion stages were classified to describe the tissue morphological changes and viral infection. The result indicates that indirect ISPCR is a more effective, cell-based diagnostic tool with good specificity to detect limited PCV2 infection in formalin-fixed and paraffin-embedded tissue specimens and it would be a useful tool for further exploring the pathogenesis of PCV2 infection.     

High-amylose corn exhibits better antioxidant activity than typical and waxy genotypes

The consumption of fruits, vegetables, and whole grains rich in antioxidative phytochemicals is associated with a reduced risk of chronic diseases such as cancer, coronary heart disease, diabetes, Alzheimer's disease, cataract, and aged-related functional decline. For example, phenolic acids are among the main antioxidative phytochemicals in grains that have been shown to be beneficial to human health. Corn (Zea mays L.) is a major staple food in several parts of the world; thus, the antioxidant activity of several corn types was evaluated. The 2,2-Diphenyl-1-picryhydrazyl free radical (DPPH*) scavenging activity, total phenolic content (TPC), antioxidant capacity of lipid-soluble substances (ACL), oxygen radical absorbance capacity (ORAC), and phenolic acid compositions of typical and mutant genotypes (typical-1, waxy, typical-2, and high-amylose) were investigated. The DPPH* scavenging activity at 60 min was 34.39-44.51% in methanol extracts and 60.41-67.26% in HCl/methanol (1/99, v/v) extracts of corn. The DPPH* scavenging activity of alkaline hydrolysates of corn ranged from 48.63 to 64.85%. The TPC ranged from 0.67 to 1.02 g and from 0.91 to 2.15 g of ferulic acid equiv/kg of corn in methanol and HCl/methanol extracts, respectively. The TPC of alkaline hydrolysates ranged from 2.74 to 6.27 g of ferulic acid equiv/kg of corn. The ACL values were 0.41-0.80 and 0.84-1.59 g of Trolox equiv/kg of corn in methanol and HCl/methanol extracts, respectively. The ORAC values were 10.57-12.47 and 18.76-24.92 g of Trolox equiv/kg of corn in methanol and HCl/methanol extracts, respectively. ORAC values of alkaline hydrolysates ranged from 42.85 to 68.31 g of Trolox equiv/kg of corn. The composition of phenolic acids in alkaline hydrolysates of corn was p-hydroxybenzoic acid (5.08-10.6 mg/kg), vanillic acid (3.25-14.71 mg/kg), caffeic acid (2.32-25.73 mg/kg), syringic acid (12.37-24.48 mg/kg), p-coumaric acid (97.87-211.03 mg/kg), ferulic acid (1552.48-2969.10 mg/kg), and o-coumaric acid (126.53-575.87 mg/kg). Levels of DPPH* scavenging activity, TPC, ACL, and ORAC in HCl/methanol extracts were obviously higher than those present in methanol extracts. There was no significant loss of antioxidant capacity when corn was dried at relatively high temperatures (65 and 93 degrees C) postharvest as compared to drying at ambient temperatures (27 degrees C). Alkaline hydrolysates showed very high TPC, ACL, and ORAC values when compared to methanol and HCl/methanol extracts. High-amylose corn had a better antioxidant capacity than did typical (nonmutant) corn genotypes.     

Comparison of the phosphorus and mineral concentrations in bran and abraded kernel fractions of a normal barley (Hordeum vulgare) cultivar versus four low phytic acid isolines

Phytic acid consists of 65-80% of the total phosphorus (P) in cereal grains. Its salts are concentrated in the germ and aleurone layers, which are typically removed during milling. We hypothesize that concentrations of different types of P and minerals in milled products will be greatly altered in low phytic acid (lpa) barleys. Seeds of cv. Harrington (control) and four lpa isolines-lpa1-1, lpa2-1, lpa3-1, and M955-were abraded by a laboratory method into five surface layer and four remaining kernel fractions. Results show that phytic acid in the four lpa lines ranged from 75% to 5% of the control. The decrease in phytic acid P concentration was matched almost equally by an increase in inorganic P, so that the rest of P (the sum of all P-containing compounds other than phytic acid P and inorganic P) and total P levels remained relatively unchanged among the five genotypes. These trends were also observed for the processed fractions. The major mineral elements in barley seeds were P, K, Mg, S, and Ca, while minor ones were Fe, Zn, Mn, Cu, and Ba. All types of P and other minerals measured were generally concentrated in the outer layers of the grain. Although there were substantial differences in mineral contents of bran fractions among genotypes, the level of phytic acid P had little effect on mineral contents in whole or abraded kernels. One major exception was Fe, which had the highest level in all tissues of M955 genotype. The above findings were all confirmed by analyzing another set of barley samples grown in a different environment. Thus, in general, breeding lpa barleys does not lead to reduced mineral contents in whole grains or elevated mineral levels in milled products.     

Simple, rapid, and inexpensive cleanup method for quantitation of aflatoxins in important agricultural products by HPLC

A chemical cleanup procedure for low-level quantitative determination of aflatoxins in major economically important agricultural commodities using HPLC has been developed. Aflatoxins were extracted from a ground sample with MeOH/H2O (80:20, v/v), and after a cleanup step on a minicolumn packed with Florisil, aflatoxins were quantified by HPLC equipped with a C18 column, a photochemical reactor, and a fluorescence detector. Water/MeOH (63:37, v/v) served as the mobile phase. Recoveries of aflatoxins B1, B2, G1, and G2 from peanuts spiked at 5, 1.7, 5, and 1.7 ng/g were 89.5+/-2.2, 94.7+/-2.5, 90.4+/-1.0, and 98.2+/-1.1, respectively (mean+/-SD, %, n=3). Similar recoveries, precision, and accuracy were achieved for corn, brown and white rice, cottonseed, almonds, Brazil nuts, pistachios, walnuts, and hazelnuts. The quantitation limits for aflatoxins in peanuts were 50 pg/g for aflatoxin B1 and 17 pg/g for aflatoxin B2. The minimal cost of the minicolumn allows for substantial savings compared with available commercial aflatoxin cleanup devices.     

Molecular characteristics of an immobilization antigen gene of the fish-parasitic protozoan Ichthyophthirius multifiliis strain ARS-6

Ichthyophthirius multifiliis (Ich), a ciliated protozoan parasite of fish, expresses surface antigens (i-antigens), which react with host antibodies that render them immobile. The nucleotide sequence of an i-antigen gene of I. multifiliis strain ARS-6 was deduced. The predicted protein of 47 493 Da is comprised of 460 amino acids (aa's) arranged into five imperfect repeats with periodic cysteine residues with the structure: CX₍₁₉₎₂₀CX₂CX₁₆₋₂₇CX₂CX₂₀₍₂₁₎CX₃. The N-terminal aa's typify a signal peptide motif while a stretch of C-terminal aa's resemble a glycosyl–phosphatidyl–inositol (GPI)-anchor addition site. The degree of deduced i-antigen aa sequence identity of strain ARS-6 (GenBank accession # ACH87654 and # ACH95659) with other I. multifiliis i-antigen sequences present in GenBank ranges from 99% to 36% identity with 52 kDa i-antigens of I. multifiliis strain G5 (accession #s AAK94941 and AAK01661 respectively). Immunoblot analysis of i-antigens following exposure of I. multifiliis theronts to catfish anti- I. multifiliis immune serum did not show any appreciable alteration in i-antigen expression. The mechanism that regulates i-antigen expression in I. multifiliis remains a puzzling question.