Development of simple PCR-based markers linked to the <Emphasis Type="Italic">Ms</Emphasis> locus,a restorer-of-fertility gene in onion (<Emphasis Type="Italic">Allium cepa</Emphasis> L.)

In order to implement reliable marker-assisted selection systems for the restorer-of-fertility locus (Ms) in onions (Allium cepa L.), simple PCR-based codominant markers linked to the Ms locus were developed. Based on the EST probe sequences of previously reported RFLP markers, full-length genomic sequences of the gene encoding putative oligopeptide transporter (OPT) was obtained by RACE. The first intron contained two 108 and 439-bp indel polymorphisms between the two Ms allele-linked OPT alleles. A simple PCR marker for OPT was developed by designing a primer pair on the flanking regions of the 108-bp indel which is created by two tandem repeats. The second simple PCR marker was developed from the EST probe encoding photosystem I subunit O (PsaO). Two 14 and 39-bp tandem repeats were identified from the 5′ upstream sequences of the PsaO-coding gene, which were isolated by genome walking. Three different compositions of these tandem repeats were identified from diverse onion germplasm. A primer set binding to the flanking sequence of these polymorphic repeats was used to amplify three different marker haplotypes. The OPT marker was tightly linked to the Ms locus at a distance of 1.5 cM, but the analysis of the linkage relationship showed little linkage disequilibrium between the marker and the Ms locus. Even so, these simple PCR markers are valuable tools for the marker-assisted selection of segregating individuals in onion F₁ hybrid breeding programs.     

Three AFLP markers tightly linked to the genic male sterility <Emphasis Type="Italic">ms</Emphasis><Subscript><Emphasis Type="Italic">3</Emphasis></Subscript> gene in chili pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) and conversion to a CAPS marker

Genic male sterility (GMS) has long been used as a tool for hybrid seed production in chili pepper (Capsicum annuum L.). We developed DNA markers linked to the GMS ms ₃ gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 512 primer combinations. Three AFLP markers (Eagg/Mccc₂₇₆, Eagc/Mctt₁₇₈, and Ecag/Mtgc₂₀₄) were identified as tightly linked to the ms ₃ locus. Among them, we converted the AFLP marker Ecag/Mtgc₂₀₄ to the cleavage amplified polymorphic sequence (CAPS) marker, named GMS3-CAPS, based on sequencing analysis of internal and flanking regions for the markers between male-fertile and sterile plants. This marker will be useful for pepper breeding using the GMS system.     

QTL mapping for capsaicin and dihydrocapsaicin content in a population of Capsicum annuum ‘NB1’ × Capsicum chinense ‘Bhut Jolokia’

Capsaicinoids are pungent compounds used for industrial and medical purposes including food, medicine and cosmetics. The Indian local variety ‘Bhut Jolokia’ (Capsicum chinense Jacq.) is one of the world's hottest chilli peppers. It produces more than one million Scoville heat units (SHUs) in total capsaicinoids. In this study, our goal was to identify quantitative trait loci (QTLs) responsible for the high content of capsaicin and dihydrocapsaicin in ‘Bhut Jolokia’. Capsicum annuum ‘NB1’, a Korean pepper inbred line containing 14 000 SHUs, was used as a maternal line. An F₂ population derived by crossing between ‘NB1’ and ‘Bhut Jolokia’ was generated to map QTLs for capsaicinoids content. A total of 234 markers, including 201 HRM, 21 SSR, 2 CAPS and 10 gene‐based markers of the capsaicinoid synthesis pathway, were mapped. The final map covered a total distance of 1175.2 cM and contained 12 linkage groups corresponding to the basic chromosome number of chilli pepper. Capsaicin and dihydrocapsaicin content were analysed in 175 F₂ pepper fruits using the HPLC method. The maximum total capsaicinoids content was 1389 mg per 100g DW (dry weight), and the minimum content was 11 mg per 100g DW. Two QTLs (qcap3.1 and qcap6.1) for capsaicin content were identified on LG3 and LG6, and two QTLs (qhdc2.1 and qdhc2.2) for dihydrocapsaicin content were located on LG2. We did not detect QTLs for total capsaicinoids content. The QTL positions for capsaicin content were different from those for dihydrocapsaicin content. These results indicate that the complexity of selecting for more pungent chilli peppers must be considered in a chilli pepper breeding programme. The QTL‐linked markers identified here will be helpful to develop more pungent pepper varieties from ‘Bhut Jolokia’, a very hot pepper.     

Methyl jasmonate alleviated salinity stress in soybean

We studied the role of methyl jasmonate (MeJA) in alleviating NaCl-induced salt stress on soybean growth and development in hydroponics medium. Soybean seedlings were exposed to 60 mM NaCl stress for 2 weeks, 24 h after the application of 20 and 30 μM MeJA. NaCl stress induced a significant reduction in plant growth, endogenous bioactive gibberellin (GA4), photosynthesis and transpiration rate, while a marked increase in the endogenous abscisic acid (ABA) and proline contents were recorded. MeJA application greatly mitigated the adverse effects of NaCl on soybean growth and endogenous hormones. MeJA significantly increased ABA levels, while the endogenous amount of GA4 was reduced by the application of NaCl. Our study revealed that MeJA counteracted the negative effects of NaCl stress on plant growth, chlorophyll content, leaf photosynthetic rate, leaf transpiration rate, and proline content.     

DNA profiling and genetic diversity of Korean soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill) landraces by SSR markers

Approximately 7,000 accessions of Korean soybean (Glycine max (L.) Merrill) landraces, largely composed of three collections, the Korea Atomic Energy Research Institute’s soybean (KAS), the Korean Crop Experiment Station’s soybean (KLS) and the Korean Agricultural Development and Technology Center’s soybean (KADTC) collections, have been conserved at the Rural Development Administration (RDA) genebank in Korea. The accessions within collections were classified based on their traditional uses such as sauce soybean (SA), sprouted soybean (SP), soybean for cooking with rice (SCR), and OTHERS. A total of 2,758 accessions of Korean soybean landraces were used to profile and to evaluate genetic structure using six SSR loci. A total of 110 alleles were revealed by at the six SSR loci. The number of alleles per SSR locus ranged from 9 to 39 in Satt187 and Satt_074, respectively. The number of alleles ranged from 87 in the KADTC collection to 96 in the KLS collection, and from 63 in the SCR group to 95 in the SP group. Nei’s average genetic diversity ranged from 0.68 to 0.70 across three collections, and 0.64 to 0.69 across the usage groups. The average between-group differentiation (G _st) was 0.9 among collections, and 4.1 among the usage groups. The similar average diversity among three collections implies that the genetic background of the three collections was quite similar or that there were a large number of duplicate accessions in three collections. The selection from the four groups classified based upon usage may be a useful way to select accessions for developing a Korean soybean landrace core collection at the RDA genebank. DNA profile information of accessions will provide indications of redundancies or omissions and aid in managing the soybean collection held at the RDA genebank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding programs and could be used to develop a core collection.     

Utilization of monolinolein as a substrate for conjugated linoleic acid production by Bifidobacterium breve LMC 520 of human neonatal origin

This study was designed to isolate bifidobacteria from human intestines that efficiently converts monolinolein, a monoglyceride form of linoleic acid, into conjugated linoleic acid (CLA), as well as to optimize culture conditions for improving CLA production during milk fermentation. Among 150 screened neonatal bifidobacteria, Bifidobacterium breve LMC 520 showed the highest CLA-producing ability and was tested with different types of fat substrates at various concentrations to determine the optimal conditions for CLA production. Monolinolein was tested as a substrate for CLA production. The incubation time optimized for CLA production was 24 h, and CLA production was proportionally increased with monolinolein concentration. The incubation of LMC 520 with commercial starter strains caused minimal reduction in CLA production. Our results demonstrate that the CLA-producing ability of B. breve LMC 520 could offer beneficial effects when utilized as a starter culture for the development of functional dairy products.     

Panax ginseng extract rich in ginsenoside protopanaxatriol attenuates blood pressure elevation in spontaneously hypertensive rats by affecting the Akt-dependent phosphorylation of endothelial nitric oxide synthase

Nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS) is a fundamental regulator of systemic blood pressure. Ginsenosides from Panax ginseng have been investigated in vitro for the molecular and biochemical mechanisms by which they stimulate NO release in vascular endothelial cells; however, little research has been done to confirm the physiological relevance of these in vitro studies. To address this research gap, the effects of a P. ginseng extract rich in ginsenosides from protopanaxatriol on spontaneously hypertensive rats (SHRs) was examined. Ginseng extract administration stimulated nongenomic Akt-mediated eNOS activation, enhanced NO production, improved vessel wall thickening, and alleviated hypertension in SHRs, confirming the physiological relevance of previous in vitro studies with ginsenosides.     

Different response of silicate fertilizer having electron acceptors on methane emission in rice paddy soil under green manuring

Cultivation of green manure plants during the fallow season in rice paddy soil has been strongly recommended to improve soil properties. However, green manuring may impact greenhouse gas emission, methane (hereafter, CH₄) in particular, under the flooded rice cultivation and thus, application of chemical amendments being electron acceptors can be an effective mitigation strategy to reduce CH₄ emissions in irrigated rice (Oryza sativa L.) field amended with green manure. To investigate the effect of iron (Fe) slag silicate fertilizer (hereafter, silicate fertilizer), which was effective in reducing CH₄ emission and increasing rice productivity, in green manure-amended paddy soil, the aboveground biomass of Chinese milk vetch (hereafter, vetch) was added at rates of 0, 10, 20, and 40 Mg (fresh weight) ha⁻¹ before the application of silicate fertilizer, which was added at rates of 0 and 2.3 Mg ha⁻¹. Silicate fertilization reduced the seasonal CH₄ flux by ca. 14.5% and increased rice yield by ca. 15.7% in the control (no vetch application) treatment. However, CH₄ production was increased by silicate fertilization in vetch-treated soil particularly at the initial rice growing stage, which was probably due to the enhanced decomposition of added organic matters by the silicate liming effect. In conclusion, silicate fertilization is not effective in reducing CH₄ production in green manure-amended rice paddy soils and its use should be properly controlled.     

Fortification of Wheat Dough with Calcium and Magnesium Ions Affects Empirical Rheological Properties

This study evaluated the influence of calcium and magnesium ions on the empirical rheological properties of wheat flour to verify possible effects of these ions on processing because, in addition to their nutritional importance, they are also responsible for water hardness. Calcium (0–1.30 g/100 g) and magnesium (0–0.34 g/100 g) ions from sulfate salts were added to wheat flour, according to a central composite rotatable design. The farinograph and extensigraph properties of wheat flour and its mixtures were evaluated. The results were analyzed by response surface methodology. Calcium ions stood out for increasing water absorption, decreasing mixing stability, and producing a delayed effect on dough extensibility (reduced at 135 min). Magnesium ions influenced most flour rheological properties in a similar manner to oxidizing agents (increased dough stability, increased resistance to extension, and reduced extensibility), thus proving to be a possible replacement agent for these additives. An interaction effect of the combined calcium and magnesium ions was observed on dough development time. The results showed that effects on processing can occur when wheat flour fortification is made, and adaptations on wheat flour specifications, product formulation, and processing parameters may be required.     

Ultrasonographic measurement of caudal vena cava to aorta ratios for determination of volume depletion in normal beagle dogs

A noninvasive method for quantifying hydration status would be helpful for clinical management and for research applications in dogs. This prospective, experimental, pilot study aimed to assess the feasibility of ultrasonographic measurement of the caudal vena cava to aorta ratio as a method for quantifying volume depletion in dogs. In 12 normal beagle dogs, furosemide was administered intravenously at a dose of 1 mg/kg, every 2 h, for 8 h, to induce consecutive volume depletion. Every 30 min after administration, ultrasonographic images of the caudal vena cava and aorta, and physical and biological parameters related to dehydration were acquired. On transverse and longitudinal planes of caudal vena cava and aorta images, the height and area of the caudal vena cava and aorta were measured to calculate the caudal vena cava/aorta ratios. All images were acquired by approaching from the right intercostal space with the dogs in left lateral recumbency. A negative correlation was present between the percentage of weight loss in dogs and all four investigated caudal vena cava/aorta ratios (transverse plane width of the caudal vena cava [TW]/aorta; transverse plane height of caudal vena cava [TH]/aorta; longitudinal plane area of the caudal vena cava [TA]/aorta; and longitudinal plane maximal height of the caudal vena cava [L]/aorta). Significant differences (P < 0.001) were seen between dogs with and without clinical signs of dehydration for all caudal vena cava/aorta ratios. Findings indicated that ultrasonographic caudal vena cava/aorta ratios are feasible methods for quantifying volume depletion and for use as an adjunct to standard subjective methods for estimating hydration status in dogs.