Biofortification of soybean meal: immunological properties of the 27 kDa γ-zein

Legumes, including soybeans ( Glycine max ), are deficient in sulfur-containing amino acids, which are required for the optimal growth of monogastric animals. This deficiency can be overcome by expressing heterologous proteins rich in sulfur-containing amino acids in soybean seeds. A maize 27 kDa γ-zein, a cysteine-rich protein, has been successfully expressed in several crops including soybean, barley, and alfalfa with the intent to biofortify these crops for animal feed. Previous work has shown that the maize 27 kDa zein can withstand digestion by pepsin and elicit an immunogenic response in young pigs. By use of sera from patients who tested positive by ImmunoCAP assay for elevated IgE to maize proteins, specific IgE binding to the 27 kDa γ-zein is demonstrated. Bioinformatic analysis using the full-length and 80 amino acid sliding window FASTA searches identified significant sequence homology of the 27 kDa γ-zein with several known allergens. Immunoblot analysis using human serum that cross-reacts with maize seed proteins also revealed specific IgE-binding to the 27 kDa γ-zein in soybean seed protein extracts containing the 27 kDa zein. This study demonstrates for the first time the allergenicity potential of the 27 kDa γ-zein and the potential that this protein has to limit livestock performance when used in soybeans that serve as a biofortified feed supplement.     

Long-Term Fertilization Effects on Rice Productivity and Nutrient Efficiency in Korean Paddy

ABSTRACT

Long-term fertilization tests evaluated rice (Oryza sativa) productivity in relation to application of nitrogen (N)-phosphorus (P)-potassium (K) (120-34.9-66.7 kg ha^{? 1}, respectively) during 1967–1972 and N-P-K (150-43.7-83.3 kg ha^{? 1}, respectively) during 1973–2000. The comparison treatments (NP, PK, and NK) and the control (not fertilized) were selected for calculating nutrient efficiency. Rice grain yield increased at a 17.78 kg ha^{? 1} yr^{? 1} in the control, mainly due to development of improved cultivars. Phosphorus management was found to be important for indigenous fertility and rice productivity in this paddy soil. Yield increased significantly with P fertilization. Without N fertilization (PK), rice productivity increased 56.85 kg ha^{? 1} yr^{? 1} from 62% of NPK at the initial stage to 74% after passing 34 years, which might be affected by increasing biological N fixation with P accumulation in soil. In NK treatment, rice yield increased at a relatively low rate (37.82 kg hr^{? 1} yr^{? 1}) from the same rice productivity with that of NPK in 1967 to 91% after 34 years. In comparison, yield increased at a high rate (62.82 kg hr^{? 1} yr^{? 1}) without K fertilization (NP) from ca. 90% of NPK and might exceed the yield of NPK after 64 years of long-term fertilization. Therefore, K fertilization level might be readjusted after long-term fertilizing in paddy soil.     

A Laboratory Column Study on the Biodegradation of Toluene and Methyl tert–Butyl Ether: The Effect of Substrate Interactions

Soil vapor extraction (SVE) is one of the most effective remediation technologies for soil and groundwater contamination. Soil particles can be mobilized by air perturbation during SVE, resulting in the differentiation of porous media, which has not been well addressed. This paper developed a numerical method to study the flow pattern and quantify the change of porous media for the first time. Based on the mass equilibrium and Darcy’s law, a two-phase water–air flow model was constructed with integration of saturation, relative permeability, and capillary pressure during SVE. Relationship between porosity and saturation was deduced and coupled with the two-phase flow model for quantifying change of porous media in real time. Results reveal that both porosity and permeability increase sharply in the early stage of SVE then gradually to a quasi-steady state. These increases in vadose zone tapered off with distance from the SVE screen and the steady period occurred later as well. The influence radius of a single SVE well and the change degree in porosity and permeability of media were proportional to the extraction vacuum and the driving coefficient C, which is more sensitive than extraction vacuum according to the simulation results. Knowledge from this modeling exercise provides a useful tool to estimate the change of remediated zone and assess the environmental risk of remedial activities at real-world contamination sites.     

Improved Calibration for Estimating Soil Properties with a Multifunctional Heat Pulse Probe

A multifunctional heat pulse probe (MFHPP) can measure soil thermal and hydraulic properties. Though its successful implementation has been documented, previous studies have reported some limitations. One specific cause of the limitations is the absorption of the generated heat pulse within the probe itself, which creates error in the measurements. The objective of this study was to develop and evaluate a new calibration method to account for measurement error due to heat loss to the probe. A MFHPP was constructed and tested in six soil types using both a traditional method and the newly developed calibration method. The new calibration utilizes heat pulse response curves from real soils with thermal conductivities similar to that of the MFHPP rather than the traditional agar-stabilized water solution. This new approach significantly reduced average measurement errors from 9.1% to 2.4% for heat capacity and 13.5% to 4.5% for volumetric water content.     

Nomenclature and genetic relationships of apples and pears from Terceira Island

Heritage apple (Malus domestica Borkh. hybrids) and pear (Pyrus communis L. hybrid) trees grow in villages throughout Terceira Island, Azores, Portugal. Some of these pears have different names but similar morphology. The objective of this study was to determine synonymy, homology, and phylogeny of apples and pears from Terceira and to examine potential relationships of the island pears with standard apples and pears of Portuguese or American descent. Nine apple microsatellite markers were used to determine genetic relationships. Distance- and parsimony-based cluster analysis grouped these genotypes into separate apple and pear clades. The Terceira apples were divided into two clades: the maçā and the reineta-reinette. Among the 17 heritage apple genotypes, seven unique accessions were identified and four groups of synonyms, or possibly clones, were detected including: ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares; ‘Reineta Castanha’ and ‘Reineta Verde Miuda’; ‘Maçā Pêra,’ ‘Maçā Calhau’, ‘Pêro Branco’ from Salga and from Terra-Chā and ‘Maçā Marmelo’; and the five genotypes ‘Maçā Sao Joao’, ‘Malápio Rosa’, ‘Maçā Gaspar’, ‘Maçā Branca’ and ‘Maçā Pato’. In addition, two homonyms were detected. ‘Pêro Vermelho’ from Terra Chā was a separate genotype from a tree from Doze Ribeiras of the same name, but Pêro Branco from Terra Chā appears to be a clone that can be distinguished by an additional allele at CH1F07a from a tree with that name from Salga. One pair of apple clones, ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares appear to be derived from an unreduced gamete of ‘Golden Delicious.’ Another apple genotype ‘Maçā Acida’ could be a sibling of the ‘Maçā Pêra’ clonal group. Other tested standard apples from the US genebank were unrelated to Terceira genotypes. Of the seven heritage pears, five unique genotypes and one pair of synonyms were detected. ‘Pêra Papo Pintassilgo’ from Raminho and ‘Pêra Vermelha’ from the nursery of Serviço de Desenvolvimento Agario da Terceira (SDAT) were synonyms. ‘Passans du Portugal’ was related to ‘Pêra Cabaca’ but other standard pears from the US genebank were unrelated to Terceira genotypes. Future studies will include additional apple and pear cultivars from other Islands of the Azores and continental Portugal, and wild Asian species to further explore genetic relationships.     

黑曲霉固体发酵产木聚糖酶的响应面优化设计及其酶学性质的研究

本研究采用响应面法的中心组合对影响黑曲霉(aspergillus niger JL 15)固体发酵产木聚糖酶的条件进行了优化.以橘皮粉为基质,补充碳源、氮源,以及含水量和发酵时间对木聚糖酶产量具有显著影响(P<0.05).模拟二次多项式回归预测模型,并建立自变量与响应值的回归方程,获得各因素的最佳水平,即:甘油、硫酸铵的添加量分别为4.2%和3.1%,含水量为61%,发酵时间为73.4 h,木聚糖酶活性最大预测值达922.9 U/g干发酵产物,验证值为917.7 U/g干发酵产物,高出基础培养基酶活3.2倍.酶学性质分析研究表明,黑曲霉木聚糖酶(XylA)的最适温度和最适pH分别为55oC和pH 5.0.XylA的K_m和V_(max)值分别为9.24 mg/mL和54.05 μmol/min/mL.Mn~(2+)、Zn~(2+)和斗和Mg~(2+)对XylA活性具有促进作用,而Fe~(3+)和cu~(2+)对XylA活性具有明显抑制作用.HPLC分析结果表明,XylA的水解桦木木聚糖和麸皮不溶性木聚糖的产物均为木糖至木六糖,主要产物为木三糖.     

SNP Markers Discovery in Soybean with Use of Degenerate Oligolucleotide Primed PCR

There are several strategies that can be applied in SNP discovery, as for example the locus-specific amplification of target genome regions （Primmer et al., 2002; Van et al., 2004） or simultaneous assembly of anonymous sequences which are the product of whole genome shotgun sequencing （Webber and Myers, 1997） or reduced representation shotgun sequencing （Altshuler et al., 2000）. The majority of these methods are highly cost-prohibitive, however, and there is a necessity for development of new strategies that would be more suitable for small and medium-scale laboratories.     

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.     

Isolation and characteristics of eight novel polymorphic microsatellite loci from the genome of garlic (Allium sativum L.)

Here we characterized eight novel polymorphic SSR markers, developed from an enriched genomic library of garlic (Allium sativum L.). These SSRs produced a total of 64 alleles across 90 garlic accessions, with an average of 8 alleles per locus. Values for observed (H_O) and expected (H_E) heterozygosity ranged from 0.16 to 0.77 (mean = 0.44) and from 0.22 to 0.86 (mean = 0.65), respectively. Six loci deviated significantly (P < 0.05) from Hardy–Weinberg equilibrium (HWE). The averages of gene diversity and PIC values were 0.65 and 0.62, respectively. The mean genetic similarity coefficient was 0.4380, indicating that among garlic accessions existed wide genetic variation. Based on 64 alleles obtained by 8 SSRs, a phenogram was constructed to understand the relationships among the 90 accessions. These newly developed SSRs should prove very useful tools for genotypes identification, assessment of genetic diversity and population structure in garlic.     

Varietal differences in phenolic content and astringency in skin and flesh of hardy kiwifruit resources in Japan

Varietal differences in the total phenolic content and astringency in the skin and flesh were determined among the cultivars and local collections of hardy kiwifruit with a ploidy variance found in Japan. The average values of the total phenolic content in the skin and flesh were 2.66 and 0.18 g 100 g⁻¹ FW, respectively. There were large varietal differences in the total phenolic content in the skin in the range of 1.3–5.0 g 100 g⁻¹ FW. Kochi (tetraploid), while Gassan and ‘Mitsuko’ (hexaploid) contained a larger amount of total phenolics. High astringency was found in Gassan, ‘Mitsuko’ and ‘Hoko’ (hexaploids) and Kochi. HPLC analysis showed that the major components of phenolics in the flesh were (+)-catechin, chlorogenic acid, rutin, (−)-epicatechin and quercetin.