Ionomic Profiling of Rice Genotypes and Identification of Varieties with Elemental Covariation Effects

Ionomic profiles are primarily influenced by genetic and environmental factors. Identifying ionomic responses to varietal effects is necessary to understand the ionomic variations among species or subspecies and to potentially understand genetic effects on ionomic profiles. We cultivated 120 rice (Oryza sativa) varieties to seedling stage in identical hydroponic conditions and determined the concentrations of 26 elements (including 3 anions) in the shoots and roots of rice. Although the subspecies effects were limited by the genus Oryza pre-framework and its elemental chemical properties, we found significant differences in ionomic variations in most elements among the aus, indica and japonica subspecies. Principal component analysis of the correlations indicated that variations in the root-to-shoot ionomic transport mechanisms were the main causes of ionomic differences among the subspecies. Furthermore, the correlations were primarily associated with the screening of varieties for elemental covariation effects that can facilitate breeding biofortified rice varieties with safe concentrations of otherwise toxic elements. The japonica subspecies exhibited the strongest elemental correlations and elemental covariation effects, therefore, they showed greater advantages for biofortification than the indica and aus subspecies, whereas indica and aus subspecies were likely safer in metal(loid) polluted soils. We also found that geographical and historical distribution significantly defined the ionomic profiles. Overall, the results of this study provided a reference for further association studies to improve the nutritional status and minimize toxicity risks in rice production.     

Antimicrobial resistance of Salmonella isolated from finishing swine and the environment of 60 Alberta swine farms

The study objective was to describe and evaluate antimicrobial resistance profiles in Salmonella isolated from Alberta swine finishing farms. Salmonella isolates (n = 322) were obtained from 192 fecal and 84 environmental samples of the 60 Salmonella-positive swine finishing farms. Isolates were classified susceptible, intermediate or resistant based on NCCLS guidelines. More than half of the isolates (53.4%) were susceptible to all of the 18 antimicrobials in the testing panel. No resistance was observed to amikacin, amoxicillin/clavulanic acid, cefoxitin, ceftiofur, ceftriaxone, cephalothin, ciprofloxacin, imipenem or nalidixic acid. Less than 1% of isolates were resistant to apramycin, gentamicin and trimethoprim/sulfamethoxazole. Higher frequencies of resistance were observed for chloramphenicol (4.7%), ampicillin (7.8%), kanamycin (11.8%), sulfamethoxazole (21.1%), streptomycin (25.5%) and tetracycline (38.8%). Eleven Salmonella serovars had isolates with resistance to > or =3 antimicrobials. The most frequently resistant serovar was Salmonella Derby, with 27 (38.0%) isolates resistant to > or =3 antimicrobials, including resistance to five and six antimicrobials. An absence of resistance to cephalosporins and fluoroquniolones and a low proportion of isolates resistant to amikacin, amoxicillin/clavulanic acid, apramycin, gentamicin and trimethoprim/sulfamethoxazole are encouraging findings from public health and animal health perspectives. Frequent resistance observed for ampicillin, kanamycin, sulfamethoxazole, streptomycin and tetracycline, antimicrobials commonly used in veterinary medicine for decades, indicates an urgent need to utilize these antimicrobials more prudently if their benefits are to be preserved.     

Interaction between the helicase domain of the tobacco mosaic virus replicase and a tobacco arginine decarboxylase

In a yeast two-hybrid screening test for tobacco proteins that interact with TMV replicase using the helicase (H) domain as bait, a cDNA clone was selected that encodes a polyamine biosynthetic enzyme, arginine decarboxylase (ADC). In yeast cells, the C-terminal internal region of ADC interacted with the H domain. This observation was confirmed in vitro by far-Western blotting. Inhibition of the binding between the H domain and the IRnHEL (I region and N-terminus of helicase domain) region by ADC using a yeast three-hybrid assay suggested possible interference of the heterodimerization of 126K and 183K by ADC.The nucleotide sequence data of pADCF reported in this study is available in the DDBJ/EMBL/GenBank databases under accession number AB110952     

DNA methylation status of bovine blastocyst embryos obtained from various procedures

DNA methylation is an important factor for the regulation of gene expression in early embryos. It is well known that the satellite I sequence is more heavily methylated in bovine somatic cell nuclear transfer (NT-SC) embryos than in embryos derived from in vitro fertilization (IVF). However, the methylation status of bovine embryos obtained by other procedures is not well known. To clarify DNA methylation levels of bovine embryos obtained from various procedures, we examined satellite I sequences in bovine blastocyst (BC) embryos derived from NT-SC, NT using embryonic blastomeres (NT-EM), in vivo (Vivo), IVF and parthenogenetic treatment (PA). Furthermore, in order to evaluate the efficacy of DNA demethylation by the NT procedure, we determined the DNA methylation levels in bovine embryos in which NT was recapitulated (Re-NT). Although the DNA methylation levels in the NT-SC embryos were higher than those in the other embryos, the NT-EM embryos exhibited lower DNA methylation levels. The satellite I sequence in the NT-SC embryos was more demethylated than that in the donor cells. Although the DNA methylation level in the individual NT-SC embryos showed variation, the full-term developmental efficacy of these embryos were not different. These findings suggest that the methylation level of the satellite I sequence at the BC stage is not related to the abnormalities of bovine embryos produced by NT-SC. There was no difference in methylation levels between Re-NT and NT-SC embryos. Our results indicated that the DNA methylation status differed among embryos produced by various methods and that at least some of the demethylation of the donor cell genome occurred in the recipient cytoplast after NT-SC, but the demethylation ability of the NT procedure was noted in the first NT but not in the second NT.     

Genetic and functional leaf traits variability of Quercus laurina along an oak diversity gradient in Mexico

The ecological literature has documented the effects of plant hybridization on phenotypic variation, and dominant, intermediate, or novel morphological, chemical and physiological traits in hybrids. It is important to understand the ecological consequences of hybridization by evaluating their impact on phenotypic expression of functional traits. We evaluated the relationship between genetic diversity of Quercus laurina and functional foliar traits along an oak diversity gradient. We selected five study sites that represent an oak diversity gradient where Q. laurina is present. Using chloroplast and nuclear microsatellites, we evaluated genetic diversity, measured functional foliar traits of Q. laurina in each site and assessed the effects of local climate variables on the oak community and functional traits. We found a greater abundance of Q. laurina in all study sites. We did not find a relationship between the number of accompanying red oak species and the population genetic diversity in Q. laurina, but higher genetic diversity was found in all study sites in comparison with European oak species. Sites with more oak species had more variation of foliar functional traits. Our results do not support the hypothesis that predicts higher levels of genetic diversity of Q. laurina in communities with greater oak diversity from the same section, but we demonstrated an increase in the foliar functional traits of Q. laurina associated with oak richness and climate variables. We highlight the need to consider environmental and ecological variables linkages as regulatory mechanisms of the phenotypic plasticity expressed in changes of some functional attributes of oaks.

     

Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

不同产地野生黑果枸杞资源果实多酚组成分析

【目的】黑果枸杞（Lycium ruthenicum Murr.）主要分布于中国西北地区。其果实在传统中药中被用来治疗心脏病、月经异常及停经等疾病。近年来,发现多酚特别是花色苷类物质是其主要的功效成分。测定并评价不同产地野生黑果枸杞果实中花色苷等多酚的组成情况,为黑果枸杞的品种选育及开发利用提供依据。【方法】采用分光光度计测定其总酚、总黄酮、总花色苷含量;采用HPLC-MS 方法分析鉴定黑果枸杞酚类物质组成及结构;并根据总多酚含量、总黄酮含量、总花色苷含量及多酚共有峰峰面积分别对26个不同产地黑果枸杞的多酚检测结果进行聚类分析。【结果】26个不同产地的黑果枸杞资源果实的总酚含量、总黄酮含量、总花色苷含量的变化幅度分别为8.25-87.77 mg GAE·g^-1、18.03-60.44 mg RE·g^-1、8.21-31.46 mg·g^-1。不同产地的材料间差异较大,其中青海格尔木乌图美仁、新疆甘河子、新疆巴仑台镇、宁夏贺兰12-01黑果枸杞总酚、总黄酮及总花色苷含量较高;通过HPLC-MS分析鉴定,19个多酚类化合物被鉴定出。其中7个为酰化类花色苷,主要花色苷为Petunidin-3-O-rutinoside (cis-p-coumaroyl)-5-O-glucoside。在所有供试黑果枸杞不同产地的材料中共检测到12种共有酚类化合物,其中Petunidin-3-O-rutinoside (cis-p-coumaroyl)-5-O-glucoside在所有供试黑果枸杞中的含量均最高,是黑果枸杞花色苷类多酚的主要成分。各产地样品HPLC特征图谱相似,有个别质量较差的情况存在;聚类参数不同,聚类结果略有差异。根据总酚含量、总黄酮含量、总花色苷含量聚类,聚为7类,甘肃民勤中渠、新疆喀什、宁夏贺兰12-1聚为第六类,新疆甘河子单独聚为第七类,该两类总酚、总黄酮及总花色苷含量均含量分别介于57.74-87.77 mg·g^-1、41.52-55.46 mg·g^-1和12.25-28.54 mg·g^-1。总酚、总黄酮及总花色苷含量都很高,特别是新疆甘河子产地,可作为以多酚和花色苷含量为指标的优质良种材料。根据共有峰面积共聚为5类,其中宁夏贺兰12-1、青海格尔木乌图美仁、新疆甘河子为一类,该类峰面积很高,其中主成分峰面积介于2 350.84-3 092.94。新疆巴轮台镇单独一类,其主成分峰面积高达为3 579.96。【结论】26个不同产地的黑果枸杞中多数富含多酚类物质,特别是酰化矮牵牛素花色苷含量丰富,是较好的开发富含花色苷类多酚的功能性食品资源。