Molecular mapping of soybean seed tocopherols in the cross ‘OAC Bayfield’ × ‘OAC Shire’

Soybean (Glycine max [L.] Merr.) is cultivated primarily for its protein and oil in the seed. In addition, soybean seeds contain nutraceutical compounds such as tocopherols (vitamin E), which are powerful antioxidants with health benefits. The objective of this study was to identify molecular markers linked to quantitative trait loci (QTL) that affect accumulation of soybean seed tocopherols. A recombinant inbred line (RIL) population derived from the cross ‘OAC Bayfield’ × ‘OAC Shire’ was grown in three locations over 2 years. A total of 151 SSR markers were polymorphic of which a one‐way analysis of variance identified 42 markers whereas composite interval mapping identified 26 markers linked to tocopherol QTL across 17 chromosomes. Individual QTL explained from 7% to 42% of the total phenotypic variation. Significant two‐locus epistatic interactions were identified for a total of 122 combinations in 2009 and 152 in 2010. The multiple‐locus models explained 18.4–72.2% of the total phenotypic variation. The reported QTL may be used in marker‐assisted selection (MAS) to develop high tocopherol soybean cultivars.     

甘蓝型油菜茎秆菌核病抗性与木质素及其单体比例的相关性分析及QTL定位

菌核病是一类非专一性的植物真菌病原菌，寄主范围广泛，严重危害农作物的生产。对高世代重组自交系群体(RIL)及F₂群体终花期茎秆进行菌核病抗性接种鉴定，根据构建的近红外模型对接种鉴定的茎秆木质素含量、单体组分比例进行测定，并进行相关性分析和QTL定位。结果表明在2013年和2014年RIL群体茎秆菌斑大小与木质素含量呈极显著负相关，相关系数分别为–0.348和–0.286，与单体G/S呈显著正相关，相关系数分别为0.198和0.167。2014年F₂群体菌斑大小与木质素含量呈极显著负相关，相关系数为–0.306，与单体G/S相关性为0.142。F_2:3家系抗(感)植株茎部切片间苯三酚染色观察表明抗性较强的材料木质素含量高于抗性较弱的材料。根据已构建的重组自交系高密度SNP遗传图谱，利用复合区间作图法对上述性状进行QTL分析，共检测到18个QTL，其中9个菌核病抗性相关QTL分布于A05、A06、C04和C06染色体，单个QTL可解释的表型变异为2.38%~12.05%；3个木质素含量QTL分别位于A04、A05和C01染色体，单个QTL可解释表型变异的2.03%~13.75%。6个木质素单体G/S QTL分布于A08、C03和C07染色体，单个QTL可解释表型变异的2.06%~8.66%。本文研究结果为油菜菌核病抗性育种提供了新的思路和理论基础。     

Association analysis for detecting significant single nucleotide polymorphisms for phosphorus-deficiency tolerance at the seedling stage in soybean [Glycine max (L) Merr.]

Tolerance to low-phosphorus soil is a desirable trait in soybean cultivars. Previous quantitative trait locus (QTL) studies for phosphorus-deficiency tolerance were mainly derived from bi-parental segregating populations and few reports from natural population. The objective of this study was to detect QTLs that regulate phosphorus-deficiency tolerance in soybean using association mapping approach. Phosphorus-deficiency tolerance was evaluated according to five traits (plant shoot height, shoot dry weight, phosphorus concentration, phosphorus acquisition efficiency and use efficiency) comprising a conditional phenotype at the seedling stage. Association mapping of the conditional phenotype detected 19 SNPs including 13 SNPs that were significantly associated with the five traits across two years. A novel cluster of SNPs, including three SNPs that consistently showed significant effects over two years, that associated with more than one trait was detected on chromosome 3. All favorable alleles, which were determined based on the mean of conditional phenotypic values of each trait over the two years, could be pyramided into one cultivar through parental cross combination. The best three cross combinations were predicted with the aim of simultaneously improving phosphorus acquisition efficiency and use efficiency. These results will provide a thorough understanding of the genetic basis of phosphorus deficiency tolerance in soybean.     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

甲状腺素对猪小肠上皮细胞miR-let-7a基因表达及细胞增殖的影响

miR-let-7a在动物细胞的分化、增殖与凋亡等方面发挥越来越重要的作用。甲状腺激素(TH)作用非常广泛,机体的每个细胞几乎都是TH作用的靶细胞,其可以促进组织分化、生长和成熟。本实验用甲状腺素(T4)浓度分别为(0、0.02、0.03、0.05、0.075、0.1、0.2μmol/L)在体外培养猪的小肠上皮细胞。结果表明:T4处理组的细胞体积形态相对于空白对照组没有明显变化;当T4添加浓度为0.03μmol/L时,细胞的增殖率显著低于其他组(P0.05);当T4浓度为0~0.03μmol/L时,let-7a的表达随着添加剂量的增加而升高,浓度从0.03~0.2μmol/L变化时,let-7a的表达呈现降低趋势,浓度为0.03μmol/L时表达量极显著高于其他组(P0.01)。let-7a的表达量与细胞增殖呈负相关。     

Soil property differences among high‐ and average‐yielding soya bean areas in Arkansas,USA

Soya bean [Glycine max (L.) Merr] yields >6719 kg/ha (100 bu/ac) have only recently and infrequently been achieved. Quantifying soil property differences between high‐ and average‐yielding areas can help to further identify non‐plant‐related properties contributing to soya bean yield potential. The objective of this study was to evaluate the effects of region and soil depth on soil property differences between high‐ and average‐yielding areas. In each of the seven regions of the ‘Grow for the Green’ yield contest in Arkansas, prior to or just after harvest in 2014 and 2015, soil samples were collected from the top 20 cm of one contest high‐yield (HY ) area that was in close proximity to an average‐yield (AY ) area. Across all regions and both years, soya bean yields differed (P  <  0.05) between yield areas, averaging 4701 and 5498 kg/ha in AY and HY areas, respectively. Averaged across soil depth and years, numerous soil properties differed (P  <  0.05) between HY and AY areas within at least one of seven regions. Total soil C content was at least 20.2% greater in the HY than in the AY area in three of seven regions. Extractable soil P content was, on average, 19.4 kg/ha greater in HY than in AY areas in three of the seven regions. Results from this study have the potential to help producers better understand soil properties that contribute to or hinder achieving ultra‐high (>6719 kg/ha) soya bean yields.     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

豆粕酶解发酵物对仔猪生长性能、血清指标及肠道菌群结构的影响

本试验旨在研究豆粕酶解发酵物对仔猪生长性能、血清指标及肠道菌群结构的影响。选用144头49日龄健康的"杜×长×大"三元杂交育肥仔猪,随机分为3组,每组6个重复,每个重复8头猪(公母各占1/2)。阴性对照组饲喂无抗生素添加的基础饲粮,抗生素对照组饲喂基础饲粮+60 g/t吉他霉素,试验组饲喂试验饲粮(基础饲粮中10%豆粕被豆粕酶解发酵物替代)。试验期为42 d。结果表明:1)试验组仔猪的末重、平均日增重、平均日采食量分别比阴性对照组高5.06%、10.07%、5.03%(P0.05),腹泻率比阴性对照组低39.59%(P0.05),与抗生素对照组相比各指标差异均不显著(P0.05)。2)试验组仔猪血清碱性磷酸酶和谷丙转氨酶活性、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性均显著高于阴性对照组(P0.05),丙二醛(MDA)含量显著低于阴性对照组(P0.05);试验组和抗生素对照组仔猪血清指标差异不显著(P0.05)。3)试验组仔猪盲肠、大肠中乳酸杆菌数量显著高于阴性对照组和抗生素对照组(P0.05),十二指肠中总细菌数显著低于阴性对照组和抗生素对照组(P0.05)。综上,豆粕酶解发酵物能提高仔猪生长性能,增强机体抗氧化能力,利于乳酸杆菌在肠道内形成优势菌群,降低腹泻率,生长性能达到使用抗生素水平。     

C语言在线评价与测试系统的开发与探讨

随着网络教育的发展,接受网络教育和考试认证的人越来越多,在线考试正在得以迅速发展.本文简要介绍在线系统工作原理和系统配置.